Annual Report 2008 - Istituto di Ricerche Farmacologiche Mario Negri

IRFMN
PREFACE
ANNUAL REPORT
MARIO NEGRI INSTITUTE, MILAN
www.marionegri.it
DEPARTMENTS
Department of Oncology ………………….………………………….……………………… 7
Department of Environmental Health Sciences ……….………….……….……………… 57
Department of Neuroscience ………………….………………………….………………… 77
Department of Cardiovascular Research ………………….…………………….………… 123
Department of Molecular Biochemistry and Pharmacology .…….………….………… 155
Department of Epidemiology……………………………..…….………….……………….. 181
Department of Public Health............................................................................. 217
LABORATORIES AND CENTERS
Laboratory of Regulatory Policies ……….………………………..……….………….……… 241
Centre of Computer Science Engineering………………………………………….………… 247
Italian Cochrane Center …….….……………………………………………………………… 251
The Catullo and Daniela Borgomainerio Center…………………………………………… 259
Library ……….….…………………………………………………………….………….………… 261
NEGRI BERGAMO LABORATORIES
DEPARTMENTS
Department of Molecular Medicine ……….……………………………………….………… 267
Department of Biomedical Engineering……….……………………………………………… 285
Laboratory of Biology and Therapy of Metastasis…………………………………….. 301
ALDO and CELE DACCO’ CENTER
DEPARTMENT
Department of Renal Medicine…………….…………………………………………………… 305
LABORATORIES AND CENTERS
Laboratory of Coordination of Diagnosis and Information on Rare Diseases …..… 325
The Transplant Research Center…………….………………………………………………… 337
EDUCATION ACTIVITIES 339
STAFF 341
All the staff of the Institute is listed on its website www.marionegri.it
PUBLICATIONS
A comprehensive list of the Institute’s publications is available on the www.marionegri.it
website – Section Publications
1
ANNUAL REPORT 2008

IRFMN
Edited by Isabella Bordogna
printed April 2009
2
ANNUAL REPORT 2008

IRFMN
PREFACE
This booklet provides a brief description of the research and training work done at the Mario
Negri Institutes in Milan and Bergamo. It is divided by departments, and in some cases by
single laboratories. Details of the results are provided in the text itself, so here we shall just
draw a few general remarks.
This is our first year in the new Milan Headquarters: during this time we devoted most of our
efforts improving the new available technologies. Particularly, we have carried out translational
research following the “mouse clinic” patterns. Nuclear magnetic resonance, micro cat scanner,
ultrasound, Doppler, photon microscope are some of the methodologies used to do research on
human diseases in mice, following the techniques utilized in medical clinic.
This will allow to transfer data in more effective ways, to study more in depth and to use a lower
number of animals in experimental research. Imaging will play an important role, thanks to the
installation of electronic microscopy, of contrast, time- lapse microscopy and atomic force
microscopy. New important technologies will also allow us to develop our proteomics research.
The tendency nowadays is towards widespread use of molecular biology techniques, especially
for studying the mechanism of action of drugs.
In vitro studies are an essential basis for thorough investigations although a significant number
of in vivo experiments are still needed as this is the only mean we have of validating in vitro
findings and establishing models that resemble human diseases as closely as possible. This
has led to a substantial increase in the use of transgenic animals.
The Institute is still concentrating on its traditional research lines: oncology, neurosciences,
cardiovascular and renal diseases, organ transplants – with strong cell biology and molecular
biochemistry connotations. Significant work has been carried out on the environment and
health as a whole. Experimental, clinical and epidemiological research on rare diseases and
orphan drugs is growing all the time.
The Mario Negri Institute strives to develop a multifaceted approach to all these research
themes, ranging from basic research, to pharmacokinetics, pharmacology, controlled clinical
trials, epidemiological analysis and – whenever possible – the epidemiology of services.
So far we have published more than 11.000 articles on peer reviewed scientific journals.
If research is to continue young scientists must be continually trained. Working in the laboratory
not only gives them an outlet for their ideas, but enables them to obtain worthwhile
qualifications by taking part in the Institute’s training schemes, which are recognised by the
Lombardy Region in Italy, or by working for a Ph.D. awarded by the Open University, UK .
Training courses are also available on biomedical statistics, for general practitioners, family
pediatricians, and clinical trial nurses.
A vital part of the Institute’s work involves providing information, at all levels. This is done, in
particular, through the Rare Diseases Information Center, the Center for Information on
Medicinal Drugs and the Website (www.marionegri.it). We also provide information to medical
doctors, nurses, patients’ associations and lay people, through the media and through a
recently developed website: www.partecipasalute.it.
Because research is going through a very difficult time it is vital to be supported by the
Government, by private and public bodies as well as by foundations and private citizens.
Silvio Garattini
Director
3
ANNUAL REPORT 2008

IRFMN
4
ANNUAL REPORT 2008

IRFMN
Mario Negri INSTITUTE FOR
PHARMACOLOGICAL RESEARCH Milan
ANNUAL
REPORT 2008
departments and laboratories
5
ANNUAL REPORT 2008

IRFMN
6
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF ONCOLOGY
STAFF
Chief
Maurizio D’INCALCI, M.D.
Oncological Studies Office and Documentation
Scientific Documentalist
Stefania FILIPPESCHI, Chemist
Laboratory of Cancer Pharmacology
Head
Biophysics Unit
Head
Flow Citometry Unit
Head
Cancer Clinical Pharmacology Unit
Head
Maurizio D’INCALCI, M.D.
Paolo UBEZIO, Phys.D.
Eugenio ERBA, Biochem.D
Massimo ZUCCHETTI, Chem.Pharm.D.
Laboratory of Molecular Pharmacology
Head
DNA Repair Unit
Head
Massimo BROGGINI, Ph.D.
Giovanna DAMIA, M.D.
Laboratory of Biology and Therapy of Metastasis
Head
Tumor Angiogenesis Unit
Head
Unit located in Bergamo
Molecular Cancer Therapeutics Unit
Head
Raffaella GIAVAZZI, Biol.Sci.D., Ph.D.
Giulia TARABOLETTI, Biol.Sci.D.
Maria Rosa BANI, Biol.Sci.D., Ph.D.
7
ANNUAL REPORT 2008

IRFMN
Laboratory for the development of new pharmacological strategies
Head
Valter TORRI, M.D.
Laboratory of Clinical Trials
Head
Irene FLORIANI, Dr.Sci.Biol., Dr.Stat., Ph.D.
Laboratory of Translational and Outcome Research in Oncology
Head
Gynecology Oncology Unit
Head
Giovanni APOLONE, M.D.
Roldano FOSSATI, M.D.
CERP: Center for the Evaluation and Research on Pain
Head
Giovanni APOLONE, M.D.
Oscar CORLI, M.D.
Laboratory of Medical Research and Consumer Involvement
Head
Paola MOSCONI, Biol.Sci.D.
8
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Maurizio D'Incalci obtained his Medical Degree cum Laude from the University of Milan in 1977. After
specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of
Genoa, he worked in the Laboratory of Molecular Pharmacology of the National Cancer Institute in
Bethesda, MD, USA. Since 1986 he has been chief of the Laboratory of Cancer Chemotherapy at the
Mario Negri Institute and since 1996 he has become chief of the Department of Oncology at the Mario
Negri Institute.
He has been President of the Pharmacology and Molecular Mechanisms Group of the European
Organization for Research and Treatment of Cancer (EORTC). From 1994 to 1997 he was Chairman of
the New Drug Development Coordinating Committee and from 1997 to 2000 he was chairman of the
Research Division of the EORTC. He has been member of the Board of the EORTC from April 2000 to
2003.
From 1997 he is the Preclinical Coordinator of the Southern Europe New Drug Organization (SENDO)
and since 2006 the Chairman of the New Agents Committee (NAC).
He is on the editorial board of many international cancer-related scientific journals and since September
2000 he is Editor for Experimental Oncology of the European Journal of Cancer.
Dr D'Incalci is author of more than 440 papers on cancer chemotherapy published in peer reviewed
international journals, and of several chapters in books on cancer chemotherapy.
Selected publications
• Grosso F., Jones R.L. Demetri G.D., Judson I.R., Blay J.Y., Le Cesne A., Sanfilippo R., Casieri P., Collini P., Dileo P.,
Spreafico C., Stacchiotti S., Tamborini E., Tercero J.C., Jimeno J., D’Incalci M., Gronchi A., Fletcher J.A., Pilotti S.,
Casali P.G. Efficacy of Trabectedin (ET-743) in advanced pre-treated myxoid liposarcomas. Lancet Oncology, 2007;
8:595-602.
• Salvati E., Leonetti C., Rizzo A., Scarsella M., Mottolese M., Galati R., Sperduti I., Stevens M., D’Incalci M., Blasco
M., Chiorino G., Horard B., Gilson E., Zupi G., Biroccio A. Telomere damage promotes antitumoral activity of the G-
quadruplex ligand RHPS4. J. Clin. Invest., 2007; 117:3236-3247.
• Zongaro S., de Stanchina E., Colombo T., D’Incalci M., Giulotto E., Mondello C. Stepwise neoplastic transformation of
a telomerase immortalized fibroblast cell line. Cancer Research, 2005; 65:(24): 11411-11418.
• Allavena P., Signorelli M., Chieppa M., Erba E., Bianchi G., Marchesi F., Garbi A., Lissoni A., de Braud F., Jimeno J.
and D’Incalci M. Anti-inflammatory properties of the novel antitumor agent Yondelis (Trabectedin): inhibition of
macrophage differentiation and cytokine production. Cancer Res., 2005; 65(7):2964-2971.
• Tavecchio M., Natoli C., Ubezio P., Erba E., D’Incalci M. Dynamics of cell cycle phase perturbations by Trabectedin
(ET-743) in nucleotide excision repair (NER)-deficient and NER-proficient cells, unravelled by a novel mathematical
simulation approach. Cell proliferation, 2007; 40: 885-904.
• Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M,
Dell'Anna T, Apolone G, Broggini M, D'Incalci M
Analysis of gene expression in early-stage ovarian cancer
Clin Cancer Res 2008 14 : 7850-7860
Giovanni Apolone, got his Medical degree in 1982 (Pavia, Italy) and his post-doctoral specializations in
Internal Medicine in 1987 (Pavia, Italy) and Pharmacological Research (1992). He is Head of the
Laboratory of Translational and Outcome Research. He is also Vice-President of the Ethics Committee
of the European Institute of Oncology in Milan (Italy) and listed as National Expert at the European
Agency for the Evaluation of Medicinal products (EMEA) in London (UK). His main fields of interest
are:
• Methodological, ethical and regulatory aspects of clinical research, with special emphasis on oncology
and the cancer pain.
• Health care evaluation with special emphasis on oncology;
• Development and validation of case-mix and patient-reported outcome measures;
• Education and health promotion research and programs.
He has authored or co-authored over 200 publications.
Selected publications
• Trotta F, Apolone G, Garattini S, Tafuri G Stopping a trial early in oncology: for patients or for industryAnn Oncol
2008 19 : 1347-1353
9
ANNUAL REPORT 2008

IRFMN
• Apolone G, Corli O, Greco M T, Zagonel V, CPOR SG Investigators Factors influencing the decision to take or reject
opioids for cancer pain: are we on target Ann Oncol 2008 19 : 1021-1022
• Deandrea S, Montanari M, Moja L, Apolone G Prevalence of undertreatment in cancer pain. A review of published
literature Ann Oncol 2008 19 : 1985-1991
• Apolone G, Tafuri G, Trotta F, Garattini S A new anti-cancer drug in the market: good news for investors or for patients
Eur J Cancer 2008 44 : 1786-1788
• Apolone G, Patarnello F The value of a drug: From innovation to the payment via Karl MarxJ Ambul Care Manage 2008
31 : 52-55
Massimo Broggini followed the faculty of Science of the University of Milan, got the specialization in
Biochemistry at Mario Negri Institute, and the PhD degree at the Open University, London,UK.
He worked for a period in the laboratory of Molecular Pharmacology of the National Cancer Institute of
Bethesda, Md, in 1986. From 1991 is the head of the Molecular Pharmacology Unit of the Mario Negri
Institute and from 1999 of the Laboratory of Molecular Pharmacology of the same Institute.
His main fields of interest are the study of the mechanism of action of new anticancer agents, the search
of proteins and genes altered in human cancer and the study of oncosuppressor genes. He is member of
the "Pharmacology and Molecular Mechanisms Group" of the European Organisation for the Research
and Treatment of Cancer (EORTC) and of the American Association for Cancer Research. He is in the
Editorial board of the European Journal of Cancer.
He is author of more than 100 articles published in international journals.
Selected publications
• Zangrossi S, Marabese M, Broggini M, Giordano R, D'Erasmo M, Montelatici E, Intini D, Neri A, Pesce M, Rebulla P,
Lazzari L. Oct-4 expression in adult human differentiated cells challenges its role as a pure stem cell marker. Stem Cells.
2007;25(7):1675-80.
• Marrazzo E, Marchini S, Previdi S, Broggini M. Questioning the oncogenic role of DeltaNp73alpha in different cell lines
expressing p53 or not. Cancer Biol Ther. 2006;5(7):794-803.
• Polato F, Codegoni A, Fruscio R, Perego P, Mangioni C, Saha S, Bardelli A, Broggini M. PRL-3 phosphatase is
implicated in ovarian cancer growth. Clin Cancer Res. 2005 11:6835-9.
• Maffucci T, Piccolo E, Cumashi A, Iezzi M, Riley AM, Saiardi A, Godage HY,Rossi C, Broggini M, Iacobelli S, Potter
BV, Innocenti P, Falasca M. Inhibition of the phosphatidylinositol 3-kinase/Akt pathway by inositol pentakisphosphate
results in antiangiogenic and antitumor effects. Cancer Res. 2005;65:8339-49.
• Marabese M, Marchini S, Sabatino MA, Polato F, Vikhanskaya F, Marrazzo E, Riccardi E, Scanziani E, Broggini M.
Effects of inducible overexpression of DNp73alpha on cancer cell growth and response to treatment in vitro and in vivo.
Cell Death Differ. 2005;12:805-14.
• Sabatino MA, Colombo T, Geroni C, Marchini S, Broggini M. Enhancement of in vivo antitumor activity of classical
anticancer agents by combination with the new, glutathione-interacting DNA minor groove-binder, brostallicin. Clin
Cancer Res. 2003;9:5402-8.
Irene Floriani got her degree in Biological Sciences at the University of Milan in 1988, her degree in
Biostatistics and Experimental Statistics at the University of Milan in 2003 and her phD in Life Sciences
at Open University of London (UK) in 2005. After ten-year experience in pharmaceutical industry, in
2002 she became Head of the Biometry and Data Management Unit of the Laboratory of Clinical
Research in Oncology and since 2006 she is Head of Laboratory of Clinical Trials. She is also member as
bio-statistician of three Italian Ethics Committees. Her main fields of interest are: statistical aspects of
methodology of clinical research with focus on Controlled Clinical Trials in Oncology; Systematic
Overview of the medical literature and Methodological aspects of diagnostic test evaluation.
Selected publications
• Graziano F, Ruzzo A, Loupakis F, Canestrari E, Santini D, Catalano V, Bisonni R, Torresi U, Floriani I, Schiavon G,
Andreuzzi B, Maltese P, Rulli E, Humar B, Falcone A, Giustini L, Tonini G, Fontana A, Masi Gianluca, Magnani M
Pharmacogenetic profiling for cetuximab/irinotecan therapy in patients with refractory advanced colorectal cancer. J
Clin Oncol 2008 ; 26 : 1427-1434
• Ludovini V, Gori S, Colozza M, Pistola L, Rulli E, Floriani I, Pacifico E, Tofanetti F R, Sidoni A, Basurto C, Rulli A,
Crino' L. Evaluation of serum HER2 extracellular domain in early breast cancer patients: correlation with
clinicopathological parameters and survival. Ann Oncol 2008 ; 19 : 883-890
• Di Costanzo F, Gasperoni S, Manzione L, Bisagni G, Labianca R, Bravi S, Cortesi E, Carlini P, Bracci R, Tomao S,
Messerini L, Arcangeli F, Torri V, Bilancia D, Floriani I, Tonato M, Italian Oncology Group for Cancer Research.
Adjuvant chemotherapy in completely resected gastric cancer: a randomized phase III trial conducted by GOIRC. J Natl
Cancer Inst 2008 ; 100 : 388-398
• Cascinu S, Berardi R, Labianca R, Siena S, Falcone A, Aitini E, Barni S, Di Costanzo F, Dapretto E, Tonini G,
Pierantoni C, Artale S, Rota S, Floriani I, Scartozzi M, Zaniboni A, GISCAD. Cetuximab plus gemcitabine and cisplatin
10
ANNUAL REPORT 2008

IRFMN
compared with gemcitabine and cisplatin alone in patients with advanced pancreatic cancer: a randomised, multicentre,
phase II trial. Lancet Oncol 2008 ; 9 : 39-44
• Santini D, Loupakis F, Vincenzi B, Floriani I, Stasi I, Canestrari E, Rulli E, Maltese P, Andreoni F, Masi Gianluca,
Graziano F, Baldi G G, Salvatore L, Russo A, Perrone G, Tommasino M R, Magnani M, Falcone A, Tonini G, Ruzzo A.
High concordance of KRAS status between primary colorectal tumors and related metastatic sites: implications for
clinical practice. Oncologist 2008 ; 13 : 1270-1275
• Cascinu S, Labianca R, Barone C, Santoro A, Carnaghi C, Cassano A, Beretta GD, Catalano V, Bertetto O, Barni S,
Frontini L, Aitini E, Rota S, Torri V, Floriani I. Adjuvant treatment of high-risk, radically resected gastric cancer
patients with 5-fluorouracil, leucovorin, cisplatin, and epidoxorubicin in a randomized controlled trial.
J Natl Cancer Institute (2007); 99: 601-7
Raffaella Giavazzi got her Biological Sciences degree (1979) at the University of Milan, and her Ph.D. in
Pharmacology at the Mario Negri Institute of Milan (1984), followed by a specialization in pharmacology
(1994) at the University of Milan. From 1981 to 1983 she was a Fellow in the Cancer Metastasis and
Treatment Laboratory, NCI-FCRDC, Frederick, MD., and from 1983 to 1985 Assistant Professor at the
Department of Cell Biology of M.D. Anderson Hospital and Tumour Institute, University of Texas System
Cancer Centre in Houston (TX).
Raffaella Giavazzi’s research interests are in the field of tumour biology and pharmacology. Specifically, she is
studying aspects related to the metastatic process and angiogenesis. She is involved in the pre-clinical
evaluation of new therapeutic strategies against cancer focusing on the angiogenesis inhibitors and combination
therapies.
From 1986 to 1993 she was Head of the Cancer Metastasis Treatment Unit and since 1993 she has been the
Head of the Laboratory of Biology and Treatment of Metastasis at Mario Negri Institute for Pharmacological
Research.
She is also adjutant Professor in Oncology, Medical School-University of Brescia, member of the Teaching
Committee for the PhD course in Physiology-Pharmacology-Molecular and Cellular Tossicology-University of
Siena, member of the Executive Committee at SENDO (South Europe New Drug Development Organization),
member of the Pezcoller Foundation Scientific Committee and member of the Executive Committee of the
European Association for Cancer Research (EACR).
She was consulting scientist for the NCI-Drug Therapeutics Program, USA (1996-2006);
She is a member of the American Association for Cancer Research (AACR), International Metastases Research
Society, EORTC-Screening and Pharmacology Group, European Association for Cancer Research (EACR),
Italian Cancer Society (SIC) of which she was President (2006-2007).
She is on the Editorial Board of the European Journal of Cancer, Journal of Clinical Experimental Metastasis,
Journal Exp. Therapeutic & Oncology, The International Journal of Biological Markers, Current Cancer
Therapy Reviews and Journal of Chemotherapy.
She is on the Editorial Board of several international scientific journals. She has published approximately
200 articles on “peer reviewed” scientific journals.
Selected publications
• Belotti D., Calcagno C., Garofalo A., Caronia D., Riccardi E., Giavazzi R., Taraboletti G. Vascular Endothelial Growth
Factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian cancer invasion. Molecular
Cancer Research, 6(4):525-34, 2008.
• Valentini G., D’Andrea C., Ferrari F., Pifferi A., Cobeddu R., Martinelli M., Natoli C., Ubezio P., Giavazzi R. In-vivo
measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. Photochemistry and
Photobiology, 84(5):1249-56, 2008.
• Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through
gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, 2008.
• Martinelli M., Bonezzi K., Riccardi E., Kuhn E., Frapolli R., Zucchetti M., Ryan A.J., Taraboletti G., Giavazzi R.:
Sequence dependent antitumour efficacy of the vascular disrupting agent ZD6126 in combination with paclitaxel.
British Journal of Cancer 97:888-94, 2007.
• Giavazzi R., Bani M.R.,Taraboletti G.: Tumor–host interaction in the optimization of paclitaxel-based combination
therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:481–88, 2007
• Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R., Giavazzi
R.: The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor, causing
apoptosis of endothelial cells and inhibition of angiogenesis. Clin. Cancer Research 12(6):1839-49, 2006.
Paola Mosconi got his Biological Science degree (Milan 1982) and the specialisation in Pharmacological
Research (Milan 1984). Her main areas of interest are:
• development of strategies to involve patients-consumer associations in health debate, and research
projects;
• assessment of quality of life, translation and cultural adaptation of questionnaires for quality of life;
11
ANNUAL REPORT 2008

IRFMN
• studies to evaluate the type of information on diseases and treatments received by patients, mainly
in cancer patients; set-up of websites targeted on consumers/patients www.partecipasalute.it,
www.paincare.it, www.fondazione Mattioli.it ;
• studies to evaluate the consumers’ level of satisfaction with the health services and cure.
Paola Mosconi has participated as a teacher, or coordinator, to the realization of training course on
“Methodological aspects of clinical research” or “Evaluation of quality of life” for health care
professionals and representatives of voluntary associations.
Selected publications
• Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M,
Tempia P, Guala A, Fasolo G, Riva E. Association of mild anemia with cognitive, functional, mood and quality of life
outcomes in the elderly: the “health and anemia” study. PlosONE April 2008, Vol 3, Issue 4, e1920: 1-8
• Mosconi P, Apolone G, Mingardi G. Quality of life assessment and instruments in end-stage renal disease. Journal of
Nephrology 2008; 21(suppl 13): S107-S112
• Mosconi P, Colombo C, Guella F, Pierotti B, Vimercati F. Are Italian medical societies bridging the distance from
citizen and patients associations Result of a survey. Journal of Preventive Medicine and Hygiene 2008: 3
• Mosconi P, Colombo C, Satolli R, Liberati A. PartecipaSalute, an Italian project to involve lay people, patients’
associations and scientific-medical representatives on the health debate. Health Expectations 10: 194-204, 2007.
• Mosconi P, Satolli R, Colombo Cinzia, Liberati A, Donati S, Mele A. Hormone replacement therapy and information: in
Italy a Consensus Conference to help woman decision. British Medical Journal, 2007
http://www.bmj.com/cgi/eletters/335/7613/239#174012
Valter Torri got his Medical degree in 1985 and the specialization in medical Oncology in 1989 at the
University of Milano.
Education: 1985: MD Degree with full honors cum Laude, University of Milano; 1988 Post-Doctoral
Degree in Pharmacological Research, Mario Negri Institute, Milano; 1989 Post-Doctoral Degree in
Medical Oncology, University of Milano; 1989-1991 Research Fellow at the Biometric Research Branch
of Cancer Treatment Evaluation Program, NCI, Bethesda, MD (USA)
Areas of Interest: Statistical aspects of clinical research methodology with focus on Controlled Clinical
Trials in Oncology; Systematic Overview of the medical literature; Methodological aspects of diagnostic
test evaluation.
Present Position: Head of Laboratory of Clinical Research In Oncology, Oncology Department, Mario
Negri Institute, Milano.
Chronology of Professional Appointments: 1983-1985: Clinical research Fellow in Internal Medicine at
the University Hospital, University of Milan; 1985-1989: Research assistant at the Clinical Trial Unit of
the Laboratory of Clinical Epidemiology, Mario Negri Institute for Pharmacological Research, Milano;
1989-1991: Research fellow at the Biometric Research Branch of Cancer Treatment Evaluation Program,
NCI, Bethesda, MD (USA); 1994: Head of Biometric Unit of the Laboratory of Cancer Clinical
Epidemiology, Oncology Department, Mario Negri Institute for Pharmacological Research, Milano, Italy;
1995 Vice Director of the Italian “Cochrane” Center; 2001: Head of Laboratory of Clinical Research In
Oncology, Oncology Department, Mario Negri Institute, Milano. 2006: Head of Laboratory for the
development of new pharmacological strategies , Oncology Department, Mario Negri Institute, Milano.
.
Selected publications
• Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M,
Dell'Anna T, Apolone G, Broggini M, D'Incalci M. Analysis of gene expression in early-stage ovarian cancer. Clin
Cancer Res. 2008 Dec 1;14(23):7850-60.
• Benedetti Panici P, Basile S, Maneschi F, Alberto Lissoni A, Signorelli M,Scambia G, Angioli R, Tateo S, Mangili G,
Katsaros D, Garozzo G, Campagnutta E,Donadello N, Greggi S, Melpignano M, Raspagliesi F, Ragni N, Cormio G,
Grassi R,
Franchi M, Giannarelli D, Fossati R, Torri V, Amoroso M, Crocè C, Mangioni C. Systematic pelvic lymphadenectomy
vs. no lymphadenectomy in early-stage endometrial carcinoma: randomized clinical trial. J Natl Cancer Inst. 2008 Dec
3;100(23):1707-16.
• Mitry E, Fields AL, Bleiberg H, Labianca R, Portier G, Tu D, Nitti D, Torri V,Elias D, O'Callaghan C, Langer B,
Martignoni G, Bouché O, Lazorthes F, Van Cutsem E, Bedenne L, Moore MJ, Rougier P. Adjuvant chemotherapy after
potentially curative resection of metastases from colorectal cancer: a pooled analysis of two randomized trials. J Clin
Oncol. 2008 Oct 20;26(30):4906-11.
• Cascinu S, Labianca R, Barone C, Santoro A, Carnaghi C, Cassano A, Beretta GD, Catalano V, Bertetto O, Barni S,
Frontini L, Aitini E, Rota S, Torri V, FlorianiI; Italian Group for the Study of Digestive Tract Cancer, Adjuvant
treatment of high-risk, radically resected gastric cancer patients with 5-fluorouracil, leucovorin, cisplatin, and
epidoxorubicin in a randomised controlled trial. J Natl Cancer Inst. 2007;99(8):601-7.
12
ANNUAL REPORT 2008

IRFMN
• Graziano F, Kawakami K, Ruzzo A, Watanabe G, Santini D, Pizzagalli F, Bisonni R, Mari D, Floriani I, Catalano V,
Silva R, Tonini G, Torri V, Giustini L, Magnani M Methylenetetrahydrofolate reductase 677C/T gene polymorphism,
gastric cancer susceptibility and genomic DNA hypomethylation in an at-risk Italian population. Int J Cancer 2006 118 :
628-632
• Torri V: Clinical trials and data management In: Oxford textbook of oncology, 2nd. ed. Vol. 1. Oxford Univ. Press,
Oxford; 2002 : 1123-1134
Maria Rosa Bani got her Biological Sciences degree at the University of Milan in 1998 attaining the
Italian Government Qualification to practice as Biologist in 1990. She obtained the specialization in
Pharmacological Research from the Department of Education of the Regional Government of Lombardia
in 1991 and the specialization in Biomedical Research from the Department of Education of the Regional
Government of Abruzzo in 1993.
In 2005 she was awarded the degree of Doctor of Philosophy (PhD), Discipline of Life Sciences of the
Open University Research School (UK).
From 1991 to 1995 she was a Post Doctoral Fellow in the Cancer Research Division, Sunnybrook Health
Science Centre, University of Toronto (Canada); from 2000 to 2001 she was Guest Scientist at the
Advance Technology Centre, National Cancer Institute, National Institute of Health (USA).
At the MarioNegri Institute for Pharmacological Research she was a Fellow Research Scientist in the
Laboratory of Biology and Treatment of Metastasis, Bergamo from 1996 to 2002 and she became a Staff
Research Scientist in 2003. In April 2004 she became Head of the Molecular Cancer Therapeutics Unit.
She is the Scientific Manager of STROMA(since 2004) and ADAMANT(since2008), two Integrated
Projects within the 6 th and 7 th Framework Programs of the European Commission.
She is a member of the American Association for Cancer Research (AACR), the European Association
for Cancer Research (EACR) and the Italian Cancer Society (SIC).
Maria Rosa Bani research interests are in the field of cancer biology and molecular therapeutics.
She is co-author of 34 peer reviewed publications, 2 book chapters and 65 proceedings of which 15
selected for oral presentation at international meetings.
Selected publications
• Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through
gene expression profiling of tumor-derived endothelium. BMC Genomics, 30(9), 201, 2008.
• Giavazzi R., Bani M.R.,Taraboletti G.: Tumor–host interaction in the optimization of paclitaxel-based combination
therapies with vascular targeting compounds. Cancer Metastasis Rev. 26:481–88, 2007.
• Naumova E., Ubezio P., Garofalo A., Borsotti P., Cassis L., Riccardi E., Scanziani E., Eccles S.A., Bani M.R. &
Giavazzi R. The vascular targeting property of paclitaxel is enhanced by SU6668, a receptor tyrosine kinase inhibitor,
causing apoptosis of endothelial cells and inhibition of angiogenesis. Clinical Cancer Research 12: 1839-1849, 2006.
• Bani M.R., Nicoletti M.I., Alkharouf N.W., Ghilardi C., Petersen D., Erba E., Sausville E.A., Liu E.T. and Giavazzi R.
Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts. Molecular Cancer Therapeutics
3: 111-121, 2004.
• Vikhanskaya F.*, Bani M.R.*., Borsotti P., Ghilardi C., Ceruti R., Ghisleni G., Marabese M., Giavazzi R., Broggini M.
& Taraboletti G. p73 overexpression increases VEGF and reduces thrombospondin-1 production: implication for tumor
angiogenesis. Oncogene 20 : 7293-7300, 2001.
• Taraboletti G., Sonzogni L., Vergani V., Hosseini G., Ceruti R., Ghilardi C., Bastone A., Toschi E., Borsotti P.,
Scanziani E., Giavazzi R., Pepper M.S., Stetler-Stevenson W.G. & Bani M.R. Post-transcriptional stimulation of
endothelial cell matrix metalloproteinases 2 and 1 by endothelioma cells. Experimental Cell Research 258 : 384-394,
2000.
Giovanna Damia obtained her Medical Degree cum Laude from the University of Milan in 1985. After
specializing in Pharmacology at the Mario Negri Institute of Milan and in Oncology at the University of
Milan, she worked as a post-doctoral fellow in the Laboratory of Experimental Immunology of the
National Cancer Institute, Frederick, USA. She worked as a research fellow in the Laboratory of Cancer
Chemotherapy at the Mario Negri Institute and since April 2003 she has become chief of the DNA Repair
Unit at the Mario Negri Institute. From 1992 to1995 she has been consultant of the General Secretariat of
the Progetto Finalizzato CNR "Applicazioni Cliniche della Ricerca Oncologica". Since September 2005
she is Deputy Editor for Experimental Oncology of the European Journal of Cancer.
Her main fields of interest are: mechanism of action of anticancer drugs, cell cycle checkpoints
and natural compounds.
Selected publications
• Simone M, Erba E, Damia G, Vikhanskaya F, Di Francesco A M, Riccardi R, Baldeyrou B, Bailly C, Cuevas C, Sousa-
Faro J M F, D'Incalci M. Variolin B and its derivate deoxy-variolin B: New marine natural compounds with cyclindependent
kinase inhibitor activity. Eur J Cancer 2005; 41: 2366-2377
13
ANNUAL REPORT 2008

IRFMN
• Carrassa L, Broggini M, Erba E, Damia G. Chk1, but not Chk2, is involved in the cellular response to DNA damaging
agents. Differential activity in cells expressing or not p53. Cell Cycle 2004; 3: 1177-1181
• Damia G, Broggini M. Improving the selectivity of cancer treatment by interfering with cell response pathways. Eur J
Cancer 2004; 40: 2550-2559
• Damia G, Broggini M. Cell cycle checkpoint proteins and cellular response to treatment by anticancer agents. Cell Cycle
2004; 3: 46-50
• Carrassa L, Broggini M, Vikhanskaya F, Damia G. Characterization of the 5' flanking region of the human chk1 gene.
Identification of E2F1 functional sites. Cell Cycle 2003; 2: 604-609
• Damia G, Sanchez Y, Erba E, Broggini M. DNA damage induces p53-dependent down-regulation of hCHK1. J Biol
Chem 2001; 276: 10641-10645
Eugenio Erba has obtained his Biological and Biochemmistry Analysis Degree at the University of
Urbino. He worked as a research fellow in the Laboratory of Cancer Chemotherapy at the Mario Negri
Institute and since 1984 he is head of the Flow Cytometry Unit in the Department of Oncology at the
Mario Negri Institute of Milan. He has worked as a visiting fellow in the Department of Istochemistry
and Cytochemistry of the University of Leiden, The Netherlands in 1983. Since 1997 he is Teacher of
Post-Graduate Studies in Cytometry at the University of Milan and Co-ordinator and Teacher of Post-
Graduate Studies in Cytometry for the Italian Cytometry Group. He has been President of the Italian
Cytometry Group from 1999 to 2001. Since 2001 he is member of the Executive Board of the Italian
Cytometry Group.
Scientific areas of interest: studies on the mechanism of action of different compounds with provided
antitumoral activity evaluating the mechanism of cell death and cell cycle phase perturbations induced
on different human cancer cell lines by using flow cytometry. Co-ordinator of working-group in a
quality control study on flow cytometric DNA content analysis in human tumors.
Selected publications
• C. Valli, G. Paroni, A. Di Francesco, R. Riccardi, M. Tavecchio, E. Erba, A. Boldetti, M. Giannì, M. Fratelli, C. Pisano,
L. Merlini, A. Antoccia, C. Cenciarelli, M. Terao, E. Garattini. Atypical retinoids ST1926 and CD437 are S-phase
specific agents causing DNA double-strand breaks: significance for the cytotoxic and antiproliferative activity. Mol. Ca.
Ther. 7(9),2941-54, 2008
• L. Roncoroni, L. Elli, E. Delfini, E. Erba, E. Dogliotti, C. Terrai, L. Doneda, MG. Grimoldi, MT. Bardella. Resveratrol
inhibits cell growth in a human cholangiocarcinoma cell line. Liver Int. 28(10),1426-36, 2008
• M.Tavecchio, M. Simone, E.Erba, I. Chiolo, G. Liberi, M. Foiani, M. D’Incalci, G. Damia. Role of homologous
recombination in trabectedin-induced DNA damage. Eur. J. Ca 44:609-618 (2008)
• Paulis M., Bensi M., Orioli D., Mondello C., Mazzini G., D’Incalci M., Falcioni C., Radaelli E., Erba E.,
Raimondi E., De Carli L. Transfer of a Human Chromosomal Vector from a Hamster Cell Line to a Mouse
Embryonic Stem Cell Line. Stem Cell , 25:2543-2550 (2007)
• Tavecchio M., Natoli C., Ubezio P., Erba E., D’Incalci M. Dynamics of cell cycle perturbations by trabectedin
(ET-743) in nucleotide excision repair (NER) –deficient and NER-proficient cells, unravelled by a novel
mathematical simulation approach. Cell Prolif., 40:885-904 (2007)
• Tognon G., Bernasconi S., Celli N., Faircloth G.T. Cuevas C., Jimeno J., Erba E., D’Incalci M. Induction of
resistance to Aplidin® in a human ovarian cancer cell line related to MDR expression. Cancer Biology and
Therapy, 4(12): 1325-1330 (2005).
Roldano Fossati got his Medical Degree cum Laude from the University of Milan in 1980, his Post-
Doctoral Degree in Endocrinolgy cum Laude from the University of Verona in 1983 and his Post-
Doctoral Degree in Medical Statistics from the University of Milan in 1992. He has been consultant at
the Mario Negri Institute since 1983 and, at present, he is head of the Gynecology and Oncology Unit of
the Laboratory of Translational and Outcome Research.
Areas of Interest: Statistical and methodologic aspects of clinical research with focus on Controlled
Clinical Trials in Oncology; Systematic Overview of the medical literature.
Selected publications
• Labianca R., Fossati R. , Zaniboni A., Torri V., Marsoni S., Nitti D., Boffi L., Scatizzi M., Tardio B., Mastrodonato N.,
Banducci S., Consani G., Pancera G. on behalf of ACOI/GIVIO/GISCAD investigators. Randomized Trial of Intraportal
and/or Systemic Adjuvant Chemotherapy in Patients with Colon Carcinoma. J Natl Cancer Inst 96:750-8;2004
• P. Benedetti Panici, A. Maggioni, N. Hacker, F. Landoni, S. Ackermann, E. Campagnutta, K. Tamussino, R. Winter, A.
Pellegrino, S. Greggi, R. Angioli, N. Manci, G. Scambia, T. Dell'Anna, R. Fossati, I. Floriani, R.S. Rossi, R. Grassi, G.
Favalli, F. Raspagliesi, D. Giannarelli, L. Martella, C. Mangioni. Systematic Aortic and Pelvic Lymphadenectomy versus
Resection of Bulky Nodes Only in Optimally Debulked Advanced Ovarian Cancer: A Randomized Clinical Trial J Natl
Cancer Inst 97:1-6;2005
• Buda A, Fossati R, Colombo N, Fei F, Floriani I, Gueli Alletti D, Katsaros D, Landoni F, Lissoni A, Calzoni C, Sartori E,
Scollo P, Torri V, Zola P, Mangioni C. Randomized trial of neoadjuvant chemotherapy comparing paclitaxel, ifosfamide,
14
ANNUAL REPORT 2008

IRFMN
and cisplatin with ifosfamide and cisplatin followed by radical surgery in patients with locally advanced squamous cell
cervical carcinoma: The SNAP01 (Studio Neo-Adjuvante Por. J Clin Oncol 2005; 23: 4137-4145.
• Maggioni A, Benedetti Panici P, Dell'anna T, Landoni F, Lissoni A, Pellegrino A, Rossi RS, Chiari S, Campagnutta E,
Greggi S, Angioli R, Manci N, Calcagno M, Scambia G, Fossati R, Floriani I, Torri V, Grassi R, Mangioni C.Randomised
study of systematic lymphadenectomy in patients with epithelial ovarian cancer macroscopically confined to the pelvis. Br
J Cancer. 2006 Sep 18;95(6):699-704.
• Maggi R, Lissoni A, Spina F, Melpignano M, Zola P, Favalli G, Colombo A, Fossati R. Adjuvant chemotherapy vs
radiotherapy in high-risk endometrial carcinoma: results of a randomised trial. Br J Cancer. 2006 Aug 7;95(3):266-71
• Fruscio R, Colombo N, Lissoni AA, Garbi A, Fossati R, Ieda' N, Torri V, Mangioni C.A phase II randomised clinical trial
comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and epirubicin in newly diagnosed advanced
epithelial ovarian cancer: long-term survival analysis. Br J Cancer. 2008 Feb 5.
Giulia Taraboletti got her degree cum laude in Biological Sciences at the University of Pavia (Pavia,
Italy) in 1983, and the specialization in Pharmacological Research at the Mario Negri Institute, Milano,
Italy in 1986. From 1986 to 1988 she was a post-doctoral fellow at the Laboratory of Pathology, NCI,
NIH, Bethesda, MD, and from 1988-1995 research scientist at Mario Negri Institute in Bergamo, Italy.
Since 1995 she is Head of the Unit of Tumor Angiogenesis, at Mario Negri Institute, in Bergamo.
Research interests include tumor angiogenesis, endogenous inhibitors of angiogenesis (thrombospondin-
1) and preclinical studies of antiangiogenic and vascular disrupting compounds, including tubulintargeting
agents. She is member of Metatasis Research Society (MRS), American Association for Cancer
Research (AACR), European Association for Cancer Research (EACR), and the Italian Society of
Oncology (SIC). She is on the editorial board of European Journal of Cancer.
Selected publications
• Margosio B, Rusnati M, Bonezzi K, Cordes B-lA, Annis DS, Urbinati C, Giavazzi R, Presta M, Ribatti D, Mosher DF,
and Taraboletti G. Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic
domain. Int J Biochem Cell Biol 40: 700-709, 2008.
• Giavazzi R., Bani M.R.,Taraboletti G. Tumor–host interaction in the optimization of paclitaxel-based combination
therapies with vascular targeting compounds. Cancer Metastasis Rev, 26:481–88, 2007.
• Martinelli M., Bonezzi K., Riccardi E., Kuhn E., Frapolli R., Zucchetti M., Ryan A.J., Taraboletti G., Giavazzi R.
Sequence dependent antitumour efficacy of the vascular disrupting agent ZD6126 in combination with paclitaxel. Br J
Cancer 97:888-94, 2007.
• Margosio B., Marchetti D., Vergani V., Giavazzi R., Rusnati M., Presta M., and Taraboletti G. Thrombospondin-1 as a
scavenger for matrix-associated fibroblast growth factor-2. Blood 102: 4399-4406, 2003.
• Taraboletti G. Micheletti G, Rieppi M, Poli M, Turatto M, Rossi C, Borsotti P, Roccabianca P, Scanziani E, Nicoletti MI,
Bombardelli E, Morazzoni P, Riva A, and Giavazzi R. Antiangiogenic and antitumor activity of IDN 5390, a new taxane
derivative. Clin Cancer Res. 8: 1182-1188, 2002
• Taraboletti G., Morbidelli L., Donnini S., Parenti A., Granger H.J., Giavazzi R., and Ziche M.The heparin binding 25
kDa fragment of thrombospondin-1 promotes angiogenesis and modulates gelatinase and TIMP-2 production in
endothelial cells. FASEB J., 14: 1674-1676, 2000.
Paolo Ubezio got his B.Sc. degree in Physics at the University of Milan, in 1982, and the specialisation
in Pharmacological Research Specialist" at the Mario Negri Institute for Pharmacological Research in
1986.
Main activities are: i) Study of cell-cycle mathematical models; ii) Development of flow cytometric
methods; iii) Optimization of anticancer drug scheduling.
Since 1991 is Head of the Unit of Biophysics at the Mario Negri Institute
Selected publications
• Basse, B., Ubezio, P. (2007) A generalised age and phase structured model of human tumour cell populations both
umperturbed and exposed to a range of cancer therapies. Bull. Math. Biol. 69:1673-90.
• Lupi, M., Matera, G., Natoli, C., Colombo, V., Ubezio, P. (2007) The Contribution of p53 in the Dynamics of Cell Cycle
Response to DNA Damage Interpreted by a Mathematical Model. Cell Cycle 6:943-950.
• Lupi, M., Matera, G., Branduardi, D., D'Incalci M. and Ubezio, P. (2004) Cytostatic and cytotoxic effects of topotecan
decoded by a novel mathematical simulation approach. Cancer Res. 64: 2825-2832.
• Matera, G., Lupi, M.,and Ubezio, P. (2004) Heterogeneous cell response to topotecan in a CFSE-based proliferation test.
Cytometry 62A:118-128.
• Ubezio, P. (2004) Unraveling the complexity of cell cycle effects of anticancer drugs in cell populations.Discrete and
Continuous Dynamical Systems-Series B 4:323-335.
• Montalenti, F., Sena, G., Cappella, P., and Ubezio, P. (1998) Simulating cancer-cell kinetics after drug treatment:
Application to cisplatin on ovarian carcinoma. Phys. Rev. E, 57:5877-5887.
Massimo Zucchetti obtained his Chem. Pharm. Degree from the University of Milan in 1982. After
specializing in Pharmacology at the Mario Negri Institute of Milan (1988), he worked in the Laboratory
15
ANNUAL REPORT 2008

IRFMN
of Clinical Pharmacology of Department of Oncology at San Giovanni Hospital, Bellinzona, Switzerland
(1988-1990). Since 1996 he has been chief of the Cancer Clinical Pharmacology Unit at the Mario Negri
Institute. He is member of the Pharmacology and Molecular Mechanisms Group of the European
Organization for Research and Treatment of Cancer (EORTC) from 1988 up to date. His main field of
interest are:
- Clinical pharmacology, phase I and Phase II studies
- Analysis of drugs, pharmacokinetic and pharmacodynamic studies in humans in GCP and GLP
conditions
- Pharmacokinetic and metabolic studies in animals
- Pharmacokinetic drug interaction
Dr Zucchetti is author of more than 80 papers on pre-clinical and clinical cancer chemotherapy published
in peer reviewed international journals.
Selected publications
• Gambacorti-Passerini CB, Tornaghi L, Marangon E, Franceschino A, Pogliani EM, D'Incalci M, Zucchetti M.. Imatinib
concentrations in human milk Blood. 2007 Feb 15;109(4):1790.
• Marangon E, Sala F, Caffo O, Galligioni E, D'Incalci M, Zucchetti M. Simultaneous determination of gemcitabine and
its main metabolite, dFdU, in plasma of patients with advanced non-small-cell lung cancer by high-performance liquid
chromatography-tandem mass spectrometry. J Mass Spectrom. 2008 Feb;43(2):216-23.
• Frapolli R., Marangon E., Zaffaroni M., Colombo T., Falcioni C., Bagnati R., Simone M., D’Incalci M., Manzotti C.,
Fontana G., Morazzoni P., Zucchetti M. Pharmacokinetics and metabolism in mice of IDN 5390 (13-(N-Boc-3-ibutylisoserinoyl)-C-7,8-seco-10-deacetylbaccatin
III), a new oral C-seco-taxane derivative with antiangiogenic property
effective on paclitaxel-resistant tumors. Drug Metabolism and Disposition, 34(12):2028-2035 (2006).
• Rizzari C., Citterio M., Zucchetti M., Conter V., Chiesa R., Colombini A., Malguzzi S., D’Incalci M. Pharmacological
study on pegylated asparaginase used in front-line treatment of children with acute lymphoblastic leukemia.
Hematologica, 91: 24-31 (2006).
• Fruscio R., Lissoni A.A., Frapolli R., Corso S., Mangioni C., D’Incalci M., Zucchetti M. Clindamycin-Paclitaxel
pharmacokinetic interaction in ovarian cancer patients. Cancer Chemother. Pharmacol., 58(3): 319-325 (2006).
• Gambacorti Passerini C, Zucchetti M, Russo D, Frapolli R, Verga M, Bungaro S, Tornaghi L, Rossi F, Pioltelli P,
Pogliani E, Alberti D, Corneo G, D'Incalci M Alpha 1 acid glycoprotein binds to imatinib (STI571) and substantially
alters its pharmacokinetics in chronic myeloid leukemia patients Clin Cancer Res 2003; 9: 625-632
16
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Oncology Department comprises three preclinical experimental laboratories (Laboratory of
Cancer Pharmacology, Laboratory of Molecular Pharmacology and Laboratory of Biology and
Therapy of Metastasis) and four laboratories dealing with clinical research and clinical trials
(Laboratory for the Development of New Pharmacological Strategies, Laboratory of Clinical
Trials, Laboratory of Translational and Outcome Research in Oncology and Laboratory for
Medical Research and Consumer Involvement).
The Oncology department hosts the coordination center of two networks of hospitals that carry
on clinical research in gynecologic cancer (MaNGO: Mario Negri Gynecologic Oncology) and
in cancer pain (CPOR-SG: Cancer Pain Outcome Research Study Group) and a center for
cancer pain assessment and research (CERP:Center for the Evaluation and Research on Pain).
In some cases research projects are carried out by single laboratories or research units, in other
cases by collaborations between different laboratories of the Oncology Department or other
departments, or other groups outside the Institute (see National and International
Collaborations).
Preclinical laboratories focus on the discovery and development of new antitumor and
antimetastatic drugs and their new combinations; on tumor biology, not only to acquire new
scientific knowledge, but particularly as a base for more selective therapeutic approaches and to
identify and evaluate experimental models for discovering and studying new drugs or
treatments.
Clinical new drug development involves close participation in the activity of SENDO (South
Europe New Drug Development Organization) and studies driven by the Laboratory of Cancer
Pharmacology, the Laboratory of Molecular Pharmacology and the Laboratory of Biology and
Therapy of Metastasis. The Laboratory for the Development of New Pharmacological
Strategies, the Laboratory of Clinical Trials, the Laboratory of Translational and Outcome
Research in Oncology and the Laboratory for Medical Research and Consumer Involvement are
involved in the evaluation of the effects of new therapeutic modalities in phase I/II and in phase
III comparative and effectiveness outcome studies.
Outcome Research implies organizing trials to clarify the results of certain health care practices
and interventions in clinical practice. Observational (surveys) and outcome research
(effectiveness) studies are carried out, in collaboration with regional and national health
authorities and other scientific associations.
At the preclinical and clinical level there are studies of various human tumors, with particular
emphasis on ovarian tumors and more recently on soft tissue sarcomas.
FINDINGS/MAIN RESULTS
At nanomolar concentrations, ET-743 (Yondelis, Trabectedin) affects the regulatory
mechanisms of the transcription. Nucleotide excision repair deficient cells that are
hypersensitive to UV rays and to other DNA damaging drugs are resistant to Trabectedin.
The selective activity of ET-743 against human myxoid liposarcoma appears related to the drug
ability to modulate the transcription of genes involved in adipocytic differentiation.
Use of mathematical models of tumor growth and anticancer treatment to interpret experimental
data and to manage the complexity of underlying biological phenomena.
17
ANNUAL REPORT 2008

IRFMN
The theoretical relationship between proliferation, quiescence and cell loss leading to growth
control of tumor cell populations was found.
Gene profiling analysis shows specific molecular signatures according to the histotype and
prognosis of stage I ovarian carcinoma.
The expression of a truncated form of p63 (DNp63) increases with the increased malignancy of
ovarian cancer. Patients expressing high levels of DNp63 have a worst prognosis. DNp63
represents therefore a new potential target for selective therapies in this malignancy.
CHK1 downregulation by specific inhibitors or siRNA, increases the antitumor activity of 5-
fluorouracil in vivo. This effect is particularly evident in p53 deficient tumors indicating that
this combination increases the selectivity of this anticancer agent.
An anthracycline derivative, Nemorubicin, has a peculiar mechanism of action and is active
against tumors resistant to drugs such as cisplatin. A mechanism of resistance against this drug
has been identified, which involves the abrogation of the expression of a nucleotide excision
repair gene.
Overexpression of a truncated form of p73 (DNp73beta) in human cancer cells lead to growth
arrest and formation of tetraploid cells, suggesting a role of this isoform in mitosis.
A new mechanism of p73 activation mediated by the protease HtrA2 has been identified. After
apoptotic stimuli this protease cleaves p73 in the C-terminal region increasing its apoptotic
potential.
Inositol pentaphosphate analogues interfere with the PI3-kinase-induced phosphorylation of akt
and possess antitumor activity in vitro and in vivo, particularly when combined with other
anticancer agents.
Human umbilical cord-derived stem cells express checkpoints proteins only in specific
differentiation stages. It is likely that this is related to the different susceptibility of the cells.
Inhibition of PLC gamma, through siRNA technology, reduces the in vivo growth of tumors and
reduces the formation of metastasis.
Identification of genes preferentially expressed by tumor associated endothelial cells.
VEGF released by cancer cells modulatesgene expression in the tumor microenvironment
(stroma).
VEGF produced by ovarian tumor cells stimulates host MMP9 expression; the anti-VEGF
antibody bevacizumab (Avastin®) inhibited MMP9 expression and abolished ovarian tumor
invasion.
Soluble human VEGFC levels in serum and ascites correlate with tumor progression and
metastatic capability of ovarian carcinoma models.
A new antiangiogenic domain of thrombospondin-1 (an endogenous inhibitor of angiogenesis)
that binds the angiogenic factor FGF-2 has been identified and characterized.
New antineoplastic compounds directed against the tumor vasculature (vascular disrupting
agents) have been identified.
18
ANNUAL REPORT 2008

IRFMN
The histone deacetylase inhibitor SAHA potentiates the cytotoxic effect of paclitaxel in human
ovarian carcinoma cells resistant to paclitaxel. The effect is mediated by the acetylation of
tubulin.
The expression of protease-activated receptor-1 (PAR-1) correlates with the malignant
phenotype of human melanomas and is accountable for their motility and invasive features
ICON4, a, randomised trial of second-line chemotherapy in advanced ovarian cancer,
coordinated by the Mario Negri Institute and by MRC, showed for the first time a reduction in
mortality in favour of platinum and paclitaxel chemotherapy.
The response to chemotherapy was a good surrogate endpoint of survival in patients with
locally advanced cervix carcinoma.
Adjuvant chemotherapy with the regimen vindesin, mitomycin C and cisplatin (MVP) did not
improve survival of non small cell lung cancer (NSCLC) patients compared with surgery alone.
The website of the project PartecipaSalute (www.partecipasalute.it) has a very innovative
character in comparison with the other health Italian sites because introduces and develops with
ad-hoc instruments the information transfer in an active way.
The project PartecipaSalute (www.partecipasalute.it) has an innovative character of the website
panorama in Italy. This web - compared to other Italian health websites - introduces and
develops ad hoc methods to transfer in an active way information on health.
A randomized phase III trial has shown that pelvic systematic lymphadenectomy in early
endometrial cancer does not improve overall survival. As pelvic systematic lymphadenectomy
is not devoid of side effects, this trial will spare patients with early endometrial cancer
important early and late surgical morbidities. A twin trial in ovarian carcinoma, published in
2005, came to similar conclusions.
Bupropion more than doubled the odds of continuous abstinence from smoking from week 4 to
7 and from week 4 to 12 months in a way similar to that observed in academic studies. The
adherence of GPs and participants to the protocol was excellent, making our findings robust and
easy to generalize to the context of primary care.
A randomized phase II trial has shown that the efficacy and tolerability of a chemotherapic
regimen containing paclitaxel and cisplatin (TP) in the neoadjuvant treatment of locally
advanced squamous cell cervical cancer is inferior to the triplet containing also iphosphamide
(TIP). Adding up to previous similar findings, the TIP regimen (TP plus ifosfamide) has been
proving to be one the best chemotherapic regimen in this setting of patients as it can adequately
reduce the cancer volume allowing the surgical ablation with curative intent in most of the
patients.
An observational longitudinal study carried out in 110 Italian centers and involving about 1800
patients with metastatic cancer and pain have documented that in Italy most patients at the time
of study inclusion were still on active anti-cancer treatment (>50%), were not aware of their
prognosis (about 70%). In addition, up to 45% did not received an appropriate analgesic
treatment (under-treatment), especially in some sub-groups.
In terms of analgesics effectiveness, although the observational design, results suggest that each
drugs prescribed by investigators (morphine, fentanyl, buprenorphine and oxycodone) were able
to reduce the intensity of pain of about 2 points on a 11-eleven point numerical rating scale
19
ANNUAL REPORT 2008

IRFMN
(p50%,
and III) endometrial carcinoma showed the substantial equivalence between radiotherapy or
chemotherapy as an adjuvant therapy after surgery. Although both radiotherapic and
chemotherapic approaches are still unsatisfactory, since the risk of progression or death remains
high, this encouraging evidence of clinical activity suggest a possible use of their concurrent or
sequential use in an adjuvant setting.
The estimates of the prevalence and impact of cancer pain in a large and representative sample
of cancer patients (1800) recruited by several Italian centers (more than 120), with the
evaluation of the actual proportion of cases that received a substantial analgesic under-treatment
(about 25%), mainly attributable to a sub-optimal utilization of opiods.
An evaluation of the activity of the EMEA over the last 10 years, has documented that most of
the new anti-cancers drugs has actually received an approval on the basis of very preliminary
findings: in 48% of case the approval was based on studies using surrogate endpoints, and in
40% of cases the design of the study was non-comparative and non-randomized.
The activity of training and information organized with the associations of citizens & patients in
the framework of the PartecipaSalute project has been finalized to the organization of the Parita
task “Participate to the research project with the associations”. Parita is organised to discuss
with the scientific community the grey areas of the medical assistance and clinical research
identified from the patients and their associations, and to develop ad hoc protocols for future
research programs. An ad hoc data collection with the cooperation of patient’s associations has
highlighted the missmacht between patients and researchers agenda.
Development and validation of a new short questionnaire, the PGWBI-short version available to
be used in large sample of citizens or patients.
20
ANNUAL REPORT 2008

IRFMN
NATIONAL COLLABORATIONS
ASR, Agenzia Sanitaria Regionale, Bologna
AIFA, Agenzia Italiana del Farmaco (Roma)
Azienda Sanitaria Locale, Rimini
Assessorato Sanità, Regione Emilia Romagna
Azienda Sanitaria Unica Regionale, Regione Marche
Casa Sollievo della Sofferenza, San Giovanni Rotondo (IRCCS)
CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano
CNR IGBE, Pavia
CNR, Istituto di Chimica del Riconoscimento Molecolare, Milano
Cochrane Collaboration
Fondazione IRCCS Istituto Nazionale dei Tumori (INT), Milano
Fondazione LUVI, Milano
Fondazione Nerina e Mario Mattioli Onlus, Milano
Fondazione Salvatore Maugeri, Pavia
Fondazione SmithKline (FSK), Milano
Fondo Edo Tempia, Laboratorio di Farmacogenomica, Biella
I.A.S.I., Roma
Istituto Clinico Humanitas, Rozzano MI
Istituto Dermopatico dell'Immacolata, Roma
Istituto FIRC per l’Oncologia Molecolare (IFOM)
Istituto Ortopedico Galeazzi, Milano
Istituti Ortopedici Rizzoli, Bologna
Istituto Europeo di Oncologia (IEO), Milano
Istituto di Fisica, Politecnico di Milano
Istituto di Genetica Molecolare CNR, Sezione di Istochimica e Citometria, Pavia
Istituto Nazionale per la Ricerca sul Cancro (IST), Genova
Istituto Nazionale Tumori Fondazione G. Pascale, Napoli
Istituto Regina Elena, Roma
Istituto Superiore di Sanità
Laboratorio Cell factory, Policlinico di Milano
Ospedale San Gerardo, Monza, Milano
Ospedale San Matteo, Pavia
Rete Oncologica Lombarda (ROL), Milano
Unità di Tossicologia e Scienze Biomediche, ENEA Centro Ricerche, Roma
Università Cattolica del Sacro Cuore, Roma
Università di Bari
Università di Brescia
Università di Chieti
Università di L’Aquila
Università di Milano
Università di Milano Bicocca
Università di Modena e Reggio Emilia
Università di Monza
Università di Catania
Università di Padova
Università di Pisa
Università di Siena
21
ANNUAL REPORT 2008

IRFMN
Università “La Sapienza”, Roma
Zadig, Agenzia di Giornalismo scientifico
INTERNATIONAL COLLABORATIONS
ADAMANT Consortium, IP 7th FP, EC
ARCAGY (Association de Recherche sur les Cancers Gynécologiques), Francia
Breakthrough Breast Cancer Center, Instutite of Cancer Reasearch, Londra, Gran Bretagna
Cancer Biomarkers and Prevention Group, University of Leicester, Gran Bretagna
Cancer Research UK, Londra, Gran Bretagna
Cancerdegradome Consortium, IP 6th FP, EC
EORTC, Bruxelles, Belgio
EUROPA DONNA
European Agency for the Evaluation of Medicinal Products (EMEA), Londra, Gran Bretagna
European Association for Palliative Care (EAPC)
Eusoma – (European Society of Breast Cancer Specialist) Firenze, Italy
Executive Board of GCIG (Gynecologic Cancer Intergroup)
Frontier science & technology Research Foundation Southern Europe (FSE)
Genome Institute of Singapore (GIS), Singapore
German Cancer Research Center, Division of Toxicology and Cancer Risk Factors, Heidelberg,
Germania
Goteborg University, Lundberg Laboratory for Cancer Research, Goteborg, Svezia
Helios Klinikum Erfurt GmbH, Institute of Pathology, Germania
Istituto Oncologico della Svizzera Italiana
Johns Hopkins University, USA
Ludwig Institute for Cancer Research, Londra, Gran Bretagna
National Cancer Center, Singapore
Stony Brook University, NY USA
Massachusetts General Hospital and Harvard Medical School, USA
MD Anderson Cancer Center, Houston, Texas, USA
MRC, Londra, Gran Bretagna
National Cancer Institute (NCI), Bethesda and Frederick, MD, USA
Ospedale San Giovanni, Bellinzona, Svizzera
Paterson Institute for Cancer Research, Manchester, Gran Bretagna
Southern Europe New Drug Organization (SENDO), Milano, Italia
Stroma Consortium, IP 6th FP, EC
Swiss Federal Institute of Technology, Zurigo, Svizzera
The Sackler Institute, University College Londra, Gran Bretagna
Tumor Biology and Metastasis Institute of Cancer Research, Sutton, Gran Bretagna
University College, London Medical School, Londra, Gran Bretagna
University of Birmingham, Gran Bretagna
University of Cincinnati, USA
University of Crete Medical School, Greece
University of Newcastle, Gran Bretagna
University of Pau, Francia
University of Wisconsin, Madison, WI, USA
Kyoto University, Giappone
22
ANNUAL REPORT 2008

IRFMN
Weizmann Institute of Science, Israele
EDITORIAL BOARD MEMBERSHIP
Attualità in Senologia (Paola Mosconi)
British Journal of Cancer (Maurizio D’Incalci)
Chemotherapy (Maurizio D’Incalci)
Clinical Experimental Metastasis (Raffaella Giavazzi)
Current Opinion in Oncologic, Endocrine and Metabolic Drugs (Maurizio D’Incalci)
Current Cancer Therapy Reviews (Raffaella Giavazzi)
European Journal of Cancer (Maurizio D’Incalci, Giovanna Damia, Raffaella Giavazzi,
Massimo Broggini e Giulia Taraboletti)
Health and Quality of Life Outcomes (Giovanni Apolone, Paola Mosconi)
International Journal of Biological Markers (Raffaella Giavazzi)
International Journal for Quality in Health Care (Giovanni Apolone)
Journal of Ambulatory Care and Management (Giovanni Apolone)
Journal of B.U.ON. (Maurizio D’Incalci)
Journal of Cancer Microenvironment (Raffaella Giavazzi)
Journal of Chemotherapy (Raffaella Giavazzi)
Journal of Experimental Therapeutics and Oncology (Raffaella Giavazzi)
Journal of Medicine and the Person (Giovanni Apolone)
Journal of Preventive Medicine anf Hygiene (Giovanni Apolone)
Molecular Cancer Therapeutics (Maurizio D’Incalci)
Oncology Research (Maurizio D’Incalci)
Tumori (Maurizio D’Incalci, Raffaella Giavazzi)
www.PartecipaSalute.it (Paola Mosconi)
www.fondazionemattioli.it (Paola Mosconi)
PEER REVIEW ACTIVITIES
American Journal of Pathology, Annals of Oncology, Anti-cancer Drugs, Biochemical
Pharmacology, BioMed Central Editorial, British Journal of Cancer, British Journal of
Pharmacology, British Medical Journal, Cancer Chemotherapy and Pharmacology, Cancer
Detection and Prevention, Cancer Letters, Cancer Research, Carcinogenesis, Chemico-
Biological Interactions, Clinical & Experimental Metastasis, Clinical Cancer Research,
Cytometry, European Journal of Cancer, European Journal of Immunology, Faseb Journal,
Gynecologic Oncology, Health and Quality of Life Outcomes, Health Expectations, European
Journal of Neurology, Intensive Care Medicine, International Journal of Biological Markers,
International Journal of Cancer, International Journal for Quality in Health Care, Journal of
Ambulatory Care and Management, Journal of Biological Chemistry, Journal of Biological
Markers, Journal of Cell Biochemistry, Journal of Clinical Oncology, Journal of Experimental
Therapeutics and Oncology, Journal of Medicine and the Person, Journal of the National Cancer
Institute, Journal of Neurology, Journal of Preventive Medicine and Hygiene, Journal of the
National Cancer Institute, Leukemia, Molecular Cancer Therapeutics, Nature, Nature
23
ANNUAL REPORT 2008

IRFMN
Biotechnology, Nature Reviews, Oncology Research, PharmacoEconomics, Quality of Life
Research, Science, Tumori, ZEG Centre for Epidemiology & Health Research.
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Ethical Committee, Centro di Riferimento Oncologico, Aviano PN, Italy
Ethical Committee, Ente Ospedaliero San Paolo, Milan, Italy
Ethical Committee, Istituto Europeo di Oncologia, Milan, Italy
Ethical Committee, Istituto Neurologico Carlo Besta, Milan, Italy
Ethical Committee, Istituto Scientifico Eugenio Medea, Bosisio Parini, Lecco, Italy
Ethical Committee, Ospedale San Gerardo, Monza, Milan, Italy
Ethical Committee, Ospedale Sant’Anna, Como, Italy
Ethical Committee, Ospedale della Valtellina e Valchiavenna, Sondrio, Italy
Ethical Committee, IRCCS MultiMedica, Sesto San Giovanni, Milan, Italy
Ethical Committee, Azienda USL di Bologna, Italy
Executive Board of GCIG (Gynecologic Cancer Intergroup)
Scientific Committee, Associazione Italiana Ematologia e Oncologia Pediatrica, Monza, Milan,
Italy
Scientific Committee, Pezcoller Foundation, Trento, Italy
Technical-Scientific Commitee, Associazione Italiana per la Ricerca sul Cancro, Milan, Italy
Board of Directors, Fondazione Nerina e Mario Mattioli Onlus, Milan, Italy
Board of Directors, Società Italiana di Cancerologia (SIC)
Board of Directors, Società Italiana di Citometria (GIC)
Directional Council Areas-CCI
National Advisory Board 8th World Congress of Psycho-Oncology
Developmental Therapeutics Program, National Cancer Institute (NCI)
Decision Network and Executive Committee, South Europe New Drug Organization (SENDO)
Executive Board, Europa Donna
Executive Committee, European Asociation for Cancer Research (EACR)
External Scientific Committee, Angiotargeting Consortium – EU Sixth Framework Programme,
University of Bergen, Norvegia
NHS R&D National Coordinating Centre for Health Technology Assessment, UK
Scientific Committee, Swiss Cancer League
University Medical School of Siena, Italy
EVENT ORGANIZATION
Investigator Meeting “Studio ITACAS” Istituto di Ricerche Farmacologiche Mario Negri,
Milan, February 18, 2008.
Investigator Meeting “Studio Head & Neck” Congresso AIOM, Verona, October 12, 2008.
Investigator Meeting “Studio Tailor” Congresso AIOM, Verona, October 12, 2008.
I° Corso di Epidemiologia Clinica e Metodologia della Ricerca, Rimini – Progetto “Il dolore nel
paziente con cancro”, Rimini, October 27 and 28, 2008
24
ANNUAL REPORT 2008

IRFMN
Simposio Satellite “Progetto Il dolore nel paziente con cancro” – XV Congresso Nazionale
Società Italiana di Cure Palliative, Giardini Naxos, November 3/6, 2008.
Workshop Progetto “Fragilità ed Equità in Sanità”, Milan, November 7, 2008
Conferenza di consenso “Quale informazione per la donna in menopausa sulla terapia ormonale
sostitutiva”, Torino, May 16-17, 2008.
International Clinical Trials’ Day. Donne & ricerca clinica, Milan, May 21, 2008.
I corso di formazione per componenti laici di Comitati Etici 2008, Milan, September 2008-
January 2009.
III edizione percorso di formazione “Orientarsi in salute & sanità per fare scelte consapevoli”,.
Milan, September-December 2008.
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
Meeting: 29th Winter Meeting of the Pharmacology and Molecular Mechanisms Group.
"Selective Modulation of Transcription Regulation by Natural Products with Antitumor
Properties". Palermo (Italy), January 30 – February 2, 2008.
Meeting: Cytosquelette microtubulaire & cancer. “Microtubule-affecting agents for vascular
targeting therapy”. Marseil (France), February 26, 2008.
Conference: Gordon Research Conference on Molecular Mechanisms in Lymphatic Function
and Disease. “Ovarian carcinoma xenografts as a model to study lymphangiogenesis”. Ventura
CA, USA, March 2 -7, 2008.
Conference: 1 st International Ovarian Cancer Action. “Angiogenesis: basic aspects”.
“Angiogenesis-preclinical development”. Londra (UK), March 7-8, 2008.
Meeting: AACR Annual Meeting. “The effect of vandetanib on tumor vasculature remodeling
and paclitaxel uptake in a xenograft model of ovarian carcinoma”. San Diego (USA), April 12-
16, 2008.
Meeting: AACR Annual Meeting. “Characterization of predictive markers and target genes of
ProLindac, a novel DACH-platinum compound, compared to oxaliplatin and cisplatin in human
cancer cell lines”. San Diego (USA), April 12-16, 2008.
Meeting: AACR Annual Meeting. “Anti-inflammatory properties of the novel anti-tumor agent
trabectedin: Reduction of ccl2, il-6 and pentraxin-3 by monocytes/macrophages and tumor
cells”. San Diego (USA), April 12-16, 2008.
Congress: XXIV International Congress ISAC: Cytometry in the Age of Systems Biology.
Budapest Sportarena, Budapest, May 17-21, 2008.
25
ANNUAL REPORT 2008

IRFMN
Conference: CancerSim 2008 – Euroconference on Modeling and Simulation of Cancer Growth
and Therapy. “Targeting the tumor vasculature: pharmacological end-points for the design of
combination treatments”. Torino, May 19-21, 2008.
Meeting: Workshop SIICA Angiogenesi: basi molecolari ed implicazioni terapeutiche II.
Certosa di Pontignano, Siena, May 21-23, 2008.
“Pharmacological end-points to develop combination therapies with vascular targeting agents”.
“Lymphangiogenesis in ovarian carcinoma xenografts”.
Meeting: 5th Research Forum of the European Association for Palliative Care (EAPC)
Trondheim, Norway, May 29-31, 2008. Pain in cancer. An Outcome Research Project to
evaluate the epidemiology, the quality and the effects of pain treatment in cancer patients.
Meeting: 9 th International Conference, Angiogenesis: Basic Science and Clinical Applications.
“Pharmacological end-points to develop combination therapies with inhibitors of angiogenesis”.
Patras (Grecia), June 22-26, 2008.
Meeting: 20 th Meeting of the European Association for Cancer Research. “Large scale
comparative proteomic study of accessible vascular proteins in mouse liver metastases and
normal liver”. Lione (Francia), July 5-8, 2008.
Conference: Joint Metastasis Research Society-AACR Conference on Metastasis “Tubulintargeting
agents as inhibitors of cell motility” Vancouver, BC, Canada, August 3-7, 2008.
Congress: IASP - 12 th World Congress on Pain, Glasgow (Scozia), August 17-22, 2008.
Pain in cancer. An Outcome Research Project to evaluate the epidemiology, the quality and the
effects of pain treatment in cancer patients.
Course: XXV Incontro di Aggiornamento e Formazione Scuola Nazionale di Citometria “ La
Citometria nella Clinica e nella Biologia: metologie di base e protocolli applicativi”, Campus
Scientifico Università di Urbino, October 1-4, 2008.
Congress: 50 th Annual Meeting of the Italian Cancer Society. Napoli, October 6-9,
2008.
“Soluble vegfr-1 expression in human melanoma cell lines established from primary or
metastatic lesions”.
“Novel markers of tumor vasculature identified through gene expression analysis of endothelial
cells”.
Meeting: Cancer Degradome Symposium. “Bevacizumab inhibits organ-specific host MMP9
expression and ovarian tumor invasion”. London, UK, October 8-9, 2008.
Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer
Therapeutics. Ginevra (Svizzera), October 21-24, 2008.
“The effects of treatment sequencing on the antitumor activity of vandetanib and paclitaxel in a
xenograft model of human ovarian carcinoma”.
Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer
Therapeutics. Ginevra (Svizzera), October 21-24, 2008.
“Tubulin acetylation and the antimotility effects of tubulin-targeting agents”.
26
ANNUAL REPORT 2008

IRFMN
Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer
Therapeutics. Ginevra (Svizzera), October 21-24, 2008.
“The complexity of cell cycle dynamic of anticancer drugs unraveled by the use of
mathematical models suitable for a quantitative assessment of G1, S and G2M checkpoint
activities”.
Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer
Therapeutics. Ginevra (Svizzera), October 21-24, 2008.
“The novel taxane derivative, IDN6140, crosses the Blood Brain Barrier and has a promising
activity in CNS tumors”.
Simposium: 20th EORTC-NCI-AACR symposium on Molecular Targets and Cancer
Therapeutics. Ginevra (Svizzera), October 21-24, 2008.
“Expression of genes involved in DNA damage response pathways in ovarian cancers”.
Workshop: 2 nd Workshop – CM 0602 Cost Action: Inhibitors of Angiogenesis – Design,
synthesis and biological exploitation. “Angiogenesis inhibitors: strategies for combination
therapies in cancer treatment”. Favignana (Trapani), October 24-26, 2008.
Congress: XV Congresso Nazionale Società Italiana di Cure Palliative, Giardini Naxos
November 3-6, 2008.
Prevalenza dell'undertreatment nel trattamento del dolore da cancro. Risultati di una revisione
sistematica e di uno studio osservazionale.
Conference: Conferenza ECTA2008 : 3 rd European Conference on Tumor Angiogenesis
and Antiangiogenic Therapy. Abano Terme (PD), November 6-8, 2008.
“Molecular profile of the stroma of human ovarian carcinoma xenografts equipped with
different angiogenic phenotypes”.
Preclinical studies on combinations with vascular-targeting and anti-angiogenic agents and
chemotherapy.
Meeting: British Gynaecologic Cancer Society, Liverpool (UK), November 13-14, 2008.
Radical (type II) vs Extrafascial Hysterectomy (type I) for Clinical Stage I Endometrial
Carcinoma: Final Results of an Italian Randomized Clinical Trial.
GRANTS AND CONTRACTS
Arcispedale Santa Maria Nuova di Reggio-Emilia
Azienda Sanitaria Locale - Rimini
Azienda Sanitaria Unica Regionale - MARCHE
CNPDS, Centro Nazionale per la Prevenzione e Difesa Sociale (CNPDS), Milano
Ministero della Salute (Sesto Progetto Integrato Oncologia), Roma
Grunenthal Italia, Milano
AIFA Agenzia Italiana del Farmaco
AIL Associazione Italiana contro le Leucemie, Padova
27
ANNUAL REPORT 2008

IRFMN
Amgem SpA, Milano
AIRC Associazione Italiana per la Ricerca sul Cancro
ASL Padova
ASL Provincia di Lodi
Astra Zeneca SpA
Astra Zeneca UK
AVAPO (Associazione Volontari Assistenza Pazienti Oncologici)
Aventis Pharma
Azienda Ospedaliera Fatebenefratelli e Oftalmico- Milano
Azienda Sanitaria Locale, Rimini
Azienda Sanitaria Unica Regionale, Marche
Azienda Ospedaliera “Spedali Civili di Brescia”
Bracco Imaging SpA, Milan
Centro Cochrane Italiano
Chiesi Farmaceutici SpA
CIPOMO (Collegio Italiano dei Primari Oncologi Medici Ospedalieri)
CNPDS, Centro Nazionale per la prevenzione e Difesa Sociale, Milano
CNR Consiglio Nazionale delle Ricerche
CNR-MIUR Ministero Istruzione Università e Ricerca
Compagnia di San Paolo
CTI Cell Therapeutics, Inc.
Cyclacel Ltd.
Dompé
Eli Lilly Italia SpA
Elsevier Science Ltd.
EORTC-European Organization for Research and Treatment of Cancer
EOS SpA
European Commission - 7th Framework Programme (ADAMANT)
FIRB-MIUR Fondo per gli Investimenti della Ricerca di Base-Ministero Istruzione Università e
Ricerca
FIRC Fondazione Italiana per la Ricerca sul Cancro
Fondazione Cassa di Risparmio delle Province Lombarde
Fondazione Lu.V.I.
Fondazione Nerina e Mario Mattioli Onlus
Fondo Edo Tempia
FSE
GISCAD(Gruppo Italiano Studi di Carcinomi Apparato Digerente)
GlaxoSmithKline, Verona
Grunenthal, Milano
Indena SpA
Institut de Recherche Pierre Fabre
Istituto Clinico Humanitas – Rozzano
Istituto Superiore di Sanità
Italfarmaco
Komen Italia Onlus
Lottomatica
Madaus Srl
Medac
Merck Sharp & Dome
Ministero della Salute
NCI –SAIC Frederick
28
ANNUAL REPORT 2008

IRFMN
Nerviano Medical Science S.r.l.
Novartis
Novartis Farma SpA
Oncoethix
Optigenex Inc.
Pfizer Global Research and Development
Pfizer Italia
Pharma Mar, SA
Pharmatex
Pharminox Ltd, UK
Policlinico di Padova / C.O.R.
PTC Pharma AG
Regione Emilia Romagna
Regione Lombardia
Regione Veneto
Sanofi-Aventis Pharma
Sara Bet, Roma
SENDO-Tech Srl
Sigma-Tau SpA
Università degli Studi di Padova
Università Federico II – Napoli (Dipartimento di Endocrinologia ed Oncologia molecolare e
clinica)
29
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Corica F, Corsonello A, Apolone G, Mannucci E, Lucchetti M, Bonfiglio C, Melchionda N, Marchesini G,
QUOVADIS Study Group
Metabolic syndrome, psychological status and quality of life in obesity: The QUOVADIS study
Int J Obes 2008 32 : 185-191
Marchini S, Marabese M, Marrazzo E, Mariani P, Cattaneo D, Fossati R, Compagnoni A, Fruscio R, Lissoni A A,
Broggini M
ΔNp63 expression associates with poor survival in ovarian cancer
Cancer Res 2008 19 : 501-507
Graziano F, Ruzzo A, Loupakis F, Canestrari E, Santini D, Catalano V, Bisonni R, Torresi U, Floriani I, Schiavon G,
Andreuzzi B, Maltese P, Rulli E, Humar B, Falcone A, Giustini L, Tonini G, Fontana A, Masi Gianluca, Magnani M
Pharmacogenetic profiling for cetuximab/irinotecan therapy in patients with refractory advanced colorectal cancer
J Clin Oncol 2008 26 : 1427-1434
Mosconi P, Apolone G, Mingardi G
Quality of life assessment and instruments in end-stage renal disease
J Nephrol 2008 21 Suppl. 13 : S107-S112
Marangon E, Sala F, Caffo O, Galligioni E, D'Incalci M, Zucchetti M
Simultaneous determination of gemcitabine and its main metabolite, dFdU, in plasma of patients with advanced nonsmall-cell
lung cancer by high-performance liquid chromatography-tandem mass spectrometry
J Mass Spectrom 2008 43 : 216-223
Cooper W N, Dickinson R E, Dallol A, Grigorieva E V, Pavlova T V, Hesson L B, Bieche I, Broggini M, Maher E R,
Zabarovsky E R, Clark G J, Latif F
Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer
Oncogene 2008 27 : 1805-1811
Tavecchio M, Simone M, Erba E, Chiolo I, Liberi G, Foiani M, D'Incalci M, Damia G
Role of homologous recombination in trabectedin-induced DNA damage
Eur J Cancer 2008 44 : 609-618
Marabese M, Marchini S, Marrazzo E, Mariani P, Cattaneo D, Fossati R, Compagnoni A, Signorelli M, Moll U M,
Codegoni A M, Broggini M
Expression levels of p53 and p73 isoforms in stage I and stage III ovarian cancer
Eur J Cancer 2008 44 : 131-141
Marabese M, Mazzoletti M, Vikhanskaya F, Broggini M
HtrA2 enhances the apoptotic functions of p73 on bax
Cell Death Differ 2008 15 : 849-858
Marchini S, Mariani P, Chiorino G, Marrazzo E, Bonomi R, Fruscio R, Clivio L, Garbi A, Torri V, Cinquini M,
Dell'Anna T, Apolone G, Broggini M, D'Incalci M
Analysis of gene expression in early-stage ovarian cancer
Clin Cancer Res 2008 14 : 7850-7860
Belotti D, Calcagno C, Garofalo A, Caronia D, Riccardi E, Giavazzi R, Taraboletti G
Vascular endothelial growth factor stimulates organ-specific host matrix metalloproteinase-9 expression and ovarian
cancer invasion
Mol Cancer Res 2008 6 : 525-534
Apolone G, Patarnello F
The value of a drug: From innovation to the payment via Karl Marx
J Ambul Care Manage 2008 31 : 52-55
Ferketich A K, Gallus S, Colombo P, Fossati R, Apolone G, Zuccaro P, La Vecchia C
Physician-delivered advice to quit smoking among italian smokers
Am J Prev Med 2008 35 : 60-63
30
ANNUAL REPORT 2008

IRFMN
Corsonello A, Lucchetti M, Corica F, Apolone G, Marchesini G
Metabolic syndrome and health-related quality of life: does psychological well-being matter
Ann Epidemiol 2008 18 : 592-593
Ludovini V, Pistola L, Gregorc V, Floriani I, Rulli E, Di Carlo L, Semeraro A, Daddi G, Darwish S, Stocchi L,
Tofanetti F R, Bellezza G, Sidoni A, Tognellini R, Crino' L, Tonato M
Biological markers and DNA flow cytometric analysis in radically resected patients with non-small cell lung cancer.
A study of the Perugia Multidisciplinary Team for Thoracic Tumors
Tumori 2008 94 : 398-405
Ludovini V, Gori S, Colozza M, Pistola L, Rulli E, Floriani I, Pacifico E, Tofanetti F R, Sidoni A, Basurto C, Rulli
A, Crino' L
Evaluation of serum HER2 extracellular domain in early breast cancer patients: correlation with clinicopathological
parameters and survival
Ann Oncol 2008 19 : 883-890
Ludovini V, Pistola L, Gregorc V, Floriani I, Rulli E, Piattoni S, Di Carlo L, Semeraro A, Darwish S, Toffanetti F R,
Stocchi L, Mihaylova Z, Bellezza G, Del Sordo R, Daddi G, Crino' L, Tonato M
Plasma DNA, microsatellite alterations, and p53 tumor mutations are associated with disease-free survival in
radically resected non-small cell lung cancer patients. A study of the Perugia Multidisciplinary Team for Thoracic
Oncology
J Thorac Oncol 2008 3 : 365-373
Trotta F, Apolone G, Garattini S, Tafuri G
Stopping a trial early in oncology: for patients or for industry
Ann Oncol 2008 19 : 1347-1353
Ganzinelli M, Carrassa L, Crippa F, Tavecchio M, Broggini M, Damia G
Checkpoint kinase 1 down-regulation by an inducible small interfering RNA expression system sensitized in vivo
tumors to treatment with 5-fluorouracil
Clin Cancer Res 2008 14 : 5131-5141
Cecchinato V, Erba E, Basile A, Scarpati B, Fazi C, Brando B, Comi P, Chiaramonte R
Hexamethylene bisacetamide inhibits malignant phenotype in T-ALL cell lines
Leuk Res 2008 32 : 791-797
Tavecchio M, Simone M, Bernasconi S, Tognon G, Mazzini G, Erba E
Multi-parametric flow cytometric cell cycle analysis using TO-PRO-3 iodide (TP3): Detailed protocols
Acta Histochem 2008 110 : 232-244
Apolone G, Patarnello F
Finance fibrillation and research rhythm: do we need a pace-maker Reply to
J Ambul Care Manage 2008 31 : 187-189
Fruscio R, Colombo N, Lissoni A A, Garbi A, Fossati R, Ieda N, Torri V, Mangioni C
A phase II randomised clinical trial comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and
epirubicin in newly diagnosed advanced ovarian cancer: long-term survival analysis
Br J Cancer 2008 98 : 720-727
Giusti I, D'Ascenzo S, Millimaggi D, Taraboletti G, Carta G, Franceschini N, Pavan A, Dolo V
Cathepsin B mediates the pH-dependent proinvasive activity of tumor-shed microvesicles
Neoplasia 2008 10 : 481-488
Apolone G, Corli O, Greco M T, Zagonel V, CPOR SG Investigators
Factors influencing the decision to take or reject opioids for cancer pain: are we on target
Ann Oncol 2008 19 : 1021-1022
Porcu L, Poli D, Torri V, Rulli E, Cropalato di Tullio M, Cinquini M, Bajetta E, Labianca R, Di Costanzo F, Nitti D,
Floriani I
Impact of recent legislative bills regarding clinical research on Italian ethics committee activity
J Med Ethics 2008 34 : 747-750
Benedetti Panici P, Basile S, Maneschi F, Lissoni A A, Signorelli M, Scambia G, Angioli R, Tateo S, Mangili G,
31
ANNUAL REPORT 2008

IRFMN
Katsaros D, Garozzo G, Campagnutta E, Donadello N, Greggi S, Melpignano M, Raspagliesi F, Ragni N, Cormio G,
Grassi R, Franchi M, Giannarelli D, Fossati R, Torri V, Amoroso M, Crocè C, Mangioni C
Systematic pelvic lymphadenectomy vs no lymphadenectomy in early-stage endometrial carcinoma: randomized
clinical trial
J Natl Cancer Inst 2008 100 : 1707-1716
Deandrea S, Montanari M, Moja L, Apolone G
Prevalence of undertreatment in cancer pain. A review of published literature
Ann Oncol 2008 19 : 1985-1991
Margosio B, Rusnati M, Bonezzi K, Cordes B A, Annis D S, Urbinati C, Giavazzi R, Presta M, Ribatti D, Mosher D
F, Taraboletti G
Fibroblast growth factor-2 binding to the thrombospondin-1 type III repeats, a novel antiangiogenic domain
Int J Biochem Cell Biol 2008 40 : 700-709
Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone M V, Detoma P, Giacomin A, Clerico
M, Tempia P, Guala A, Fasolo G, Riva E
Association of mild anemia with cognitive, functional, mood and quality of life outcomes in the elderly: The Health
and Anemia" study
PLoS One 2008 3 : e1920
Mosconi P, Colombo C, Guella F, Pierotti B, Vimercati F.
Are Italian medical societies bridging the distance from citizen and patients associations Result of a survey.
Journal of Preventive Medicine and Hygiene 2008 9: 112-115
Di Costanzo F, Gasperoni S, Manzione L, Bisagni G, Labianca R, Bravi S, Cortesi E, Carlini P, Bracci R, Tomao S,
Messerini L, Arcangeli F, Torri V, Bilancia D, Floriani I, Tonato M, Italian Oncology Group for Cancer Research
Adjuvant chemotherapy in completely resected gastric cancer: a randomized phase III trial conducted by GOIRC
J Natl Cancer Inst 2008 100 : 388-398
Cascinu S, Berardi R, Labianca R, Siena S, Falcone A, Aitini E, Barni S, Di Costanzo F, Dapretto E, Tonini G,
Pierantoni C, Artale S, Rota S, Floriani I, Scartozzi M, Zaniboni A, GISCAD
Cetuximab plus gemcitabine and cisplatin compared with gemcitabine and cisplatin alone in patients with advanced
pancreatic cancer: a randomised, multicentre, phase II trial
Lancet Oncol 2008 9 : 39-4
Apolone G, Tafuri G, Trotta F, Garattini S
A new anti-cancer drug in the market: good news for investors or for patients
Eur J Cancer 2008 44 : 1786-1788
Rossi A, Torri V, Gridelli C
Paclitaxel plus bevacizumab for metastatic breast cancer
N Engl J Med 2008 358 : 1637
Labianca R, Garassino M, Torri V
Predicting response of molecular targeted therapies: a still possible challenge
Ann Oncol 2008 19 : 829-830
Santini D, Loupakis F, Vincenzi B, Floriani I, Stasi I, Canestrari E, Rulli E, Maltese P, Andreoni F, Masi Gianluca,
Graziano F, Baldi G G, Salvatore L, Russo A, Perrone G, Tommasino M R, Magnani M, Falcone A, Tonini G, Ruzzo
A
High concordance of KRAS status between primary colorectal tumors and related metastatic sites: implications for
clinical practice
Oncologist 2008 13 : 1270-1275
Valli C, Paroni G, Di Francesco A M, Riccardi R, Tavecchio M, Erba E, Boldetti A, Gianni M, Fratelli M, Pisano C,
Merlini L, Antoccia A, Cenciarelli C, Terao M, Garattini E
Atypical retinoids ST1926 and CD437 are S-phase-specific agents causing DNA double-strand breaks: significance
for the cytotoxic and antiproliferative activity
Mol Cancer Ther 2008 7 : 2941-2954
Ferketich A K, Fossati R, Apolone G
An evaluation of the Italian version of the Fagerstrom Test for Nicotine Dependence
32
ANNUAL REPORT 2008

IRFMN
Psychol Rep 2008 102 : 687-694
Casciani E, Polettini E, Carmenini E, Floriani I, Masselli G, Bertini L, Gualdi G F
Endorectal and dynamic contrast-enhanced MRI for detection of local recurrence after radical prostatectomy
AJR Am J Roentgenol 2008 190 : 1187-1192
Torri V, Floriani I, Poli D, Rulli E, Ocular Hypertension Treatment Study Group, European Glaucoma Prevention
Study Group (EGPS)
The accuracy and clinical application of predictive models for primary open-angle glaucoma in ocular hypertensive
individuals
Ophthalmology 2008 115 : 2030-2036
Lecchi C, Ceciliani F, Bernasconi S, Franciosi F, Bronzo V, Sartorelli P
Bovine alpha-1 acid glycoprotein can reduce the chemotaxis of bovine monocytes and modulate CD18 expression
Vet Res 2008 39 : 50
Cazzaniga M E, Pronzato P, Mustacchi G, De Matteis A, Di Costanzo F, Rulli E, Floriani I
The anthracyclines and the clinical practice: do all breast cancer patients benefit Results from the NORA study
Ann Oncol 2008 19 : 1811-1818
Pignon J-P, Tribodet H, Scagliotti G V, Douillard J Y, Shepherd F A, Stephens R J, Dunant A, Torri V, Rosell R,
Seymour L, Spiro S G, Rolland E, Fossati R, Aubert D, Ding K, Waller D, Le Chevalier T
Lung adjuvant cisplatin evaluation: a pooled analysis by the LACE Collaborative Group
J Clin Oncol 2008 26 : 3552-355
Ceresoli G L, Castagneto B, Zucali P A, Favaretto A, Mencoboni M, Grossi F, Cortinovis D, Del Conte G, Ceribelli
A, Bearz A, Salamina S, De Vincenzo F, Cappuzzo F, Marangolo M, Torri V, Santoro A
Pemetrexed plus carboplatin in elderly patients with malignant pleural mesothelioma: combined analysis of two
phase II trials
Br J Cancer 2008 ; 99 : 51-56
Floriani I, Rotmensz N, Albertazzi E, Torri V, De Rosa M, Tomino C, de Braud F
Approaches to interim analysis of cancer randomised clinical trials with time to event endpoints: a survey from the
Italian National Monitoring Centre for Clinical Trials
Trials 2008 9 : 46
Broggini M. Nemorubicin. In: Anthracycline Chemistry and Biology II, Mode of Action, Clinical Aspects and New
Drugs. Topics in Current Chemistry, Ed. K. Krohn, Springer-Verlag Berlin Heidelberg 2008 283: 191-206.
Valentini G., D’Andrea C., Ferrari R., Pifferi A., Cubeddu R., Martinelli M., Natoli C., Ubezio P., Giavazzi R. “In
vivo” measurement of vascular modulation in experimental tumors using a fluorescent contrast agent. Photochemistry
and Photobiology 2008 84: 1249-1256.
Ubezio P., Cameron D. Cell kiling and resistance in pre-operative breast cancer chemotherapy. BMC Cancer, 2008 8:
201-214.
Sala G., Dituri F., Raimondi C., Previdi S., Maffucci T., Mazzoletti M., Rossi C., Iezzi M., Lattanzio R., Piantelli M.,
Iacobelli S., Broggini M., Falasca M. Phospholipase C gamma1 is required for metastasis development and
progression. Cancer Res. 2008 68(24): 10187-10196.
Leonetti C., Scarsella M., Riggio G., Rizzo A., Salvati E., D’Incalci M., Staszewsky L., Frapolli R., Stevens M.F.,
Stoppacciaro A., Mottolese M., Antoniani B., Gilson E., Zupi G., Biroccio A. The G-quadruplex ligand RHPS4
potentiates the antitumor activity of camptothecins in preclinical models of solid tumors. Clin. Cancer Res. 2008
14(22): 7284-7291.
Ghilardi C., Chiorino G., Dossi R., Nagy Z., Giavazzi R., Bani M.R. Identification of novel vascular markers through
gene expression profiling of tumor-derived endothelium. BMC Genomics 2008 30(9): 201.
Mitry E, Fields AL, Bleiberg H, Labianca R, Portier G, Tu D, Nitti D, Torri V, Elias D, O'Callaghan C, Langer B,
Martignoni G, Bouché O, Lazorthes F, Van Cutsem E, Bedenne L, Moore MJ, Rougier P. Adjuvant chemotherapy
after potentially curative resection of metastases from colorectal cancer: a pooled analysis of two randomized trials. J
Clin Oncol. 2008 26(30): 4906-11.
33
ANNUAL REPORT 2008

IRFMN
Giavazzi R, Bonezzi K, Taraboletti G
Microtubule targeting agents and the tumor vasculature
In: Cancer drug discovery and development: the role of microtubules in cell biology, neurobiology, and oncology
Humana Press, Totowa, NJ, 2008; 519-530
LAY PRESS SELECTION PUBLISHED IN 2008
Porcu L, Floriani I
GISCAD DIDATTICA. Punta sul cavallo vincente..
Medical Oncology Progress & Perspectives 2008 ; 29 : 23-24
Costato A, Braun C, D'Incalci M, Pasina L, Nobili A
Malati e ricercatori alleati contro il tumore gastrointestinale
Partecipasalute 2008 ;
Klotchenko S A, Tsymbalenko N V, Solov'ev K V, Skvortsov A N, Zatulovskii E A, Babich P S, Platonova N A,
Shavlovskii M M, Puchkova L V, Broggini M
The effect of silver ions on copper metabolism and expression of genes encoding copper transport proteins in rat liver
Dokl Biochem Biophys 2008 ; 418 : 24-27
Mangano S, Negri Emanuele, Apolone G
Guida all'uso del programma. ; Il dolore nel paziente con cancro;
In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica,
Pavia, 2008; 123-141
Apolone G, Corli O
Prefazione
In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica,
Pavia, 2008; 5-10
Apolone G, Negri Emanuele, Mangano S, Greco M T, Montanari M
Dolore nel paziente con cancro: un progetto multidisciplinare per valutare e migliorare la qualità del trattamento
In :Il dolore nel paziente con cancro. Riflessioni e approfondimenti dalla valutazione alle terapie Selecta Medica,
Pavia, 2008; 109-121
Zagonel V, Apolone G
Cancro, curare humanum est
Sole 24 Ore Sanita 2008 11 : 37
Apolone G, Mosconi P
Le politiche che regolano il mercato dei farmaci: il ruolo delle agenzie regolatorie
In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli IRFMN, Milano, 2008; 93-
98
Apolone G, Corli O, Mosconi P
Oncologi: Cure migliori se c'è la comunicazione"
Sole 24 Ore Sanità 2008 n.42 : 14-15
Mosconi P
La Commissione oncologica nazionale e la rappresentanza dei pazienti: occasione persa
Partecipasalute 2008
Mosconi P, Liberati A, Satolli R
Il progetto Partecipasalute
In :La dispensa di Partecipasalute: orientarsi per fare scelte consapevoli IRFMN, Milano, 2008; IX-X
Braun C, Mosconi P
DHEA: l'integratore da consigliare al posto della terapia ormonale sostitutiva
Partecipasalute 2008
Pasina L, Colombo Cinzia, Mosconi P, Nobili A, Perego L, Taddei G C
34
ANNUAL REPORT 2008

IRFMN
Fare chiarezza sui farmaci equivalenti
Ricerca & Pratica 2008 n.141 : 113-116
Braun C, Mosconi P
E' tempo di vacanza: la profilassi antimalarica
Partecipasalute 2008
Simi S, Mosconi P
Così l'Italia della salute si allea con i pazienti
Sole 24 Ore Sanita 2008 11 : 10-11
Colombo Cinzia, Mosconi P
L'associazionismo sta cambiando: dall'assistenza alla nascita di un progetto
In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli IRFMN, Milano, 2008; 99-
104
Mosconi P, Satolli R, Colombo Cinzia, Liberati A, Mele A
L'informazione sulla terapia ormonale post menopausale
Bollettino Informazione Farmaci 2008 15 : 166-171
Mosconi P, Colombo Cinzia, Satolli R, Donati S, Mele A, Liberati A
Quando usare la terapia ormonale sostitutiva
Ricerca & Pratica 2008 n.143 : 209
Mosconi P
L'advocacy, voce di chi non ha voce
Janus 2008 31 : 103-106
RESEARCH ACTIVITIES
Laboratory of Cancer Pharmacology
Mode of action of Ecteinascidins
A project ongoing since several years is about the characterization of marine natural products
possessing antitumor activity. In particular we carried on the studies on the effects of ET-743 in
cells defective for some DNA repair mechanisms. Cells deficient for Homologous
Recombination (HR) are very sensitive to the drug, while cells deficient for Non Homologous
End-Joining (NHEJ) are only slightly more sensitive, but surpraisingly cell lines defective for
Nucleotide Excision Repair (NER) are less sensitive to ET-743. Flow cytometric analysis
coupled to a software of computer simulation, developed in our laboratory, has demonstrated
that NER defective cells showed, after ET-743 treatment, cell cycle perturbations different than
those occurring in NER proficient cells, probably for the activation of different and more
efficient repair mechanisms.
We study also a functional evaluation of the DNA repair mechanisms by the cell capacity to
recognize and repair double helix breaks with a recently introduced test that is very sensitive to
detect the phosphorylation of histone H2AX. An in vitro study is ongoing with flow cytometry
and immunofluorescence techniques to evaluate in different tumor cell lines the phosphorylation
level of histone H2AX in relation to the distribution of the cells in the different phases of the
cell cycle and the cytotoxic effect induced after treatment with ET-743.
Studies are in progress on the mechanism of action of new ET-743 derivates compounds that
have shown antitumoral activity on cell lines with different DNA repair mechanisms.
A new project is the study of the selective action of ET-743 on mixoid lyposarcoma, a
pathology representing 10% of all soft tissue sarcomas, trying to understand if the significative
35
ANNUAL REPORT 2008

IRFMN
antitumor effect is due to a selective action of the compound on pathogenetic alterations
characteristic of this pathology. In particular we are trying to evaluate how ET-743 interact with
the transcriptional modifications of specific genes due to the translocation FUS-CHOP that
characterizes mixoid sarcomas or those caused by the interaction host-tumor, modifying
inflammatory and angiogenetic processes. Studies are in progress to obtain cell lines and
xenografts of mixoid lyposarcomas exhibiting the same molecular features of the patients’
tumors.
Combinations of natural products of marine origin with other anticancer
drugs
We have observed additive or synergistic activity of ET-743 combined with other anticancer
drugs such as cisplatin, doxorubicin, campthotecin and inhibitors of telomerase.
Flow Cytometric analysis of the DNA content in human ovarian cancer:
clinical correlations
Conflicting results have been published on the prognostic significance of DNA aneuploidy on
advanced ovary carcinoma (stage III or IV). The Citometry unit has reported one of the largest
studies of the scientific literature indicating that the aneuploidy in the advanced ovary
carcinoma is not an independent prognostic factor. In a large number of cases of stage I and II
ovary tumors DNA content and the percentage of cells in S phase of the cell cycle, has been
measured with flow cytometry, demonstrating that in the early stages of the disease DNA
content is a prognostic factor important for ovary tumor.
Analysis of cell cycle data and interactions of different drugs
The Biophysics Unit is engaged in theoretical and methodological studies aimed at a critical
evaluation of current techniques of investigation of drug effects on heterogeneous cell
populations. Several computing tools have been produced to simulate the cell proliferation at
different levels (from molecular interactions to in vivo growth of solid tumours) and the process
of measure.
Collaborations are ongoing with other research groups for design and data analysis of drug
combination studies in vitro. In this field, a number of computer programs have been developed,
allowing comparative data analysis with the most common models of drug interaction.
Evaluation of the complexity of the response of cell populations to
treatment with anticancer drugs
This project of the Biophysics Unit addresses the issue of establishing a connection between the
intracellular drug interactions and the resulting cell cycle perturbations. It starts from the singlecell
level of investigation to reach the cell-population level where the relevant end points of
treatment efficacy are evaluated by flow cytometry and growth inhibition/cytotoxicity assays.
The model adopted for data analysis and interpretation is the result of the merging of two
mathematical models. One model describes the cell cycle, exploiting the results of the theory of
age-structured cell population dynamics. The second model describes the response to the drug's
challenge, using distinct parameters ("effect descriptors") measuring either the strength of cell
cycle arrest, damage repair or cell death in every phase (G1, S and G2M). In this way, it is
possible to reach a quantitative interpretation of the experimental results, overcoming the
current qualitative and partial approaches to this problem, which are unable to resolve the
overlapping of cytostatic and cytotoxic effects, and to establish a connection with phase-related
events.
Applying this procedure we demonstrated complex but biologically consistent patterns of time
and dose-dependence for each cell cycle effect descriptor, following a short treatment with
melphalan on a reference cell line of ovarian carcinoma. These results add to the previously
36
ANNUAL REPORT 2008

IRFMN
reported studies on topotecan, cisplatin and taxol. Eventually, this project will produce a
database containing the values associated to the new effect descriptors, related to few
compounds but rich of information about them, especially in the dose and time dependence of
the effects. This database will be used to compare the treatment response of the most common
drugs adopted in the ovarian carcinoma.
Cell cycle dysregulation in erlotinib-based treatments decoded by flow
cytometry and mathematical modeling
Epidermal growth factor receptor (EGFR) inhibitors represent one of the most promising class
of anticancer compounds, some of them, like erlotinib, are already used for clinical therapy.
Nevertheless, so far, the research has focused on molecular interaction of these compounds,
somewhat neglecting the study of the dynamics of cell cycle perturbations and underscoring the
importance of this issue for the optimization of both single and multidrug therapies.
In order to fill this gap, the Biophysics Unit will apply the multidisciplinary approach,
exploiting cell cycle simulation tools, to the study of cell cycle perturbations induced by
erlotinib as a representative EGFR inhibitor. Studies of the time- and dose-dependence of
perturbations induced by treatments are ongoing for single erlotinib treatment or for erlotinib
combined with gemcitabine, irinotecan and oxaliplatin. The appreciation and the quantification
of these effects will provide an important contribution to the comprehension of the mode of
action of erlotinib alone or in combination. Such perspectives are vital if the events are to be
decoded and used in predictive models for the exploration of pharmacodynamic actions and in
understanding the origins of treatment failure.
Anticancer Drug Effects Decoded by Time-Lapse Imaging, Flow Cytometry
and Modeling
We aim to use flow cytometric (cell-population based analysis) and time-lapse imaging (single
cell lineage based analysis) techniques to generate data that will be used to predict drug
responses in term of the two major determinant of cytostatic/cytotoxic actions of anticancer
drugs: specific cell cycle perturbations (detecting accumulation or depletion of cells in G1, S
and G2M phases) and the commitment to cell death (apoptosis).
The response to a treatment with the anticancer drugs will be investigated in a human
osteosarcoma cell line U2OS. engineered in order to express fluorescent reporter (a fusion
cyclin B1-GFP protein), so that it is now possible to follow the cells through G1, S and G2M
using time-lapse microscopy. The working hypothesis is that quantitative analysis of time-lapse
microscopy data could be integrated with the information provided by flow cytometry -
allowing for the first time a joint interpretation of both kind of experiments through a common
mathematical model that simulate the underlying phenomena. The final goal is to provide new
levels of understanding and simulation tools to the cancer research community.
Timing the changes of the cellular content of specific proteins inside G1,
in exponentially growing cells
We developed a method for measuring the content of immunocytochemically detected proteins
in individual cells progressing through G1 phase. The feasibility was demonstrated in the
analysis of cyclin E levels. The sequence of G1 events is tracked in unaltered cycling
conditions, in a cell line in the phase of balanced growth in vitro, to avoid the pitfalls of
synchronization.
The method is based on i) a bromodeoxyuridine (BrdUrd) pulse-and-chase experimental plan;
ii) triparametric flow cytometric detection of DNA, BrdUrd and cyclin E; iii) data analysis
supported by the basic mathematical theory of asynchronous growing populations with variable
cell cycle phase durations.
37
ANNUAL REPORT 2008

IRFMN
Establishing a CFSE-based method for a quantitative measure of
cytostatic effects of drugs on tumor cell populations
Carboxyfluorescein diacetate succinimidyl ester (CFSE) is currently used to investigate
migration and proliferation of hematopoietic cells. Several technical problems had precluded
until now its use in the studies of the antiproliferative activity of anticancer drugs. We analysed
several critical steps of the procedure, providing the way to overcome potential pitfalls. The
project was eventually successful and the previous limitations were overcome. The outcome
was a new standardised procedure of cytometry and data-analysis allowing a measure of the
dynamics of cell cycle blockades after drug treatment. The new method was applied in the study
of the drug topotecan, measuring the variability of response to treatment in terms percentage of
cells immediately blocked, divided once or more times and then blocked or unaffected by the
treatment.
Pharmacokinetic of new taxane derivates
Preclinical studies have been done on new taxane derivatives, chemically and biologically
different from the conventional ones. For two of these compounds we evaluated the
bioavailability after oral administration and the kinetic and metabolic profile has been entirely
characterized by applying analytical methods based on HPLC/MS/MS technique developed in
the Cancer Clinical Pharmacology Unit. They showed biological activity even in tumors with
low susceptibility to other taxanes (cerebral tumor), suggesting a potential clinical interest.
According to the hypothesis that these drugs act through an antiangiogenic mechanism, put
forward by the Laboratory of the Biology and Therapy of Metastasis, it would be important to
investigate prolonged chronic treatments and therefore we are investigating the pharmacokinetic
properties after oral administration and after prolonged daily treatment.
An interesting pk profile was found for a third compound, the 14-β-hydroxy-10-
deacetylbaccatin III derivative (IDN 6140). He showed good bioavailability and high
distribution in brain and glioblastoma achieving high concentration in comparison to that of
plasma, demonstrating high ability to cross the blood-brain-barrier. These data make this
compound of great potential interest for the therapy of Central Nervous System tumors and
metastasis.
Pharmacokinetic of sanguinarine
The pharmacokinetic of SA, a quaternary phenanthridine alkaloid with potent antiinflammatory,
antibacterial and antitumor activity, was studied in mice after single and
repeated administrations, given the drug both by oral and intravenous route. Firstly, we
developed an analytical methods based on HPLC/MS/MS technique to measure SA and its main
circulating metabolite DHSA in plasma of mice. The drug showed low absorption, being the
bioavailability lower than 10% and achieved only after the intravenous treatment,
concentrations potentially able to exert cytotoxic activity. No accumulation of drug was found
in plasma after repeated treatments.
DHSA was the main metabolite of SA, present in plasma only at concentrations less than 10%
of those of the parent drug.
Clinical pharmacokinetics of gimatecan
we are conducting a study in collaboration with SENDO on the clinical pharmacokinetics and
pharmacodynamic of a new proteosome inhibitor, CEP18770, in cancer patients during a phase
I investigation. The study is still in progress and shows that this compound is highly available,
has a long half life, with consequent long exposure to the drug.
Pharmacokinetic parameters will be correlated to the pharmacological data, i.e. inhibition of the
proteosome activity, to define the pharmacodinamy of this compound.
38
ANNUAL REPORT 2008

IRFMN
Other relevant clinical studies
ST 1926. The clinical pharmacokinetic of ST1926 a new oral retinoid derivative was
investigated in women with ovarian cancer. In particular we studied the pharmacokinetics, the
bioavailability of the drug and its metabolism during a Phase I study, providing for the first
time data on the main plasma metabolite of ST 1926 (the glucuronide conjugate). Our data show
variable absorption of the drug and high conversion to the glucuronide.
Antitumoral activity and pharmacokinetic properties of new drugs and
combinations
The antitumor activity, pharmacokinetic properties and toxicity of novel anticancer drugs with
specific targets (e.g. different kinase inhibitors), conventional anticancer drugs (camptothecin)
and combinations is being investigated using rodent tumors and human tumor xenografts.
Development of a software framework for a rationalized process of
Microarrays analysis
It is currently active in the Institute a multidisciplinary group involved in the rationalization of
the various microarray analysis aspects, with many external collaborations.
The different possible analysis procedures have been discussed, compared and formalized in
order to obtain a common work flow to be accepted from the scientific community.
Thanks to this activity it is now possible to automate some aspects of the analysis making them
faster end better reproducible for people managing the analysis itself.
This activity has involved the development of the necessary software for data analysis and the
implementation of a Beowulf computer cluster, using the old computers dismissed by the
desktop users in order to obtain a sufficient power.
Internal collaborations:
Department of Biochemistry and Molecular Pharmacology (ref.: Maddalena Fratelli, Mario
Salmona)
Department of Oncology (ref.: Sergio Marchini, Mariarosa Bani, Maurizio D'Incalci)
External collaborations:
Fondo Edo Tempia (ref.: Giovanna Chiorino)
Istituto Toscano Tumori (ref: Duccio Cavalieri)
Istituto Weizmann (ref. Eytan Domany)
Università di Birmingham (ref. Francesco Falciani)
Università di Aberdeen (ref. Tony Travis)
Microarray data analysis for the Oncology Department
The data analysis activities concern the biology of sensible versus resistant to therapy human
ovarian carcinoma and the study of the ET-743 working mechanism in human sarcoma, sensible
and resistant.
Related to the projects:
Department of Oncology (ref. Sergio Marchini, Maurizio D'Incalci, Massimo Broggini, Mirko
Marabese) –Agilent platform
Metastasis/Relapses Project (phase III ovarian)
LPS2 Project (cell lines, response to ET-743)
Department of Oncology (ref. Mariarosa Bani, Raffaella Giavazzi) –Affymetrics platform
Stroma Project
Parenchyma Project
־
־
־
־
External collaborations:
Osp. S.Gerardo, Monza (ref. Robert Fruscio)
39
ANNUAL REPORT 2008

IRFMN
Development of the new database handling software for the ovarian
tumors bio-bank, compliant with the new privacy related laws for data
handling for clinical trials and with a new reporting engine for a better
data extration
Internal collaborations:
Department of Oncology (ref.: Sergio Marchini, Mariarosa Bani, Raffaella Giavazzi, Maurizio
D'Incalci, Massimo Broggini, Mirko Marabese)
External collaborations;
Ospedale S.Gerardo, Monza (ref. Robert Fruscio)
Implementation of the data management engine for the Italian Registry
Domiciliar Artificial Nutrition, in collaboration with the Scientific Society
SINPE (ref. Loris Pironi, S.Orsola, Bologna)
This activity involves all aspects of the data management, statistical analysis, help desk,
informatic infrastructures and software for handling an observational clinical study.
For this purpose a new ibrid online/offline database has been developed, compliant with the new
privacy related laws about the data protection issues.
External collaborations:
Osp. S. Orsola, Bologna (ref. Loris Pironi)
Società Scientifica SINPE
Laboratory of Molecular Pharmacology
G2 checkpoint and cell cycle
A new system able to specifically inhibit CHK1 expression in vivo in nude mice transplanted
with human tumors has been developed. The system has been proved to be able to reduce the
expression only in cancer cells. This plasmid allows the expression of the siRNA only after
induction with tetracycline and is therefore a unique tool to determine the effect of CHK1
inhibition in human tumors growing in nude mice following treatment with anticancer agents.
The use of these cells derived from the human colocarcinoma cell line HCT116 with an
inducible expression of siRNA directed against CHK1, allowed us to demonstrate that in vivo
too we have a nice downregulation of chk1 only when tetracycline is added in the water. Chk1
downregulation does not induce a reduction in tumor growth but induce a strong sensitisazion of
the tumors to treatment with the anticancer agent 5-fluorouracil. This effect is particularly
evident in p53 deficient tumors, indicating that the combination is likely to result in increased
selectivity.
Characterization of new potential oncosuppressor genes
DRAGO gene, identified and cloned in our laboratory is one of the most interesting projects of
the group. The characterization of the response of KO mice for DRAGO to ionising radiation is
similar to normal mice. The characterization of the transcriptional regulation of DRAGO
indicated that the gene is not only a p53-responsive gene, but, inside the p53 family, p73 has a
strong ability to induce the transcription. Drago therefore, represents a new p73 responsive
gene.
Molecular characterization of ovarian carcinoma
The molecular characterization of stage I ovarian carcinomas has been further studied.
The gene expression profile analysis has showed interesting results.
40
ANNUAL REPORT 2008

IRFMN
It is possible to identify genes able to discriminate in ovarian cancer the different histotypes,
suggesting that specific biological and molecular characteristics are responsible for the
differences in morphology and clinical behaviour. These studies can help in identifying new
specific molecular targets for the different subclasses of ovarian cancer that have a different
clinical outcome.
It is possible to identify patients with higher probability of relapse because there are genes able
to differentiate these two classes of patients.
We have demonstrated that stage I borderline patients have a gene expression profile similar to
that of grade 1 patients, while they are well distinguishable from grade 2 and grade 3 patients.
This can have important clinical implications for the treatment of this particular subgroup of
patients.
We are currently evaluating the differential expression of genes, microRNA and proteins in
tumor biopsies of patients at first surgery and after relapse, with the idea of finding genes
associated with response to treatment.
Expression of gene involved in DNA repair in human ovarian cancer
By Real Time PCR, the expression of genes involved in DNA repair has been evaluated in 90
stage I and 90 stage III ovarian cancer. The genes analysed include those belonging to the
nucleotide excision repair, in the fanconi anemia repair, in the base excision repair. In addition,
genes important for the cellular response to damage, such as chk1 and claspin have been
studied. The analysis that is still ongoing, suggests that there are difference in the expresison of
some of the genes between early (stage I) and late (stage III) ovarian tumors. Thes information
will be useful for the determinatin of the mechanisms responsible for tumor progression. We are
also attempting to identify correlation between gene expression and response to chemotherapy.
These data will help in identifying patients more likely to respond to platinum based therapy,
which represents so far the standard therapy for this type of malignancy.
Inhibition of the signal mediated by PI3K/akt
In ovarian carcinoma cells PI3K gene is often overexpressed or mutated and this kinase is then
constitutively activated. The consequence is the presence of proteins involved in cell survival,
like akt, constitutively phosphorylated and then active. This project evaluated the capacity of
tetra and pentaphosphate inositols and of some analogues to inhibit akt recruitment to the
membrane and its further phosphorylation. IP5 and some new synthetic molecules, were shown
to induce a reduction of the phosphorylation of akt and therefore to reduce cell growth. The
activity of these molecules is not restricted to ovarian cancer, but has been shown also for other
tumors such as breast and prostate. The possibility to combine PI3K inhibitors and mTOR ( a
kinase downstream to PI3K and akt) inhibitors has been evaluated. It has in fact been shown
that some mTOR inhibitors induce an aberrant phosphorylation, and hence activation of akt, and
the combination of these inhibitors can block these undesired effects. The in vitro results show
that the combination has at least an additive effect and open the way to find out new treatment
schedules to verify whether the sequence of combination can be important for the
antiproliferative effects.
Role of phospholipase C γ1 in the development of metastasis
We have evidentiated an important role for phospholipase C γ1, in determining metastasis
formation. By generating tumor model with inducible downregulation of phospholipase C γ1,
we have demonstrated in vitro a reduced motility and migration of cancer cells and in vivo a
reduced formation of metastasis. The results have been obtained in different cell lines of human
and murine origin.
Our result strongly suggest phospholipase C γ1 as a target for the development of new
molecules with antimetastatic activity.
41
ANNUAL REPORT 2008

IRFMN
Oncosuppressors p53 and p73
The p53 analogue, p73 is present in different isoforms derived from alternative splicing of the
C-terminal. Among these isoforms there is one called DNp73 in which the transactivation
domain in the N-terminal of the protein is absent. This DN form of p73 is an antagonist of p53.
The DNp73 isoforms can be further processed through alternative splicing, to generate a number
of isoforms with a not yet clarified biological activity. We have previously shown that the alpha
form of DNp73 does not modify either the growth in vitro and in vivo of cancer cells or the
response to treatment with anticancer agents.
We have therefore generated clones derived from the human lung cancer derived cell line
H1299 which express DNp73 beta following induction. These clones show, upon induction,
interesting effects on cell growth suggesting that the presence of high levels of this isoform can
have unpredicted effects. Since the data generated on the expression of p73 isoforms in cancer
patients indicate that the beta isoform of DNp73 is indeed present in human cancer, the results
obtained have a particular relevance and will be further analysed to verify which are the effects
linked to the overexpression of this isoform.
Identification of a new proteolytic mechanism of activation of p73
Another member of the p53 family, p73, has been characterised in the laboratory for its ability
to modify the growth and resposne to therapy of cancer cells. We have further studied this
potential tumor suppressor by studying new possible mechanisms of activation. We have found
an additional regulation mechanism for the p73-alpha form that occurs through proteolytic
cleavage connected to the activity of the serine protease HtrA2. Following apoptotic stimuli,
HtrA2 accumulates in the nucleus and cleaves p73alpha in the C-terminal portion, enabling the
protein to increase its transactivation activity on the apoptotic gene bax but not on the cell-cycle
regulator gene p21. In the presence of HtrA2, p73 is more prone to cause caspase activation and
nuclei fragmentation: p73 needs HtrA2 to activate and enhance its apoptotic functions. This new
relation between p73 and HtrA2 may help to understand the different behavior of the p73
protein in cell physiology and in the responses of cancer cells to chemotherapy.
Mechanisms of action of new antitumor drugs
The mechanism of action of a new anthracycline derivative, nemorubicin (methoxy morpholino
doxorubicin) has been characterised. Nemorubicin presents a pattern of antitumor activity in
vitro and in vivo different from that of doxorubicin.
We have found that cell lines selected for resistance to Nemorubicin have a reduced DNA repair
ability particularly they show defects in the nucleotide excision repair (NER).
In particular, all the cell lines selected fro resistance do not have detectable levels of XPG
protein and have a collateral sensitivity to UV light, while they are maintaining the same
sensitivity to ionising radiations. Furthermore, the mechanism of resistance to nemorubicin
seems similar to that observed for the molecule of marine origin ET-743 and in fact cells
resistant to nemorubicin are also resistant to ET-743.
Cells resistant to nemorubicin maintain their resistance also when transplanted in nude mice and
the tumor growing in vivo has the same molecular characteristics of the cells growing in vitro.
The combintin studies between the distamycin derivative brostallicin and the demethylating
agent zebularin have been completed. The rational for this combination derives from previous
data generated in the lab indicating that brostallicin activity depends on the presence of GST
and GSH. In tumors like prostate cancer, the GST gene (and particularly the pi isoform) is
methylated and hence the protein is not expressed. In these cells the pretreatment with
compounds able to revert the methylation increases the in vitro activity of brostallicin.
The study has been conducted in vivo using different treatment schemes with the demethylating
agent zebularin and brostallicin. We have found that zebularin, which does not have antitumor
activity per se, increases the activity of brostallicin. This effect is associated with an increased
42
ANNUAL REPORT 2008

IRFMN
GST activity. However, from a molecular point of view, we did not find re-expression of the pi
isoform of GST, probably because in these cells the gene is heavily methylated. We have found
that another isoform of GST, the GST-M, is also methylated (much less than the pi isoform) and
hence the increased GST activity observed could be due to re-expression of this specific
isoform.
Generation of new cellular systems for in vivo imaging
We have generated new cell clones derived from human cancer cells growing in vitro, which
stably express fluorescent or luminescent probes which can allow us to follow in vivo the
growth of primary tumors and metastasis in mice. These systems generated in human ovarian ,
breast and prostate cancer cell lines, can be implanted in nude mice and the growth and response
to therapy followed by either optical and luminescent imaging or microTAC analysis.
We have in particular set up models derived from human breast cancer, which are able to
metastasis to the bone which can be evidentiated by optical imaging and microTC techniques in
laboratory animals. Utilising different reporter genes, we have generated fluorescent and
luminiscent human cancer cell lines which can be transplanted in immunodeficient mice. These
cells can then be visualised in organs such as peritoneum and lungs were these cells were
previously be observed only after sacrifice of the animal. The cells generated to be fluorescent
or bioluminiscent will also have specific gene defects which will be useful for understanding the
mechanism of action of new molecules. These systems will be particularly useful to study the
antimetastatic potential of new drugs.
Characterization of response of stem cells to damage
The therapeutic use of stem cells is continuously increasing. This project aims to investigate the
ability of stem cells isolated from umbilical cord to respond to stress induction with particular
emphasis on their ability to activate checkpoint proteins. Stem cells isolated and maintained in
vitro with specific cytokine cocktails able to induce partial differentiation, have shown a
peculiar expression of proteins controlling the cell cycle. In particular, there is a specific time
point in the differentiation process in which cell cycle checkpoints proteins are present at high
levels. This could imply that this represents the time point in which these cells are more
susceptible and must be “protected” from external damages. The characterization of these
important molecular aspects will be further studied.
Identification of cancer stem cells from ovarian cancer
This project is aimed at isolating and characterizing a possible cancer stem cell from ovarian
cancers. There are increasing evidences supporting the idea that few important multipotent
cancer cells, termed cancer stem cells, are among the most relevant cells to be killed in a tumor.
Normally present as quiescent cells inside the tumors, they are able to rapidly generate dividing
and growing cancer cells. The current hypothesis is that normally dividing cancer cells can be
preferentially killed by chemotherapy while the cancer stem cells would be more difficult to kill
and would be responsible for the relapse following treatment. The possibility to identify and
characterize the cancer stem cell would theoretically open the way to the selection of new
generation molecules able to preferentially kill these cells. Cancer stem cells have been already
identified in different human tumors including breast and hematopoietic tumors. We will focus
our attention on human ovarian cancer by the use of antibodies directed against surface marker
proteins to identify such potentially relevant cancer sub populations.
43
ANNUAL REPORT 2008

IRFMN
Determination of the impact of EGFR mutations in the activity of tyrosine
kinase inhibitors in patients with NSCLC
We have started a multicenter, three year project aimed at identifying those patients who can
have the chance to respond to tyrosine kinase inhibitors.
The study will define whether patients without mutations in the EGFR have more chance to
respond to conventional therapy rather than to treatment with TKI.
Our laboratory is involved in the detection of mutations of EGFR in patients with NSCLC either
in the tumor tissue or in the blood of patients with a secondary aim of defining whether is
possible to detect mutations in the blood circulating DNA which are representative for the
tumor.
DNA sequencing and scorpion arms-based PCR methods are used for these studies.
Laboratory of Biology and Therapy of Metastasis
Physiologic regulation of angiogenesis
Angiogenesis, the formation of blood vessels from existing ones is a fundamental process in
tumor progression. A delicate balance between pro- and antiangiogenic factors finely tunes this
process. In the last years we have been interested in endogenous angiogenesis-regulatory
factors. During 2008 we have continued the study of trombospondin-1 (TSP-1), an endogenous
inhibitor of angiogenesis. TSP-1 directly binds to angiogenic factors, in particular FGF-2
(Fibroblast Growth Factor-2), inhibiting their bioavailability and activity. In a structure/function
relationship analysis of different active domains of TSP-1, we have identified the FGF-2
binding site of TSP-1. This sequence is now used as a model to design new antiangiogenic
compounds based on the active sequence of TSP-1.
We are studying the involvement of matrix metalloproteinases (MMPs) in angiogenesis and
tumor progression. In 2008, we have shown the cross-talk between MMPs and Vascular
Endothelial Growth Factor (VEGF), a factor that stimulates angiogenesis and vessel
permeability, during ovarian carcinoma progression. In addition we have evaluated the
possibility of using VEGF inhibitors to affect MMP-dependent ovarian tumor invasion.
We are currently analyzing modifications in metastasis/invasion-related gene expression profile
in stroma cells induced by tumor VEGF.
Lymphangiogenesis in ovarian carcinoma
Lymphatic spread in early ovarian cancer is a predictor of outcome with potential clinical value.
With the aim to clarify the molecular mechanisms involved in the process of lymphangiogenesis
in ovarian cancer, the expression of VEGFC, a factor that stimulates lymphangiogenesis, has
been measured in serum and ascites of mice bearing human ovarian carcinoma xenografts and
correlated with lymphonode infiltration by neoplastic cells.
To better represent the clinical features of ovarian neoplasia, an orthotopic model of human
ovarian carcinoma xenograft has been created by injecting ovarian tumor cells under the bursa
of the ovary. Infection of ovarian carcinoma cells with lentivirus vectors carrying the coding
sequence of VEGFC, the firefly luciferease gene and fluorescent probes are ongoing; the
invasive and metastatic potential of tumor cells producing VEGFC will be evaluated using
optical imaging techniques, in mice bearing tumor.
How the microenvironment affects endothelial cell gene expression
It is fundamental to understand qualitative and functional differences between tumor and normal
tissue endothelial cells (EC) and the molecular mechanisms that drive the angiogenic process.
This could lead to the identification of selective markers of the vascular endothelium associated
to pathologies and/or of target molecules for the development of novel therapeutic strategies. To
this purpose we analysed the gene expression profile of endothelial cells isolated from ovarian
44
ANNUAL REPORT 2008

IRFMN
carcinoma and adrenal glands exposed or not to an “angiogenic/tumor” environment
reconstituted in vitro. We have found that:
i) the “angiogenic/tumor” environment is able to modulate EC gene expression
ii) few genes are preferentially expressed by tumor associated endothelial cells.
Preliminary results suggest that those genes might be of interest as markers of tumor
endothelium. Their expression is higher in endothelial cells from tumor specimens than from
normal tissues and are not expressed by tumor cells. The putative new tumor endothelial
markers have been further validated as anti-cancer / vascular targets by in situ hybridization
analysis of normal and tumor tissues. The putative targets (identified as mRNA transcript) are
being produced as recombinant proteins, with the aim to produce antibodies, directed against the
recombinant proteins, suitable for immunohistochemical analyses.
Preclinical models: role of Vascular Endothelial Growth Factor (VEGF) on
tumor growth, vascularization and response to therapy
To study the role of VEGF induced angiogenesis in the response of cancer to chemotherapy we
have generated a variant of the human ovarian carcinoma A2780/1A9 by stable transfection
with the VEGF121 isoform (1A9-VS1) or the antisense (1A9-VAS3). In mice 1A9-VS1
xenografts i) show dilated blood vessels sc ii) produce ascites when implanted orthotopically in
the peritoneal cavity, iii) release human VEGF in the plasma, iv) the level of circulating VEGF
correlates with tumor burden.
We have found that the response to chemotherapy (e.g. paclitaxel) differ in xenografts
producing high levels of VEGF. The blockade of VEGF, by the administration of Avastin®,
greatly improved the antitumor activity by paclitaxel treatment of 1A9-VS1.
Studies are ongoing to elucidate the mechanism, associated to modification of the tumor
microenvironment, that are responsible of these differences.
This model is therefore being used to study gene expression. Based on circulating VEGF levels
and morphological analysis of the tumor vasculature, samples were chosen and tissue slices of
the 1A9-VS1 and 1A9-VAS3 were microdissected (PALM Microlaser system, in collaboration
with the Institute of Pathology at Helios Klinikum, Germany) in order to isolate the stroma
compartment to be evaluated by mean of Affymetrix’s GeneChip® Arrays technology. The
microarray hybridisation data were analyzed with the aid of specialized software (i.e.
GeneSpring and Rosetta Resolver) and differentially expressed genes were identified.
Specifically, we have discovered that in the stroma of tumors 294 genes were up- and 162 were
down-regulated in consequence of the VEGF produced by the cancer cells. Gene Ontology
(GO) enquiry (using Expression Analysis Systematic Explorer EASE), identified overrepresented
categories of potentially biologically relevant genes in the stroma of 1A9VS1 (high
VEGF) tumors. Structural molecule activity, cell organization and biogenesis, and basement
membrane were among the up-regulated categories.
Preclinical evaluation of inhibitors of angiogenesis and vascular targeting
agents
Antineoplastic therapies directed against the tumor vascular system may be designed with two
different strategies. Antiangiogenic therapy prevents the formation of new vessels, while
vascular disrupting agents (VDA) aim to selectively destroy the already formed tumor vessels.
In 2008 we have investigated the activity of several inhibitors of angiogenesis and VDA.
We have investigated the antiangiogenic activity and mechanism of action of new molecules –
peptides or non-peptidic small molecules – which mimic endogenous inhibitors of angiogenesis,
including thrombospondin (TSP-1).
Among the VDA, we have studied the properties of novel tubulin binding agents (analogues of
colchicines and combretastatins), which cause microtubules depolymerization, selective damage
45
ANNUAL REPORT 2008

IRFMN
to tumor blood vessels and tumor necrosis in experimental models in vivo. In collaboration with
Prof. Bellina at the Department of Chemistry, University of Pisa (Prof. Bellina and Prof. Rossi),
we have screened classes of compounds with such properties. The lead compound/s will be
further characterized for pharmacological and vascular targeting/antineoplastic properties.
The challenge of combination
The optimization of biological therapies against selective targets in combinations with
chemotherapy is one of the interest of this laboratory. We are investigating small molecule
receptor tyrosine kinase inhibitors, that affects angiogenesis. In 2008 we studied vandetanib, an
inhibitor of VEGFR, EGFR and Ret in combination with pacltaxel (PTX) on a model of human
ovarian carcinoma xenograft. In a study designed to determine the relationship between the
effects of vandetanib on tumor vascular morphological and functional changes and paclitaxel
uptake i)we did not find significant change in vessel number, while a reduced number of large
vessels, together with an increase in the percentage of mature vessels were particularly evident
after 5 days of treatment; ii) pre-treatment with vandetanib resulted in decreased tumor levels of
paclitaxel within one hour from its injection, though at 24 hours the levels were comparable to
controls.
The combination therapy, following two different sequences of PTX administration (PTX
before and after vandetanib), was more efficacious compared to each of the single agent alone.
However the greatest efficacy was obtained with PTX administered before vandetanib; this
outcome was enforced by enhanced fibrotic tissue reorganization in the PTXvandetanib
tumors. These findings imply that the analysis of vascular changes and paclitaxel uptake in
vandetanib treated tumors may assist to guide the schedule of this combination.
High PAR-1 expression is associated to a malignant phenotype
Protease-activated receptor-1 (PAR-1) over-expression has been associated to a variety of
human cancers, and increasing evidence implicates PAR-1 as a contributor to human melanoma
malignancy. We investigated human melanoma cells, isolated from lesions representing various
stages of disease progression, for the expression of PAR-1 (in collaboration with Prof. Naldini
at the University of Siena) and evaluated their migratory and metastatic capabilities. Cells from
advanced stage melanomas expressed higher levels of PAR-1 than those from early stages. The
metastatic capability showed by the melanoma cells which overexpressed PAR-1 allowed these
cells to colonize the lungs in 70-100% of the mice. Accordingly, melanoma cells overexpressing
PAR-1 had higher migrated (chemotaxis assay) and invading (invasion assaymatrigel)
cell counts than those expressing low PAR-1. Migration and invasion were decreased
by silencing PAR-1 (siRNA) and treating with SCH9797, a PAR-1 specific inhibitor.
Laboratory for the development of new pharmacological
strategies
The laboratory was born out of the consideration that the advent of oncological drugs endowed
with mechanisms of action different from those of traditional chemiotherapics, introduces new
treatment opportunities. At the same time, new problems arise concerning the choice of the
most appropriate and effective design for research into the clinical activity profile of these new
treatments.
The traditional paradigm where the choice of dose is based on the maximal tolerated toxicity,
and the screening of therapeutic activity focus on tumor mass reduction, may not necessarily be
suitable for the evaluation of new agents whose targets may include the extracellular
compartment or specific molecular targets.
46
ANNUAL REPORT 2008

IRFMN
The clinical development of ‘non toxic’ anti tumor molecules requires a critical review of the
existing models as well as of all the aspects relative to the conduction of clinical trials
including: dose selection criteria, methods for determination and confirmation of
pharmacological activity, and the validation of new technologies and laboratory methods.
This is where the need for a profound integration of the ‘clinical screening’ and the preclinical
research lies. It is a prerequisite for the construction of the pharmacological rationale for the
identification of the most interesting molecules, the choice of dose, the hypotheses of
combination with other drugs, and of the most appropriate indicators of clinical activity.
The acquisition of know how and the development and application of new designs for clinical
activity studies, including the use of randomization, the introduction of groups of patients
treated with placebo, and new discontinuation designs, proceed in parallel to the above.
Another fundamental issue in laboratory research is the recognition that the genomic
characterization of any single tumor may now play a more relevant role in drug development
and treatment identification.
This notwithstanding, numerous uncertainties remain regarding the role of biomarkers in drug
development and in the implementation of genomic technologies in clinical trials. It is therefore
necessary to improve the methodology and more biomarkers evaluation already in the early
stages of research, thus shifting translational research from a simple process of correlation
search to one producing knowledge regarding the predictive role of the clinical activity of the
investigational treatments.
Therefore, the primary focus of the laboratory is the optimization of the methods for evaluating
the activity of cytotoxic drugs, but mostly for those therapies aimed at specific molecular
targets, as well as the identification of factors predictive of therapeutic response.
In 2008 two phase II studies exploring the activity of sorafenib in patients with pancreatic
cancer and colorectal cancer have been initiated.
Laboratory of Clinical Trials
The Laboratory of Clinical Trials is involved in the planning, coordination and analysis of
randomized clinical trials in oncology, conducted in cooperation with a network of medical
oncologists. Main covered research areas are gastric, colorectal, breast and lung cancer.
Moreover the laboratory works on a single arm, prospective, experimental study in children
with affected by primary glaucoma, refractory to surgical procedures. The study is sponsored by
the Azienda Ospedaliera “Spedali Civili Di Brescia” and supported by the Agenzia Italiana del
Farmaco (AIFA).
Gastric cancer
ITACAS ”Intergruppo Nazionale Adiuvante Gastrico” study is a multicenter, randomized,
controlled, open-label, superiority, phase III trial aimed at assessing the role of adjuvant
chemotherapy in the treatment of gastric cancer. It compares the efficacy and safety of a
sequential treatment (irinotecan plus flurouracil/leucovorin, followed by docetaxel and
cisplatin) versus flurouracil/leucovorin regimen, used as standard reference in patients with
radically resected adenocarcinoma of the stomach or of the gastro-esophageal junction. The
study, sponsored by Mario Negri Institute, involves 11 Italian oncologic collaborative groups
and is being conducted in more than 110 Italian experimental centers. From February 2005,
more than 950 patients out of the 1100 planned have been enrolled and it is expected to
conclude the recruitment in the first half of 2009.
Lung cancer
On November 2007 a a phase III, Italian, multicentre, open label, randomized trial was started.
47
ANNUAL REPORT 2008

IRFMN
Aim of the study is to assess whether it is possible to optimize second-line treatment in
advanced non small cell lung cancer (NSCLC) patients using biological and clinical markers.
The trial compares the efficacy in terms of overall survival of erlotinib vs. docetaxel, given as
second line therapy in pts without EGFR mutations. In particular, the predictive value of K-
RAS mutation, EGFR protein expression and EGFR gene amplification in determining the
effect of erlotinib as compared to chemotherapy will be assessed. Even if erlotinib is registered
in all patients affected by NSCLC in second and subsequent lines with a small benefit, recent
evidence suggest that it should be possible to select patients according with clinical and
biological features. Most of these proofs are drawn from case series or post hoc analyses and not
from properly planned randomized clinical trials making their interpretation controversial.
EGFR mutations, EGFR copy number and EGFR expression should positive select responders,
while K-RAS mutations should predict negative outcome. All these data are suggesting that a
"tailored therapy" based on individual molecular features may result in better responses and
optimization of sources and costs.
The study, sponsored by Azienda Ospedaliera Fatebenefratelli e Oftalmico of Milan and
supported by AIFA, will enroll approximately 1500 patients in 3 years.
Colon cancer
3 studies are being conducted in this area
On June 2007 started the accrual of a randomised, controlled, with a factorial design, phase III
clinical trial aimed at identifying the best therapeutic adjuvant strategy in radically resected
colon cancer patients. The study, sponsored by Fondazione Giscad per la Cura dei Tumori and
supported by AIFA, will address the following two questions:
1) Optimal duration of FOLFOX-4 regimen (3 vs 6 months), by means a non-inferiority
trial between 3-month FOLFOX-4 vs. a 6-month FOLFOX-4, considered as the standard
treatment.
2) benefit of the addition of bevacizumab to FOLFOX-4 regimen, only in high risk stage
III patients.
For both questions, primary efficacy endpoint will be recurrence free survival. On February
2009 more than 120 Italian centres have been involved with the enrollment of approximately
700 patients out of the targeted 3500.
A multicenter, randomized, controlled, open-label, superiority, phase III study is now starting
aimed at compare the efficacy of the addition of cetuximab to FOLFIRI vs. FOLFIRI alone
given as first line therapy in patients K-RAS wild type with advanced colorectal cancer. In
particular, the predictive value of PTEN mutation will be assessed in determining the effect of
cetuximab+FOLFIRI as compared to chemotherapy alone. The efficacy will be evaluated in
terms of progression free survival. This study foresees the involvement of approximately 30
experimental centers and the enrollment of 300 K-RAS non mutated patients.
Another open-label, randomized, parallel group, phase III, multicenter trial, sponsored by
Fondazione Giscad per la Cura dei Tumori and supported by AIFA, started. Aim of this study is
to compare the efficacy and safety of two different sequences of chemotherapeutic agents
(Irinoteca/Cetuximab followed by FOLFOX-4 vs. FOLFOX-4 followed by
Irinotecan/Cetuximab) in order to optimize the treatment of patients with metastatic colorectal
cancer progressed to a first line chemotherapy with FOLFIRI and bevacizumab. Primary
endpoint will be overall survival. The maximum estimated study duration is approximately 52
months and about 350 patients will be enrolled.
Breast cancer
The TOP (Trastuzumab Optimisation trial) study is aimed at increasing the knowledge on the
efficacy of herceptin in the treatment of locally advanced or metastatic breast cancer patients. It
includes two randomised, open label, phase III clinical trials. The first is aimed at assessing
48
ANNUAL REPORT 2008

IRFMN
whether a maintenance therapy with herceptin is superior in terms of progression free survival
to no further herceptin treatment in patients with locally advanced and/or metastatic breast
cancer over expressing HER2 and not progressed to a first line chemotherapy containing
trastuzumab. The second evaluates whether a second line chemotherapy with trastuzumab after
failure of a first line regimen of chemotherapy plus trastuzumab is superior in terms of overall
survival to chemotherapy alone in patients with locally advanced and/or metastatic breast cancer
over expressing HER2. The results of TOP study will allow to evaluate the cost/benefit ratio of
herceptin treatment and to optimize the efficacy of such a drug, already approved in Italy, but
used without clear data supporting evidence of benefit in the considered setting. This study is
sponsored by Regione Lombardia and supported by AIFA.
Head and neck
On March 2008 started the accrual of a randomized, multicenter, open-label, with a factorial
design, phase III trial evaluating the overall survival in patients with locally advanced
squamous cell carcinoma of head and neck treated with loco-regional treatment (radiotherapy
plus concomitant chemotherapy or cetuximab) with or without neo-adjuvant chemotherapy.
Patients will be randomized to receive 3 cycles of neo-adjuvant chemotherapy (TPF - docetaxel,
cisplatin e 5-flurouracil) followed by radiotherapy plus concomitant chemo or cetuximab, or
radiotherapy plus concomitant chemo or cetuximab alone. The primary objectives of the study
are to compare the overall survival between neo-adjuvant and no neo-adjuvant arm and to
compare the toxicity between concomitant chemo-radiotherapy and radiotherapy plus
cetuximab. This study, sponsored by AVAPO (Associazione Volontari Assistenza Pazienti
Oncologici) Ricerche of the Ospedale Civile SS. Giovanni e Paolo, Venezia, will be conducted
by a multidisciplinary team, composed by oncologists, radiotherapists and othorinolaryngologists
and will enroll approximately 350 patients in Italian centers.
Laboratory of Translational and Outcome Research in Oncology
The Laboratory is mainly aimed at documenting, by using either Systematic Literature Review,
Randomized or Outcome Research studies, the value of new diagnostic and therapeutic
interventions in oncology, paying particular attention to two critical steps: the passage from
early to late clinical research (from the activity to efficacy evaluation) and from phase III to
clinical practice (from efficacy to effectiveness). The principal lines of research are three:
cancer pain evaluation, clinical research on gynecologic cancers and evalution of the
effectiveness of complex clinical programs in oncology care. In order to facilitate the research
activities and optimize the outputs, the Laboratory hosts the Coordination Centers of two multidisciplinary
Groups (MANGO: Mario Negri Gynecologic Oncology and the CP-OR: Cancer
Pain Outcome Research Study Groups). As from 2007 on, all the activities of research and
training in the field of chronic pain has been coordinated by a dedicated center (CERP:Center
for the Evaluation and Research on Pain).
CERP
The objective of this newly set up center is in line with the Mario Negri Institute’s purpose:
CERP is aimed at advancing the scientific knowledge of chronic pain and particularly the cancer
pain and at improving the quality of palliative care. Activities concerns three fields: preclinical
and clinical research, education and information. Multidisciplinary and multiinstitutional
approach are used with special care to pharmacologic therapy. Multidiscipinary
teams have been created to value any contribution from either physicians or patients and with
the involvement of scientific societies and patients’ associations. Several educational and
clinical activities focusing on patients with cancer pain are currently on going and are being
conducted with both private and public funds. Major activities the following:
49
ANNUAL REPORT 2008

IRFMN
- Analysis and dissemination of results from the Outcome Research study carried out
between 2006-2007
- Design of a large 4-arm multicenter RCT comparing the efficacy of 4 strong opioids
that will be launched in 2009
- Implementation of several educational courses for members of the CPOR-SG
addressing epidemiological statistical and methodological topics
- Implementation of new experiments in vivo, in collaboration with the Laboratory of
Experimental Psychology to test new approaches in animal models
- Collaboration with the Laboratory of Medical Research and Consumers Involvement in
the implementation and coordination of a population-based information and education
project to change the knowledge, opinions, attitudes and utilization of opioid drugs in
cancer patients that is at present ongoing in an Italian Region (Le Marche).
- Activation of collaborations with European Research Networks
As to the first point, in the context of a wide multidisciplinary project, a nationwide
multicenter, prospective outcome research study was launched in Italy in 2006 to investigate the
epidemiology of cancer pain, the pattern and quality of analgesic-drug therapy, and the
evolution of health outcomes over time.
In a large, prospective, cohort of advanced cancer patients reporting pain, investigators collected
predictive and prognostic variables, information about type of care, as well as several patientreported-outcomes,
such as pain, quality of life, satisfaction with analgesic care using
standardized questionnaires and data collections forms. 110 centers recruited 1801 patients
from February 2006 to March 2007.
Preliminary results were presented to investigators during a meeting held at the Mario Negri
Institute on December 2007. Final results, reported on 4 papers already published or
forthcoming, document that most of patients did not receive exhaustive information about
prognosis, the prevalence of undertreatment was quite high at the time of study inclusion (up to
45%), and that analgesic drugs prescribed by physicians to control cancer pain awere effective
(reducing the average pain reported by patients of about 35%) but with some differences in
terms of dosages and side effects. These results are the basis for the design of the next RCT that
has the objective to confirm these preliminary findings. The study will involve 80 centers and
about 1000 patients.
The collaborative group in clinical gynecologic oncology named MaNGO
The Mario Negri Gynecologic Oncology group (MaNGO) is a new name for a collaborative
group that has been active in clinical gynecologic oncology for several years. Infact, this group
consolidated its network and logistics while running the ICONs studies which were conducted
in very close partnership with researchers at the Medical Research Council, Clinical Trial Unit,
UK. MaNGO was formally set up in May 2006 and is mainly representative of the northern part
of Italy, although there are important sites in the central and southern part of the country too.
Participating centers are either general public and private hospitals or university clinics. One of
MaNGO’s main statutory objectives was to foster an active collaboration with the Gynecologic
Cancer Intergroup (GCIG), a true International Forum that circulates the scientific proposals
from fifteen collaborative groups through ten countries. In 2008, two randomized clinical trials
in ovarian cancer completed the inclusion phase, one sponsored by a French group (CALYPSO
study) and the other sponsored by EORTC (TARCEVA study). MaNGO launched the PORTEC
3 study in Italy: this is a randomized phase III trial in endometrial cancer promoted by the Dutch
collaborative. In 2008 MaNGO was involved in finalizing the protocols of two randomized
clinical trials in ovarian cancer with new antiangiogenic drugs. These trials should be
internationally coordinated by the German onco-gynecologic group named AGO
50
ANNUAL REPORT 2008

IRFMN
During 2008, MaNGO’s Technical-Scientific Committe met quarterly while MaNGO affiliates
were conveyed in one General Assembly and new representatives of the TS Commitee were
elected.
Colo-rectal cancer
The assessment of efficacy of screening for relapses of colorectal carcinoma has been debated
for a long time, with controversial results. GILDA is an open label, international, randomised
study comparing two different strategies of post surgical surveillance in colorectal cancer
(Dukes B2-C stage): minimalist versus intensive. Primary endpoints of this trial are disease free
survival (which is used to assess diagnostic anticipation of metastases), overall survival, health
related quality of life, direct and indirect costs evaluation. At present, GILDA trial is the largest
randomised study evaluating the efficacy of two follow-ups in colorectal carcinoma. The trial
was closed to patient entry in September 2006 when a total of 1200 patients had been enrolled.
At the end of 2008 a manuscript for an Italian journal has been prepared to present the progress
of the trial and in 2009 the final analyses will be submitted to an international peer-review
journal.
Ovarian cancer: clinical and translational studies
During 2008, the Unit of Gynecology-Oncology has coordinated the participation of a selected
network of Italian hospitals to two international randomized clinical trials. The CALYPSO trial
was sponsored by ARCAGY, France and compared two chemotherapy regimens (carboplatintaxol
vs carboplatin- pegylated liposomal doxorubicin ) in patients with epithelial ovarian
cancer in late relapse. The TARCEVA trial was sponsored by EORTC and evaluated the impact
of adding erlotinib for two years in patients with no evidence of progression after first line,
platinum-base chemotherapy for ovarian cancer. Both studies reached their target sample size
well in advance to the anticipated date of recruitment closure, thanks to really succesful
international collaboration. Results will be available in about two years.
During 2008, the Gynecology-Oncology Unit has started the collection of retrospective data
aimed to describe the therapeutic approaches used to manage the ovarian cancer recurrence that
develop between 6 and 12 months from the end of a first line platinum-based chemotherapy.
This study should give clues to optimize the choice of drug combination in this specific clinical
setting. MaNGO will be a partner in the European network involved in two randomized clinical
trials sponsored by the German onco-gynecologic group AGO. These two trial wills be
exploring the effect of two angiogenic drugs, BIBF 1120 and pazopanib, in combination with
the standard first line chemotherapic treatment (carboplatinum-taxol) of ovarian carcinoma. The
Unit of Gynecology-Oncology has prepared an ancillary translational proposal to assess a panel
of circulating proteins (VEGFC, VEGFB, VEGFA and their soluble receptors VEGFR2 and
3)that are possible critical signalling effectors of the angiogenic pathway.
Endometrial cancer: clinical studies
In 2008, the Gynecology Oncology Unit has started randomization for the PORTEC 3 protocol.
This is an international randomized phase III sponsored by the Dutch Cooperative Gynecologic
Oncology Group and it is aimed at comparing concurrent chemoradiation and adjuvant
chemotherapy with pelvic radiation alone in high risk and advanced stage Endometrial
Carcinoma. During 2008 a prospective observational study, assessing the value of trans-vaginal
ultrasound imaging in predicting the myometrial infiltration of endometrial carcinoma, has been
launched and recruitment is expected to last for 12 months . Should this diagnostic procedure
show satisfactory concordance rate with final histopathological examination, clinicians could
use it to improve surgical planning. In 2008 the results of a coordinate effort with the Sapienza
University, Rome was published in the Journal of the National Cancer Institute. It was a
randomized clinical trial that demonstrated that the systematic pelvic lymphadenectomy does
51
ANNUAL REPORT 2008

IRFMN
not impact on disease free survival and overall survival of patients with early endometrial
cancer. In the end of 2008, A draft of a manuscript of a prospective pooling of the data with a
trial of the Nordic gynaecologic oncology group (chemoradiotherapy versus radiotherapy alone
in high risk endometrial cancer) has been prepared. This manuscript will be finalized in early
2009.
Outcome research project in cancer pain
In the context of a wide multidisciplinary project (J. Ambulatory Care Manage 29:332-
341,2006), a nationwide multicenter, prospective outcome research study was launched in Italy
in 2006 to investigate the epidemiology of cancer pain, the pattern and quality of analgesic-drug
therapy, and the evolution of health outcomes over time.
In a large, prospective, cohort of advanced cancer patients reporting pain, investigators collected
predictive and prognostic variables, information about type of care, as well as several patientreported-outcomes,
such as pain, quality of life, satisfaction with analgesic care using
standardized questionnaires and data collections forms.
110 centers recruited 1801 patients from February 2006 to March 2007. Subjects were
monitored for 28 days and then with a simplified scheme for further 8 weeks.
At inclusion, 50% had bone metastasis, 73% a level of pain classified as moderate-severe, 48%
reported episodes of breakthrough pain, 49% were still on active anti-cancer treatments and
60% were already on treatment with strong opioids. When the Index of Pain Management was
computed to provide a rough estimate of how pain was treated (Cleeland et al, NEJM 330:592-
596, 1994), up to 45% had negative values suggesting a possible analgesic under-treatment,
with large variations according to a selected list of clinical variables. In the sub-sample of
patients with a complete follow-up at 28 days (no.=1461), all pain and palliative outcomes did
significantly improve on average, with variations according to case mix, type of treatments and
type of recruiting centers. Outcomes based on worst and average pain intensity showed the
higher effect size estimates (0.84 and 0.69) when compared to patients' satisfaction and quality
of life (0.44, 0.35). Up to 26% of patients were classified as non-responders.
This outcome research study carried out at national level produced data to help implement
future educative and research activities in Italy.
Other research activities
During 2008, other activities on patient-oriented clinical translational research in oncology were
carried out. The focus was on the improvement of transferring information from the pre-clinical
to clinical and research setting, and from clinical research to clinical practice and public health .
Most of this work involves data-entry, storage and other computerized applications for the
management of large and complex databases of biological and clinical data. In addition to the
methodological and bio-informatics issues that are relevant in this area, particular attention was
also given to issues related to the ethical and legal issues pertaining the collection, storage and
utilization of biological samples from patients and citizens.
Finally, we would like to mention two research projects that deal with the problem of testing the
effectiveness of complex clinical programs in the context of public health using formal RCT.
Evalaution of the effectiveness of long-term follow-up for early stage
breast cancer
Despite the lack of evidence that post-operative surveillance programs improve the quality of
care and eventually the outcomes of breast cancer patients after primary curative therapy,
variations in practice patterns exist (between and across nations and clinical settings), with a
significant use of quite intensive programs. Most physicians favor intensive surveillance
programs assuming that detecting disease recurrence as its earliest stage would offer the chance
52
ANNUAL REPORT 2008

IRFMN
of cure and improve survival and quality of life. As a final result, in addition to frequent visits
and useless routine blood tests, complex and costly interventions such as imaging studies, tumor
markers, PET and breast RMI are required for unselected women, thus consuming medical
resources, increasing costs and creating long waiting lists.
The Project is based on the hypothesis that to change the current attitude to use intensive
follow-up programs, it is necessary to involve as many as possible stakeholders and to produce
new pieces of evidence addressing the issues related to all relevant barriers, including nonscientific
or non-medical drivers, such as organization and health-care factors.
In particular, the Project has the main objective to develop specific follow-up/surveillance
strategies that are targeted to specific sub-groups of patients having different risk of
experiencing disease recurrence, and to assess, using a comparative randomized approach, their
yield in terms of diagnostic anticipation, recurrence-related clinical events, quality of life and
satisfaction.
In this context, in addition to carry out a RCT to assess the yield of a very intensive follow-up
regime in a high risk women with breast and endometrial cancer, the Project will test in the
Italian setting the hypothesis that a routine minimal follow-up program carried out by family
physicians (General Practitioner,) is a safe and effective alternative to follow-up carried out by
specialist (basically, oncologists), in low risk women.
The project, sponsored by The Italin Ministry of Health, involve 5 teams in 5 Italian Regions,
started in 2008 and is coordinated by the Laboratory of Translational and Outcome Research.
Laboratory of Medical Research and Consumer Involvement
This Laboratory promotes activities of research in the field of involvement of citizens and
patients and their associations in decision making in health and medical issues. Moreover the
activities of this laboratory include: researches on information conveyed to patients on illness
and treatment, implementation of web site on the topics of the health and the information
(www.partecipasalute.it; www.paincare.it; www.fondazionemattioli.it); strategy of involvement
of groups of patients for the publication of educational material; research on the evaluation of
the quality of the life and satisfaction with care through studies on ad hoc selected groups, and
implementation on validated ad hoc questionnaires.
PartecipaSalute (“Participate in Health Care”) - fostering a strategic
alliance between consumers’ associations and medical community
The project was born in 2003. It is coordinated by Mario Negri Institute, with the collaboration
of the Italian Cochrane Centre and Zadig, an editorial and publishing company. It is supported
by Compagnia di San Paolo, a non profit private-law foundation.
During 2008, training courses for consumers, surveys and the website were developed.
Training courses: The 3 th edition of the training course Informed decision making in healthcare,
targeted to patients’ associations representatives started in September 2008. Thirty people
attended the course. A course for lay members of ethic committees was also organized, targeted
to the ethic committees of the Regione Lombardia. Twenty people attended the course.
The duplicated lecture notes of the course were published and printed.
Surveys: In 2007 a 14 items questionnaire has been posted to 147 patients’ associations entitled
Does clinical research answer to patients needs Considering the results of this survey, a
questionnaire specifically targeted to pediatric associations about the relevance of the research
was developed in 2008 together with the Laboratorio per la salute materno infantile.
53
ANNUAL REPORT 2008

IRFMN
The website: Three areas with access registration restricted to different working groups were set
up. The working groups are: the jury of the Consensus Conference “Informing women about
hormone replacement therapy”; lay members of ethic committees attending the course; patients’
associations representatives attending the course. New tools to critically evaluate healthcare
information and issues were discussed and developed: the misura-campagne, to evaluate public
awareness campaigns; the misura-medico, to evaluate medical practitioners. This tool will be
reviewed by a sample of medical practitioners and lay people. We set up a channel on You tube
(http://it.youtube.com/Partecipasalute) and another on Slideshare.net
(http://www.slideshare.net/Partecipasalute) in order to share videos and power point
presentations. The monthly visits to the websites were on average 30.000.
Consensus conference “Informing women about hormone replacement
therapy”
The Partecipasalute project together with the National Guidelines System (SNLG) based at the
Istituto Superiore di Sanità, organized the Consensus conference “Informing women on
hormone replacement therapy” in order to assess the current status of the quality of information
on hormone replacement therapy (HRT) and re-visit recent research findings on its
risks/benefits.
We chose a structured approach based on the traditional consensus conference method
combined with a structured preparatory work supervised by an organizing committee and a
scientific board. The organizing committee and scientific board chose the members of the CC’s
jury and appointed three multidisciplinary working groups composed by healthcare
professionals, journalists, citizens and patients’ representatives. Before the CC, the three
working groups carried out: a literature review on the risk/benefit profile of HRT and two
surveys on the quality of information on lay press and booklets targeted to women. A
population survey on women’s knowledge, attitude and practice was also carried out. The jury
received the documents in advance, listened the presentations during the two-day meeting of the
CCs, met immediately after in a closed-door meeting and prepared the final document, available
at http://www.partecipasalute.it/cms/files/Documento-definitivo-consenso.pdf. The public
session of the conference took place in Turin on 16 and 17 May 2008. The project was
supported by Compagnia di San Paolo, a non profit private-law foundation.
Programma 1, WP5 -Alleanza contro il cancro – Servizio nazionale di accoglienza
e informazione sul cancro. Gruppo per l'Informazione ACCP1 WP5
The project is coordinated by the Istituto superiore di sanità. Other partners are the Centro di
Riferimento Oncologico - Aviano, AIMaC Associazione italiana malati di cancro, Istituto
Nazionale dei Tumori, Istituto Europeo di Oncologia. The Laboratory of Medical research on
consumer involvement planned to evaluate the quality of websites dealing with breast cancer,
colon rectal cancer, cervical cancer. The focus on internet is due to the increasing number of
people searching for information about cancer, treatments and diagnosis on the web. The
protocol was draft on June 2007 and the evaluation form was defined on October 2008. The
websites included in the sample were collected through a research by key words on the search
engine google (November 2008). Each website will be evaluated by two reviewers.
SNAP project - Smoke, Nutrition, Alcohol and Physical Activity
SNAP is a campaign for the health addressed to employees in a firm in Brianza. This project,
for want of FSE - Frontier Science & Technology Research Foundation, Southern Europe, a
foundation for the support to the independent search - in collaboration with Istituto Mario
Negri, has been launched with the attempt to increase knowledge and to modify the opinions,
the attitudes and the behaviors of the people on the four topics examined, with an active process
of information addresses to increase to the knowledge and the information. The formationinformation
plan is structured through the circulation of paper material and a public event of
54
ANNUAL REPORT 2008

IRFMN
discussion. In order to estimate the effectiveness of this plan, a form on knowledge, opinions
and behaviors has been carried out before and three months after the discussion.
The data of the first survey have been already analyzed and have been introduced during the
event in October 2008, to notice the high rate of participation: 313 employees on 522 have
answered the form. The data on the follow-up of the second form will be analyzed and
confronted with a before-after analysis methodology.
A population based evaluation of an intervention to improve cancer pain
management
In collaboration with the ASUR of the Marche region, it has been promoted a project that aimed
to improve the use of opioids cancer patients. The project - carried out in collaboration with the
laboratory of Giovanni Apolone - has been sponsored by the Italian Agency for Drug evaluation
(AIFA). The aim of this project is to pull down the existing barriers on the use of these drugs
and to help patients to deal, in the better way, with cancer pain. The population, practitioners,
oncologists, chemists, psychologists, nurses, patients and cancer voluntary associations will be
involved in this project. In the course of the project, formative and informative participations
and distribution of popular and scientific material (depending on the target) are previewed and
surveys on opinions, attitudes and behaviors of people will be organized. Throughout this year,
all the operative phases have been activated: from the draft of informative pamphlets addressed
to health workers and those for patients and their families, as well as patients’ organizations, to
the preparation of training courses, to the draft of a pocket book addressed to doctors,
concerning interactions between opioids and other drugs, to draft of forms for surveys,
involving patients and general practitioners.
Follow-up in oncology setting
Two studies on follow-up have been designed and carried out in collaboration with the
laboratory of Giovanni Apolone.
The first in collaboration with the Network Oncologica Piemontese regards the follow-up of
patients with endometrial cancer organization for which the evidence available is not sufficient
to draw a path of sure effectiveness. TOTEM study that has the characteristics of an open
randomized multicenter study comparing two different modulations of visits and examinations
and in 2008 was discussed and developed the protocol and all related forms, including from the
point of fair and comprehensive information to patients who will be invited to participate in the
study.
The second study that takes place in the context of the 6th Integrated Project Oncology (Health
Ministry) provides for the comparative assessment of two follow-up for women at moderatelow
risk with a diagnosis of breast cancer and lead to a randomization minimalist follow-up
coordinated by the oncologist or by general practitioner. The study will be operational from
2009.
Quality of life projects
No specific research projects have been carried out on quality of the life evaluation. However
are on going the activities of support and coordination of other groups using the instruments of
quality of life translated and validated by our research group, SF-36, SF-12, PGWBI. During the
year it has been periodically up-to-date the specific website http://crc.marionegri.it/qod.
55
ANNUAL REPORT 2008

IRFMN
56
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF ENVIRONMENTAL
HEALTH SCIENCES
STAFF
Head
Roberto FANELLI, Biol.Sci.D.
Laboratory of Analytical Biochemistry
Head
Chiara CHIABRANDO, Biol.Sci.D.
Laboratory of Environmental Chemistry and Toxicology
Head
Emilio BENFENATI, Chem.D.
Industrial and Environmental Health Unit
Head
Laboratory of Food Toxicology
Marco LODI, Chemist
Head
Ettore ZUCCATO, M.D.
Laboratory of Mass Spectrometry
Head
Enrico DAVOLI, Anim.Sci.D.
Laboratory of Molecular Toxicology
Head
Protein and Gene Biomarkers Unit
Head
Luisa AIROLDI, Pharm.D.
Roberta PASTORELLI, Biol.Sci.D
Department’s Units
Environmental Pollutants Risk Assessment Unit
Head
Elena FATTORE, Biol.Sci.D
Analytical Instrumentation Unit
Head
Renzo BAGNATI, Chem.D.
57
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Roberto Fanelli, Head of the Environmental Health Sciences Department since 1997, Laboratory Head
1978-97, Researcher 1975-78, Research fellow 1969-74 at the Mario Negri Institute.
Doctoral Degree in Biological Sciences (University of Milan, 1973), Assistant Professor in Biochemistry
at Baylor College of Medicine (Houston, Texas). Member of the Commissione Consultiva Prodotti
Fitosanitari (Ministero Salute), Member of the Scientific Panel on Contaminants in the Food Chain
(European Food Safety Authority, 2003-2006), Certified Italian Toxicologist. Member of the Comitato
Scientifico Ente Risi.
Research areas: Sources, diffusion, toxicology, human exposure and risk assessment of persistent
environmental pollutants. Environmental risk of plant protection products. Development of analytical
methods for identification and measurement of biomarkers in toxicology. Mechanisms of toxic action by
proteomic techniques.
Selected publications:
• Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater
and surface water. Mass Spectrom Rev 2008 ; 27 : 378-394
• Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis.
Environ Health Perspect 2008 ; 116 : 1027-1032
• Hodgson S, Thomas Laura, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L.
Bone mineral density changes in relation to environmental PCB exposure. Environ Health Perspect 2008 ; 116 : 1162-
1166
• Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey
A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation
with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: 882-894
• Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new
evidence-based tool to monitor community drug abuse. Environ Health 2005; 4: 14
(http://www.ehjournal.net/content/4/1/14 2005)
• Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the
identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: 4936-4945
Luisa Airoldi, Head of the Molecular Toxicology Laboratory since 1994, Unit Head 1987-94, Researcher
1978-87, Technician 1967-75 at the Mario Negri Institute.
Doctoral Degree in Pharmacy (University of Milan, 1975), Postdoctoral fellow at the Massachusetts
Institute of Technology (Cambridge, MA, 1976) and at the Northwestern University Medical School
(Chicago, Il, 1977), Researcher at the Yale University Medical School (New Haven, CT, 1980-81).
Research areas: Proteomics in toxicology with particular interest on the study of proteome changes in
tissues and biological fluids from animals and humans after exposure to toxic compounds; clinical
proteomics aimed at the identification of protein biomarkers as diagnostic tools; molecular epidemiology
focused on the identification and measurement of biomarkers of exposure to environmental carcinogens
and disease susceptibility.
Selected publications:
• Peluso M, Airoldi L, Colombi A, Munnia A, Veglia F, Autrup H, Dunning A, Garte S, Gormally E, Malaveille C,
Matullo G, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh
V, Tumino R, Panico S, Bueno-De-Mesquita H B, Peeters P H, Kumle M, Gonzalez C A, Martinez C, Dorronsoro M,
Barricarte A, Tormo M J, Quiros J R, Berglund G, Janzon L, Jarvholm B, Day N E, Key T J, Saracci R, Kaaks R, Riboli
E, Bingham S, Vineis P. Bulky DNA adducts, 4-aminobiphenyl hemoglobin adducts, diet and air pollution in a healthy
European population. Br J Nutr 2008 100: 489-495.
• Veglia F, Loft S, Matullo G, Peluso M, Munnia A, Perera F, Phillips D H, Tang D, Autrup H, Raaschou-Nielsen O,
Tjonneland A, Vineis P, Genair-EPIC Investigators. DNA adducts and cancer risk in prospective studies: a pooled
analysis and a meta-analysis. Carcinogenesis 2008 ; 29 : 932-936.
• Vineis P, Hoek G, Krzyzanowski M, Vigna-Taglianti F, Veglia F, Airoldi L, Overvad K, Raaschou-Nielsen O, Clavel-
Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D, Krogh V, Tumino R, Panico S, Bueno-De-Mesquita HB,
Peeters PH, Lund E E, Agudo A, Martinez C, Dorronsoro M, Barricarte A, Cirera L, Quiros JR, Berglund G, Manjer J,
Forsberg B, Day NE, Key TJ, Kaaks R, Saracci R, Riboli E. Lung cancers attributable to environmental tobacco smoke
and air pollution in non-smokers in different European countries: a prospective study. Environ Health. 2007 15; 6:7.
• Pastorelli R, Saletta F, Campagna R, Carpi D, Dell'Osta C, Schiarea S, Vineis P, Airoldi L, Matullo G Proteome
characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl Proteome Sci 2007 5:
6
58
ANNUAL REPORT 2008

IRFMN
• Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey
A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation
with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: 882-894
• Airoldi L, Vineis P, Colombi A, Olgiati L, Dell'Osta C, Fanelli R, et al. 4-Aminobiphenyl-hemoglobin adducts and risk
of smoking-related disease in never smokers and former smokers in the European Prospective Investigation into Cancer
and Nutrition Prospective study. Cancer Epidemiol Biomarkers Prev 2005; 14: 2118-2124
Emilio Benfenati, Head of the Laboratory of Environmental Chemistry and Toxicology since 1997, Unit
Head 1987-97, Researcher 1986-87, Research fellow 1981-86 at the Mario Negri Institute. Researcher at
Istituto Biochimico Italiano 1979-1981.
Doctoral Degree in Chemistry (University of Milan, 1979).
Member of Commissione Consultiva Prodotti Fitosanitari (Ministero Salute 1997-99), Certified Italian
Chemist.
Resarch areas: Computer-based models for chemistry and toxicology; Molecular descriptors; QSAR;
Toxicity prediction; Metabolism studies; Characterization and assessment of wastes, industrial effluents,
emissions from landfill and incinerator; Integration of chemical analysis and eco-toxicological data;
Chemical analysis of organic compounds by mass spectrometry.
Principali pubblicazioni:
• Zhao C, Boriani E, Chana A, Roncaglioni A, Benfenati E, A new hybrid QSAR model for the prediction of
bioconcentration factors (BCF), Chemosphere 2008 73 : 1701-1707
• Fjororova N, Novich M, Vrachko M, Kharchevnichova N, Zholdakova Z, Sinitzyna O, Benfenati E,
Regulatory assessment of chemicals within OECD Member Countries, EU and in Russia, J Environ Sci Heal
C 2008 26: 40-88
• Roncaglioni A, Benfenati E, In silico-aided prediction of biological properties of chemicals: oestrogen
receptor-mediated effects, Chem Soc Rev 2008 37: 441-450
• Porcelli C, Boriani E, Roncaglioni A, Chana A, Benfenati E, Regulatory perspectives in the use and validation
of QSAR. A case study: DEMETRA model for daphnia toxicity, Environ Sci Technol 2008 42 : 491-496
• Pandelova M, Henkelmann B, Roots O, Simm M, Jarv L, Benfenati E, Schramm K-W, Levels of PCDD/F and
dioxin-like PCB in Baltin fish of different age and gender, Chemosphere 2008 71: 369-378
• Benfenati E (Ed.), Quantitative Structure-Activity Relationships (QSAR) for Pesticide Regulatory Purposes,
Elsevier Science Ltd, Amsterdam, The Netherlands (2007), 1-510
Chiara Chiabrando, Head of the Analytical Biochemistry Laboratory since 1997, Unit Head 1987-97,
Researcher 1978-87, Research fellow 1975-78 at the Mario Negri Institute.
Doctoral degree in Biological Sciences (University of Milan, 1974), Postdoctoral fellow at the Baylor
College of Medicine (Houston, Texas, 1974-75). Postgraduate degree in Pharmacological Research,
Mario Negri Institute (1977).
Research areas: Development and application of bio-analytical methods based on mass spectrometry in
the fields of biochemistry, metabolism, clinical chemistry and pharmacology. Identification and
characterization of proteins and peptides of biomedical interest by proteomic approaches and mass
spectrometry. Proteomics in oncology.
Selected publications
• Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele C,
Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic
peptides. J Am Soc Nephrol. 2009; 20:123-30. Epub 2008 Dec 17.
• Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis.
Environ Health Perspect 2008; 116: 1027-1032.
• Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometric analysis of illicit drugs in wastewater
and surface water. Mass Spectrom Rev 2008; 27: 378-394.
• Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs
and their metabolites in urban wastewaters by liquid chromatography tandem mass spectrometry (HPLC-MS-MS). Anal
Chem 2006;78: 8421-8429.
• Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the
identification of in vivo estrogen-regulated proteins in bone. Proteomics. 2005; 5:4936-4945.
• Chiabrando C, Rivalta C, Bagnati R, Valagussa A, Durand T, Guy A, Villa P, Rossi JC, Fanelli R. Identification of
metabolites from type III F2-isoprostane diastereoisomers by mass spectrometry. J Lipid Res. 2002;43:495-509.
59
ANNUAL REPORT 2008

IRFMN
Enrico Davoli, Head of the Mass Spectrometry Laboratory since 1997, Unit Head 1994-97, Researcher
1989-94, Research Fellow 1985-87 at the Mario Negri Institute. Fellow at USDA, Beltville, MD 1977-78.
Doctoral Degree in Animal Sciences (University of Milan, 1983), Postdoctoral fellow at the University of
Nebraska (Lincoln, NE, 1987) and at the University of Colorado Health Sciences Center (Denver, CO,
1988). Postgraduate degree in Pharmacological Research, Mario Negri Institute (1988). Member of the
American Association for Mass Spectrometry (ASMS) of the Environment and Safety Commission of
IGQ and of the ETS (Emission Trading System) commission. Member of the National Biomass Research
Center Scientific Committee. Environmental Applications Interest Group Coordinator (ASMS).
Research areas: Development of methodology, instrumentation and software for environmental research.
Studies of urban air pollution and characterization of environmental odor annoyance.
Selected publications
• Davoli E, Bianchi G. Odour emission rates from a waste treatment plant: results from a multi year follow-up study.
Chemical Engineering Transactions 2008; 15 : 95-102.
• Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from
European countries. Food Chem Toxicol 2008, 46: 1062-1067.
• Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in
milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21
: 1998-2002
• Riservato M, Rolla A, Davoli E . An isotopic dilution approach for 1,3-butadiene tailpipe emissions and ambient air
monitoring. Rapid Commun Mass Spectrom 2004; 18: 399-404
• Davoli E, Gangai L, Morselli P, Tonelli D. Characterisation of Odorant emissions from Landfills by SPME and GC/MS.
Chemosphere 2003; 51: 357-368
• Davoli E, Cappellini L, Fanelli R, Bonsignore M, Gavinelli M. On-Site Analysis of World War II Cylinders and Barrels
with Unknown Contents. Field Anal. Chem. Technol. 2001; 5: 313-319
Ettore Zuccato, Head of the Food Toxicology Laboratory since 2005, Unit Head 1997-2005, Researcher
1986-97, Technician 1975-86 at the Mario Negri Institute.
Doctoral degree in Medicine (University of Milan, 1986), Postdoctoral degree in Human Nutrition
(1999), Postdoctoral fellow at the King’s College School of Medicine (London, UK, 1988-89).
Member of the ANSISA, EMEA expert, member of the Commissione Consultiva per i Prodotti
Fitosanitari, and expert for the evaluation of plant protection products for registration within the EU.
Research areas: Food safety, including the study of dietary chemical contaminants, safety assessment of
GMO in human nutrition, food allergens and toxicants, emerging issues in food toxicology, risk
perception and risk communication to the consumers, and evaluation of plant protection products for
registration within the European Union. Environmental pollution by pharmaceuticals, and monitoring of
illicit drugs in surface waters to estimate community drug abuse.
Selected publications
• Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater analysis.
Environ Health Perspect 2008, 116: 1027-1032.
• Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometry analysis of illicit drugs in wastewater
and surface water. Mass Spectrom Rev, 2008, 27: 378-394.
• Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from
European countries. Food Chem Toxicol 2008, 46: 1062-1067.
• Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs
and their metabolites in urban wastewaters by liquid chromatography tandem mass spectrometry (HPLC-MS-MS). Anal
Chem 2006, 78: 8421-8429.
• Zuccato E, Calamari D, Castiglioni S, Chiabrando C, Bagnati R, Fanelli R. Cocaine in surface water: a new evidencebased
tool to monitor community drug abuse. Environmental Health: A Global Access Science Source 2005, 4:14
• Zuccato E, Calamari D, Natangelo M, Fanelli R. Presence of therapeutic drugs in the environment. Lancet 2000; 355:
1789-1790
60
ANNUAL REPORT 2008

IRFMN
Renzo Bagnati, Head of the Analytical Instrumentation Unit since 2005, Researcher 1992-2005,
Research fellow 1986-92 at the Mario Negri Institute.
Doctoral degree in Chemistry (University of Turin, 1985), Postgraduate degree in Pharmacological
Research, Mario Negri Institute (1989).
Research areas: Mass spectrometry applied to the analysis of biological and environmental relevant
substances (proteins, peptides, hormones, pharmaceuticals, drugs of abuse, pesticides).
Selected publications
• Zuccato E, Castiglioni S, Bagnati R, Chiabrando C, Grassi P, Fanelli R. Illicit drugs, a novel group of environmental
contaminants. Water Res 2008 ; 42 : 961-968.
• Analysis of phosphoinositides and their aqueous metabolites. Berrie CP, Iurisci C, Piccolo E, Bagnati R, Corda D.
Methods Enzymol. 2007;434:187-232.
• Bagnati R, Bianchi G, Marangon E, Zuccato E, Fanelli R, Davoli E. Direct analysis of isopropylthioxanthone (ITX) in
milk by high-performance liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2007 ; 21
: 1998-2002.
• Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and measurement of illicit drugs
and their metabolites in urban wastewater by liquid chromatography-tandem mass spectrometry. Anal Chem 2006; 78:
8421-8429.
• Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, Fanelli R. Cocaine in surface waters: a new
evidence-based tool to monitor community drug abuse. http://www.ehjournal.net/content/4/1/14 2005.
• Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C. Proteome analysis for the
identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: 4936-4945.
Elena Fattore, Head of the Environmental Pollutants Risk Assessment Unit since 2005, Researcher
2001-2004, Research fellow 1991-1997 at the Mario Negri Institute.
Doctoral Degree in Biological Sciences (University of Milan, 1991), Postgraduate degree in
Pharmacological Research, Mario Negri Institute (1994), Postdoctoral fellow at the National Institute of
Environmental Medicine, Karolinska Institutet, Stockholm (1998-2000).
Research areas: Environmental chemistry, toxicology, assessment of human exposure and risk from
environmental pollutants with emphasis on dioxins and dioxin-like compounds.
Selected publications
• Fattore E, Fanelli R, Dellatte E, Turrini A, Di Domenico A. Assessment of the dietary exposure to non-dioxin-like PCBs
of the Italian general population. Chemosphere 2008, 73: S278-S283.
• Hodgson S, Thomas L, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup L Bone
mineral density changes in relation to environmental PCB exposure. Environmental Health Perspective 2008, 116: 1162-
1166.
• Carubelli G, Fanelli R, Mariani G, Nichetti S, Crosa G, Calamari D, Fattore E. PCB contamination in farmed and wild
sea bass (Dicentrarchus labrax L.) from a coastal wetland area in central Italy. Chemosphere 2007 ; 68 : 1630-1635.
• Fattore E, Fanelli R, Turrini A, Di Domenico A. Current dietary exposure to polychlorodibenzo-p-dioxins,
polychlorodibenzofurans, and dioxin-like polychlorobiphenyls in Italy. Mol Nutr Food Res 2006; 50: 915-921
• Fattore E, Guardo A, Mariani G, Guzzi A, Benfenati E, Fanelli R. Polychlorinated Dibenzo-p-Dioxins and
Dibenzofurans in Air of Seveso, Italy, 26 years after the accident. Environ Sci Technol 2003; 37: 1503-1058
• Fattore E, Fanelli R, La Vecchia C. Persistent organic pollutants in food: Public health and implications. J Epidemiol
Community Health 2002; 56: 831-832
61
ANNUAL REPORT 2008

IRFMN
Marco Lodi, Head of the Industrial and Environmental Unit since 2002, Consultant 1997-2002 at the
Mario Negri Institute.
General Certificate of Education in Industrial Chemistry (Milan, 1974).
Member of AIDII (Italian Industrial Hygiene Association), certified by ACGIH (American Conference of
Governmental Industrial Hygienist).
Research areas: Emission sources, environmental diffusion, toxicology, human exposure and risk
assessment of persistent environmental pollutants. Environmental risk of chemical pollution products.
Development of sampling methods for environmental toxic compounds.
Selected publications
• Benfenati E, Azimonti G, Auteri D, Lodi M Environmental and ecological toxicology: computational risk assessment.
Computational toxicology. Risk assessment for pharmaceutical and environmental chemicals John Wiley, Hoboken, NJ,
2007; 625-650
• Grosso M, Cernuschi S, Palini E, Lodi M, Mariani G. PCDD/Fs release during normal and transient operation of a full
scale MSWI plant. Organohalogen Compounds 2004; 66: 1243-1249
• Benfenati E, Mariani G, Lodi M, Reitano G, Fanelli R. Is bioexsiccation releasing dioxins Organohalogen Compounds
2004; 66: 955-961
• Fattore E, Mariani G, Guzzi A, Di Guardo A, Benfenati E, Lodi M, Fanelli R. Air dioxin concentrations in Seveso area.
In: Halogenated Environmental Organic Pollutants, 18th. Symp., Stockholm, Sweden, august 17-21, 1998. 1998 : 237-
240
• Benfenati E, Mariani G, Schiavon G, Lodi M, Fanelli R. Diurnal, weekly and seasonal air concentrations of PCDD and
PCDF in an industrial area. Fresenius Journal Analytical Chemistry 1994; 348: 141-143
• Benfenati E, Pastorelli R, Castelli M G, Fanelli R, Carminati A, Farneti A, Lodi M. Studies on the tetrachlorodibenzo-pdioxins
(TCDD) and tetrachlorodibenzofurans (TCDF) emitted from an urban incinerator. Chemosphere 1986; 15: 557-
561
Roberta Pastorelli, Head of Protein and Gene Biomarkers Unit since 2004, Researcher 1992-2003,
Research fellow 1983-92 at the Mario Negri Institute.
Doctoral Degree in Biological Sciences (University of Milan, 1982), Postgraduate degree in
Pharmacological Research, Mario Negri Institute (1986), Postdoctoral fellow at the Massachusetts
Institute of Technology, Cambridge, MA (1987-89 and 1991).
Research areas: Toxicoproteomic investigation of global protein expression profiles and their modulation
evoked by environmental compounds in different biological compartments. Critical targets and pathways
in toxicology. Pharmacogenetics: effects of genetic polymorphisms in the human population on the
individual susceptibility to environmental xenobiotic and carcinogen effects.
Selected publications:
• Pastorelli R, Saletta F, Campagna R, Carpi D, Dell’Osta C, Schiarea S, Vineis P, Airoldi L, Matullo G. Proteome
characterization of a human urothelial cell line resistant to the bladder carcinogen 4-aminobiphenyl. Proteome Science
2007.
• Moretti M, Dell'omo M, Villarini M, Pastorelli R, Muzi G, Airoldi L, Pasquini R. Primary DNA damage and genetic
polymorphisms for CYP1A1, EPHX and GSTM1 in workers at a graphite electrode manufacturing plant. BMC Public
Health. 2007 7: 270
• Pastorelli R, Carpi D, Campagna R, Airoldi L, Pohjanvirta R, Viluksela M, Hakansson H, Boutros P C, Moffat I D, Okey
A B, Fanelli R. Differential expression profiling of the hepatic proteome in a rat model of dioxin resistance: correlation
with genomic and transcriptomic analyses. Mol Cell Proteomics 2006; 5: 882-894
• Pastorelli R, Carpi D, Airoldi L, Chiabrando C, Bagnati R, Fanelli R, Moverare S, Ohlsson C.Proteome analysis for the
identification of in vivo estrogen-regulated proteins in bone. Proteomics 2005; 5: 4936-4945
• Airoldi L, Magagnotti C, Pastorelli R, Fanelli R. Enzyme polymorphisms influencing the metabolism of heterocyclic
aromatic amines. J Chromatogr B Analyt Technol Biomed Life Sci 2004; 802: 175-181
• Vineis P,V eglia F, Anttila S, Benhamou S, Clapper M L, Dolzan V, Ryberg D, Hirvonen A, Kremers P, Le Marchand L,
Pastorelli R, Rannug A, Romkes M, Schoket B, Strange R C, Garte S, Taioli E. CYP1A1, GSTM1 and GSTT1
polymorphisms and lung cancer: a pooled analysis of gene-gene interactions. Biomarkers 2004; 9: 298-305
62
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department works to investigate environmental factors and their effects on human health.
The main research lines focus on the survey of environmental contaminants, the assessment of
human exposure with related health risks, and toxicity mechanisms of pollutants.
The assessment of environmental contamination is carried out not only for well-known and
widespread compounds, like dioxins and PCBs, but also for new classes of "unconventional"
pollutants, e.g., endocrine disruptors, potentially toxic "natural" compounds, and drugs entering
the environment after human or veterinary use. The identification –for the first time– of illicit
drugs in urban waste and river waters, led to a new original tool for the evidence-based
monitoring of community drug abuse. For all these survey activities sophisticated analytical
methods based on advanced mass spectrometric techniques are developed.
The Department is active in the assessment of human exposure to toxic compounds in the
atmosphere and the diet, which is the main source of priority pollutants (PCBs, dioxins and
other endocrine disruptors). Assessment of the risk associated to contamination in real-life
scenarios has recently gained much importance. In order to respond to the growing demand for
information, the Department is more and more involved in toxicological and ecotoxicological
risk analysis, based on studies in field and predictive models of toxicity.
Molecular epidemiology studies are used to identify genetic and/or environmental factors
posing risks to human health. By this approach, we search for new useful “biological markers"
to identify susceptible subjects, in view of finding appropriate preventive strategies.
The Department has implemented an advanced technological proteomic platform, in order to
identify proteins differentially expressed in biological compartments in various experimental
and clinical conditions. This approach is particularly relevant in toxicology, since it can
contribute to find new biomarkers of toxicity or pathology, and to identify molecular targets and
toxic effect mechanisms of pollutants and drugs. To integrate our proteomic studies, we have
now introduced among our activities metabolomics, i.e., the study of small molecules, such as
amino acids, carbohydrates, lipids, hormones etc., the final products of protein expression and
activity which contribute to define the biochemical phenotype of a biological system.
Mass spectrometry (MS) is a central analytical technique at the Department, where a complete
set of state-of-the-art instrumentation is available, from GC-MS and LC-MS to MALDI-TOF-
MS. These instruments are provided with modern solutions for sample introduction (chip-based
nanoLC), sample ionization (ESI, DESI and MALDI), tandem MS (MSn) by triple quadrupole
and TOF-TOF instruments, high mass resolution analysis (hybrid ion trap/orbitrap).
FINDINGS/MAIN RESULTS
The lack of retinoic acid (knock-out mice for the retinal dehydrogenase 1 gene, RALDH1)
modifies the proteome profile of the bone,through changes in the expression of proteins
involved in chondrogenesis and osteoclastogenesis.
Resistance to the bladder carcinogen 4-aminobiphenyl in human urothelial cells is mediated by
deregulation of apoptosis and membrane trafficking proteins.
Bone protein profile in a murine model of osteoporosis.
Identification of novel protein targets responsive to the effects of estrogens in bone.
TCDD's effect on the liver proteome profile of exposed rats. Determination of a subset of rat
hepatic proteins indicative of differences in dioxin susceptibility.
The presence of 4-aminobiphenyl-hemoglobin adducts may help identify nonsmokers at high
risk of cancers related to environmental tobacco smoke exposure.
63
ANNUAL REPORT 2008

IRFMN
Reference values of allele and genotype frequency of several metabolic genes in 15,000 control
subjects.
CYP1A1 polymorphism affects lung tumor risk.
Identification of CYP2C9 genetic polymorphism as a determinant of severe adverse reactions to
phenytoin.
On-line in silico models to predict ecotoxicity of pesticides for regulatory purposes.
A new model for identification of endocrine disruptors using molecular docking.
A method aimed at characterizing environmental odors to identify odor sources in complex
environments.
Albumin can become a source of potentially antigenic peptides in proteinuric nephropaties. An
acid protease released by rat renal proximal tubular cells selectively cleaves an albumin N-
terminal fragment (ALB1-24). Kidney infiltrating dendritic cells (DC) can capture ALB1-24
and process it, through the proteasome, to shorter peptides capable of activating syngeneic
CD8+ T cells.
Moderate-to-high fish consumption can result in exceeding the daily intake safety levels of
PCBs and dioxin-like compounds established by the European Commission.
The same food type may contain significantly different concentrations of PCBs and dioxin-like
compounds, depending on the geographic origin (this may help lower the risk for the consumers
by understanding and controlling the causes of the differences).
The environmental levels of several drugs exceed the “safety limits” established for them.
Environmental pollution from pharmaceuticals is a general phenomenon that can be described
by controllable variables (environmental load and mass balance).
Illicit drug residues and their metabolites were found in urban waste and river waters.
Environmental levels can be used as a new tool to estimate illicit drugs consumption in the
population.
The distribution of dietary intake values of dioxins, dioxin-like PCBs and non dioxin-like PCBs
was characterized for the general Italian population.
The higher intake of PCBs due to consumption of farmed fish vs. wild fish is mainly due to the
higher fat content in farmed fish.
Development of novel mass spectrometric methods for the selective measurement of therapeutic
and illicit drugs in environmental samples.
NATIONAL COLLABORATIONS
ARPA Emilia Romagna
ARPA Veneto
ASL di Brescia
ASL di Como
ASL di Cagliari
ASL di Napoli
CNR - IRSA
Comune di Rosignano Marittimo (LI)
CSRA-Asti
Fondazione 'S. Maugeri'
Fondazione ISI, Torino
INRAN-Istituto Nazionale di Ricerca sugli Alimenti e la Nutrizione
ISPO, Firenze
Istituto Clinico Humanitas, Milano
64
ANNUAL REPORT 2008

IRFMN
Istituto Superiore di Sanità
I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana
Ministero dell'Ambiente
Politecnico di Milano
Politecnico di Torino
Provincia di Vercelli
Provincia Pordenone
Università degli Studi di Cagliari
Università degli Studi di Genova
Università degli Studi di Milano
Università degli Studi di Napoli "Federico II"
Università degli Studi di Palermo
Università degli Studi di Pavia
Università degli Studi di Perugia
Università degli Studi di Roma "La Sapienza"
Università degli Studi di Torino
Università dell’Insubria, Varese
Università degli Studi di Verona
65
ANNUAL REPORT 2008

IRFMN
INTERNATIONAL COLLABORATIONS
BASF Agricultural Centre, Limburgerhorf, Germany
Central Science Laboratory, York, UK
Centre for Environmental Policy, Imperial College, London, UK
Danish Institute of Agricultural Sciences, Research Centre Foulum, Tjele, Denmark
Department of Analytical and Pharmaceutical Chemistry, The Royal Danish School of
Pharmacy, Denmark
Department of Anatomy and Cell Biology, University of Oulu, Oulu, Finland
Department of Chemistry, Loyola University, Chicago, IL, USA
Department of Computer Science and Engineering, University of Galati, Romania
Department of Electrical and Computer Engineering, University of Patras, Greece
Department of Environmental Health, National Public Health Institute, Kuopio, Finland
Department of Epidemiology & Public Health, Imperial College, London, United Kingdom
Department of Inland Fisheries, Institute of Freshwater Ecology and Inland Fisheries, Germany
Department of Molecular Biology, University of Bergen, Bergen, Norway
Department of Organic Chemistry, Universidad de Cadiz, Spain
Division of Endocrinology, Department of Internal Medicine, Sahlgrenska University Hospital,
Gothenburg, Sweden
Environmental Chemistry, IIQAB-CSIC, Barcelona, Spain
Environmental Hygiene and Chemistry Department, Institute of Environmental Medicine and
Hospital Epidemiology, University of Freiburg, Germany
Environmental Protection Agency, US EPA - National Risk Management Research Laboratory
(NRMRL), Cincinnati OH, USA
European Monitoring Centre for Drugs and Drug Addiction (EMCDDA), Lisbon, Portugal
Faculté de Médicine et de Pharmacie, Université de Mons-Hainaut, Mons, Belgium
Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
Forschungzentrum Jülich Gmbh, Jülich, Germany
Gruppo Collaborativo sulla Suscettibilità Genetica ai Cancerogeni Ambientali (GSEC), Italia
Institute of Environmental Medicine. Karolinska Institute, Stockholm, Sweden
Institute of Pharmaceutical Chemistry, University of Pécs, Pecs, Hungary
Institute of Phytomedicine, Biological Control, Horticulture and Nematology, Wien, Austria
Institute of Soil Science and Plant Cultivation, Pulawy, Poland
Interuniversitaeres Forchunginstitut fuer Agrarbiotechnologie, Tulln, Austria
Istituto di Chimica di São Carlos, Università di São Paulo, Brazil
KnowledgeMiner Software, Berlin, Germany
In Vitro Testing Industrial Platform, Tres Cantos (Madrid), Spain
Laboratory of Chemometrics & Bioinformatics, University of Orléans, Orléans, France
Laboratory of Neurobiology, Centro de Investigation Principe Felipe, Valencia, Spagna
Lithuanian Institute of Agricultrure, Vilnius, Lithuania
Liverpool John Moores University, Liverpool, United Kingdom
National Institute of Chemistry, Kemijski Institut Ljubljana, Ljubljana, Slovenia
Natural Resources Research Institute, University of Minnesota, Duluth, MN, USA
National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands
Pesticide Safety Directorate, York, UK
Plant Protection Institute, Hungarian Academy of Sciences, Budapest, Hungary
School of Biomedical Sciences, University of Ulster, Coleraine, Northern Ireland
Syngenta Crop Protection AG, Basel, Switzerland
UFZ Leipzig, Germany
University of Tartu, Tartu, Estonia
66
ANNUAL REPORT 2008

IRFMN
EDITORIAL BOARD MEMBERSHIP
Journal of Environmental Science and Health, Part B (Emilio Benfenati), Journal of
Environmental Science and Health, Part C (Emilio Benfenati), International Journal of
Computational Intelligence (Emilio Benfenati), International Journal of Information Technology
(Emilio Benfenati), International Journal of Signal Processing (Emilio Benfenati), Chemistry
Central Journal (Emilio Benfenati), Current Computer Aided Drug Design (Emilio Benfenati),
Advances in Chemoinformatics and Computational Methods (Emilio Benfenati), The Open
Biomarkers Journal (Luisa Airoldi).
PEER REVIEW ACTIVITIES
Addiction, Analytical and Bioanalytical Chemistry, Chemical Biology & Drug Design,
Chemical Research Toxicology, Chemometrics and Intelligent Laboratory Systems,
CHEMOLAB, Chemosphere, Clinical Chemistry and Laboratory Medicine, Environment
International, Environmental Pollution, Environmental Modeling & Software, Environmental
Science & Technology, Journal of Chemical Information and Modeling, International Journal of
Molecular Science, Journal Computer-Aided Molecular Design, Journal of Hazardous
Materials, Molecular Diversity, Proteomics, Royal Society's Philosophical Transactions, Stroke,
Toxicology Letters, Waste Management, Water Research.
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
CCPF - Commissione Consultiva Prodotti Fitosanitari (Ministero della Salute, Ministero
dell'Ambiente)
ECCO - European Commission Coordination
EFSA - European Food Safety Authority
IGQ - Commissione Ambiente e Sicurezza
IGQ - Commissione ETS
EVENT ORGANIZATION
Workshop “Il triplo quadrupolo: trent'anni di successi!”, Istituto di Ricerche Farmacologiche
“Mario Negri”, Milan, Italy, December 18, 2008.
Thermo Proteomics Seminar, Istituto di Ricerche Farmacologiche “Mario Negri”, Milan, Italy,
November 13, 2008.
Seminario “Procedure per l’analisi e la caratterizzazione delle proteine: panoramica delle
soluzioni”, Istituto di Ricerche Farmacologiche “Mario Negri”, Milan, Italy, October 27, 2008.
NOSE2008. International Conference on Environmental Odour Monitoring and Control. Rome,
Italy, July 6-8, 2008.
67
ANNUAL REPORT 2008

IRFMN
Workshop “I modelli predittivi per il REACH: Istruzioni per l’uso”, Istituto di Ricerche
Farmacologiche “Mario Negri”, Milan, Italy, July 1, 2008.
Workshop “Il Monitoraggio dei Microinquinanti”, Istituto di Ricerche Farmacologiche “Mario
Negri”, Milan, Italy, May 29, 2008.
Workshop “From classical Qualitative Proteomics to Imaging and Quantitative Proteomics: an
Overview of Technologies and Applications”, Istituto di Ricerche Farmacologiche “Mario
Negri”, Milan, Italy, April 8, 2008.
Workshop of the SCARLET EC project on in silico methods for carcinogenicity and
mutagenicity, Istituto di Ricerche Farmacologiche “Mario Negri”, Milan, Italy, April 2-4, 2008.
1 st Assembly of the OSIRIS EC project, Istituto di Ricerche Farmacologiche “Mario Negri”,
Milan, Italy, March 11-13, 2008.
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
The Belgian Society for Toxicology & Ecotoxicology (BELTOX) and The Luxembourg
Toxicology Society (BLT) Joint Meeting, Brussels, Belgium, November 28, 2008.
Venice 2008. Second International Symposium on Energy from Biomass and Waste. Venice,
Italy, November 17-20, 2008.
EPAA Annual Conference "Research into alternative approaches (3Rs) in regulatory testing:
Are we on the right track", Brussels, Belgium, November 3, 2008.
1st SETAC Europe Special Science Symposium - "Integrated Testing Strategies for REACH:
From science to practical implementation", Brussels, Belgium, October 23-24, 2008.
5th European Conference on Pesticides and Related Organic Micropollutants in the
Environment, Marseille, France, October 22-25, 2008.
Workshop of the ATHON EC project, Midterm review, Amsterdam, The Netherlands,
September 28-30, 2008.
Convegno “Interferenti endocrini: valutazione e prevenzione dei possibili rischi per la salute
umana”. Istituto Superiore di Sanità, Rome, Italy, October 15, 2008.
Dioxin 2008, 28 th International Symposium on Halogenated Persistent Organic Pollutants
(POPs), Birmingham, UK, August 17-22, 2008.
HUPO 2008, 7 th world congress, Amsterdam, The Netherlands, August 16-20, 2008.
56th American Society for Mass Spectrometry Conference, Denver, CO, USA, June 1-5, 2008.
Workshop “Il Monitoraggio dei Microinquinanti”, a c. di Environnement Italia Spa, Istituto di
Ricerche Farmacologiche “Mario Negri”, Milan, Italy, May 29, 2008.
SETAC Europe 18th Annual Meeting, Warsaw, Poland, May 25-29, 2008.
68
ANNUAL REPORT 2008

IRFMN
Corso “Problems & approaches in computational chemistry”, Politecnico di Milano, Milan,
Italy, April 21-22, 2008.
Workshop of the SCARLET EC project on in silico methods for carcinogenicity and
mutagenicity, Istituto di Ricerche Farmacologiche “Mario Negri”, Milan, Italy, April 2-4, 2008.
KNAPPE International Conference, Nimes, France, February19-20, 2008.
4th International Workshop on Liquid Chromatography in Environmental and Food samples,
Barcelona, Spain, February 7-8, 2008.
.
GRANTS AND CONTRACTS
A2A Brescia
ACEGAS S.p.A, Trieste
AIIPA (Associazione Italiana Industrie Prodotti Alimentari)
AMA, Roma
ASL Como
ASL Mantova
ASL Napoli 2
Associazione Italiana Ricerca sul Cancro
COOP Italia
CSRA
Comune di Lomello (PV)
Comune di Mazzano e Rezzato (BS)
Consorzio Quadrifoglio S.p.A.
ECODECO, Pavia
European Commission (MEBFOOD, HERBICBIOREM, ATHON, CASCADE, CAESAR,
CHEMOMENTUM, CHEMPREDICT, OSIRIS, SCARLET)
Federchimica, Milano
FIAT Auto S.p.A.
Fondazione CARIPLO, Milano
Fondazione Italo Monzino, Milano
HERA S.p.A. (Holding Energia Risorse Ambiente)
I.Z.S.L.T - Istituto Zooprofilattico Sperimentale del Lazio e Toscana
Lucart, Cartiera Lucchese S.p.A. Porcari, Lucca
Ministero della Salute, Italia
Ministero dell'Ambiente, Italia
Oxon Italia S.p.A., Pero (MI)
Provincia di Pordenone
Provincia di Vercelli
SO.GE.NU.S. S.p.A
Telethon
Tenacta Group
TM.E. S.p.A
69
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Peluso M, Airoldi L, Colombi A, Munnia A, Veglia F, Autrup H, Dunning A, Garte S, Gormally E, Malaveille C,
Matullo G, Overvad K, Raaschou-Nielsen O, Clavel-Chapelon F, Linseisen J, Boeing H, Trichopoulou A, Palli D,
Krogh V, Tumino R, Panico S, Bueno-De-Mesquita H B, Peeters P H, Kumle M, Gonzalez C A, Martinez C,
Dorronsoro M, Barricarte A, Tormo M J, Quiros J R, Berglund G, Janzon L, Jarvholm B, Day N E, Key T J, Saracci
R, Kaaks R, Riboli E, Bingham S, Vineis P. Bulky DNA adducts, 4-aminobiphenyl hemoglobin adducts, diet and air
pollution in a healthy European population. Br J Nutr 2008 100: 489-495.
Veglia F, Loft S, Matullo G, Peluso M, Munnia A, Perera F, Phillips D H, Tang D, Autrup H, Raaschou-Nielsen O,
Tjonneland A, Vineis P, Genair-EPIC Investigators. DNA adducts and cancer risk in prospective studies: a pooled
analysis and a meta-analysis. Carcinogenesis 2008 ; 29 : 932-936.
Roncaglioni A, Piclin N, Pintore M, Benfenati E. Binary classification models as screening tools for endocrine
disrupter effects mediated through the estrogen receptor. SAR QSAR Environ Res 2008 19 : 697-733
Viganò L, Benfenati E, van Cauwenberge A, Eidem J K, Erratico C, Goksøyr A, Kloas W, Maggioni S, Mandich A,
Urbatzka R. Estrogenicity profile and estrogenic compounds determined in river sediments by chemical analysis,
ELISA and yeast assays. Chemosphere 2008 73 : 1078-1089
Casalegno M, Sello G, Benfenati E. Definition and detection of outliers in chemical space. J Chem Inf Model 2008
48 : 1592-1601
Zhao C, Boriani E, Chana A, Roncaglioni A, Benfenati E. A new hybrid QSAR model for the prediction of
bioconcentration factors (BCF). Chemosphere 2008 73 : 1701-1707
Toropov A A, Toropova A P, Benfenati E. QSPR modeling for enthalpies of formation of organometallic
compounds by means of SMILES-based optimal descriptors. Chemical Physics Letters 2008 461 : 343-347
Slavov S, Gini G, Benfenati E. QSAR trout toxicity models on aromatic pesticides. J Environ Sci Heal B 2008 43 :
633-637
Colombo A, Benfenati E, Karelson M, Maran U. Definition of a univocal architecture for chemical splitting to
optimise QSAR models for aquatic toxicity. Chemosphere 2008 72 : 772-780
Fjodorova N, Novic M, Vracko M, Smirnov V, Kharchevnikova N, Zholdakova Z, Novikov S, Skvortsova N,
Filimonov D, Poroikov V, Benfenati E. Directions in QSAR Modeling for Regulatory Uses in OECD Member
Countries, EU and in Russia. J Environ Sci Heal C 2008 26 : 201-236
Toropov A A, Benfenati E. Additive SMILES-based optimal descriptors in QSAR modelling bee toxicity: Using
rare SMILES attributes to define the applicability domain. Bioorg Med Chem 2008 16 : 4801-4809
Bursztyka J, Perdu E, Tulliez J, Debrauwer L, Delous G, Canlet C, De Sousa G, Rahmani R, Benfenati E, Cravedi
J-P. Comparison of genistein metabolism in rats and humans using liver microsomes and hepatocytes. Food Chem
Toxicol 2008 46: 939-948
Porcelli C, Roncaglioni A, Chana A, Benfenati E. A comparison of DEMETRA individual QSARs with an index
with an index of evaluation of uncertainty. Chemosphere 2008 71: 1845-1852.
Fjororova N, Novich M, Vrachko M, Kharchevnichova N, Zholdakova Z, Sinitzyna O, Benfenati E. Regulatory
assessment of chemicals within OECD Member Countries, EU and in Russia. J Environ Sci Heal C 2008 26: 40-88
Roncaglioni A, Benfenati E. In silico-aided prediction of biological properties of chemicals: oestrogen receptormediated
effects. Chem Soc Rev 2008 37: 441-450
Porcelli C, Boriani E, Roncaglioni A, Chana A, Benfenati E. Regulatory perspectives in the use and validation of
QSAR. A case study: DEMETRA model for daphnia toxicity. Environ Sci Technol 2008 42 : 491-496
Pandelova M, Henkelmann B, Roots O, Simm M, Jarv L, Benfenati E, Schramm K-W. Levels of PCDD/F and
dioxin-like PCB in Baltin fish of different age and gender. Chemosphere 2008 71: 369-378
70
ANNUAL REPORT 2008

IRFMN
Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele
C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic
peptides. J Am Soc Nephrol. 2009; 20:123-30. Epub 2008 Dec 17.
Davoli E, Bianchi G. Odour emission rates from a waste treatment plant: results from a multi year follow-up study.
Chemical Engineering Transactions 2008 ; 15 : 95-102.
Castiglioni S, Pomati F, Miller K, Rossetti C, Zuccato E, Calamari D and Neilan BA. Exchange of the multiple
resistance gene marA in antibiotic-contaminated environments involving Bacillus sp. Water Research, 2008,
42:4271-4280.
Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug abuse by wastewater
analysis. Environmental Health Perspectives, 2008, 116: 1027-1032.
Castiglioni S, Zuccato E, Chiabrando C, Fanelli R, Bagnati R. Mass spectrometry analysis of illicit drugs in
wastewater and surface water. Mass Spectrometry Reviews, 2008, 27: 378-394.
Farré M, Petrovic P, Gros M, Kosjek T, Martinez E, Heath E, Osvald P, Loos R, Le Menach K, Budzinski H, De
Alencastro F, Müller J, Knepper T, Fink G, Ternes TA, Zuccato E, Kormali P, Gans O, Quintana JB, Pastori F,
Gentili A, Barceló D. First interlaboratory exercise on non-steroidal anti-inflammatory drugs analysis in
environmental samples. Talanta, 2008, 76: 580-590.
Pomati F, Orlandi C, Clerici M, Luciani F, Zuccato E. Effects and interactions in an environmentally relevant mixture
of pharmaceuticals. Toxicol Sci 2008, 102(1): 129-137.
Zuccato E, Castiglioni S, Bagnati R, Chiabrando C, Grassi P, Fanelli R. Illicit drugs, a novel group of environmental
contaminants. Water Research 2008, 42: 961-968.
Zuccato E, Grassi P, Davoli E, Valdicelli L, Wood D, Reitano G, Fanelli R. PCB concentrations in some food from
European countries. Food Chem Toxicol, 2008, 46: 1062-1067.
Fattore E, Fanelli R, Dellatte E, Turrini A, Di Domenico A. Assessment of the dietary exposure to non-dioxin-like
PCBs of the Italian general population. Chemosphere 2008, 73: S278-S283.
Hodgson S, Thomas Laura, Fattore E, Lind P M, Alfven T, Hellstrom L, Hakansson H, Carubelli G, Fanelli R, Jarup
L Bone mineral density changes in relation to environmental PCB exposure. Environmental Health Perspective 2008,
116: 1162-1166.
LAY PRESS SELECTION PUBLISHED IN 2008
Colombo A, Benfenati E, Lodi M. Diossine, fonti di emissione e conseguenze. Laboratorio 2000 2008 : 46-48
Fattore E, Paiano V. Il rischio sanitario in relazione alla qualità dell'aria. Ricerca & Pratica 2008, 144: 231-241
OTHER PRODUCTS PUBLISHED IN 2008
Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug use. In: EMCDDA Insight
9: Assessing illicit drugs in wastewater; potential and limitation of a new monitoring approach. European Monitoring
Centre for Drug and Drug Addiction, Lisbon 2008: 21-34.
Davoli E, Fattore E, Paiano V, Fanelli R, Rossi AN, Il Grande M. Integrated Risk Assessment of Waste Management:
A Case Study of a Solid Waste Landfill in South Italy. Proceedings Venice 2008, Second International Symposium
on Energy from Biomass and Waste, Venice, Italy, 17-20 November 2008. Available only on CD.
71
ANNUAL REPORT 2008

IRFMN
RESEARCH ACTIVITIES
Laboratory of Molecular Toxicology
Proteome Analysis
Proteome analysis includes protein separation by one- and two-dimensional gel electrophoresis,
protein excision from the gel, their digestion with proteolytic enzymes and their identification
by mass spectrometry (MALDI-TOF-MS, LC-ESI-MS/MS) coupled to the use of existing
databases. Alternatively, peptides resulting from the digestion of protein mixtures with specific
proteases are separated by two-dimensional liquid chromatography.
Toxicoproteomics
Studies are ongoing on the characterization of changes in the proteome profile induced by
environmental toxic compounds, with the aim of obtaining protein biomarkers with the ability
to differentiate two or more biological states. Proteome changes in tissues and target organs of
animals, and cells treated with endocrine disruptors, estrogens, or environmental carcinogens,
are related to functional changes during toxicological processes.
Clinical Proteomics
Qualitative and quantitative proteome changes resulting from the exposure to environmental
toxic compounds or in pathological conditions are monitored in human biological plasma and
urine. Ongoing studies aim at the characterization of protein biomarkers for early diagnosis of
diseases and for the identification of therapeutic targets.
Metabolomics
Metabolites are the biological endpoints of gene expression and enzyme activity and include
small molecules such as amino acids, carbohydrates, fatty acids, hormones, etc., that provide the
metabolic phenotype of individuals. Metabolomic research focuses on the quantitative analysis
of metabolites in biological fluids to link human metabolic profile variations to endogenous or
exogenous pathophysiological stimuli and to genetic modifications.
Selected metabolites are extracted from plasma or urine by solid phase extraction and analyzed
by HPLC coupled to high-resolution mass spectrometry.
The integration of proteomic and metabolomic studies will provide information that can help to
better understand disease development and to identify preventive interventions.
Molecular Epidemiology
The laboratory works mainly on the measurement of biological markers used to assess human
exposure to environmental toxic compounds. Our studies include DNA- and blood proteinadduct
formation by several environmental carcinogens. In addition, we study whether the
polymorphism of genes coding for enzymes involved in the activation and detoxification of
carcinogens are determinants of adduct formation. Genotypes are detected by restriction
fragment length polymorphism analysis, after the amplification by polymerase chain reaction of
specific nucleotide sequences of the genes under study.
The laboratory participates in an international cooperation study aimed at the collection of
reference values on allele and genotype frequency of the most common metabolic enzyme
polymorphisms in control populations.
72
ANNUAL REPORT 2008

IRFMN
Laboratory of Analytical Biochemistry
Identification and characterization of proteins by mass spectrometry
Our laboratory is developing different analytical and instrumental techniques, based on
chromatography, electrophoresis and mass spectrometry, for the identification and
characterization of proteins and peptides in biological samples. This activity is mainly aimed at
1) global characterization and comparison of secretomes from human cancer cell lines to
identify proteins that may be functionally relevant for tumor growth and spread; 2) profiling
proteins in biological fluids for discovery and identification of biomarkers of physiopathological
and toxicological relevance, 3) identifying and characterizing endogenous degradation products
of proteins, 4) identifying proteins produced by cells in vitro in response to given stimuli, 5)
selectively isolating biologically relevant proteins by immunoaffinity-based micro-techniques.
Ongoing projects include the study of exogenous protein degradation in renal tubular cells in
relation to antigen presentation mechanisms, and the characterization of the secretome of cancer
cell lines in vitro to identify factors affecting immune cells.
Method development in proteomics
The laboratory works on the optimization of various analytical methodologies for proteomics,
i.e. various complementary techniques for protein isolation and identification by mass
spectrometry (MALDI-TOF-MS, LC-ESI-MS/MS).
Laboratory of Environmental Chemistry and Toxicology
Development and use of analytical methods to evaluate contamination in
water bodies, soil, biota, human samples in exposed population
Analytical methods are developed to study environmental pollutants in water ecosystems,
landfills, contaminated sites. Qualitative and quantitative analyses of organic pollutants are done
by mass spectrometry (GC-MS, LC-MS, LC-MS/MS). Typical analyses include PCDD/F, PCB,
PAH, polybrominated diphenylethers, pesticides, endocrine disruptor chemicals, and industrial
pollutants.
Studies on environmental, toxicological and ecotoxicological properties
of chemicals
Research is carried out on pollutant properties, searching literature data, comparing and
evaluating different sources, and mainly developing predictive models to cope with the lack of
experimental data. Thus, we develop models starting merely from the chemical structure. The
research addresses the different kinds of chemical descriptors and chemical fragments, obtained
with different software. Then, we develop models using algorithms such as neural network,
fuzzy logic, genetic algorithms, classifiers, multivariate analysis, etc. Different methods are
compared and integrated within a structured ensemble. Standardized methods for pesticides
were developed and validated according to OECD guidelines.
Risk assessment of pollutants
Studies are aimed at assessing the risk of pollutants for human population and environment. For
this we model transport and diffusion of pollutants, to obtain a predicted concentration on given
space and time scales. Such an activity is integrated with those above described on chemical
analyses and toxicity prediction, to achieve a continuous transfer of data and research.
73
ANNUAL REPORT 2008

IRFMN
Research on pollutants emitted in the atmosphere (Unit of Industrial and
Environmental Hygiene)
Studies address different aspects of atmospheric pollution. Research deals with: sampling areas
around the pollution source, chemical analyses, transport modeling depending on
meteorological conditions and orography, risk assessment for population and environment.
Qualitative and quantitative analyses are done by gas chromatography-mass spectrometry using
high resolution for PCDDs/PCDFs, and negative ion-chemical ionization for PCBs.
Laboratory of Mass Spectrometry
Particulate air pollution
Epidemiological studies consistently show an association between an increasing number of
pathologies, both acute and chronic, and particulate air pollution. This has been shown not only
in respiratory, but in cardiovascular diseases as well. Airborne particulate sampling and analysis
strategies are developed to characterize both adsorbed compounds and exposition in different
activities.
Method development in environmental sciences
Methods, analytical methodologies, instrumentation and software for data acquisition and
reduction, are developed for environmental studies. High-sensitivity instrumentation, mainly
based on mass spectrometry, is developed for trace and ultra-trace analysis. Also, transportable
instrumentation is developed for field studies or continuous monitoring.
Characterization of environmental odor annoyance
Characterization of odors poses several analytical problems because they result from a complex
mixture of compounds (odorants) stimulating receptors in the nasal cavity. Most odorants are
volatile organic compounds (VOC) generated by bacterial degradation of organic matter. They
are often present at trace levels, while numerous sources can contribute to the total odor. Using
sampling techniques specifically developed for olfactometry, solid phase microextraction and
GC/MS analysis, we can detect traces (low ppb to high ppt) of a wide polarity/volatility range of
airborne VOC odorant compounds. With a chemometric approach, we can characterize the
sources of emissions, assess odor control methods, and identify emissions that contribute to
odors in ambient air.
Laboratory of Food Toxicology
Chemical contaminants in food
We are studying human exposure to dietary PCBs and dioxins. PCBs were measured in food
items in different European countries, showing differences in PCBs exposure of European
consumers. Further studies were aimed at measuring PCBs and dioxins in samples of fish
caught in Italy and in food items from an Italian area at high risk of contamination. Ongoing
studies are focused to assess levels of PCBs and dioxins in samples of human milk collected
from mothers living in highly contaminated areas.
Therapeutic and illicit drugs in the environment
Pharmaceuticals are a class of emerging environmental pollutants. We have organized a
campaign to detect the presence of pharmaceuticals and their metabolites in Italian rivers and
sewage treatment plants, with the aim of better characterizing the contamination and assessing
related risks.
74
ANNUAL REPORT 2008

IRFMN
Human and environmental risks are evaluated by studying the toxic effects of pharmaceuticals
at environmental levels, on cultures of human and zebra fish cells. Further ongoing studies are
aimed at investigating a possible relationship between antibiotic occurrence and resistance in
environmental bacteria.
The possible presence of illicit drugs in water samples from sewage treatment plants and rivers
was investigated, starting with cocaine and its metabolites. Their levels, used to estimate drug
abuse in the local population, revealed that cocaine consumption greatly exceeds official
estimates. This approach has been subsequently extended to include other common drugs of
abuse such as cannabis, opiates (heroin, morphine), and amphetamines (amphetamine,
methamphetamine, ecstasy). Consumption of these drugs have been subsequently estimated in
some European cities. Our evidence-based method allows monitoring of patterns and trends of
drug abuse in local communities, and is able to detect qualitative and quantitative consumption
changes in real time. This tool can therefore complement survey methods in more realistically
describing the drug abuse phenomenon.
Regulatory activities
On behalf of the Ministry of Health, we carried on the evaluation of the dossiers required for
pesticide registration within the European Union.
Unit of Environmental Pollutants Risk Assessment
Exposure to environmental pollutants
Research activities include focus both on quantitative measurement of contaminants in
environmental samples, and assessment of human exposure. Specific projects projects are the
following:
Measurements of persistent organic pollutants such as polybrominated dioxins and furans,
perfluorooctanoic acid (PFOA), perfluorooctane sulphonate (PFOS), and identification of new
pollutants, in farmed and wild fish coming from Mediterranean sea. Exposure assessment to
these persistent pollutants for human population due to fish consumption.
on dietary exposure of the general Italian population include
: an exposure assessment to PCBs thorough dietary farmed and wild fish coming from
Mediterranean sea; a study combining available data from food consumption surveys in Italy
with data on contamination by polychlorinated dibenzo-p-dioxins (PCDDs), furans (PCDF),
dioxin- and non-dioxin-like PCBs in European foodstuffs; an investigation of the contamination
level by PCDD, PCDF and PCB in different food coming from a suspected polluted area.
A study on occupational exposure deals with measurements of PCBs and DDE in human blood
with the aim of assessing the correlation between exposure to these organochlorine compounds
and the bone mineral density of the population investigated.
Toxicological risk assessment
The unit activities also include risk assessment studies related to specific environmental
conditions or human activities which can pose a risk for human health. During 2008 studies of
risk assessment related to air quality in a industrialised area, and to the emissions from an
incinerator and a landfill have been carried out.New methods for exposure assessment are
developed, employing probabilistic approaches and more refined statistical models, starting
from real cases of contamination. A current study deals with the exposure of the Seveso
population to dioxin from contaminated soil.
Evaluation of toxicological data
Toxicological data resulting from in vivo sub-chronic studies in rats exposed to individual
dioxin-like and non dioxin-like PCBs are evaluated in detail, in order to investigate the dose-
75
ANNUAL REPORT 2008

IRFMN
response relationship and the applicability of the “benchmark dose” approach.
Unit of Analytical Instrumentation
Development and application of analytical methods for compounds of
biological and environmental interest.
Methods are developed mainly using solid phase extraction (SPE) followed by liquid
chromatography - mass spectrometry (LC-ESI-MS/MS) or gas chromatography - mass
spectrometry (GC-MS). Available intruments include mass spectrometers equipped with
different analyzers: magnetic fields, time of flight (TOF), quadrupoles (single and triple), ion
traps and high resolution orbitrap. The main ionization techniques are electron ionization (EI),
chemical ionization (CI), MALDI, ACPI and Electrospray (conventional and nanoSpray).
Substances of interest include proteins, peptides, hormones, pharmaceuticals, drugs of abuse,
pesticides, and other environmental contaminants (PCBs, hydroxy-PCBs, perfluorinated
compounds).
Work has been started for the development of imaging methods in biological tissues with the
MALDI-TOF technique.
76
ANNUAL REPORT 2008

IRFMN
77
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF NEUROSCIENCE
STAFF
Head
Gianluigi FORLONI, Biol.Sci.D.
Laboratory of Biology of Neurodegenerative Disorders
Head
Cell Death and Neuroprotection Unit
Head
Gianluigi FORLONI, Biol.Sci.D.
Tiziana Borsello, Biol.Sci.D.
Laboratory of Drug Metabolism
Head
Silvio CACCIA, Farm.D.
Laboratory of Experimental Neurology
Head
Annamaria VEZZANI, Biol.Sci.D.
Laboratory of Experimental Psychopharmacology
Head
Luigi CERVO, Ph.D.
Laboratory of Geriatric Neuropsychiatry
Head
Ugo LUCCA, MSc
Epidemiology and Social Psychiatry Unit
Head
Barbara D’AVANZO, Philos.D.
Geriatric Epidemiology Unit
Head
Geriatric Pharmacology Unit
Head
Mauro TETTAMANTI, Biol.Sci.D.
Emma RIVA, M.D.
Laboratory of Inflammation and Nervous System Diseases
Head
Pharmacology of septic shock Unit
Head
Maria Grazia DE SIMONI, Biol.Sci.D.
Pia VILLA, Biol. Sci. D
Laboratory of Molecular Neurobiology
Head
Caterina BENDOTTI, Farm.D.
78
ANNUAL REPORT 2008

IRFMN
Laboratory of Neurochemistry abd Behavior
Head
Roberto William INVERNIZZI, Biol. Sci D
Pharmacology of Cognitive Behavior Unit
Head
Mirjana CARLI, Ph.D.
Laboratory of Neurological Disorders
Head
Ettore BEGHI, M.D.
Laboratory of Quality Assessment of Geriatric Services Unit
Head
Alessandro NOBILI, M.D.
79
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Gianluigi Forloni, obtained the Degree of Biological Science at the University of Milan in 1985. After
two years of post doc at the Department of Neuroscience and Psychiatry at Johns Hopkins University in
Baltimore, USA, he came back to the Mario Negri Institute and between 1992 and 1996 he was the head
of the Neurobiology of Alzheimer's disease Unit; since 1996 he is the Head of the Biology of
Neurodegenerative Diseases Lab and since 2002 the Head of the Neuroscience Department. His scientific
interest is focused on the biological and genetic bases of aging-related disorders in particular Alzheimer’s
disease, Prion-related encephalopathies and Parkinson’s disease. He has been member of several
European committees for the examination of projects in the neuroscience field. He is now member of the
coordination group of the European IMI Consortium Pharmacog. He is President of the Italian
Association on Brain Aging Research (AIRIC) and member of the European Academy of Sciences. He is
the author of more than 160 peer-reviewed scientific articles and about 30 reviews or book chapters.
Selected publications
• Forloni G. Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O.,Tagliavini F. Neurotoxicity of a prion protein
fragment. Nature 362: 543-546 (1993)
• Forloni, G., Tagliavini, F.,Bugiani, O. and Salmona, M. Amyloid in Alzheimer’s disease and prion-related
encephalopathies: Studies with synthetic peptides. Progr. Neurobiol. 49: 287- 315 (1996)
• Forloni, G., Bertani, I. Calella, AM., Thaler, F.Invernizzi. R. Alpha-synuclein and Parkinson's disease selective
neurodegeneration effect of alpha synuclein fragment on dopaminergic neurons in vitro. Ann. Neurol. 47: 632-640
(2000)
• Forloni G. Iussich, S. Awan T. Colombo L. Angeretti, N. Girola, L. Bertani, I. Poli, G. Caramelli, M. Bruzzone,
MG.Farina, L. Limido, L. Rossi, G. Giaccone G. Ironside, JW. Bugiani, O.Salmona M. and Tagliavini, F. Tetracyclines
affect prion infectivity Proc. Natl. Acad. Sci . New York 99: 10849-10854 (2002)
• Pesaresi M, Lovati C, Bertora P, Mailland E, Galimberti D, Scarpini E, Quadri P, Forloni G, Mariani C. Plasma levels of
beta-amyloid (1-42) in Alzheimer's disease and mild cognitive impairment. Neurobiol Aging., 27:904-5 (2006)
• Fioriti, L. Angeretti, N.. Colombo, L., De Luigi A., Manzoni, C., Colombo A., Morbin, M., Tagliavini, F., Salmona, M.
Chiesa, R. Forloni, G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-Sträussler-
Scheinker disease amyloid protein. J. Neurosci. 27: 576-83 (2007)
• Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M,
Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa Mutant prion
protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model.
Neuron ; 60 : 598-609 (2008)
Ettore Beghi (EB) graduated in Medicine in 1972 and received his specialty in neurology in 1976 at the
University of Milan. He trained in epidemiology with a fellowship at the Department of statistics and
Epidemiology of the Mayo Clinic in Rochester, MN (USA). He is Head of the Laboratory of
Neurological Disorders at the Mario Negri Institute, Director of the Neurophysiology/Epilepsy Unit and
Professor of Neuroepidemiology at the University of Milano-Bicocca, Monza. He is member of the
editorial board of the journals Epilepsia, Neuroepidemiology, Inpharma, Drugs in R & D, Clinical Drug
Investigation, Neurological Sciences and is a referee of several national and international medical
journals. The main areas of interest and research include studies on the descriptive, analytic, and
experimental epidemiology in the field of epilepsy, peripheral neuropathies, headache, and amyotrophic
lateral sclerosis.
Selected publications
• Leone, MA. Solari, A.,Beghi, E. for the FIRST Group. Treatment of the first tonic-clonic seizure does not affect longterm
remission of epilepsy. Neurology 2006; 67: 2227-2229
• Millul, A., E. Beghi, G. Logroscino, A. Micheli, E. Vitelli, A. Zardi, for the “Registro Lombardo SLA”(SLALOM).
Survival of patients with amyotrophic lateral sclerosis in a population-based registry. Neuroepidemiology 2005; 25: 114-
119.
• Tonini, C., E. Beghi, A.T. Berg, G. Bogliun, L. Giordano, R.W. Newton, A. Tetto, E. Vitelli, D. Vitezic, S. Wiebe.
Predictors of epilepsy surgery outcome: a meta-analysis. Epilepsy Res 2004; 62: 75-87.
• Van den Broek, M., and Beghi E., for the RESt-1 Group. Morbidity in patients with epilepsy: type and complications. A
European Cohort Study. Epilepsia 2004; 45: 71-76.
• Van den Broek, M. and Beghi E. for the RESt-1 Group. Accidents in patients with epilepsy: type and complications. A
European Cohort Study. Epilepsia 2004; 45: 667-672.
80
ANNUAL REPORT 2008

IRFMN
• Musico, M., E. Beghi, A. Solari, F. Viani for the First Seizure Trial Group. Treatment of the first tonic-clonic seizure
does not improve the prognosis of epilepsy. Neurology 1997; 49: 991-998.
Caterina Bendotti, got her degree in Pharmacy at the University of Milano in 1984; In 1986 -1988 she was
post doc at the Genetic developmental Lab, Dept. of Physiology of the Johns Hopkins University, Baltimore,
USA. In 1988 -1992 she was research fellow in the laboratory of Neuropharmacology and in the 1992, she
became head of the Molecular Neurobiology Unit in Institute, since 1998 she is head of laboratory. The
major research interest is the study of pathogenetic mechanisms of familial Amyotrophic Lateral Sclerosis..
Since 2002 she is a member of the editorial board of Journal of Neurochemistry. In 2002-2003 has been
Member of Scientific Committees of the International Symposia on ALS held in Milano, 17-19
Novembre,2003. In 2003-2007 has been member of the Italian Ministry of Health Committees for the
diagnosis, cure, care and assistance of patients with ALS. Since 2005 is member of the Board of Directors of
the Italian Society of Neuroscience. Since 2006 is member of the Research Advisory Panel of the MND
Association, UK. Scientific reviewer of 11 international scientific journals. In 2007 she has co-organised the
first international meeting on” Mutant SOD1 and familial ALS:from the molecule to man” held in
Milano(13-16 September). She is author and co-author of 110 articles 100 of which with peer-review.
Rapporteur of many communications in national and international meetings.
Selected publications
• Sau D, De Biasi S, Vitellaro-Zuccarello L, Riso P, Guarnieri S, Porrini M, Simeoni S, Crippa V, Onesto E, Palazzolo I,
Rusmini P, Bolzoni E, Bendotti C, Poletti A. Mutation of SOD1 in ALS: a gain of a loss of function. Hum Mol Genet.
16(13):1604-18, 2007.
• Massignan T, Casoni F, Basso M, Stefanazzi P, Biasini E, Tortarolo M, Salmona M, Gianazza E, Bendotti C, Bonetto V.
Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse.Biochem Biophys
Res Commun. 353(3):719-25, 2007
• Peviani M, Cheroni C, Troglio F, Quarto M, Pelicci G, Bendotti C. Lack of changes in the PI3K/AKT survival pathway
in the spinal cord motor neurons of a mouse model of familial amyotrophic lateral sclerosis.Mol Cell Neurosci. 34:592-
602, 2007
• Carri MT, Grignaschi G, Bendotti C. Targets in ALS: designing multidrug therapies. Trends Pharmacol Sci. 27(5):267-
73, 2006
• Cheroni C., Peviani M., Cascio P., Debiasi S., Monti C. and Bendotti C. Accumulation of human SOD1 and
ubiquitinated deposits in the spinal cord of SOD1G93A mice during motor neuron disease progression correlates with a
decrease of proteasome. Neurobiol. Disease. 18(3): 509-522, 2005
• Bendotti C and MT Carri. Lessons from models of SOD1-linked familial ALS. Trends Mol Med. 10(8):393-400, 2004
• Bendotti C., Tortarolo M., Suchak S.K., Calvaresi N., Carvelli L., Bastone A., Rizzi M., Rattray M. and Mennini T.
Transgenic SOD1 G93A mice develop reduced GLT-1 in spinal cord without alterations in cerebrospinal fluid glutamate
levels. J. Neurochem.,79, 737-746, 2001
• Migheli A., Atzori C., Piva R., Tortarolo M., Girelli M., Schiffer D. and Bendotti C. Lack of apoptosis in mice with
ALS. Nature Medicine: 5, 966-967, 1999.
Silvio Caccia. Laurea in Pharmacy at the University of Milan. Diploma in Industrial Chemistry at the L.
Cobianchi Technical Institute (Verbania, NO) and Diploma in Biochemical Research at the Istituto di
Ricerche Farmacologiche “Mario Negri” (Milan).
Research fellow, Laboratory of General Pharmacology at the Mario Negri Institute, 1970-1973;
Permanent Researcher, Laboratory of Neuropharmacology, 1975; Head of Pharmacokinetic Unit, 1983;
Head of Drug Metabolism Laboratory, 1988 to date, doing research in the field of pharmacology and
toxicology with particular focus on pharmacokinetic aspects, both at the pre-clinical and clinical level.
He has been member of the scientific assessment teams (acting as expert) for the evaluation of marketing
authorisation applications submitted to the European and Italian Agencies.He is author and co-author of
more than 200 articles, including reviews, monographs and book chapters.
Selected publications
• Caccia S. N-dealkylation of arylpiperazine derivatives: disposition and metabolism of the 1-aryl-piperazines formed.
Curr Drug Metab 2007 ; 8 : 612-622.
• Caccia S. Main active components of St. John's Wort (Hypericum Perforatum) extracts: current analytical procedures for
pharmacokinetics and concentration-response studies. Curr Pharm Anal 2006; 2: 59-68.
• Caccia S. Antidepressant-like components of Hypericum perforatum extracts: An overview of their pharmacokinetics
and metabolism. Curr Drug Metab 2005; 6: 531-543
• Caccia S. Metabolism of the newest antidepressants: Comparisons with related predecessors IDrugs 2004; 7: 143-150.
• Caccia S. Biotransformation of post-clozapine antipsychotics. Pharmacological implications. Clin Pharmacokinet
2000; 38: 39.
• Caccia S. Metabolism of the newer antidepressants. An overview of the pharmacological and pharmacokinetic
implications. Clin Pharmacokinet 1998; 34: 281-302.
81
ANNUAL REPORT 2008

IRFMN
Luigi Cervo was awarded the degree of Doctor of Philosophy (Ph.D.) from the Open University, Milton
Keynes, U. K. in 2005. Since 2006 he has been the head of the Experimental Psychopharmacology
Laboratory. From 1978 to 2001 he has been a research fellow and then Chief of the Behavioural
Pharmacology Unit in the laboratory of Neuropharmacology and in 1981 he was awarded the degree in
Biochemical Research from the “M. Negri” Institute. Between 1981 and 1983 he spent two years as a
research fellow in the Department of Psychiatry at the Chicago University, Illinois, U.S.A. His main
research interests concern Neuropsychopharmacology and the mechanism of action of psychotropic
drugs. In particular the role of receptors subtypes for serotonin, dopamine, noradrenaline and glutamate
in drug dependence and drug craving, depression, anxiety. Author and co-author of several peer-review
articles, author of communications in international meetings, he is scientific reviewer of several
international scientific journals. Member of Society for Neuroscience and Italian Society of
Neuropsychopharmacology.
Selected publications
• Cervo L, Carnovali, F, Stark JA, Mennini T. Cocaine-seeking behavior in response to drug-associated stimuli in rats:
involvement of D 3 and D 2 dopamine receptors. Neuropsychopharmacology 2003; 28: 1150-1159
• Cervo L, Cocco A, Carnovali F. Effects on cocaine and food self-administration of (+)-HA-966, a partial agonist at the
glycine/NMDA modulatory. Psychopharmacology (Berl) 2004; 173: 124-131
• Grignaschi G, Burbassi S, Zennaro E, Bendotti C, Cervo L. A single high dose of cocaine induces behavioural
sensitization and modifies mRNA encoding GluR1 and GAP-43 in rats. Eur J Neurosci 2004; 20:2833-2837
• Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi R.
Deficits of serotonin synthesis cause resistance to antidepressants, J Neuroscience 2005; 25: 8165-8172
• Cervo L, Cocco A, Putrella C, Heidbreder CA. Selective antagonism at dopamine D3 receptors attenuates cocaineseeking
behaviour in the rat. Int J Neuropsychopharmacol. 2007; 10: 167-181.
• Burbassi S, Cervo L. Stimulation of serotonin(2C) receptors influences cocaine-seeking behavior in response to drugassociated
stimuli in rats. Psychopharmacology (Berl). 2008; 196: 15-27.
• Burattini C, Burbassi S, Aicardi G, Cervo L. Effects of naltrexone on cocaine- and sucrose-seeking behaviour in
response to associated stimuli in rats. Int J Neuropsychopharmacol. 2008; 11, 103-109.
• Marchesi F, Piemonti L, Fedele G, Destro A, Roncalli M, Albarello L, Doglioni C, Anselmo A, Doni A, Bianchi P,
Laghi L, Malesci A, Cervo L, Malosio M, Reni M, Zerbi A, Di Carlo V, Mantovani A, Allavena P. The chemokine
receptor CX3CR1 is involved in the neural tropism and malignant behavior of pancreatic ductal adenocarcinoma. Cancer
Res. 2008; 68, 9060-9069.
• Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi RW. Strain differences in
paroxetine-induced reduction of immobility time in the forced swimming test in mice: role of serotonin. Eur J
Pharmacol. 2008; 594, 117-124.
Maria Grazia De Simoni got the Doctoral Degree in Biological Sciences in 1977 at the University of
Milano, Italy. 1981: Research Specialist in Pharmacology (PhD), Mario Negri Institute, Milan, Italy.
1981-1982: European Community fellowship for "Advanced Professional Training", INSERM U 171,
Universitè Claude Bernard, Lyon, France; 1984 Department of Histology, Karolinska Institute,
Stockholm. Working experience:1987-1997: Chief of the Neurochemistry Unit, Mario Negri Institute,
Milano; 1998-present: Chief of the Laboratory of Inflammation and Nervous System Diseases, Mario
Negri Institute. Scientific interests: pathogenesis of cerebral ischemia/reperfusion and traumatic brain
injury; inflammatory response and apoptotic mechanisms as targets of therapeutic strategies; animal
models and clinical studies. She is member of the board of “Master in Tecnologie Avanzate Applicate
alle Patologie Neurodegenerative", University of Milan and member of the board of “Associazione
Italiana per la Ricerca sull’Invecchiamento Cerebrale” (AIRIC).
Selected pubblications
• De Simoni MG, Storini C, Barba M, Catapano L, Arabia AM, Rossi E, Bergamaschini L. Neuroprotection by
complement (C1)-inhibitor in mouse transient brain ischemia. J Cereb Blood Flow Metab, 23: 232-239, 2003.
• De Simoni M G, Rossi E, Storini C, Pizzimenti S, Echart C, Bergamaschini L. The powerful neuroprotective action of
C1-inhibitor on brain ischemia-reperfusion injury does not require C1q. Am J Pathol., 164: 1857-1863, 2004.
• Bergamaschini L, Rossi E, Storini C, Pizzimenti S, Distaso M, Perego C, De Luigi A, Vergani C and De Simoni MG.
Peripheral treatment with enoxaparin, a low-molecular weight heparin, reduces plaques and β-amyloid accumulation in a
mouse model of Alzheimer’s disease. J. Neurosci. 24: 4181-4186, 2004
82
ANNUAL REPORT 2008

IRFMN
• Troglio F, Echart C, Gobbi A, Pawson T, Pelicci PG, De Simoni MG & Pelicci G. The neuron-specific Rai (Shc C)
adaptor regulates the PI3K-Akt pathway in vivo and protects against cerebral ischemia. Proc Natl Acad Sci U S A
101(43): 15476-15481, 2004.
• Storini C, Bergamaschini L, Gesuete R, Rossi E, Maiocchi D, De Simoni MG. Selective inhibition of plasma kallikrein
protects brain from reperfusion injury. JPET 318: 849-854, 2006
• Capone C, Fabrizi C, Piovesan P, Principato MC, Marzorati C, Ghirardi O, Fumagalli L, Carminati P and De Simoni
MG. 2-Aminotetraline derivative protects from ischemia/reperfusion brain injury with a broad therapeutic window,
Neuropsychopharmacology, 32: 1302-1311, 2007
• Capone C, Frigerio S, Fumagalli S, Gelati M, Principato M C, Storini C, Montinaro M, Kraftsik R, De Curtis M, Parati
E, De Simoni MG. Neurosphere - derived cells exert a neuroprotective action by changing the ischemic
microenvironment. PLoS ONE 2 e373, 2007.
• Pastori C, Librizzi L, Breschi GL, Regondi C, Frassoni C, Panzica F, Frigerio S, Gelati M, Parati E, De Simoni MG, de
Curtis M.Arterially perfused neurosphere-derived cells distribute outside the ischemic core in a model of transient focal
ischemia and reperfusion in vitro.PLoS ONE. 3(7):e2754. 2008
Roberto W. Invernizzi started his career in the laboratory of Neuropharmacology of the “Istituto di
Ricerche Farmacologiche “Mario Negri” in 1976, where, at present, he heads the Laboratory of
Neurochemistry and Behavior. In 1986 he got his degree in Biological Sciences at the Università Statale di
Milano and in 1996 he was nominated head of the Intracerebral Microdialysis Unit. Of particular interest
to Invernizzi’s research team is the study of the neurochemical mechanisms and neuronal circuitries
involved in the pathology of the main psychiatric diseases, such as depression and schizophrenia and in
the mechanism of action of psychotropic drugs. Since 1987 he applied the intracerebral microdialysis
technique to study the in vivo release of monoamines. Using this technique, Invernizzi’s team first
contributed to clarifying the role of serotonergic and adrenergic autoreceptors in the effect of
antidepressant drugs suggesting new hypotheses on their mechanism of action. Currently, Invernizzi’s
laboratory is involved in two main collaborative projects aimed at clarifying the neurochemical
mechanisms involved in the “resistance” to antidepressant drugs and the role of glutamatergic and
serotonergic mechanisms in attentional processes. Reviewer for various international journals in the field
of pharmacology and neurochemistry. Author and co-author of more than 60 peer-reviewed articles.
Member of the Italian Society of Neuroscience and the Italian Society of Pharmacology.
Selected publications
• Baviera M, Invernizzi RW, Carli M. Haloperidol and clozapine have dissociable effects in a model of attentional
performance deficits induced by blockade of NMDA receptors in the mPFC. Psychopharmacology 2008; 196: 269-280.
• Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi RW Strain differences in
paroxetine-induced reduction of immobility time in the forced swimming test in mice: Role of serotonin. Eur. J.
Pharmacol. 2008; 594: 117-124
• Calcagno E, Canetta A, Guzzetti S, Cervo L, Invernizzi RW. Strain differences in basal and post-citalopram extracellular
5-HT in the mouse medial prefrontal cortex and dorsal hippocampus: relation with tryptophan with tryptophan
hydroxylase-2 activity. J Neurochem 2007; 103 : 1111-1120
• Invernizzi RW, Pierucci M, Calcagno E, Di Giovanni G, Di Matteo V, Benigno A, Esposito E. Selective activation of
5HT2C receptors stimulates GABA-ergic function in the rat substantia nigra pars reticulata: a combined in vivo
electrophysiological and neurochemical study. Neuroscience 2007 144 : 1523-1535
• Calcagno E, Carli M, Invernizzi RW The 5-HT1A receptor agonist 8-OH-DPAT prevents prefrontocortical glutamate
and serotonin release in response to blockade of cortical NMDA receptors J Neurochem 2006; 96: 853-860
• Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi
RWGenotype-dependent activity of tryptophan hydroxylase-2 determines the response to citalopram in a mouse model
of depression J Neurosci 2005; 25: 8165-8172
• Greco B, Invernizzi RW, Carli M Phencyclidine-induced impairment in attention and response control depends on the
background genotype of mice: reversal by the mGLU2/3 receptor agonist, LY379268 Psychopharmacology (Berl) 2005;
179: 68-76
Ugo Lucca got his Master of Science, University of Aberdeen - UK, 1999. At the Mario Negri Institute
he was investigator from 1986- 1995, head of the "Clinical Evaluation of Antidementia Drugs Unit"
(1995-1996) and, since 1996, head of the "Laboratory of Geriatric Neuropsychiatry".
The main areas of interests include epidemiology and clinic features of dementia; natural history of
dementia; neuropsychiatric disorders of the elderly; instruments for the screening diagnosis and clinical
course assessment of dementia; clinical evaluation of anti dementia treatments and CNS active drugs
(phase I, II, III, IV and observational studies).
83
ANNUAL REPORT 2008

IRFMN
Selected publications
• Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-L-carnitine
treatment in Alzheimer's disease. Neurology 1991; 41:1726-1732
• Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimer’s
disease: a prospective study. J Am Geriats Society 1993; 41: 45-49.
• Lucca U, Tettamanti M, Forloni G, Spagnoli A. Nonsteroidal anti-inflammatory drug use in Alzheimer’s disease.
Biological Psychiatry 1994; 36: 854-856.
• Imbimbo BP, Martelli P, Troetel WM, Lucchelli F, Lucca U, Thal LJ, and the Eptastigmine Study Group. Efficacy and
safety of eptastigmine for the treatment of patients with Alzheimer’s disease. Neurology 1999; 52: 700-708.
• Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B-
12 in mild cognitive impairment, Alzheimer’s disease and Vascular Dementia. Am J Clinical Nutr 2004; 80: 114-122.
• Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. New England Journal of
Medicine 2006; 355: 1390.
• Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M,
Tempia P, Guala A, Fasolo G, Riva E.Association of mild anemia with cognitive, functional, mood and quality of life
outcomes in the elderly: the "Health and Anemia" study. PLoS ONE. 3(4):e1920 (2008)
Alessandro Nobili got his degree in Medicine (Milan, 1990). Master in Biotechonological Research,
Regione Lombardia, Milan 1988. International School of Pharmacology, 31° Course on: Drug
Epidemiology and Post-marketing Surveillance, Erice, September 1990. Course on: Methods in
Epidemiological Research, Milan, October 1990. Course: Long Term Clinical Trials, Cogne January
1991.
Main areas of interest Methodology of Randomized Clinical Trials; Pharmacoepidemiology and postmarketing
surveillance research; Drug utilization studies; Quality assessment of geriatric services;
Qualitative studies on caregiver role in the care of patients with dementia; Methodological evaluation of
the Special Care Unit for Alzheimer Disease patients; Methodology of drug information. Employment
and research experience Chief of the Unit of Quality Assessment of Geriatric Services Chief of the Drug
Information Services for the Elderly, Laboratory of Geriatric Neuropsychiatry, Istituto di Ricerche
Farmacologiche “Mario Negri”, Milan. Editorial Board of the MICROMEDEX Inc., Englewood,
Colorado 80111-4740 USA. National Expert accredited by Italian Ministry of Health for The Italian
(AIFA) and European Agency for the Evaluation of Medicinal Products (EMEA). Head of the
Laboratory of the Quality Assessment of Geriatric Services at the Mario Negri Institute since 2007.
Selected publications
• Nobili A, Tettamanti M, Frattura L, et al. Drug use in the elderly in Italy. Ann Pharmacother 1997; 31:416-422.
• Nobili A, Gebru F, Rossetti A, et al. Doctorline a private toll-free telephone medical information service. Ann
Pharmacother 1998; 32:120-5.
• Nobili A, Riva E, Tettamanti M, et al. The effect of a structured intervention on caregivers of patients with dementia and
problem behavior: a randomized controlled pilot study. Alzheimer Dis Assoc Disord 2004; 18: 75-82.
• Lucca U, Nobili A, Riva E, Tettamanti M. Low level of B vitamins and the risk of cognitive and functional decline in the
very-old: results from the Monzino 80-Plus Study. Neurobiol Aging 2004; 25: 31.
• Lucca U, Nobili A, Riva E, Tettamanti M Cholinesterase inhibitor use and age in the general population Arch Neurol
2006; 63: 154-155.
• Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U Low folate and the risk of cognitive and functional deficits in the
very old: The Monzino 80-plus study J Am Coll Nutr 2006; 25: 502-508
• Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M.Alzheimer
special care units compared with traditional nursing home for dementia care: are there differences at admission and in
clinical outcomesAlzheimer Dis Assoc Disord. 22:352-6 (2008).
Annamaria Vezzani got her Degree in Biological Science at the University of Milan in 1978 and she
specialized in Neuropharmacology at the Mario Negri Institute in 1982. She spent her post-doctoral
period in Baltimore at the University of Maryland in 1983-1984 working on the mechanisms of
epileptogenesis in experimental models of epilepsy. She spent additional post-doctoral periods at the
University of Stockholm and at the Karolinska Institute between 1985 and 1999. She was on sabbatical at
the Albert Einstein College of Medicine in 2002 in the laboratory of Developmental Epilepsy. She is
involved in studies on the biochemical and molecular mechanisms involved in the etiopathogenesis of
seizures disorders using experimental models of epilepsy. The present research is focused on the
functional role of neuroactive peptides and inflammatory mediators in the modulation of neuronal
excitability and seizure-related neurodegeneration. Focus of the research is also on the mechanisms of
pharmacoresistance. Since 1997 she is the Head of the Laboratory of Experimental Neurology at the
Mario Negri Institute. She is member of the Editorial Board of Epilepsy Currents and Neuroscience and
84
ANNUAL REPORT 2008

IRFMN
Associate Editor for Exp Models of Epilepsia. She is appointed of the Chair of the Commission on
Neurobiology of International League Against Epilepsy which is promoting initiatives for improving
translational research in epilepsy.
Selected publications
• Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional
pathway mediates the proconvulsive effects of interleukin-1 beta (2008) Brain, 131: 3256-65
• Balosso S, Ravizza T, Perego C, Peschon J, Campbell I, De Simoni MG, Vezzani A. TNF-alpha inhibits kainic acidinduced
seizures in mice via p75 receptors (2005) Ann Neurol, 57, 804
• Dube’ C., Vezzani A., Behrens M., Bartfai T., Baram TZ. (2005) Interleukin-1beta contributes to the generation of
experimental febrile seizures. Ann Neurol, 57,152.
• Richichi C, E-J. D. Lin, D. Stefanin, D. Colella, T. Ravizza,G. Grignaschi, G. Sperk, M. J. During and A. Vezzani “
Anticonvulsant and antiepileptogenic effects mediated by adeno-associated virus vector neuropeptide Y expression in
the rat hippocampus” (2004) J Neurosci, 24,3051
• Rizzi M, Caccia S, Guiso G, Richichi C, Gorter JA, Aronica E, Aliprandi M, Bagnati R, Fanelli R, D'Incalci M,
Samanin R, Vezzani A.“Limbic seizures induce P-glycoprotein in rodent brain: functional implications for
pharmacoresistance” (2002) J Neurosci, 22, 5833
• Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S.,
Lundkvist J., Iverfeldt K. and Bartfai T. (2000) "Powerful anticonvulsant action of IL-1 receptor antagonist upon
intracerebral injection and astrocytic overexpression in mice" Proc. Natl. Acad. Sci. USA, 97, 11534.
• Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S.,
Lundkvist J., Iverfeldt K. and Bartfai T. (2000) "Powerful anticonvulsant action of IL-1 receptor antagonist upon
intracerebral injection and astrocytic overexpression in mice" Proc. Natl. Acad. Sci. USA, 97, 11534.
Tiziana Borsello got her Degree in Biological Science at the University of Torino in 1990 and she then
obtained a PhD in Neuroscience at the University of Turin Medical School. She won 1 year fellow of the
European Science Foundation scholarship for work at the Netherlands Research Institute of Amsterdam. From
1997 to 1999 she was a Researcher at the Institute of Neurobiology, CNR, Rome Italy. In the period 1999-2003
she was Premier Assistant, Département de Biologie Cellulaire et de Morphologie, Université de Lausanne,
Switzerland, and then became Maitre Assistant and group leader in the same institute. In 2004 joined the Biol.
Neurodeg. Disorders Lab at the "Mario Negri” Institute. In 2005 won the Prize of the Pfizer Foundation,
Neuroscience and Diseases Nervous System. Since 2006 she is the Head of the Unit: Neuronal Death and
Neuroprotection. Her main scientific interest is understanding the role of signaling pathways in neuronal death
after different stress-stimuli and the neuroprotection. In particular, the present research is focused on the study
of the mechanisms of excitotocic stress, ischemia, Traumatic Brain Injury and the cell death pathways in
neurodegenerative diseases as Alzheimer, with the challenge to define the neuronal death pathways to design
more specific methods of neuroprotection.
Selected pubblications
• Repici M, Centeno C, Tomasi S, Forloni G, Bonny C, Vercelli A, Borsello T.Activation After Cerebral Ischemia And
Effect Of D-Jnki1 On C-Jun And Caspase-3 Activation. Neuroscience. 2007, 150: 40-9
• Borsello T., Centeno C., Riederer IM, Haeflinger JA and Riedere BM. Phosphorylation-dependent dimerization and
subcellular localization of islet-brain 1/c-Jun N-terminal kinase-interacting protein 1. J Neurosci Res. 2007, 85:3632-41.
• Borsello T Ed “Neuroprotection: Methods In Molecular Biology” Published By Humana Press, Usa Humana Press,
USA, Methods in Molecular Biology, June 2007
• Colombo A, Repici M, Pesaresi M, Santambrogio S, Forloni G, Borsello T. The Tat-Jnk Inhibitor Peptide Interferes With
Beta Amyloid Protein Stability Cell Death Differ. 2007, 14:1845-8.
• Borsello T and Forloni G. JNK signalling: a possible target to prevent neurodegeneration. Current Pharmaceutical
Design 2007, 13, 1875-1886
• Centeno C., Repici M., Chatton J. Y., Riederer B. M., Bonny C., Nicod P., Price M., Clarke P. G., Papa S., Franzoso G.
and Borsello T. Role of the JNK pathway in NMDA-mediated excitotoxicity of cortical neurons. Cell Death Differ ,
2007, 14: 240-253.
• Borsello T. and Bonny C.Use of cell-permeable peptides to prevent neuronal degeneration. Trend in Mol. Med. 2004, 10:
239-44
• Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C. A peptide inhibitor of
c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nature Med. 2003, 9: 1180-6
Mirjana Carli started his scientific career in the laboratory of Neuropharmacology of the “Istituto di
Ricerche Farmacologiche Mario Negri” Milan in 1977, where, at present, she is head of the
Pharmacology of Cognitive Behaviour Unit. She spent a few years in the laboratory of Cognitive
Neuroscience, Dept. of Experimental Psychology, University of Cambridge (UK) directed by Prof.
85
ANNUAL REPORT 2008

IRFMN
Trevor W. Robbins. Here she took interest in the role of brain monoamines in attention, and for this
purpose developed several behavioral tests for rats. In 1986 she returned to the laboratory of
Neuropharmacology of the “Istituto di Ricerche Farmacologiche Mario Negri”. Here she devoted her
efforts to the study of the role played by neuronal mechanisms in cognitive processes such as memory,
attention and executive functions. Her work has improved the knowledge of the role played by some
serotonin receptors in cognitive processes.
Selected publications
• Carli M, Baviera M, Invernizzi R, Balducci C Dissociable contribution of 5-HT1A and 5-HT2A receptors in the medial
prefrontal cortex to different aspects of executive control such as impulsivity and compulsive perseveration in rat
Neuropsychopharmacology 2006; 31: 757-767
• Greco B, Carli M Reduced attention and increased impulsivity in mice lacking NPY Y2 receptors: Relation to anxiolyticlike
phenotype Behav Brain Res 2006; 169: 325-334
• Carli M, Baviera M, Invernizzi R, Balducci C The serotonin 5-HT2A receptors antagonist MI00907 prevents impairment
in attentional performance by NMDA receptor blockade in the rat prefrontal cortex Neuropsychopharmacology 2004;
29: 1637-1647
• Balducci C, Nurra M, Pietropoli A, Samanin R, Carli M Reversal of visual attention dysfunction after AMPA lesions of
the nucleus basalis magnocellularis (NBM) by the cholinesterase inhibitor donepezil and by a 5-HT(1A) receptor
antagonist WAY 100635 Psychopharmacology (Berl) 2003; 167: 28-36
• Carli M, Balducci C, Samanin R. Stimulation of 5-HT1A receptors in the dorsal raphe ameliorates the impairment of
spatial learning caused by intrahippocampal 7-chloro-kynurenic acid in naive and pretrained rats Psychopharmacology
(Berl) 2000; 158: 39-47
• Carli M, Samanin R The 5-HT1A receptor agonist 8-OH-DPAT reduces rats' accuracy of attentional performance and
enhances impulsive responding in a five-choice serial reaction time task: Role of presynaptic 5-HT1A receptors
Psychopharmacology (Berl) 2000; 149: 259-268
Barbara D’Avanzo obtained her master in Philosophy in 1989 at the University of Milan. Her main field
of interest is epidemiologic research in mental health. She was involved in the analysis of the
implementation of the psychiatric reform in Italy and quality evaluation of services and their recent
modifications with specific attention to the role of psychiatric residential facilities in the community
service networks; evaluation of effectiveness of the most common psychosocial interventions; suicide
trend monitoring and study of suicide prevention programs and initiatives. More recently, she is working
on issues like recovery-oriented services, consumers’ empowerment, and methods of participation of
consumers to evaluation of services, and acknowledgment of the value of the consumers’ point of view
about psychiatric treatments and services.
She worked as researcher in the Laboratory of General Epidemiology between 1991 and 1996, and she is
Chief of the Unit of Epidemiology and Social Psychiatry since 2002. Member of the Scientific National
Board of WAPR Italy and of the World Head Office of the WAPR.
Selected publications
• Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q. 2008,
79:121-32.
• Barbato A, D'Avanzo B. Marital therapy for depression. Cochrane Database Systematic Reviews 2006; Issue 2.
• Parabiaghi A, Barbato A, D'Avanzo B, Erlicher A, Lora A. Assessing reliable and clinically significant change on Health
of the Nation Outcome Scales: method for displaying longitudinal data. Aust N Z J Psychiatry 2005; 39: 719-725.
• Barbato A, D'Avanzo B. Involuntary placement in Italy. Br J Psychiatry 2005; 186: 542-543.
• Guaiana G, Andretta M, Corbari L, Mirandola M, Sorio A, D'Avanzo B, Barbui C. Antidepressant drug consumption and
public health indicators in Italy, 1955-2000. J Clinical Psychiatry 2005; 66: 750-755.
• D'Avanzo B, Battino R N, Gallus S, Barbato A. Factors predicting discharge of patients from community residential
facilities: A longitudinal study from Italy. Aust N Z J Psychiatry 2004; 38: 619-628.
• D'Avanzo B, Barbato A, Barbui C, Battino N, Civenti G, Frattura L. Discharges of patients from public psychiatric
hospitals in Italy between 1994 and 2000. Int J Social Psychiatry 2003; 49: 27-3
Emma Riva, Medical Doctor degree in 1984 University of Milan, PhD in 1990 in Cardiovascular
Pathophysiology at the University of London (UK) Training: Research Assistant, Department of
Pharmacology, Medical School, University of Ottawa, Canada; Internship in Internal Medicine, Ospedale
Luigi Sacco, Milan; Cardiac Fellow, St Thomas' Hospital, London, UK. Field of interest: Prevalence and
effects of anemia on cognitive, functional and clinical variables in the elderly; Problem behaviors in
dementia; Burden for care-givers of Alzheimer Disease patients; End of life care. Present and past roles
in Institute Head of the Geriatric Pharmacology Unit, Istituto "Mario Negri", Milan; Scientific Director
of the hospice “Via di Natale Franco Gallini”, Aviano, Italy; Consultant Istituto Geriatrico “Pio Albergo
86
ANNUAL REPORT 2008

IRFMN
Trivulzio”, Milan: Project member of PREDICT (Policy Review and Evaluation of Dementia and
Institutional Care Trends): a Transnational Comparison.
Selected publications
• Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M,
Tempia P, Guala A, Fasolo G, Riva E.Association of mild anemia with cognitive, functional, mood and quality of life
outcomes in the elderly: the "Health and Anemia" study. PLoS ONE. 3(4):e1920 (2008)
• Tettamanti M, Garrì MT, Nobili A, Riva E, Lucca U. Low folate and risk of cognitive and functional deficits in the very
old: The Monzino 80-plus study. J Am Coll Nutr 2006;25:502-508
• Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol
2006;63:154-155
• Nobili A, Riva E, Tettamanti M, Lucca U, Liscio MR, Petrucci B, Salvini Porro G. The effect of a structured intervention
on caregivers of patients with dementia. Results of a Randomized Controlled Study. Alzheimer Dis and Associated
Disorders 2004;18:75-82
• Il malato terminale oncologico. Esperienze dall’hospice. Ed. Emma Riva. Il Pensiero Scientifico, 2001
• Riva E, Tettamanti M, Gallini C. Il ruolo del medico di medicina generale nella gestione dei malati terminali oncologici.
Indagine svolta tra i medici di medicina generale in Friuli Venezia Giulia. Ricerca & Pratica 2001
• Riva E, Nobili A, Trecate F. Per un impiego "ragionato" dei neurolettici, per la gestione dei disturbi del comportamento
in corso di Malattia di Alzheimer. Rec Prog Med 1998;89:598-603
Mauro Tettamanti got his Biology Degree at the Università degli Studi di Milano in 1986, and the
specialisation in Epidemiology and Medical Statistics in 1993, at the Università degli Studi di Pavia.
Teaching experience Introduction course to statistics, Master in Ergonomy, Politecnico di Milano, years
2001-2004 Areas of interest: Planning, conduction and analysis of clinical trials and epidemiologic
researches in the geriatric field: Phase I, II, III and observational studies on the efficacy of drugs on
neurologic disorders, with special emphasis on dementia; Effects of multi-disciplinary interventions on
geriatric/dementia patients; Epidemiology and risk factors of dementia; Care of patients with terminal
illness; Association of anemia with prevalence of diseases and cognitive problems Scholarship between
1989 and 1998, Senior Researcher since 1999 and Head of the Unit of Geriatric Epidemiology at the
Mario Negri Institute since 2001.
Selected publications
• Spagnoli A, Lucca U, Menasce G, Bandera L, Cizza G, Forloni G, Tettamanti M, et al. Long-term acetyl-L-carnitine
treatment in Alzheimer's disease. Neurology 1991; 41:1726-1732.
• Lucca U, Comelli M, Tettamanti M, Tiraboschi P, Spagnoli A. Rate of progression and prognostic factors in Alzheimer's
disease: A prospective study. J Am Geriatr Soc 1993; 41:45-49
• Quadri P, Fragiacomo C, Pezzati R, Zanda E, Forloni G, Tettamanti M, Lucca U. Homocysteine, folate, and vitamin B-
12 in mild cognitive impairment, Alzheimer disease, and vascular dementia. Am J Clin Nutr 2004; 80: 114-122
• Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol
2006; 63:154-155
• Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. N Engl J Med 2006; 355:1390
• Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U. Low folate and the risk of cognitive and functional deficits in the
very old: The Monzino 80-plus study. J Am Coll Nutr 2006; 25: 502-508
• Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, Trevisan S, Riva E, Lucca U, Tettamanti M.Alzheimer
special care units compared with traditional nursing home for dementia care: are there differences at admission and in
clinical outcomes Alzheimer Dis Assoc Disord. 22:352-6 (2008).
87
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department of Neuroscience is formed by ten Laboratories; the activities of research are
devoted to the study of neurological and psychiatric diseases, evaluated by the biological point
of view, clinical and epidemiological aspects and the quality of care. Together with these
activities, in the Department other more general expertise are present. Pharmacokinetics studies,
drug information service and preparation of protocols for clinical trial and epidemiological
studies are activities in charge of the Neuroscience Department. Traditionally part of the
Department was devoted to the creation of experimental models for the pharmacological,
neurochemical and pathogenetic studies in Alzheimer or prion's diseases, epilepsy, depression
and cognitive impairment. More recently, consolidated expertise were created in the
pathogenesis of amyotrophic lateral sclerosis (ALS), cerebral stroke and drug abuse. Some of
these disorders, like epilepsy, ALS and Alzheimer's disease are investigated from the clinical
and epidemiological points of view for the evaluation of drug and care efficacy. The activities of
the Department are aimed to an integration of the different expertise to develop
multidisciplinary approaches. The purpose is to address at different levels, knowledge, therapy
and clinical practice to the numerous questions, largely unresolved, proposed by the disorders of
nervoussystem.
FINDINGS/MAIN RESULTS 2008
In a cellular model it has been shown that the peptide of D-JNK-1-TAT inhibiting the JNK
phosphorylation activity mediated by the enzyme JNK, exerts a control on β amyloid
production, these data indicate possible therapeutic perspectives in Alzheimer’s disease
The intracerebral application of synthetic β amyloid 1-40 e 1-42 in oligomeric form is
associated with a cognitive damage that does not occur when the pepetides are applied in
monomeric form of fibrils species.
The exposure of cultured hippocampal cells to β amyloid 1-42 in oligomeric form induces an
alteration of dendritic spines.
In the transgenic mice overexpressing a mutated form of human prion protein associated to CJD
generated in the Institute exhibit some behavioral features reminiscent of the clinical condition
in humans carrying the same mutation.
In the same model has been found a significant alteration of endoplasmic reticulum in specific
neuronal cells
In cellular model the presence of α synuclein mutations associated to Parkinson’s disease, does
not influence the neuroprotective activity of the protein, this indicates that these mutations
affected the aggregation state more than physiological activity of α synuclein.
Polymorphisms in gene encoded for serotonin transporter protein influenced the risk to develop
Parkinson’s disease
88
ANNUAL REPORT 2008

IRFMN
In a prospective population-based study in the very old (Monzino 80-plus Study), behavioral
disturbances are uncommon prior to clinical dementia onset. After dementia diagnosis,
behavioral disturbances affect a large proportion of subjects, increasing with disease severity
while decreasing in the oldest-olds.
In the same prospective population-based study (Monzino 80-plus Study), use of antipsychotics
in the very old did not seem to be associated with an increased risk of mortality over a follow up
period of four years.
In a prospective ambulatory population of cognitively normal or mildly cognitively impaired
elderly, individuals with baseline elevated homocysteine concentrations have an almost 3-fold
increased risk of developing dementia in the following 3-4 years.
More than 1 out of 10 elderly persons (65-84 years) were anemic and most of the cases had a
mild grade anemia. Hemoglobin concentrations decreased and prevalence of (mild) anemia
increased with increasing age. After controlling for many potential confounders, mild anemia
was found to be associated with poorer health conditions and with increased risk of clinically
relevant outcome such as hospitalization and mortality.
According to the survey conducted in the frame of GISAS Study the most important difficulty
psychiatrists face in participating to clinical trials is related to application of experimental
protocols to daily clinical practice.
Patients with severe mental disorders included in the Natural Social Networks improved
moderately but significantly on general wellbeing according to the HoNOS, and personal and
social functioning according to the GAF, whereas improvements in quality of life were absent
or not significant, particularly in relational and social dimensions.
In a sample of 22368 family members of patients followed by the mental health services there
was an overall moderately good evaluation of the services. Most areas related to accessibility
and support offered were satisfying according to 60-80% of the respondents, whereas the
efficacy in improving health conditions of the patient was satisfying according to less than 50%.
Other aspects needing more efforts were: information about the treating plan, possibility of
treatments delivered by the service and other agencies; family active involvement in the
evaluation of the course of treatment and its effects; difficulties in reaching the service by phone
and waiting lists; effective management of poor compliance of the patient; answer to emergency
needs during closing hours of the service.
Loss of ambulation, percutaneous gastrostomy and non-invasive mechanical ventilation are
outcome measures to be used in epidemiological surveys and therapeutic trials.
The probability of late remission of epilepsy is not uncommon. Partial seizures and having more
seizures prior to treatment predict late remission
The long-term prognosis of epilepsy is the same in patients treated at the first seizure and those
treated at the recurrence. These findings suggest that treatment should be started at the first
seizure only on a case-by-case basis. The use of a third drug in children refractory to two
anticonvulsants does not affect the chance of seizure remission, suggesting that drug resistance
in epilepsy can be identified at the time of failure of two drugs.
89
ANNUAL REPORT 2008

IRFMN
The incidence of acute symptomatic seizures in patients with a firstly diagnosed stroke followed
prospectively is low. Cerebral hemorrhage is the only independent predictor of acute
symptomatic seizures.
We have demonstrated the crucial involvement of some pro- and anti-inflammatory cytokines in
seizures using experimental models of epilepsy in rodents, thus describing a new
etiopathological mechanism which may be relevant for human epilepsy
The membrane-bound drug transport proteins are functionally activated by seizures and have a
significant role in decreasing the brain concentrations of antiepileptic drugs in experimental
models. Pharmacological intervention to block the activity of these proteins may contribute to
reverse multidrug resistance in epilepsy
Gene therapy studies highlight the possibility to significantly reduced spontanerous seizure that
are refractory to anticonvulsant drugs opening the perspective of using gene therapy in
pharmacoresistant forms of epilepsy.
Complement C1-inhibitor (C1-INH) has powerful neuroprotective actions in brain
ischemia/reperfusion injury and in traumatic brain injury
Microglia can explain protective actions in the ischemic environment
Neural stem cell reduce ischemia/reperfusion injury by changing the ischemic
environment
Blood-brain barrier cells can be preconditioned and induced to express a protective phenotype
A dysfunction of proteasome is found in the motor neurons of SOD1 mutant mice, which might
contribute to the accumulation of intracellular protein aggregates. This discovery open a route
for a possible therapeutic strategy based on the application of substance that may increase the
activity of proteasome.
Genetic differences in serotonin synthesis contribute to the efficacy of SSRIs in mice
5-HT 1A and 5-HT 2C receptor antagonists restore the antidepressant-like effect in mice nonresponder
to the SSRI alone
SSRI-induced serotonin release is strongly suppressed in mice carrying the allele 1473G of
TPH-2, the limiting step enzyme in brain serotonin synthesis
The blockade of NMDA receptors of the rat prefrontal cortex induces an increase of glutamate
release and is deleterious for prefrontal cortex-dependent cognitive functions
5-HT 2A receptor antagonists and 5-HT 1A and 5-HT 2C receptor agonists prevent the increase of
glutamate release and attentional deficits caused by NMDA receptors blockade suggesting that
some serotonin receptor subtypes might constitute a molecular target for the development of
drugs for the treatment of cognitive deficits of schizophrenia
90
ANNUAL REPORT 2008

IRFMN
We show that in a glutamate NMDA model of cognitive deficit of schizophrenia antipsychotics
may be differentiated by a selective effect of typical antipsychotics on compulsive
perseveration, and atypical antipsychotics on impulsivity.
A single session of cocaine self-administration is sufficient to shape rat behavior towards goaldirected
behaviors and selectively up-regulate Arc expression in mPFC. This is the first
evidence that the mPFC's function is already profoundly influenced by the first voluntary
cocaine exposure
Naltrexone, a non-selective opioid receptor antagonist, selectively modulates, in abstinent rats,
the drug seeking behaviour induced by the environmental stimuli predictive of cocaine
availability
NATIONAL COLLABORATIONS
Associazione Familiari Insonnia Familiare Fatale malattie da prioni, Treviso
Associazione Italiana GIST A.I.G.
Associazione per la Ricerca Neurogenetica, Lamezia Terme (CS) e ASL 6, Regione Calabria
Agenzia di Sanità Pubblica del Lazio, Regione Lazio
Assessorato alla Salute, Comune di Milano
Azienda Ospedaliera Ospedali Riuniti di Bergamo
Azienda Sanitaria Locale di Bergamo
CEND, Centro Eccellenza per le Malattie Neurodegenerative, Università di Milano
Centro Fatebenefratelli San Giovanni di Dio, Cernusco sul Naviglio
Centro di Neurofarmacologia, Dipartimento di Scienze Farmacologiche, Università di Milano
Centro Studi in Psichiatra, ASL 2, Torino
Centro Parkinson-Istituti Clinici di Perfezionamento
Clinica IRCSS S. Maria Nascente, Milano
Clinica Neurologica III Università di Milano, Azienda Ospedaliera S. Paolo, Milano
Clinica Psichiatrica, Università Milano Bicocca
Consorzio Ricerche Luigi Amaducci, CRIC, Arcugnano (Vc)
Consorzio MIA, Milano
DIBIT, San Raffaele Scientific Insitute, Milano.
Dipartimento di Chimica Biologica, Università di Padova
Dipartimento di Chimica, Università egli Studi di Firenze
Dipartimento Endicronologia, Università di Milano
Dipartimento Farmaco Chimico Tecnologico, Università di Siena
Dipartimento di Farmacologia Medica, Università di Milano
Dipartimento di Fisiologia Umana, Facoltà di Medicina, Università di Milano
Dipartimento di Medicina e Sanità Pubblica, Sezione di Psichiatria e Psicologia Clinica,
Università di Verona
Dip. di Morfofisiologia, Scuola di medicina Veterinaria, Università di Torino, Grugliasco (TO).
Dip. Neurologia, IRCCS Fondazione Maugeri, Pavia
Dipartimento Neurologia, Ospedale Molinette, Torino
Dipartimento di Neurologia Università di Milano, Ospedale Luigi Sacco.
Dipartimento di Neuroscienze, Università di Parma, Parma
Dipartimento di Salute Mentale di Niguarda, Milano
91
ANNUAL REPORT 2008

IRFMN
Dipartimento di Salute Mentale ASL 3 ”Genovese”, Genova
Dipartimento di Salute Mentale ASL 4, Torino
Dipartimento di Salute Mentale San Carlo, Milano
Dip. di Scienze Biomolecolari e Biotecnologie, Università di Milano
Dipartimento Scienze Neurologiche, Università di Genova, Genova
Dipartimento Scienze Neurologiche, Ospedale Maggiore Policlinico di Milano
Direzione Generale Famiglia e Solidarietà Sociale, Regione Lombardia, Milano
Direzione Generale Sanità, Regione Lombardia, Milano
Direzione Regionale Sanità e Servizi Sociali, Regione Umbria
Divisione Neurologica, Università di Bologna
Evidentia Medica, Grottaferrata, Roma
Federazione Alzheimer Italia, Milano
Franco Calori Cell Factory, Centro Trasfusionale e di Immunologia dei Trapianti,
IRCCS Ospedale Maggiore, Milano
Fondazione Clelio Angelino
Fondo Edo Tempia
Hospice “Via di Natale Franco Gallini”, Aviano (PN)
IRCSS "Casa Sollievo della Sofferenza", San Giovanni Rotondo
IRCCS Istituto Auxologico Italiano, Milano
Istituto di Ricovero e Cura a Carattere Scientifico IRCCS (I.N.R.C.A.), Ancona
IRCSS "San Raffaele", Milano
Istituto Europeo di Oncologia, IRCCS, Milano
Istituto di Farmacologia e Farmacognosia, Università di Urbino
Istituto di Farmacologia, Università di Milano
Istituto “G. Ronzoni”, Milano
Istituto Nazionale Neurologico “Carlo Besta”, Milano
Istituto Scientifico Humanitas
Istituto di Scienze e Tecnologie della Cognizione, CNR, Roma
Istituto "Stella Maris", IRCCS, Calambrone (PI)
Istituto Superiore di Sanità, Roma
Istituto Zooprofilattico Piemonte Liguria Val D'Aosta,Torino
Laboratorio di Immunopatologia Renale, Ospedale San Carlo, Milano
Laboratorio di Neuroscienze, Centro Dino Ferrari, Università di Milano
Lega Italiana per la Lotta contro i Tumori
Ospedale del Bambin Gesù, Roma
Ospedale Regionale Ca Fondello, Treviso
Ospedale "Molinette", Torino
Polo Oncologico, ASL 12, Biella
Provincia Lombardo-Veneta Ordine Ospedaliero San Giovanni di Dio, Fatebenefratelli di
Cernusco sul Naviglio
Scuola di Specializzazione in Psicoterapia IRIS-Insegnamento e Ricerca Individuo e Sistemi,
Milano
Scuola di Terapia Cognitiva “Studi Cognitivi”, Milano
Società Italiana Medicina Interna, Roma
Unione Nazionale delle Associazioni per la Salute Mentale (UNASAM), Milano
Unità di Geriatria, Ospedale Maggiore IRCCS, Università di Milano
Unità Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano
Unità Operativa di Psichiatria, Azienda Ospedaliera San Gerardo di Monza, Monza
Unità Operativa di Psichiatria di Garbagnate, Azienda Ospedaliere Salvini di Garbagnate,
Garbagnate Milanese
92
ANNUAL REPORT 2008

IRFMN
Unità Operativa di Psichiatria, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli
e Regina Elena di Milano, Milano
Università degli Studi di Foggia
Università Cattolica del Sacro Cuore di Roma
Università del Piemonte Orientale, Novara
Università di Milano, IRCCS Ospedale Maggiore, Milano
Università Milano-Bicocca, Monza
Università La Sapienza, Roma
U.O. Neurologia, Clinica S. Maria, IRCCS, Castellanza (VA).
UNASAM, Unione Nazionale delle Associazioni per la Salute Mentale
Unità Operativa di Psichiatria, Azienda Ospedaliera Luigi Sacco di Milano, Milano
93
ANNUAL REPORT 2008

IRFMN
INTERNATIONAL COLLABORATIONS
Albert Eistein College of Medicine, Bronx, NY, USA
Atomic Energy Commission, Service de Neurovirologie, Fontenay-aux-Roses, France
Beaumont Hospital, Dublin, Ireland
Brain Repair Centre, University of Cambridge, Cambridge, UK
Cambridge Centre for Brain Repair, University of Cambridge, UK
Centre for Neuroscience Research and Division of Biomolecular Sciences, GKT School, King’s
College, London, UK
Centre National de la Recherche Scientifique, Paris. France
Chorley & South Ribble General Hospital, Chorley,
Columbia Univ, Haverstraw, NY, USA
Department of Anatomy and Physiology, Laval University, Quebec
Department of Cell Biology, Washington University, St Louis, USA
Department of Chemistry,The Australian National University, Canberra City, Australia
Department of Experimental Psychology, University of Cambridge, UK
Department of Pathology and Infectious Diseases Royal Veterinary College, Herts, UK
Department of Psychiatry, Medical Center University of Mississippi, Jackson, USA
Directorate General for the Health and Consumer Protection, European Commission,
Luxembourg
Division of Medical Genetics, CHUV Lausanne, Switzerland
European Union of Family Associations of People with Mental Illness (EUFAMI)
Geriatric Division and Department of Metabolic Diseases, Ospedali Regionali of Lugano and
Mendrisio, Switzerland
HSPH Harvard University, Boston, USA
IBCM, University of Lausanne, Lausanne, Switzerland
INSERM U 751, Marseille, France
Institut de Génétique Humaine du CNRS, Montpellier, France
Jefferson Med Coll, Philadelphia, USA
Karolinska Institutet, Stockholm, Sweden
King’s College Hospital, London, UK
Lancaster University, Lancaster, UK.
Max-Delbrück-Center for Molecular Medicine, Berlin, Germany
National Insitute on Aging, NIH, Baltimore, USA
Neuroprion, Network of Excellence, WP VI, EC
Neurological Department of the University of Tirana, Albania
Neurology, GlaxoSmithKline, New Frontiers Science Park North, Harlow UK
Ninewells Hospital and Medical School, Dundee, Scotland UK
Northern Illinois University, DeKalb, IL, USA
Novartis Pharma, Basel, Switzerland
NYU, NY, USA
Ohio State Univ, Columbus, Ohio, USA
Robarts Research Institute, London, Ontario, Canada
Royal Manchester Children's Hospital, Manchester, UK
Royal Preston Hospital, Preston, UK
Sergievsky Center, Columbia University, New York, NY, USA
Servizio di Geriatria, Ospedale della Beata Vergine, Mendrisio, Switzerland
The Scripps Research Institute, Jupiter, Florida, USA
University of Alberta, Canada
University of Bristol, Frenchay Hospital, Frenchay, Bristol, UK.
94
ANNUAL REPORT 2008

IRFMN
University of Bristol, School of Medical Sciences, UK
University of California at Irvine, Irvine, CA, USA
University of Cardiff, United Kingdom
University of Chicago, Chicago, IL , USA.
University of Colorado, Denver, USA
University Hospital, London, ON, Canada
University of Innsbruck, Innsbruck, Austria
University of Lausanne, Lausanne Switzerland
University of Maryland, Baltimore, USA
University of Maastricht, the Netherlands
University of Rijeka Medical School, Rijeka, Croatia
University of Szeged, Ungary
Université de la Méditerranée ‐Hôpital de la Timone Marseille, France
Université Victor Segalen, Bordeaux, France
Unit of Molecular Genetics, CHUV Lausanne, Switzerland
Virtanen Institute for Molecular Sciences, University of Kuopio, Finland
Vrije Universiteit Medical Center, Amsterdam, The Netherlands
Walton Hospital, Liverpool, UK
WAPR (World Association for Psychosocial Rehabilitation)
Washington University, St Louis, MI,USA
Weill Cornell Medical College, New York, USA
World Mental Health, Department of Mental Health and Substance Abuse, Geneva,
Switzerland
EDITORIAL BOARD MEMBERSHIP
Annals Pharmacotherapy (Nobili)
Biochemical Journal (Chiesa)
Brain Aging (Forloni)
Clinical Drug Investigation (Beghi)
Clinical Neurology and Neurosurgery (Beghi)
Cochrane Collaboration, Epilessia (Beghi)
Dialogo sui Farmaci (Nobili)
Drugs in the R&D (Beghi)
Early Intervention in Psychiatry (Barbato)
Epidemiologia e Prevenzione (Lucca)
Epilepsia (Beghi, Vezzani. Assistant editor)
Epilepsy Current (Vezzani)
Epilepsy Research (Vezzani)
Inpharma (Beghi)
International Journal of Mental Health (Barbato)
Journal of Neurochemistry (Bendotti)
Neurological Sciences (Beghi)
Neuroepidemiology (Beghi)
Neuroscience (Vezzani)
Open Aging Journal (Forloni)
Open Drug Metabolism Journal (Caccia)
Open Geriatric Medicine Journal (Forloni)
Psichiatria di Comunità (Barbato)
95
ANNUAL REPORT 2008

IRFMN
Quality of Life Research (Barbato)
Ricerca & Pratica (Nobili)
Stroke (De Simoni, Associate editor)
PEER REVIEW ACTIVITIES
Acta Neurologica Scandinavica
Acta Psychiatrica Scandinava
Alzheimer Disease and Associated Disorders
American Journal of Clinical Nutrition
American Journal of Human Genetics
American Journal of Pathology
American Journal of Physiology
Annals of Neurology
Annals of Pharmacotherapy
Behavioural Brain Research
Behavioural Neuroscience
Biochimica et Biophysica Acta
Biochemical Journal
Biochemistry
BioMed Central Neurology
Biological Psychiatry
Brain Research
Brain Research Bulletin
Brain Research Review
Clinical Drug Investigation
Clinical Neurology and Neurosurgery
Clinical Pharmacokinetics
Clin Pharm Therapy
CNS Drugs
Dialogo sui farmaci
Drugs
Epidemiologia e Psichiatria Sociale
Epilepsia
Epilepsy & Behavior
European Journal of Immunology
European Journal of Neuroscience
European Journal of Pharmacology
European Journal of Public Health
Experimental Neurology
European Neuropsychopharmacology
Expert Opinion on Pharmacotherapy
FASEB Journal
FEBS letters
Fundamental Clinical Psychopharmacology
Future Drugs
Giornale di Neuropsichiatria dell’Età Evolutiva
Glia
International Journal of Neuropsychopharmacology
96
ANNUAL REPORT 2008

IRFMN
JAMA
Journal of the American Board of Family Practice
Journal of Biological Chemistry
Journal of Cell. Biology
Journal of Cerebral Blood Flow and Metabolism
Journal of Chemical Neuroanatomy
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Science
Journal of Headache and Pain
Journal of Histochemistry and Cytochemistry
Journal of Immunology
Journal of Internal Medicine
Journal of Neurochemistry
Journal of Neuroimmunology
Journal of Neurology, Neurosurgery and Psychiatry
Journal of Neuroscience
Journal of Pharmacy and Pharmacology
Journal of Psychopharmacology
Journal of Psychosomatic Research
Journal of Structural Biology
Life Sciences
Lancet
Lancet Neurology
Molecular Brain Research
Molecular and Cellular Neuroscience
Molecular Therapy
Nature Neuroscience
Neuroepidemiology
Neurology
Neurological Sciences
Nerobiology of Aging
Neurobiology of Diseases
Neuropharmacology
Neuropsychopharmacology
Neuroscience
Neuroscience Letters
N.S. Archives Pharmacology
Parkinsonism & Related Disorders
Pharmacological Research
Pharmacoepidemiology and Drug Safety
Pharmacology Biochemistry & Behavior
PlosONE
Proc Natl Acad Sci, USA
Psychopharmacology
Synapse
Trends Molecular Medicine
97
ANNUAL REPORT 2008

IRFMN
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Advisory Board of the Italian League Against Epilepsy
Board of "Master in Advanced Technologies for the Study of Neurodegenerative Diseases",
University of Milan
Commission on Health Care Policy of the International League Against Epilepsy (ILAE)
Commission of Italian Minister of Health for the study of the problems associated to diagnosis,
therapy and assistance ALS patients
Coordination Group of NoE Neuroprion EC
Coordination Group of the European project ”Quelles professionnalités en santé mentale.
Perspectives croisées, usagers, élus professionnels”.
Council of Italian Association on Brain Aging Research (AIRIC)
Expert for European Agency for the Evaluation of Medicines (EMEA)
Expert reviewer for the Medical Research Council (MRC), UK
Expert of the Minister of Health to EMEA
Italian Association of Neuroepidemiology (Past President)
Italian Association on Brain Aging Research (AIRIC, President)
Italian Society of Neuroscience (Council)
International Committee on “Epilepsy and the Law”
International Organizing Committee e coordinator della segreteria al Global Forum for
Community Mental Health, Department of Mental Health,WHO
International Subcommittee of the American Academy of Neurology
Medical Research Council Strategic Grant Application
Mental Health Working Party of DG-SANCO, Directorate General – Public Health and
Consumer Protection – of the European Union, Brussels, Belgium
National expert accredited by AIFA (Italian Medicines Agency) for the European Agency for
the Evaluation of Medicinal Products (EMEA)
Neurobiology Commission of the International League against Epilepsy
Neuroepidemiology Section of the American Academy of Neurology (past Chair)
Research Advisory Panel, MND Association, UK
Scientific Advisory Board of Sheffield Institute Foundation for MND
98
ANNUAL REPORT 2008

IRFMN
EVENT ORGANIZATION
6 a Giornata di studio sulla malattia di Alzheimer
La depressione nella persona anziana
1 March 2008, Ateneo Veneto, Venezia
ASL Provincia di Bergamo, Bergamo
Interazioni tra farmaci: una valutazione della rilevanza clinica.
1 April 2008 – ASL Bergamo (BG)
ASL Provincia di Bergamo, Bergamo
La prescrizione di FANS e IPP nel rispetto dell’appropriatezza e della continuità terapeutica
ospedale-territorio 23 April 2008 - ASL Bergamo.
59th Annual Meeting of American Academy of Neurology – Breakfast Seminar – How to
manage a patient with a first epileptic seizure: An evidence-based approach – 28 April – 5 May
2007, Boston, MA, USA.
Join Meetin AINP-AIRIC, Istituto di Ricerche Farmacologiche Mario Negri, Milano 18-21 June
2008
8th European Congress on Epileptogenesis, Berlin, September 2008
8th Neuroprion Meeting, Madrid, 26-28 September 2008
ASL Provincia di Bergamo, Bergamo La prescrizione di antibiotici nel rispetto
dell’appropriatezza e della continuità terapeutica 22 October 2008- ASL Bergamo.
Primo GiSAS Investigators’ Meeting 18 November 2008, Istituto di Ricerche Farmacologiche
Mario Negri, Milano
GRANTS AND CONTRACTS
Abbott GmbH & Co. KG
Amgen, Milano
ASL 2 Piemonte.
Assessorato alla Salute, Comune di Milano
Association pour la recherché sur la SLA, France
Bristol-Myers Squibb
Boehringer Ingelheim
CURE Epilepsy
Dipartimento di Salute Mentale, Azienda Ospedaliera Niguarda Ca’ Granda, Milano
Dana Foundation
Evidentia Medica, Grottaferrata (Roma)
Fondazione Cariplo, Milano
Fondazione Mariani, Milano
Fondazione Italo Monzino, Milano
99
ANNUAL REPORT 2008

IRFMN
FP6, European Union
Glaxo-SmithKline, Italy
Hospice "Via di Natale Franco Gallini", Aviano (PN)
Human Frontiers Scientific Programme
IMPHA II, DG-SANCO, Public Health and Consumers' Protection (Directorate General)
Istituto Comprensivo Statale "G.D. Romagnosi", Carate Brianza, Milano
I.R.I.S
Istituto Superiore di Sanità
Janssen-Cilag
H. Lundbeck A/S, Danimark
Ministero della Ricerca Scientifica
Ministero della Salute
MND Association, UK
Newron
Ospedale “Casa Sollievo” di San Giovanni Rotondo
Pharming
Regione Lombardia, Assessorato alla Famiglia e Solidarietà Sociale e Assessorato alla Sanità,
Milano
Rimoldi e Bergamini
Sanofi-Aventis
SELECTA MEDICA, Pavia
Servier Laboratories, Parigi
Sigma-Tau
Telethon
Unione Nazionale Associazioni per la Salute Mentale – UNASAM
Vertex
100
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Albani D, Prato F, Fenoglio C, Batelli S, Dusi S, De Mauro S, Polito L, Lovati C, Galimberti D, Mariani C, Scarpini
E, Forloni G. Association study to evaluate the serotonin transporter and apolipoprotein E genes in frontotemporal
lobar degeneration in Italy. J Hum Genet. 2008; 53:1029-33
Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional
pathway mediates the proconvulsive effects of interleukin-1 beta 2008 Brain, 131: 3256-65
Batelli S, Albani D, Prato F, Polito L, Franceschi M, Gavazzi A, Forloni G. Early-onset Alzheimer disease in an
Italian family with presenilin-1 double mutation E318G and G394V. Alzheimer Dis Assoc Disord. 2008; 22:184-7.
Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q 2008;
79: 121-132.
Baviera M, Invernizzi RW, Carli M Haloperidol and clozapine have dissociable effects in a model of attentional
performance deficits induced by blockade of NMDA receptors in the mPFC Psychopharmacology (Berl), 2008; 196:
269-280.
Beghi, E. The management of a first seizure – Still a major debate. Epilepsia 2008; 49 (Suppl.1): 1.
Beghi, E. Management of a first seizure. General conclusions and recommendations. Epilepsia 2008; 49 (Suppl.1):
58-61.
Beghi, E. Beghi, M. Epidemiology of epilepsy in women. In: Educational Kit on the Epilepsies 2008; Vol. 4: 26-30.
Beghi, E., Millul, A., Logroscino, G., Vitelli, E., Micheli, A. for the SLALOM Group. Outcome measures and
prognostic indicators in patients with amyotrophic lateral sclerosis. Amyotrophic Lateral Sclerosis 2008; 9: 163-167.
Beghi, E:, Atzeni, L. Garattini, L. Economic analysis of newer antiepileptic drugs. CNS Drugs 2008; 22: 861-875.
Bernardino L, Balosso S, Ravizza T, Marchi N, Ku G, Randle JC, Malva JO, Vezzani A. Inflammatory events in
hippocampal slice cultures prime neuronal susceptibility to excitotoxic injury: a crucial role of P2X(7) receptormediated
IL-1beta release (2008) J Neurochem 106: 271-280
Biasini E, Seegulam ME, Patti BN, Solforosi L, Medrano AZ, Christensen HM, Senatore A, Chiesa R, Williamson
RA, Harris DA. Non-infectious aggregates of the prion protein react with several PrP(Sc)-directed antibodies. J
Neurochem. 2008, 106: 2190-2204
Biasini E, Medrano AZ, Thellung S, Chiesa R, Harris DA. Multiple biochemical similarities between infectious and
non-infectious aggregates of a prion protein carrying an octapeptide insertion. J Neurochem. 2008, 104:1293-308
Cheroni C, Marino M, Tortarolo M, Veglianese P, De Biasi S, Fontana E, Vitellaro Zuccarello L, Maynard CJ,
Dantuma NP, Bendotti C. Functional alterations of the ubiquitin proteasome system in motor neurons of a mouse
model of familial Amyotrophic Lateral Sclerosis. Hum Mol Genet. 2008 Sep 29. [Epub ahead of print]
Chiesa R, Piccardo P, Biasini E, Ghetti B, Harris DA. Aggregated, wild-type prion protein causes neurological
dysfunction and synaptic abnormalities. J Neurosci. 2008, 28:13258-67.
Colovic M, Caccia S. Liquid chromatography-tandem mass spectrometry of I3,II8-biapigenin, the major
biflavone in Hypericum perforatum extracts. J Chromatogr B Biomed Appl 2008 ; 863 : 74-79
Colovic M, Fracasso C, Caccia S. Brain-to-plasma distribution ratio of the biflavone amentoflavone in the
mouse. Drug Metabolism Letters 2008, 2 : 90-94
Cosentino U, Pitea D, Moro G, Saracino GA, Caria P, Varì RM, Colombo L, Forloni G, Tagliavini F, Salmona M.
The anti-fibrillogenic activity of tetracyclines on PrP 106-126: a 3D-QSAR study. J Mol Model. 2008, 14: 987-94.
Del Bo R, Ghezzi S, Scarlato M, Albani D, Galimberti D, Lucca U, Tettamanti M, Scarpini E, Forloni G, Bresolin N,
Comi GP. Role of VEGF gene variability in longevity: a lesson from the Italian population. Neurobiol Aging. 2008,
29:1917-22.
101
ANNUAL REPORT 2008

IRFMN
de Falco, FA., Sterzi, R.,Toso, V., Consoli, D. Guidetti, D., Provinciali, L., Leone, MA., Beghi, E. The neurologist
in the emergency department. An Italian nationwide epidemiological survey. Neurol Sci 2008; 29: 67-75.
De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Limido L, Forloni G, Tagliavini F,
Salmona M. The efficacy of tetracyclines in peripheral and intracerebral prion infection.
PLoS ONE. 2008, 3(3):e1888.
Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M,
Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa Mutant
prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model.
Neuron 2008; 60 : 598-609
Ferrarin M, Rabuffetti M, Tettamanti M, Pignatti R, Mauro A, Albani G. Effect of optical flow versus attentional
strategy on gait in Parkinson's Disease: a study with a portable optical stimulating device. J. NeuroEnging. &
Rehabil. 2008; 5: 3.
Gironi, M., Guerini, FR., Beghi, E., Antonimi, G., Martinelli-Boneschi, F., Ceresa, L., Morino, S., Agliardi, C.,
Ferrante, P., Nemni, R. HLA-DRB1 polymorphisms distribution in chronic dysimmune polyneuropathy.
Neuromuscul Disord 2008; 18: 967-969.
Guzzetti S, Calcagno E, Canetta A, Sacchetti G, Fracasso C, Caccia S, Cervo L, Invernizzi R Strain
differences in paroxetine-induced reduction of immobility time in the forced swimming test in mice: role of
serotonin. Eur J Pharmacol 2008 ; 594 : 117-124
Hauser, WA ande Beghi, E.. First seizure definitions and worldwide incidence and mortality. Epilepsia 2008; 49
(Suppl.1): 8-12.
Lehnert M, Relja B, Sun-Young Lee V, Schwestka B, Henrich D, Czerny C, Froh M, Borsello T, Marzi I. A peptide
inhibitor of C-jun N-terminal kinase modulates hepatic damage and the inflammatory response after hemorrhagic
shock and resuscitation. Shock. 2008, 30:159-65.
Leopoldo M, Lacivita E, De Giorgio P, Fracasso C, Guzzetti S, Caccia S, Contino M, Colabufo N A, Berardi F,
Perrone R Structural modifications of N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4-Aryl-1-piperazinehexa namides:
Influence of lipophilicity and 5-HT7 receptor activity. Part III. J Med Chem 2008; 51 : 5813-5822
Lucca U, Tettamanti M, Quadri P. The Italian version of Consortium to Establish a Registry of Alzheimer’s Disease
(CERAD). Alzheimer’s & Dementia 2008; 4: 310.
Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico
M, Tempia P, Guala A, Fasolo G, Riva E. Association of Mild Anemia with Cognitive, Functional, Mood, and
Quality of Life Outcomes in the Elderly: The “Health and Anemia” Study. PLoS ONE 2008; 3(4): e1920.
Logroscino, G., Traynor, BJ., Hardiman, O., Chiò, A:, Couratier, P., Mitchell, JD., Swingler, RJ., Beghi, E. and for
EURALS. Descriptive epidemiology of amyotrophic lateral sclerosis: new evidence and unsolved issues. J Neurol
Neurosurg Psychiatry 2008; 79: 6-11.
Montes, J., Bendotti, C., Tortarolo, M., Cheroni, C., Hallak, H., Speiser, Z., Gore, S:, Blaugrund, E., and Gordon, PH.
Translational Research in ALS. in :Animal and Translational Models of Behavioral Disorders. Volume 2 –
Neurologic Disorders RA McArthur and F Borsini (Eds.), Elsevier, NY, 2008, 275-318,
Natalello A, Prokorov VV, Tagliavini F, Morbin M, Forloni G, Beeg M, Manzoni C, Colombo L, Gobbi M, Salmona
M, Doglia SM. Conformational plasticity of the Gerstmann-Sträussler-Scheinker disease peptide as indicated by its
multiple aggregation pathways. J Mol Biol. 2008 Sep 19;381(5):1349-61.
Nobili A, Piana I, Balossi L, Pasina L, Matucci M, Tarantola M, trevisan S, Riva E, Lucca U, Tettamnti M.
Alzheimer Special Care Units compared wuth traditional nursing home for dementia. Are there differences at
admission and in clinical outcomes Alzheimer Dis Assoc Disord 2008; 22: 352-361.
102
ANNUAL REPORT 2008

IRFMN
Noè F, Pool AH, Nissinen J, Gobbi M, Bland R, Rizzi M, Balducci C, Ferraguti F, Sperk G, During MJ, Pitkänen A,
Vezzani A. Neuropeptide Y gene therapy decreases chronic spontaneous seizures in a rat model of temporal lobe
epilepsy 2008 Brain, 131:1506-1515
Pastori C, Librizzi L, Breschi GL, Regondi C, Frassoni C, Frigerio S, Gelati M, Parati E, De Simoni MG, de Curtis
M. Arterially perfused neurosphere-derived cells do not cluster in the ischemic area in a model of transient focal
ischemia and reperfusion in vitro. 2008 PLoS ONE 3(7):e2754,
Piazzini, A., Beghi, E., Turner, K., Ferraroni, M. the LICE Quality of Life Group. Health-related quality of life in
epilepsy: findings with a new Italian instrument. Epilepsy & Behavior 2008; 13: 119-126.
Pugliatti M., Sobocki, P., Beghi, E., Pini, S., Cassano, GB., Altamura, AG, Pozzoli, S., Rosati, G. Cost of disorders of
the brain in Italy. Neurol Sci 2008; 29: 99-107.
Repici M, Zanjani HS, Gautheron V, Borsello T, Dusart I, Mariani J. Specific JNK inhibition by D-JNKI1 protects
Purkinje cells from cell death in Lurcher mutant mouse. Cerebellum. 2008, 7: 534-8.
Ravizza T, Noé F, Zardoni D, Vaghi V, Sifringer M, Vezzani A. Interleukin Converting Enzyme inhibition impairs
kindling epileptogenesis in rats by blocking astrocytic IL- 1β production Neurobiol Dis, 2008, 31: 327-332
Ravizza T, Gagliardi B, Noè F, Boer K, Aronica E and A. Vezzani. Innate and adaptive immunità during
epiletogenesis and spontaneous seizures: evidence from experimental models and human temporal lobe epilepsy
(2008) Neurobiol. Dis, 29: 142-150
LAY PRESS SELECTION PUBLISHED IN 2008
Barbato A. La riabilitazione in psichiatria: cosa è cambiato e cosa dovrebbe cambiare. Psichiatria di Comunità 2008,
7:193-195.
Barbato A. Mettere tra parentesi la malattia mentale. Nuove (e vecchie) ipotesi per la cura della sofferenza psichica.
Animazione Sociale 2008; 8/9: 3-13.
Barbato A, D'Avanzo B, Ferrannini L, Parabiaghi A, Vaggi M. Un'occasione per la ricerca clinica in Italia: lo studio
GISAS su aripiprazolo, olanzapina e aloperidolo nel trattamento dei disturbi schizofrenici. Psichiatria di Comunita
2008; 7: 46-54.
Barbato A. Confrontation entre professionels et usagers: vers une modification de la pratique in Italie. Rhizome 2008;
33:41-43.
D'Avanzo B, Aliprandini E, Beghi M, Cornaggia C M, Erlicher A, Frova M, Mascarini A, Miragoli P, Righi A.
Strutture residenziali e semiresidenziali nei servizi di salute mentale. Dove sta la differenza Epidemiologia e
Psichiatria Sociale 2008; 17: 57-64.
Invernizzi R, Caccia S. Gli antidepressivi. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e rilevanza
clinica. Selecta Medica, Pavia, 2008; 59-215
Mennini T, Caccia S. Le benzodiazepine. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e
rilevanza clinica. Selecta Medica, Pavia, 2008; 397-459
Pasina L, Caccia S. Gli antipsicotici. In: Le interazioni dei farmaci neuropsichiatrici tra farmacologia e
rilevanza clinica Selecta. Medica, Pavia, 2008; 265-395
Pasina L, Caccia S, Nobili A. Gli analgesici oppioidi. In: Le interazioni dei farmaci neuropsichiatrici tra
farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; 35-58
103
ANNUAL REPORT 2008

IRFMN
Pasina L, Caccia S, Nobili A. Antiparkinsoniani In: Le interazioni dei farmaci neuropsichiatrici tra
farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; 217-264
Pasina L, Caccia S, Nobili A. I farmaci antiepilettici. In: Le interazioni dei farmaci neuropsichiatrici tra
farmacologia e rilevanza clinica. Selecta Medica, Pavia, 2008; 461-533
RESEARCH ACTIVITIES
Laboratory of Biology of Neurodegenerative Disorders
Alzheimer's disease: genetic studies and clinical investigations
In collaboration with different neurological centers and the laboratory of Geriatric
Neuropsychiatry it has been created a bank of blood samples for DNA of patients with
Alzheimer’s disease (AD), in familial (FAD) or sporadic form (SAD), and patients with
vascular dementia (VD). In all subjects the diagnosis of dementia is performed according to the
international guidelines. Since 2005 we started also the collection of blood samples from
subjects with fronto-temporal dementia. The genetic studies are aimed to the identification of
causal factors in FAD and risk factors in SAD. Mutations on genes encoding proteins involved
in the physiopathology of AD were investigated. The pathogenic role of these mutations is
under investigation using fibroblasts obtained from skin biopsy. Furthermore, we continued the
screening of FAD samples for the genes encoding for presenilin 1 and 2 (PS-1 and PS-2) and
APP, missense mutations in these three genes were associated with AD.
Alzheimer's disease: preclinical studies
The formation of β amyloid (Aβ) deposits in brain parenchyma and on the wall of cerebral blood
vessels is an early event in AD and there are now numerous genetic, biochemical and
neuropathological studies pointing to a causal role of Aβ in the pathogenesis of AD. Thus,
prevention the formation of Aβ aggregates or their elimination once formed is a potential
therapeutic approach to the disease. This aim is strongly persecuted with different strategies
including the regulation of enzymes responsible of the synthesis and degradation of Aβ and the
enzymes influencing the metabolism of amyloid precursor protein (APP). In the lab, we developed
the idea to interfere directly with the Aβ deposits formation using anti-amyloidogenic drugs. The
experimental studies have shown the potential therapeutic activity of these drugs in AD, and now
they will be tested in a clinical setting.
The role of oligomers in the Alzheimer pathogenesis
Recent data have shown the essential role plays by oligomers, small and soluble aggregates of
Aβ, in the Alzheimer pathogenesis and in particular in the cognitive decline associated to the
disease. In collaboration with the Department of Biochemistry an Molecular Pharamacology we
developed some in vivo models to analyze the neuronal dysfunction induced by Aβ 1-40 e 1-
42 but not in monomeric or fibrillar species. The intracerebral application of these different
forms confirmed that Aβ oligomers induced behavioral impairment while monomeric or fibrillar
forms of Aβ did not affect the cognitive behavior,
The intracellular signaling pathways, by which Aβ oligomers induce synaptic failure and
consequently neuronal degeneration are poorly understood. Nevertheless increasing evidence
indicate the involvement of kinases-dependent signalling pathways, and more specifically the
JNK signalling pathway in these early degenerative events.
104
ANNUAL REPORT 2008

IRFMN
The JNK kinase phosphorylates APP (amyloid precursor protein) and its relevance in both
neuronal death and brain plasticity is well established. We recently demonstrated, by using the
specific cell penetrating JNK inhibitor peptide (D-JNKI1) charcterized by dr. Borsello, that JNK
is responsible for APP phosphorylation at Thr668, and that its specific inhibition reduced the
βAPPs and Aβ fragments production in primary cortical neurons. In addition, we could show
that JNK inhibition leads to a shift from the amyloidogenic to the non-amyloidogenic pathway,
a result with potentially important therapeutical implications.
Synaptic Dysfunction
Nowadays it is assumed that AD is a synapse-related pathology leading to synaptic dysfunction
and loss, a phenomenon that precedes extensive amyloid deposition in the brain. At the same
time, soluble diffusible forms of Aβ can perturb in an early stage of the disease the synaptic
function causing a reduction of dendritic spines density in the cortex and hippocampus, an acute
inhibition of long term potentiation (LTP) and the loss of critical spine proteins (e.g. membrane
expression of NMDA receptors).
However, the relationship between Aβ and synapses loss remains unclear and more efforts are
necessary to better understand the mechanisms underlying Aβ synaptic toxicity. Our aim is to
study the effects of Aβ oligomers on MAPKs pathways and elucidate the link between
synaptopathies and the activation/inhibition of the JNK, p38 and ERK cascades. This study can
potentially be a breakthrough in the comprehension of AD pathogenesis: understanding the
cellular and molecular alterations that lead to AD will help in developing effective and
preventive therapeutic strategies in order to counteract or nullify the degenerative processes
activated by Aβ.
MAPKs (ERK, p38 and JNK) are also implicated in the regulation of the immediate early
signaling events that modulate synaptic plasticity by controlling LTP, LTD and the recycling of
glutamate receptors (NMDAR and AMPAR) as well as their expression. Nevertheless, MAPKs
involvement in the regulation of synaptic function and dysfunction and the mechanisms by
which they trigger the synaptic loss induced by Aβ oligomers are largely unknown.
The cargo strategy as a key tool in neuroprotection
The possibility to target protein complexes and enzymes involved in intracellular signaling
pathways by means of cell permeable peptide (CPP) conjugates represents a novel, versatile and
extremely powerful way of blocking the propagation of intracellular signaling events or
intracellular processes, with an unprecedented specificity allowing for reduction of side effects.
D-JNKI1 is a cell-permeable, biologically-active peptide consisting of a carboxyl terminal
sequence derived from the JNK binding domain of the scaffold protein JIP-1/IB1 (JBD20), and
an amino terminal portion containing the HIV-TAT 48-57 transporter sequence. D-JNKI-1 has
been designed to block the interaction, mediated by JBD domain, between JNK and its targets.
D-JNKI1 afforded powerful protection against NMDA excitotoxicity in cortical neurons and
against cerebral ischemia in vivo, as well as in two other neurodegenerative models (hair-cell
loss in animal models of sudden deafness and retinal ganglion cell loss following optic nerve
crush in vivo. The peptide progressed in clinical trails for preventing brain ischemia/stroke (see
CHUV/Xigenpharma Lausanne web-link). Among the possible targets for neuroprotection there
is MKK7, that is activated, unlikely MKK4, during NMDA-stress,. To inactivate MKK7 we use
lentiviruses because of the particular capability of integrating genetic material into the genome
of non-dividing cells stably. Our lentiviral vector will carry a single si-RNA duplex of MKK7.
A second approach is to test in vitro the specific inhibitor: Gadd45, a molecule active on
MKK7. Gadd45β binds to MKK7 directly and blocks its catalytic activity, the binding between
Gadd45β/MKK7 being tighter than JIP1/MKK7. The endogenous Gadd45 interacts to MKK7
through direct, high-affinity contact but not with the other JNK upstream kinase, MKK4. For
this study we initially plan to use a viral system that will allow us to observe the role of this
105
ANNUAL REPORT 2008

IRFMN
pathway in excitotoxicity, with a final goal of producing a cell-permeable peptide with a more
specific effect in the prevention of neuronal death.
SUMOlization in acute and chronic diseases
Small ubiquitin-like modifier (SUMO) is a group of proteins responsible for post translational
modifications influencing protein function, localization and stability. Recently, protein
Sumoylation has attracted neuroscientists since it is implicated in the altered protein dynamics
that are associated with various aspects of neurodegenerative disease, including stroke amd
Alzheimer's Disease (AD). Our hypothesis is that SUMOylation confers neuroprotection against
stressful stimuli through regulation of important stress signaling pathways. The aim of this
study is to investigate the role of SUMOylation in models of acute (ischemia) and chronic brain
diseases (AD). At present we are characterising the changes in expression and localisation of
SUMO1-2/3 in an in vitro model of ischemia (NMDA application) as well as in a model of AD
(oligomers application).
Genetics of aging
In collaboration with Geriatric Neuropsychiatry Lab for the Monzino 80-plus study and with dr.
Maurizio Gallucci from the ARGel Association in Treviso for Trelong study we collected a
large number of blood samples from subjects over seventy. In these samples we are performing
a genetic analysis to identify genetic profiles associate to the longevity and /or to the agingassociated
pathologies with specific attention to the dementias. The aim is to cross the
genotype/phenotype profile with pathologies and environmental aspects including style of life,
diet and economical conditions to identify risks and protective factors. Initially the subjects
were genotypized for ApoE, whom allele E4 is a well-known risk factor for Alzheimer’s disease
and several other disorders and sirt-1 a gene codified for protein member of a enzymatic family
of sirtuins associated to the longevity in several experimental models. The results are interesting
but before drawing any conclusion we need to considere the numerous other parameters
collected in our database.
Prion's disease: in vitro studies
Prion’s diseases (TSE) are neurodegenerative disorders of sporadic inherited origin but also
transmissible, they have different clinic and neuropathological features but all are characterized
by the cerebral accumulation of an altered form of prion protein (PrPsc). TSE are rare diseases
but the transmission from bovine (BSE) to humans induced a public health alarm in UK and
successively in all Europe. PrPsc is involved in the pathogenesis of the disease and it is also an
essential component of the infective agent. In the lab numerous projects were developed to
understand the association between the presence of PrPsc and the neurodegenerative process.
The pathogenetic role of PrP was investigated not only through the external application of
peptides but also by the evaluation of intracellular mechanisms potentially involved in the
formation of PrP aggregates.
Prion’s diseases: studies in vivo
The lab has the facilities to study the experimental scrapie, mice and hamsters are inoculated
with infected brain homogenate. The hamsters after intracerebral inoculation develop the
disease in 60-70 days and died within a month. The histopathological analysis of the brain show
the presence of PrPsc deposits, neuronal damage, diffuse astrogliosis and the typical spongiosis
at cortical and thalamic level. The anti-amyloidogenic activity of tetracyclines has been
investigated also in this experimental contest. After the ex-vivo approach, where the
homogenate was treated before the inoculation, the curative effect of tetracyclines was tested in
collaboration with the lab of Biochemistry and Chemistry of Proteins by treatment the
experimental scrapie in hamsters with intramuscular doxycycline, the treatment prolonged the
106
ANNUAL REPORT 2008

IRFMN
survival of the animals. Other drugs are now under investigation to verify the curative effects in
TSE.
In the lab are available transgenic mouse developed by dr. Chiesa. These mice express the
mutated forms of PrP associated to the familial TSE. The homozygotes exhibit at different level
of severity the symptoms reminiscent of the pathologies associated with the mutations. From the
neuropathological point of view some lines exhibit an accumulation of PrP and clear
neurodegeneration of the cerebellar granular cells, in the other brain regions no evident
alterations were found. In the brain tissue of the mice was found a PrP with some of the
characteristic of PrPsc, however the brain material is not infected. The pathogenesis of TSE is
investigated in these models through different approaches including proteomics and electronic
microscopy (EM). The EM analysis have shown that in transgenic mice there was an alteration
at the neuronal level of the Golgi apparatus and/or of endoplasmic reticulum associated to the
different distribution of mutated PrP compared to native protein. Furthermore, there are in
progress studies to investigate the molecular mechanisms responsible of the neurodegenerative
processes occurring in cerebellar granular cells of the insertional prion transgenic mice
(PG14).
Parkinson’s Disease: genetic studies
Parkinson’s disease (PD) is the second more diffuse neurodegenerative disorder with an
unknown pathogenesis, however for PD several therapies are available and, although at the
symptomatic level, their efficacies is well-established. In the etiological studies on PD the
genetic component has been traditionally considered with scarce interest whereas the
environmental causes were carefully evaluated. This orientation was based on the evidence that
the exposure to several toxins can mimic the PD pathology. However the genetic studies in the
last few years have completely changed the perspective with the identification of mutations on
two genes, encoding for alpha-synuclein and parkin, associated to the juvenile forms of the
disease. A mutation on alpha synuclein gene is an event extremely rare, only three mutations
identified until now, the parkin mutations are numerous ether in puntiform or in deletion form.
The mutations on alpha-synuclein gene are dominant while the parkin mutations are associated
with PD in recessive form. We collected, in collaboration with several neurological centers,
blood samples from PD subjects and the screening of the samples involved genes like alphasynuclein,
parkin, DJ-1 and other factors potentially involved in PD. Recently, an assocation
between polymorphisms occurring in gene for serotonin transporter and the appearance of
depression in PD subjects has been investigated. The results indicate no association between the
serotonin traporter gene polymorphisms and depression in PD, but a direct association between
these polymorphisms and PD itself. This indicate a more relevant involvement o serotonergic
system in PD pathogenesis compared to whom is generally considered.
Parkinson’s disease: studies in vitro
The identification of the mutations associated to Parkinson’s disease (PD) gave a substantial
contribute to understand the disease and allowed the develop of cellular models to investigate
the pathogenesis of the disease. In past we showed the potential neurotoxic activity of alphasinuclein
using the synthetic peptide homologous to the fibrillogenic fragment 61-95 (NAC) of
the protein. Successively with help of dr. Negro at the Department of Biochemistry at the
University of Padova we prepared cDNA vectors including the sequence of wild type and
mutated alpha-synuclein Their transfection to the PC12 cells induced in specific conditions a
cellular damage. More recently alpha-synuclein was associated to a TAT sequence capable to
transport inside the cells the protein. With this method the intracellular concentration of alphasinuclein
was better controlled. In a micromolar range alpha-synuclein was toxic, but in
nanomolar range, it exerted neuroprotective effect against oxidative stress induced by hydrogen
peroxide. This double effect dose-dependent was confirmed in an “inducible” model. More
107
ANNUAL REPORT 2008

IRFMN
recently again in collaboration with Dr. Negro (Padua University), we obtained the recombinant
form of DJ-1 associated with TAT (TAT-DJ-1). This protein is similar to alpha-synuclein,
mutations of its sequence has been associated to PD. TAT-DJ-1 silencing by small interference
RNA (siRNAi) were used to study the interaction between DJ-1 and alpha synuclein..
Laboratory of Neurological Disorders
Epidemiological studies on amyotrophic lateral sclerosis (ALS)
Included are studies on the incidence, risk factors and mortality of ALS. The data are obtained
from a regional registry of the disease activated in 1998 and including all patients with newly
diagnosed ALS identified in eight provinces of Lombardy. Using similar study protocols, the
same data are collected in two additional regional registries (from Piemonte and Puglia) included
in a network with the Lombard registry. Information obtained from patients enrolled in the
Lombard registry and from cases examined by members of the Italian ALS Study Group has
been used to assess the validity and reliability of diagnostic criteria for ALS and selected
disability scales. Based on the data recorded, the annual incidence of ALS is comparable to that
obtained in other Western countries where ALS registries have been activated, and is among the
highest ever published (1.9 per 100,000). Mortality of ALS has been found to be comparable to
that of studies from similar populations studied with the same protocol. The study on the
validation of the current diagnostic criteria for ALS (the El Escorial criteria) showed that to be
considered valid and reliable, the criteria should be used after proper training of the
investigators.
In October 2004, the Laboratory of Neurological Disorders has started a European collaborative
group for the ALS registries (EURALS) with the intent to create a common database (completed
in the year 2005) with the participation of the existing regional and national disease registries.
Three major scientific activities are in course: 1. A comparative study of the clinical
characteristics of patients with ALS enrolled in the registries as compared to those seen in
secondary and tertiary health care facilities; 2. A meta-analysis of the incidence of ALS,
performed by pooling data from the 1998-99 cohorts of patients enrolled in the population-based
registries. The scientific reports of these studies have been submitted to international scientific
journals for publication. Other studies are ongoing under the coordination of the Laboratory of
Neurological Disorders: 1. A case-control study on trauma and risk of ALS (in collaboration
with the Italian registries); 2. A case-control study on ALS, physical exercise and sport (in
collaboration with the EURALS Consortium). Study # 3 is ongoing only in Italy; A survey of the
prevalence of cognitive impairment and extrapyramidal signs in patients with newly diagnosed
ALS (Italian registries); 4. A study of the mortality of ALS in the 1998-99 cohort of patients
from the European population-based registries (EURALS Consortium).
In 2008 a scientific report was published on the progression of ALS (marked by loss of
ambulatiom, percutaneous gastrostomy and non-invasive assisted ventilation).
Early and late remission of epileptic seizures
During the calendar year 2008, the analysis of the data was completed for a retrospective
observational study in a cohort of adolescents and adults with newly diagnosed epilepsy seen in
two epilepsy centers (Monza, Bari) and followed for several years. The aim of the study was
twofold: 1. To assess the long-term prognosis of epilepsy defined by the number of cases with
late remission (ie, 2+ year seizure freedom with onset after 24 months of treatment; 2.
Identification of the predictors of late remission as compared to early remission (ie, 2+ year
remission immediately after treatment start). The cohort included 372 cases, 23% of which
achieved early remission and 11% late remission. The chance of late remission increased
significantly in patients with partial seizures and an increasing number of seizures prior to
starting treatment
108
ANNUAL REPORT 2008

IRFMN
Innovative therapeutic strategies in patients with epilepsy
A cohort of patients with a first unprovoked seizure, randomised since 1988 by several Italian
centers to immediate treatment or to treatment only at the time of a seizure relapse, was
followed to verify the impact of the two therapeutic strategies on the long-term prognosis of
epilepsy, measured by the chance of achieving 5-year remission.
To provide a pragmatic definition of drug resistance in childhood epilepsy, children refractory
to two antiepileptic drugs (in sequence or in combination) were randomised to the use of a third
drug or to the optimization of the existing treatment and followed for up to three years.
Therapeutic response was measured by the achievement of a six-month period of remission. The
study has been conducted in collaboration with the IRCCS “Stella Maris” of Calambrone (PI).
The study has been concluded prior to completing patient recruitment because the eligible
patients could not be easily traced. The available data have processed and analyzed and a
scientific manuscript is in preparation
Epidemiology of neurological disorders in Albania
With the collaboration of the Fondazione Mariani and the Neurological Department of the
University of Tirana, an epidemiological survey has been started to assess the prevalence and
incidence of several neurological conditions (stroke, epilepsy, headache, dementia, peripheral
neuropathy, multiple sclerosis) comparing an urban and a rural community (Tirana and
Saranda). In 2005, a study on the validation of the diagnostic criteria was conducted and the
recruitment of the cases to include in the incidence and prevalence studies is in course.
Quality of life in children with neuromuscular disorders
With a financial support of the Telethon organization, a study has been completed on the
validation of a questionnaire on the quality of life in children and adolescents with different
neuromuscular disorders. This is an Italian multicenter study coordinated to the Child
Neurology Clinic of Pavia. The data analysis has been completed and two scientific manuscripts
have been prepared and are ready for submission to ad-hoc scientific journals.
Cerebrovascular disorders and risk of epilepsy
Epilepsy is a frequent complication of stroke. Acute symptomatic seizures (i.e. seizures
occurring in the seven days after stroke) can occur in up to two-thirds of cases and epilepsy (i.e.
repeated unprovoked seizures) in 2-4%. There are no consistent findings on the risk factors for
acute symptomatic seizures, unprovoked seizures and epilepsy in patients with stroke. For these
reasons, in 2007 a multicenter national prospective survey has been started to assess the risk of
seizures and epilepsy (and the main risk factors) in a cohort of patients with a first ischemic or
hemorrhagic stroke followed for a maximum period of 24 months. The study was also
implemented to assess the feasibility of a pragmatic therapeutic trial on the prophylaxis of
seizures and epilepsy in stroke. Based on a preliminary analysis of 583 patients, the incidence of
acute symptomatic seizures (5.7%) and the independent predictors of seizures (cerebral
hemorrhage).
Therapeutic trials in neurological disorders
During the year 2008 three therapeutic trials sponsored by the Italian Drug Agency (AIFA) and
a therapeutic trial sponsored by the Italian Ministry of Health were started or continued.
Included are: 1. A randomized double-blind parallel-group placebo-controlled trial on the
efficacy and tolerability of L-acetylcarnitine in ALS; 2. A randomized open-label parallel-group
trial comparing Erythropoietine to Metyl-prednisolone in patients with acute spinal cord injury;
3. A randomized double-blind parallel-group placebo-controlled trial on the efficacy and safety
of valproate in medication-overuse headache; 4. A randomized trial of the efficacy of a
comprehensive rehabilitation program for the prevention of falls in Parkinson’s disease. The
109
ANNUAL REPORT 2008

IRFMN
first trial aims at finding a potentially effective drug in a clinical condition for which there is
only one product (Riluzole) with at best modest efficacy on survival. L-acetylcarnitine has been
found to improve survival in experimental models of motor neuron disease. The second trial
intends to verify the efficacy of erythropoietin, a drug shown to mitigate the effects of traumatic
spinal shock and accelerate recovery in experimental animals. The drug chosen for comparison
(Methylprednisolone at high doses) has been selected for being the present gold standard in
clinical practice. The third trial aims at verifying whether valproate (a drug commonly used for
the prophylaxis of migraine) abates symptoms occurring in drug-overuse headache, a common
and frequently invalidating variety of chronic idiopathic headache. The fourth trial aims at
assessing whether a comprehensive rehabilitation program compared to usual care is follone by
a reduction in the incidence of falls in patients with Parkinson’s disease at risk of falls.
The first three trials are multicenter and nationwide. The laboratory of neurological disorders is
the coordinator of the first trial and a partner in the other two trials, where the main tasks
include protocol and CRF preparation, statistical analysis, and preparation of the final scientific
report. The fourth trial is coordinated by the Laboratory and is conducted in Lombardy in
conjunction with the Fondazione Don Gnocchi.
Last, a randomized trial is in advanced preparation on the efficacy of an educational program for
physicians working in nursing homes. The aim of the study is to verify a reduction in the
number of inappropriate prescriptions compared to usual care.
Laboratory of Drug Metabolism
1-Aryl-piperazine as active metabolites of centrally acting drug
Starting from the previously reported 5-HT 7 receptor compounds with N-(1,2,3,4-
tetrahydronaphthalen-1-yl)-4-aryl-1-piperazineexanamide structure (Leopoldo et al., J. Med. Chem.,
2007, 50, 4214), a new series of 1-(2-methythiophenyl)-, 1-(2-biphenyl)-, 1-(2-isopropylphenyl)- and
1-(2-methoxyphenyl)-piperazine derivatives were designed with the aim to obtain new potent
derivatives endowed with suitable physicochemical properties for rapid and extensive penetration into
the brain. The newly synthesized compounds underwent radioligand binding assays to assess their
affinities for 5-HT 7 , 5-HT 1A , and D 2 receptors. The intrinsic activities at 5-HT 7 receptor as well as
those for serotonin 5-HT 1A and dopamine D 2 of the most potent compounds were determined
(Leopoldo et al., J. Med. Chem., 2008; 51: 5813).
The proposed structural modifications on the tetrahydronaphthalenyl ring of the original compounds
were, in most cases, detrimental for 5-HT 7 receptor, whereas had limited impact on the affinity for 5-
HT 1A and D 2 receptors. Nonetheless, the pursued strategy led to the identification of the 1-(2-
biphenyl)piperazine derivatives which retained nanomolar affinity at 5-HT 7 receptor, still showing
lipophilicity within the target range. Among these, the N-(4-cyanophenylmethyl)-4-(2-diphenyl)-1-
piperazinehexanamide derivative also demonstrated high selectivity over 5-HT 1A and D 2 receptors
(324- and 245-fold, respectively) and showed full competitive agonism at 5-HT 7 receptor in an
isolated guinea-pig ileum assay. It rapidly reached the systemic circulation and entered the brain,
achieving concentrations in the low micromolar range after intraperitoneal doses in mice. Its brain
concentration-time profile paralleled that in plasma, indicating that it rapidly and freely distributes
across the blood-brain barrier. Similar studies were performed on the most potent and selective 5-HT 7
agonist derivative of the previous series - which had lipophilicity comparable to the new derivative -
which, however, was cleared more rapidly from mouse plasma. Moreover, the potential formation of
the 1-aryl-piperazine metabolite and its brain-to-plasma concentration ratio were also examinated,
because arylpiperazine derivatives generally undergo CYP3A-mediated N-dealkylation of the aliphatic
chain attached to the piperazine nitrogen. While the plasma concentrations of the metabolites were
always below the detection limit of the analytical procedure, their brain concentrations exceeded that
of their parent compound, by about 1.6 and 4 times for the new and original derivative, respectively.
This was not surprising as previous studies have shown that 1-arylpiperazines concentrate in brain
110
ANNUAL REPORT 2008

IRFMN
tissue. Future studies will focus on characterization of the neuropharmacological profile of the 1-arylpiperazine
metabolite compared with its parent compound.
Pharmacological role of the constituents of Hypericum perforatum
extracts
The chemical composition of extracts of hypericum perforatum L. (St. John’s wort) is essentially
known but is still not clear which constituent(s) account, wholly or in part, for the antidepressant
activity of the extracts, and through what neurochemical mechanism(s). The phloroglucinol hyperforin
shares most of the in vitro and in vivo pharmacological properties of the extracts, and is possibly a
main “antidepressant” component, but there is also evidence for other pharmacologically active
components. However, identifying the roles of the various derivatives and the mechanism(s) of their
activity is complicated by the scarcity of information about their ability to cross the blood-brain
barrier and the concentrations reached in brain after administration of the extracts. This is also true for
the biflavone biapigenin and particularly its I3’,II8 analog amentoflavone which, although present in
smaller amounts in extracts, shows a multitude of pharmacological actions in vitro and in vivo in
animal models. The lack of pharmacokinetic data in man and animals and questions about the brain
uptake of amentoflavone and biapigenin prompted us to examine their brain uptake and concentrations
and the relationships with plasma concentrations after pharmacologically effective doses in mice.
After doses of Hypericum perforatum extracts the brain concentrations of biapigenin and
amentoflavone were below the limit of quantification. The same was true for amentoflavone after a
biflavone-enriched extract of Ginkgo biloba. Levels were consistently detected only after
intraperitoneal biapigenin or amentoflavone but were low and mostly related to the residual biflavone
in the circulation. Poor brain-to-blood permeability is common to other polar components of
Hypericum perforatum, resulting in brain concentrations generally too low for any direct interaction
with neurotransmitter transporters and receptors which are obviously important for the action of
conventional antidepressants. Likewise, the in vitro interactions of biapigenin and amentoflavone with
known central mechanisms are apparently not relevant for the in vivo effects of the extracts because
they occur at biflavone concentrations far exceeding those found in the brain after pharmacologically
effective doses. However, this does not exclude that tissues other than brain may concentrate
biapigenin or amentoflavone sufficiently to exert beneficial effects after daily intake of the extracts.
Resistence to antidepressant drugs: studies in animal models
The selective serotonin reuptake inhibitors are the drugs of choice in the treatment of depression.
However, they are not or only partially effective in a fraction of depressed patients. The reasons are
substantially unknown, though pharmacogenetic studies have linked the response to serotonin reuptake
inhibitors to polymorphisms in various genes coding for serotonin mechanisms, particularly the
promoter of the serotonin transporter molecule. These studies are therefore aimed to investigate the
neurobiological mechanism(s) of resistance to antidepressant drugs in strains of mice carrying
different isoforms of tryptophan hydroxylase-2, the enzyme responsible for the synthesis of brain
serotonin.
These studies are conducted in collaboration with the laboratory of Experimental Psycopharmacology
(L. Cervo) and the laboratory of Neurochemistry and Behaviour (R.W. Invernizzi), who will provide a
brief description of recent results with the potent serotonin reuptake inhibitors citalopram and
paroxetine.
111
ANNUAL REPORT 2008

IRFMN
Laboratory of Experimental Neurology
Role of inflammatory molecules in ictogenesis and epileptogenesis
We are studying the role of IL-1beta and TNF-alpha systems in the genesis and propagation of
seizures and in the associated neurodegenerative phenomena. We have demonstrated that
epileptic activity induces the synthesis of these cytokines and related molecules involved in
inflammatory processes. IL-1beta has proconvulsant actions while its naturally occurring
antagonist (IL-1Ra) or its syntheis inhibitors have anticonvulsant activities. We are actively
studying the role of these molecules in epilepsy models with the intent of promoting their
clinical applications in drug-resistant epileptic patients. This possibility is encouraged by their
clinical application in chronic inflammatory and autoimmune diseases in humans (e.g. anakinra,
the IL-1R antagonist). We are studying pharmacological approaches to block IL-1beta-signaling
involved in the proconvulsant effects of this cytokine.
Epilepsy and postnatal development.
Seizure susceptibility is higher in early infancy although the immature brain appears to be less
susceptible to epileptogenesis. Using experimental models of seizures induced during postnatal
development in rodents, we study the mechanisms involved in age-dependent seizure
susceptibility and the associated neuronal injury. Our studies are primarily focused on
inflammatory pathways, angiogenic processes and blood-brain barrier damage.
Blood-brain barrier and epileptogenesis
We are studying BBB permeability and microvasculature changes induced in the brain by
seizures or by neurotrauma or infection and how these modifications may affect the process of
epileptogenesis. Experimental models of symptomatic epilepsy are used.
New therapeutic approaches of In vivo gene transfer
This study concerns the use of adeno-associated viral vectors to introduce genes with therapeutic
potential in the brain, thus increasing the synthesis of specific proteins to produce long-lasting
anticonvulsant effects. We have demonstrated that adeno-associated viral vector carrying the
human neuropeptide Y gene, significantly increases the brain concentration of this peptide after
its intrahippocampal injection for a prolonged time (at least up to 5 months after a single
intracerebral injection). The rats overexpressing this peptide are less susceptible to limbic
seizures and to epileptogenesis. Future development of this study concerns the optimization of
the transgene transfer technology to inhibit spontaneously recurring seizures and envisaging a
possible clinical application.
Laboratory of Geriatric Neuropsychiatry
Population study on the prevalence of dementias in the older-old
Parallel to the progressive increase of individuals aged 80 years or older within the elderly
population (65+), the number of demented patients of 80 years or older makes up an ever
increasing fraction of the total population affected by dementia. As very often happens,
the exclusion from studies of subjects in the oldest age classes tends to inevitably
underestimate the total number of individuals affected by dementia present in the
population. To fill this gap, a door-to-door population study on the prevalence, incidence,
risk factors and evolution of dementias and age-associated cognitive deficits has been set
up in an elderly population aged 80 years or older living in eight small towns of Varese
Province. The study is funded by a grant from the Fondazione Italo Monzino, Milano.
112
ANNUAL REPORT 2008

IRFMN
Effects of anemia in the elderly
A previous large survey in old resident of Biella (65-84 years old) has been conducted in
collaboration with the Local Health Authority of Biella (ASL 12) to determine the
prevalence of anemia. We have now extended the investigation to the oldest old residents
(about 1500 85 years or older individuals) in order to estimate the prevalence and impact
of mild anemia also in this segment of the elderly population.
Evaluating risk profiles in hospitalised elderly subjects
In collaboration with the Geriatric Division of the Ospedali Regionali of Lugano and
Mendrisio, Switzerland, hospitalized and ambulatory patients are evaluated from a
neuropsychological, functional and mobility point of view to estimate the impact of these
factors on heath-related outcomes and disease progression.
Longitudinal follow-up of individuals with mild cognitive impairment
(MCI)
In collaboration with the Geriatric Unit of Ospedali Regionali of Lugano and Mendrisio,
Switzerland, the follow-up study of all Mild Cognitive Impairment or Questionable
Dementia (CDR 0.5) patients seen at the Memory Clinic of the Hospitals is continuing to
estimate the rate of conversion to dementia and to evaluate the possible risk factors
associated with conversion.
Quality of care of terminally ill oncological subjects
In 1999 we started a collaborative programme with the hospice “via di Natale
Franco Gallini” in Aviano (PN). The aim of the research project was to assess the
quality of care given in hospice to terminally ill oncological patients at the end of
life. Present aim of the collaboration is the assessment of the hospice activities
after its opening and to provide nurses with continuous training on use of databank
Global Forum for Community Mental Health
The Unit of Epidemiology and Social Psychiatry is in charge of the general bureau of the Global
Forum for Mental Health, a working group promoted by the Department of Mental Health of the
World Health Organization and aiming at creating a network supporting those who are active in
mental health and care for people with mental disorders in low income countries, moving from
asylum-centred to community-centred models of care. The Global Forum identifies experiences
and practices which deserve to be known and promote and spread them; gives information and
technical assistance to those interested in delivering and evaluating sustainable and promising
interventions in underserved or disadvantaged populations; identifies barriers to the
implementation of good quality interventions and discuss how to remove them, supports the
development of research expertise based on real needs of the target populations, and through
integration of service deliverers, researchers, policy makers, users and their families, citizens.
Randomised controlled trial of the Italian Group for the Study of the
Second Generation Antipsychotics (GISAS)
The study compares three antipsychotics, aloperidole, olanzapine, aripiprazole, according to
efficacy and tolerability through a pragmatic experimental design on 800 patients, identified and
randomized in mental health centres of all Italian regions, and representing the first attempt to
conduct a study on such large numbers in Italy on this topic. A toital of 35 mental health centres
were recruited, among which 26 are active and have randomised 80 patients.
Ibn the framework of this study, a survey was conducted on the point of view of the
psychiatrists of the participating services on usefulness and feasibility of clinical trials in
113
ANNUAL REPORT 2008

IRFMN
psycopharmacology, degree of uncertainty in the use of antipsychotics and more important
sources of information supporting their prescriptive attitudes.
Natural Social Networks
The project connects patients to identified volunteers willing to accompany the patients in part
of their everyday activities. The evaluation of this intervention is based on the longitudinal
employment of tools measuring quality of life, general wellbeing and social and personal
functioning.
Suicides
In the framework of a wider project financed by the Ministry of Health, various literature
reviews were conducted of topic related to the relationship between suicide attempt/deliberate
self-harm and suicide, and treatment and assessment of suicide attempt and self-harm in the
Emergency Department. A form for the systematic registration of these events filled by the
psychiatrist during the consultation of the patient in the Emergency Department or in hospital
was prepared and introduced in the ordinary assessment of suicide attempts of various hospitals
and health trusts in North and Central Italy.
UNASAM Project – Quality improvement and evaluation in mental health
with the active participation of the associations
During the second year of the project, data collection on family members satisfaction with
mental health services was completed in 41 services in four Italian regions. The questionnaire
was developed with the participation of groups and associations of family members, who also
organized the questionnaires distribution and data collection. Data were analysed and results
were presented to the associations.
European Network of Bipolar Research Export Centres – EMBREC
Bipolar disorder is characterised by recurrent depressive and manic episodes, and affects about
1% of the European population. Delay and difficulties in diagnosis have consequences on
treatment and care, and it is worth integrating efforts and expertises of various groups. In this
framework, the Unit of Epidemiology and Social Psychiatry has the following tasks: review
evidences of effective strategies in information delivery, self-help and psychoeducation;
development, in collaboration with a sample of users, of a package of practical strategies and
tools for users information on relevant topics and self-management. The package will be tested
and implemented in one or more mental health services in Italy.
Self-accreditation and quality of the housing facilities in the Health Trusts
1 and 2 in Turin
In two Health Trusts groups composed by users, family members, professionals working in the
housing facilities and professionals working in other services have developed a tool for the
systematic evaluation and accreditation of the housing facilities. The tool was tested. Lessons
were held throughout the year to teach and train the participants to evaluation principles and
organization of the visits and evaluating activities.
114
ANNUAL REPORT 2008

IRFMN
Laboratory of Inflammation and Nervous System Diseases
Inhibition of selected aspects of the inflammatory response powerfully
reduces ischemia/reperfusion injury
Previous studies of ours have indicated that complement and related inflammatory systems such
as contact/kinin system may represent novel targets for reducing ischemia/reperfusion injury.
We have shown that C1-INH, a serine-protease inhibitor that acts as a major regulator of both
complement and kinin systems, markedly improves neurological deficits and reduces infarct
volume in mice with focal transient as well as permanent ischemia induced by middle cerebral
artery occlusion. We have further extended this finding defining its effectivness on different
strains of mice (displaying different levels of complement expression), the time-window and the
dose-response curves. Since C1-INH may act on several substrates, we have also evaluated the
specific involvement of the different complement pathways and of the other inflammatory
systems. To explore the mechanisms of C1-INH neuroprotection, we have also investigated the
expression (protein and mRNA) of inflammatory cytokines, adhesion molecules, NO synthase
isoforms, apoptosis markers.
The results obtained show that: i) C1-INH effectively and markedly reduces brain
ischemia/reperfusion injury, inducing a decrease up to 90% of the ischemic lesion; ii) C1-INH
actions lead to inhibition of cell recruitment, inflammation and apoptosis; iii) C1-INH
neuroprotection is independent from the complement classical pathway and inhibition of other
complement pathways or other inflammatory systems such as the contact/kinin system are
involved in its powerful protective action. Moreover recent data have shown C1-INH protective
actions also in a model of traumatic brain injury.
Thus C1-INH, which is presently used as replacement therapy in patients with C1-INH
deficiency, possesses potent, multi-faceted neuroprotective actions that may be beneficial in
acute brain injury. Ongoing studies are aimed at clarifying the mechanism of neuroprotection by
C1-INH.
Stem cells as a therapeutic approach in stroke and traumatic brain injury
The aim of the project is to verify the conditions for the effectivenss of stem cells (SC) in
reducing acute brain injury and to investigate the mechanisms triggered by their infusion in the
lesioned brain. SC, isolated from newborn mice or obtained from human stem banks, are
infused to mice in which transient ischemia is induced by middle cerebral artery occlusion or
traumatic brian injury by controlled cortical impact. At different time points several parameters
are evaluated: distribution and phenotype of injected cells, neurodegeneration, behavioral
deficits, cytokine and trophic factor gene expression, microglia activation. The results obtained
up to now have shown that: i) SC effectively counteract brain injury: they can decrease neuronal
loss and reverte functional impairments related to exploratory behaviour and sensory/motor
activity; ii) they may elicite an early response: in selected conditions 24 h after infusion,
chemokines, angiogenic and neurotrophic factor transcripts are activated; iii) while in selected
experimental conditions the protective mechanism of SC seems to be mainly due to the
induction of beneficial factors, in others a longer time is required for effective neuroprotective
effects; iv) activation of microglial cells is required for SC protective action; v) SC presence in
the brain tissue is favoured by the ischemic environment. Thus the reciprocal interaction
between SC and ischemic environment is crucial for stem cells protective actions Ongoing
studies include: 1) definition of the mechanisms of homing of stem cells in the injuried brain; 2)
analysis of the protective/toxic role of microglia; 3) investigation of the early events triggered
by stem cells in the ischemic brain. (Capone et al, 2007; Pastori et al. 2008).
115
ANNUAL REPORT 2008

IRFMN
Identification of ischemic tolerance mediators in cerebral ischemia
Recent studies indicate that cell death resulting from ischemic injury can be reduced when a
sublethal ischemic episode occurs hours or days before a severe ischemic insult. This
phenomenon is known as Ischemic PreConditioning (IPC) and the induced neuroprotection is
called Ischemic Tolerance (IT). Several models of induction and maintenance of tolerance have
been described, but the molecular mechanisms of the IPC-induced neuroprotection are not
identified yet and a clear view of the mechanisms responsible for ischemic preconditioning is
still lacking. Specific aims of the project are: i) to define the characteristics of IT induced by
small transient ischemic attack (TIA) in experimental models of cerebral ischemia; ii) to
elucidate the pathways and/or the mediators involved in ischemic tolerance; iii) to identify
endogenous protective molecules/pathways that may represent potential therapeutic targets for
stroke; iv) to assess if IPC can be induced also in traumatic brain injury (TBI). The project plan
includes the characterization of TIA model in mouse, the study of the effect of TIA on
subsequent ischemia and time window of ischemic tolerance, the identification of mediators
involved in IT by protein and mRNA assays, the study of direct effect of relevant protective
molecules in “in vitro” and “in vivo” ischemia models, the set up of protocols for IPC in TBI
brains. Thus ischemic preconditioning provides an opportunity to identify the putative candidate
that can confer neuroprotection against acute brain injury. A major goal is to identify the
underlying endogenous protective cellular receptor/signaling cascades, with the long-term goal
to allow therapeutic augmentation of the endogenous protective mechanisms in cerebral
ischemia.
Blood-brain barrier and ischemic preconditioning
The endothelial cells belonging to the cerebral microvasculature are the main component of the
Blood-Brain Barrier (BBB). These cells are attached to each other by intercellular Tight
Junctions and are closely associated with the endfeet of astrocytes, with pericytes and with
microglia, resulting in a complex network of cellular interactions. The integrity of this structure
is essential for the maintenance of the ischemic environment. We have recently established a
bidimensional BBB model by means of co-cultures of mouse brain endothelial cells and mixed
glial cells. These coltures retain the BBB typical features, namely they express thight junctions,
they present typical transendothelial electrical resistence (TEER), and paracellular and
transcellular permeability values. To mimic the ischemic insult, the BBB coltures are exposed to
oxygen-glucose deprivation (ODG) protocols. The ongoing project is aimed at studying the
involvement of BBB in ischemic preconditiong (IPC, see previous research topic). Albeit IPC
has been reported to reduce edema formation and thus BBB disruption following ischemia, no
information about a direct role of BBB cells in IPC is presently available. The major goal of the
research has been to assess if BBB cells may be preconditioned. To this purpose a brief OGD
stimulus was delivered before a severe OGD exposure. The results obtained have shown that a
mild OGD stimuls may actually dampen the effects of a subsequent severe OGD exposure
showing that BBB cells can be effectively preconditioned. Ongoing studies are presently aimed
at identifying mediators and/or pathways involved in activation and maintenance of IPC in the
cerebrovascular unit with the final aim of selecting new pathways and molecular targets useful
for a therapeutical stroke strategies.
116
ANNUAL REPORT 2008

IRFMN
Laboratory of Molecular Neurobiology
Study on pathogenic mechanisms of Amyotrophic Lateral Sclerosis
Role of protein aggregation
A pathological feature of ALS is the accumulation of protein aggregates in the perykaria and
axons of motor neurons. Our hypothesis is that this may be due to an impairment of the
ubiquitin/proteasome system (UPS). To functionally investigate the UPS in ALS motor neurons
in vivo, we crossed SOD1G93A mice with transgenic mice that express a fluorescently-tagged
reporter substrate of the UPS (Ub G76V -GFP, from now on indicated as GFP mice). In double
transgenic GFP/SOD1G93A mice an increase in Ub G76V -GFP reporter, indicative of UPS
impairment, was detectable in a few spinal motor neurons and not in reactive astrocytes or
microglia, at symptomatic stage but not before symptom onset. These data suggest that UPS
impairment occurs in motor neurons of mutant SOD1-linked ALS mice and may play a role in
the disease progression.
Another major route for intracellular protein degradation is the autophagy−lysosomal pathway.
The rat microtubule-associated protein 1 light chain 3 (LC3), plays a critical role in the
formation of autophagosomes and its conversion from LC3I into LC3II is accepted as a simple
method for monitoring authophagy. Recently, we have observed that levels of LC3II which is
known to be correlated with the extent of autophagosome formation, was increased in
SOD1G93A mice with at an advanced stage of disease compared with non transgenic mice.
This indicates that the activation of autophagy may be an alternative mechanism of cell
defense to eliminate the proteins misfolded when the proteasome is partially inhibited as
demonstrated above. We are now examining the autophagy at earlier times of the pathology.
In collaboration with the Molecular Biochemistry and Pharmacology department we carried out
a proteomic analysis of the protein composition of the Triton-insoluble fraction (TIF), as a
model of protein aggregates, from the SOD1G93A mice at different disease stages. We
identified several proteins enriched in TIF of ALS mice already at preclinical stage, including
intermediate filaments, chaperones and mitochondrial proteins. Some of them, HSP90,
aconitase, HSC70 and cyclophilin A, were also analyzed in TIF of spinal cord of ALS patients
and found significantly enriched. Interestingly, the majority of proteins in mice and at least
HSP90 in patients were tyrosine nitrated. We therefore investigated the role of nitrative stress in
aggregate formation in a cellular model of ALS. We could demonstrate that by inhibiting nitric
oxide synthesis it is possible to substantially reduce the amount of insoluble proteins and in
particular of aconitase, HSC70, cyclophilin A and SOD1. In conclusion, the analysis of the
insoluble fractions from cellular/mouse models and human tissues could reveal novel aggregateprone
proteins in ALS and suggest that nitrative stress may contribute to protein aggregate
formation. These results are in a manuscript submitted to Brain.
Role of glutamate AMPA receptors in the pathogenesis of ALS
To further study the role of glutamate AMPA receptors in the susceptibility of motor neurons
we have examined the expression and distribution of GluR2 subunit in motor neurons still
maintaining a functional connection with muscle fibers. We found a significant decrease of
Glur2 suggesting that this is a very early event that may play an important role in the
pathogenesis of the disease. The analysis of all these data are under completion.
In vitro studies on neuron-glia interaction
To investigate further the role of inflammatory mechanisms, we have set up an in vitro coculture
of spinal neurons and astrocytes derived from SOD1G93A mice embryos. We are
verifying in this model the alterations observed in vivo such as the activation of TNFalphap38MAPK
pathway. This model, hopefully, will allow to examine more rapidly the protective
117
ANNUAL REPORT 2008

IRFMN
effect of various strategies interfering with this pathway and will provide a model to test new
pathogenic mechanisms.
Altered axonuclear communication in motor neurons of a mouse model
of familial amyotrophic lateral sclerosis (European collaborative project
FP6 program EU-NES AXON SUPPORT)
Impairment in the maintenance of axon-nuclear communication, and viceversa, due to motor
proteins defects may play a primary role in the ALS.
To assess this hypothesis we examined the expression and the distribution of different
components of the nucleocytoplasmic-transport system, such as importins and vimentin, in the
ventral horn spinal cord and in the peripheral nerves of transgenic mice and rats carrying human
SOD1G93A. We found reduction of importin beta and slight increase of vimentin protein levels
in homogenates from ventral horn spinal cord concomitantly with the appearance of first
symptoms of the pathology. Using confocal miscroscopy, we detected abnormal accumulation
of vimentin in the perikaria of motor neurons at the presymptomatic stage; moreover we
observed a reduction of importin beta staining in the cytoplasm of motor neurons showing
accumulation of phosphorylated neurofilaments, a hallmark of neuronal damage, and defects in
retrograde transport. Based on these evidences we suggest that alterations of nucleocytoplasmic
transport-related proteins are linked with the degenerative process of motor neurons and may
play a role in ALS pathology.
Therapeutical interventions in mouse model of ALS
Development of target genes-based therapies for the protection of motor
neurons
It is emerging evidence that in the motoneurons of patients with sporadic ALS and in animal
models of the disease, there is a remarkable activation of pro-degenerative pathways (like p38
MAP-Kinase). On the other side, the mechanisms involved in the modulation of cell survival
(like PI3K/Akt pathway) are not activated, thus suggesting an impairment in the induction of
neuroprotective responses.These pathways may be considered as potential therapeutic targets.
Based on these evidences, with this project we propose: 1) to develop gene-targeted strategies
aimed at counteracting p38 pro-degenerative pathway and activating Akt pro-survival cascade
inside motorneurons of spinal cord; 2) to evaluate efficacy and safety of these potential
therapeutic interventions in an animal model of ALS. We have developed lentiviral vectors
expressing candidate p38-targeted shRNA sequences. These shRNAs were tested in primary
mouse astrocyte-motoneuronal cell co-cultures or in cultures of rat cortical neurons. They were
able to prevent activation of p38 and its downstream targets after TNFalpha stimulation, and to
reduce neuronal loss after toxic stimuli. In parallel, we have developed constructs that express
constitutively active forms of Akt1 and Akt3 (caAkt1 and caAkt3). Preliminary experiments in
cell lines showed that either caAkt1 or caAkt3 efficiently phosphorylates and inhibits
downstream pro-apoptotic targets, such as GSK3beta. Current studies aim to selectively drive
the expression of p38-shRNA and Akt1 to motor neurons and to increase the efficiency of their
cellular expression.
Studies on the effects of the long-chain omega-3 polyunsaturated fatty acids
eicosapentaenoic acid and docosahexaenoic acid in the G93A SOD1 mouse
(This is a project in collaboration with Dr. A. Michael-Titus del Queen Mary University of
London supported by MND Association) Based on the observations by Dr. A. Michael-Titus
that a diet enriched of omega 3 is neuroprotective in a model of spinal cord trauma in rat we
118
ANNUAL REPORT 2008

IRFMN
decided to investigate if such treatment could have a beneficial effect on SOD1G93A mice. The
study is in progress.
Treatment with lithium carbonate does not improve disease progression in two
different strains of SOD1 mutant mice
Female SOD1G93A mice on different genetic background and different phenotype of disease
severity were treated daily with Li 2 CO 3 37mg/kg (1 mEq /kg) i.p. starting from age 75 days
until death. We observed a significant anticipation of the onset and reduced survival in
129Sv/G93A and no effect in C57G93A mice treated with lithium as compared to vehicle
treated mice. Moreover, lithium neither exerted neuroprotective effects not increased the
expression of LCII and the activity of mitochondrial complex IV in the spinal cord. The present
study does not identify any therapeutic or neuroprotective effect of lithium in SOD1G93A
female mice. This study is in press in Amyotrophic Lateral Sclerosis.
Studies aimed to identify biomarkers for the diagnosis and progression of
the disease in ALS patients
In collaboration with the department of Neurology of the Fondazione Salvatore Maugeri, IRCCS,
of Pavia, we have started a series of studies aimed to investigate the immune system and the
oxidative stress products in the PBMC of sporadic ALS patients in comparison to healthy age
matched controls. The results show that sporadic ALS patients exhibit immunological alterations
in their blood, in respect to healthy controls. This study strengthens the hypothesis of an
involvement of the adaptive immune system associated with a neuroinflammatory process in the
pathobiology of ALS. The manuscript describing these data is under revision for the J. of
Neuroimmunology.
In parallel, we are examining the levels and the characterisation of the nitrated protein in the
PBMC of ALS patients compared to healthy controls. We also evaluated the PBMC of
SOD1G93A transgenic rats. The protein nitration on tyrosine is an oxidative mechanism that
alters the function of proteins inducing their inactivation or a gain of toxic functions. Using a
proteomic approach we have observed that a series of proteins are overnitrated in ALS patients
and in SOD1G93A rats in respect to controls. Some proteins are the same in rat and patients
suggesting that they could be a reliable biomarkers for the diagnosis and prognosis of the
disease. These data are in a manuscript under revision for the Antioxidant and Redox
Signalling.
Another approach to identify potential specific diagnostic and prognostic markers of the disease
has been set up in collaboration with the department of neurology of the Istituto Auxologico ,
IRCCS of Milano. In particular, we have used genomic and proteomic analyses to identify and
characterize genes and proteins specifically modified in the muscles of ALS transgenic mouse
models, at the onset of disease, in respect to control mice. We have completed the examinations
and the data are now under analysis.
119
ANNUAL REPORT 2008

IRFMN
Laboratory of Experimental Psychopharmacology
Drug Abuse
Neural basis of drug self-administration
To separate the direct pharmacological effects of cocaine from those associated with active drug
self-administration we employed a yoked control-operant paradigm and investigated the
expression of well established markers of the rapid action of cocaine, i.e. the inducible early
genes and trophic factors, in rats after a single intravenous (i.v.) cocaine self-administration
session. Animals self-administering cocaine did more active lever-presses than yoked-cocaine
(YC) and yoked-vehicle (YV) animals. This goal-oriented behavior was accompanied by a
selective increase in Arc mRNA levels in the medial prefrontal cortex (mPFC). These findings
demonstrate that a single session of cocaine i.v. self-administration is sufficient to shape rat
behavior towards goal-directed behaviors and selectively up-regulate Arc expression in mPFC
(of SA animals), providing the first evidence that the mPFC's function is already profoundly
influenced by the first voluntary cocaine exposure.
Neural basis of “drug craving” and “relapse” in the drug abuse assumption
Drug craving, defined as “the desire to experience the effect(s) of a previously experienced
psychoactive substance” is a cardinal feature of drug addiction and is clinically significant
because of its potential link to relapse. To provide useful indications to the development of
novel therapeutic approaches to prevent the use and abuse and the relapse of drug assumption
following the outcome of “craving”, we elaborated experimental models of self-administration
and “relapse” induced by cocaine, nicotine and alcohol-associated cues, after a period of
abstinence. It was found that naltrexone, a non selective antagonist at opioids receptors,
selectively modulate rats’ seeking behaviour induced by cocaine-associated cues after a long
period of abstinence and in the absence of any further cocaine. Ongoing studies are evaluating
whether this modulation is peculiar for cocaine or could be generalized to other abused drugs.
The role of several other neurochemical mechanisms potentially involved in the drug-seeking
behaviour are also in progress.
Resistance to antidepressant drugs: experimental and clinical studies
This project arises from a collaboration between the laboratories of Neurochemistry and
Behavior (R.W. Invernizzi), Drug Metabolism (Silvio Caccia), Biology of Neurodegenerative
Disorders (GianLuigi Forloni) and focus on behavioral and biochemical characterization of an
experimental model of resistance to the antidepressant drugs. Using an animal model predictive
of the antidepressant activity, the effects of selective serotonin reuptake blockers (SSRI) was
evaluated in several mice strains. It was found that DBA/2J and BALB/c do not respond to the
antidepressant-like activity of the SSRI. The lack of effect was attributed to genotypedependent
impairment of 5-HT synthesis since DBA/2J and BALB/c carring a single nucleotide
polymorphism (C1473G mice) in the gene for the brain-specific isoform of tryptophan
hydroxylase-2, the rate-limiting enzyme in the synthesis of serotonin are characterized by a
decreased serotonin synthesis. This hypothesis seems to be supported by the observation that
DBA/2J and BALB/c mice had less dialysate 5-HT in the medial prefrontal cortex and dorsal
hippocampus than C57BL/6J mice. Moreover, in DBA/2J and BALB/c the SSRI raised
significantly less extracellular 5-HT when compared to C57BL/6J mice. More recently it was
found that 5-HT 1A and 5-HT 2C receptor antagonists restored the SSRIs’ effect on either the
antidepressant-like activity and the extracellular 5-HT.
120
ANNUAL REPORT 2008

IRFMN
Laboratory of Neurochemistry and Behavior
“Resistance” to antidepressant drugs
Despite the wide range of antidepressant drugs available for the treatment of mood disorders the
delayed onset of the antidepressant effect and the partial or no response in a considerable
portion of patients still limits their efficacy.
Ongoing studies in collaboration with the Laboratories of “Experimental Psychopharmacology”
and “Drug Metabolism” are aimed at assessing the neurobiological mechanisms involved in the
resistance to antidepressant drugs in mice. The gene for the brain-specific isoform of tryptophan
hydroxylase-2 (TPH-2), the rate-limiting enzyme in the synthesis of serotonin, is mutated in
DBA/2J and BALB/c mice (C1473G). These mice have a low 5-HT synthesis rate and do not
respond to antidepressants inhibiting selectively the reuptake of serotonin (SSRI) in the forced
swimming test, a procedure used to screen compounds for antidepressant activity. In addition,
the effect of SSRI on the extracellular concentrations of brain serotonin is attenuated in mice
carrying the 1473G allele of TPH-2. 5-HT 1A and 5-HT 2C receptor antagonists enhanced the
effect of SSRI on extracellular 5-HT and restored the antidepressant-like effect of SSRI.
These results suggest that genetic differences in serotonin synthesis contribute to determine the
efficacy of SSRI and identify pharmacological strategies that may enhance the response in
treatment-resistant depressed patients.
Animal model of cognitive deficit of schizophrenia; typical and atypical
antipsychotics
The cognitive deficit is a core symptom of schizophrenia, which has been linked to functional
outcome and is relatively independent of psychotic symptoms. The antipsychotics, either typical
or atypical, are able to control positive symptoms such as delirium, hallucinations and paranoia.
However, the currently available atypical antipsychotics when compared to conventional
antipsychotics show somewhat superior efficacy for the management of cognitive deficits in
patients with schizophrenia.
The cognitive deficit of schizophrenia was modeled in rats and mice, by using a test of attention
such as the 5-choice serial reaction time task (5-CSRTT) and injections of glutamate NMDA
receptor antagonists into the medial prefrontal cortex (mPFC). This model makes clear links
with psychopathology as dysfunctional glutamate neurotransmission in the mPFC has been
implicated in cognitive deficits of schizophrenia and the 5-CSRTT is the rat analogue of the
continuous performance test used to assess attention and vigilance in schizophrenic patients.
Antipsychotics possess a complex pharmacology across the biogenic amine receptor families as
shown by affinity constants derived from radioligand-binding techniques. The ability to
antagonise the DA D 2 receptor function is shared by the conventional and by the atypical
antipsychotics. However, atypical antipsychotics show a high affinity also for serotonin 5-
HT 2A , 5-HT 2C and 5-HT 1A receptors. Our studies compared the effects of conventional and
atypical antipsychotics in this model of cognitive deficit of schizophrenia. The results show that
antipsychotics may be differentiated by a selective effect of typical antipsychotics on
compulsive perseveration, and atypical antipsychotics on impulsivity. Intracerebral
microdialysis studies show that attentional deficits induced by NMDA receptor antagonists is
associated with excessive glutamate in the medial prefrontal cortex of the rat and this effect was
prevented by atypical antipsychotics.
121
ANNUAL REPORT 2008

IRFMN
122
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF CARDIOVASCULAR
RESEARCH
STAFF
Head
Maria Grazia FRANZOSI, Biol.Sci.D.
Laboratory of Cardiovascular Clinical Pharmacology
Head
Roberto LATINI, M.D.
Bio-imaging Unit
Head
Cardiovascular Endocrine Unit
Head
Tissue Culture Unit
Head
Fabio FIORDALISO, Biol.Sci.D.
Serge MASSON, Ph.D.
Giovanna BALCONI, Univ.Dipl.
Laboratory of Clinical Drug Evaluation
Head
Maria Grazia FRANZOSI, Biol.Sci.D.
Bioinformatics Unit
Head
Enrico NICOLIS, Comp.Sci.Stud.
Laboratory of General Practice Research
Head
Maria Carla RONCAGLIONI, Biol.Sci.D.
Laboratory of Medical Statistics
Head
Simona BARLERA, Dr.Sci.Pol., MSc.
Laboratory of Clinical Pharmacology
Head
Nursing Research Unit
Head
Gianni TOGNONI, M.D.
Paola DI GIULIO, R.N., MSc
123
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Maria Grazia Franzosi got her Biological Science degree in 1972 at the University of Milan.
Education
1972 Doctoral degree in Biological Sciences, University of Milan, Italy
1978 Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche "Mario
Negri” di Milano, Italy
Main fields of activity
Coordination of multicentric randomised clinical trials. Relationship between genetic and environmental risk
factors in coronary events. Pharmacogenetics. Pharmacoeconomics. Drug Epidemiology and Post-Marketing
Surveillance.
Position
from 2002 Director of the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche
"Mario Negri", Milano, Italy
from 2004 Member of Steering Committee, Studio GISSI-AF Study, Milano, Italy
from 2001
from 1998
Member of Steering Committee, Studio GISSI-HF Study, Milano, Italy
Member of Steering Committee of the PROCARDIS Research Programme - A genome-wide
strategy to identify susceptibility loci in precocious coronary artery disease - University of
Oxford, UK
from 1997 Member of “Antithrombotic Trialists’ Collaboration”, Oxford, UK
from 1996 Membro dello Steering Committee e National Coordinator per l’Italia della Organization to
Assess Strategies for Ischemic Syndromes (OASIS-2, OASIS-4 CURE, Michelangelo
OASIS-5 e OASIS 6, , CURRENT OASIS-7, FUTURA OASIS-8), dello studio INTER-
HEART e degli studi ACTIVE, RELY, AVERROES, Population Health Research Institue,
McMaster University, Hamilton, Canada
1994-1996 Director of European Coordinating Centre and Member of Steering Committee, Collaborative
from 1993
from 2002
Organization for RheothRx Evaluation (CORE), McMaster University, Hamilton, Canada
Member of Steering Committee, Studio GISSI-Prevenzione, Milano, Italy
Member of “Fibrinolytic Therapy Trialists’s Collaboration”, Oxford, UK e del “Collaborative
Group on Angiotensin Converting Enzyme Inhibitors Trials”, National Institutes of Health,
Bethesda, Washington, USA
1989-2001 Head of the Laboratory of Clinical Drug Evaluation, Istituto di Ricerche Farmacologiche "Mario
Negri"
1985-1988 Head of the Clinic Drug Evaluation Unit of the Laboratory of Clinical Pharmacology, Istituto di
Ricerche Farmacologiche "Mario Negri"
from 1984
Member of the Scientific and Organising Secretariat, Gruppo Italiano per lo Studio della
Sopravvivenza nell'Infarto Miocardico (GISSI-1, GISSI-2, GISSI-3 studies) Milano, Italy
1975-1984 Researcher at the Laboratory of Clinical Pharmacology, Istituto di Ricerche Farmacologiche
"Mario Negri" and at the Regional Center for Drug Information of the Lombardy Region
Selected publications
• Franzosi MG. Should we continue to use BMI as a cardiovascular risk factor Lancet 2006; 368: 624-625
• Farrall M, Green FR, Peden JF, Olsson PG, Clarke R, Hellenius ML, Rust S, Lagercrantz J, Franzosi MG, Schulte H,
Carey A, Olsson G, Assman G, Tognoni G, Collins R, Hamsten A, Watkins H, on behalf of the PROCARDIS
Consortium. Genome-wide mapping of susceptibility to coronary artery disease identifies a novel replicated locus on
chromosome 17. PLoS Genet 2006 - http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.0020072
• Chiodini B, Franzosi MG, Barlera S, Signorini S, Lewis CM, D'Orazio A, Mocarelli P, Nicolis E, Marchioli R, Tognoni
G, GISSI, SIBioC-GISSI Prevenzione Group. Apolipoprotein E polymorphisms influence effect of pravastatin on
survival after myocardial infarction in a Mediterranean population: the GISSI-Prevenzione study. Eur Heart J 2007 ; 28:
1977-1983
• Anand SS, Islam S, Rosengren A, Franzosi MG, Steyn K, Yusufali AH, Keltai M, Diaz R, Rangarajan S, Yusuf S,
INTERHEART Investigators. Risk factors for myocardial infarction in women and men: insights from the
INTERHEART study. Eur Heart J 2008 29 : 932-940
• Broadbent HM, Peden JF, Lorkowski S, Goel A, Ongen H, Green F, Clarke R, Collins R, Franzosi MG, Tognoni G,
Seedorf U, Rust S, Hamsten A, Farrall M, Watkins H, PROCARDIS Consortium. Susceptibility to coronary artery
disease and diabetes is encoded by distinct, tightly linked, SNPs in the ANRIL locus on chromosome 9p. Hum Mol
Genet 2008; 17: 806-814
• GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R,
Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart
failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230
124
ANNUAL REPORT 2008

IRFMN
• GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R,
Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF
trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372 : 1231-1239
Simona Barlera got her degree in Political Science, area Statistics at the “Università degli Studi di
Milano” in Milano in 1992, followed by a master in Medical Statistics at the London School of Hygiene
and Tropical Medicine, “University of London” in 1998.
Education
1987-1992 Degree in Political Science, area Statistics at the “Università degli Studi di Milano” in
Milano.
1997-1998 Master post-lauream in Medical Statistics at the London School of Hygiene and Tropical
Medicine, University of London, London.
1998-1999 Visiting Scientist in the Department of Statistical Genetics, Wellcome Trust Centre for
Human Genetics, University of Oxford (UK).
Main fields of activity
Methodology of Clinical Trials in the cardiovascular field. Preparation and viewing of research protocols,
planning and conduct of statistical analyses and the reporting of findings on scientific journals.
Genetic epidemiology: genome-wide strategies (linkage analysis) to identify susceptibility genes in
coronary artery disease; case-control studies in order to identify candidate genes involved in the
cardiovascular pathology.
Position
from Oct 2006
Head of the Laboratory of Medical Statistics, Department of Cardiovascular Research,
Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy
May 99-Sept 06 Head of the Medical Statistics Unit, Department of Cardiovascular Research, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano, Italy
1998-1999 Visiting Scientist in the Department of Statistical Genetics, Wellcome Trust Centre for
Human Genetics, University of Oxford (UK).
1992-1997 Researcher in the Unit of Applied Statistics and Information Technology, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano, Italy
Selected publications
• Chiodini B, Barlera S, Franzosi MG, Labarta V, Introna M, Tognoni G.APO B gene polymorphisms with coronary artery
disease: A meta-analysis. Atherosclerosis 2003; 167: 355-366
• Latini R, Masson S, Anand I, Salio M, Hester A, Judd D, Barlera S, Maggioni AP, Tognoni G, Cohn J N, Val-HeFT
Investigatore. The comparative prognostic value of plasma neurohormones at baseline in patients with heart failure
enrolled in Val-HeFT. Eur Heart J 2004; 25: 292-299
• Maggioni AP, Latini R, Carson PE, Singh SN, Barlera S, Glazer R, Masson S, Cere` E, Rognoni G, Cohn JN. Valsartan
reduces the incidence of atrial fibrillation in patients with heart failure: Results from the Valsartan Heart Failure Trial
(Val-HeFT). Am Heart J 2005; 149: 1-10
• Masson S, Latini R, Anand IS, Vago T, Angelici L, Barlera S, Missov ED, Clerico A, Tognoni G, Cohn JN, Val-HeFT
Investigators. Direct comparison of B-type natriuretic peptide (BNP) and amino-terminal proBNP in a large population
of patients with chronic and symptomatic heart failure. The Valsartan Heart Failure (Val-HeFT) data. Clin Chem 2006;
52: 1528-1538
• Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins
R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a
genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: 221-227
• GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R,
Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart
failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230
125
ANNUAL REPORT 2008

IRFMN
Roberto Latini got his Medical Doctor degree in 1978 at the University of Milan.
Education
1970-1978 University of Milan School of Medicine, degree in Medicine
1981-1983 Merck Sharp & Dohme International Fellow in Clinical Pharmacology
Main fields of activity
Mechanisms of cardiac damage following ischemia, with focus on eurohumoral activation. Use of stem
cells for cardiac repair. Biohumoral investigations within large scale clinical trials in heart failure and
atrial fibrillation.
Positions
from 1990 Head of the Cardiovascular Clinical Pharmacology Laboratory (Cardiovascular Research
Department) Istituto di Ricerche Farmacologiche “Mario Negri”, Milan, Italy
from 2001 Member of the GISSI-HF Steering Committee
from 2004 Member of the GISSI-AF Steering Committee
from 2005 Member of the CandHeart Steering Committee
from 1999 Visiting Professor Dept of Medicine, New York Medical College, Valhalla, NY, USA
1981-1983 Cardiology Fellow (Dr. R. E. Kates, Laboratory) Stanford University Medical Center,
CA, USA
1976-1981 Member of the Sub-Group RMs for Drugs (Community Bureau of Reference,
Commission of the European Communities)
1973-1990 Fellow at the Laboratory of Clinical Pharmacology of the Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
Selected publications
• Latini R, Masson S, Anand I S, Missov E, Carlson M, Vago T, Angelici L, Barlera S, Parrinello G, Maggioni A P,
Tognoni G, Cohn J N, Val-HeFT Investigators. Prognostic value of very low plasma concentrations of troponin T in
patients with stable chronic heart failure. Circulation 2007; 116: 1242-1249
• Sarto P, Balducci E, Balconi G, Fiordaliso F, Merlo L, Tuzzato G, Pappagallo G L, Frigato N, Zanocco A, Forestieri C,
Azzarello G, Mazzucco A, Valenti M T, Alborino F, Noventa D, Vinante O, Pascotto P, Sartore S, Dejana E, Latini R.
Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure. J Card Fail 2007; 13:
701-708
• Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo
I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G. Cardiac mesoangioblasts are committed, self-renewable
progenitors, associated with small vessels of juvenile mouse ventricle. Cell Death Differ 2008; 15: 1417-1428
• Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators.
Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial).
J Am Coll Cardiol 2008; 52: 997-1003
• Salio M, Chimenti S, De Angelis N, Molla F, Maina V, Nebuloni M, Pasqualini F, Latini R, Garlanda C, Mantovani A.
Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction. Circulation 2008; 117: 1055-1064
Maria Carla Roncaglioni got her Biological Science degree in 1987 at the University of Milan.
Education
1987 Doctoral degree in Biological Sciences, University of Milan, Italy
1982-1983 “Research Fellow” at the Dept. of Biochemistry, Faculty of Medicine, Rijksuniversiteit of
Limburg, Maastricht , The Netherland (Prof. C.Hemker);
1998-1999 “Visiting Scientist” at the Cardiovascular Research Unit, Hammersmith Hospital, London,
UK (Prof. A. Maseri)
Main fields of activity
Coordination of multicenter clinical trials and observational studies in different cardiovascular areas
(neurological, angiological, cardiological). Coordination of a network of more than 1000 GPs actively
involved in epidemiological and experimental studies in the prevention of cardiovascular diseases.
Position
from 2001 Head of the Laboratory for General Practice Research, Istituto di Ricerche Farmacologiche
"Mario Negri", Milano, Italy
from 1989 Senior Researcher in the Clinical Pharmacology Laboratory, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
from 1974 Researcher in the Laboratory for the Study of Haemostasis and Thrombosis, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano, Italy
126
ANNUAL REPORT 2008

IRFMN
Selected publications
• Tognoni G, Avanzini F, Pangrazzi J, Roncaglioni M C, Bertele V, de Gaetano G, Caimi V, Tombesi M, Colombo Fabio,
Barlera S, PPP - Primary Prevention Project. Low-dose aspirin and vitamin E in people at cardiovascular risk: A
randomized trial in general practice. Lancet 2001; 357: 89-95
• Sacco M, Pellegrini F, Roncaglioni MC, Avanzini F, Tognoni G, Nicolucci A, PPP - Primary Prevention Project.
Primary prevention of cardiovascular events with low-dose aspirin and vitamin E in type 2 diabetic patients. Results of
the Primary Prevention Project (PPP) trial. Diabetes Care 2003; 26: 3264-3272
• Roncaglioni MC, Avanzini F, Roccatagliata D, Monesi L, Tamayo-Benitez D, Tombesi M, Caimi V, Longoni P, Lauri
D, Barlera S, Tognoni G, Collaborative Group Risk Prevention Study. How general practitioners perceive and grade the
cardiovascular risk of their patients Eur J Cardiovasc Prev Rehabil 2004; 11: 233-238
• Monesi L, Avanzini F, Barlera S, Caimi V, Lauri D, Longoni P, Roccatagliata D, Tombesi M, Tognoni G, Roncaglioni
MC. Appropriate use of antiplatelets: is prescription in daily practice influenced by the global cardiovascular risk Eur J
Clin Pharmacol 2005; 61: 595-601
• Berger JS, Roncaglioni MC, Avanzini F, Pangrazzi J, Tognoni G, Brown DL. Aspirin for the primary prevention of
cardiovascular events in women and men: A sex-specific meta-analysis of randomized controlled trials. JAMA 2006;
295: 306-313
• Montalvo G, Avanzini F, Anselmi M, Prandi R, Ibarra S, Marquez M, Armani D, Moreira J M, Caicedo C, Roncaglioni
MC, Colombo Fabio, Camisasca P, Milani V, Quimi' S, Gonzabay F, Tognoni G. Diagnostic evaluation of people with
hypertension in low income country: cohort study of "essential" method of risk stratification. BMJ 2008;
337: a1387
Gianni Tognoni got his Medical Doctor in 1970, University of Milan.
Main areas of methodology
Randomized clinical trials; outcomes studies; pharmacoepidemiology; pharmacoeconomics;
epidemiological monitoring and assessment of health care systems, drug policy; genetic epidemiology;
community epidemiology; transfer of technology; health and human rights.
Main clinical areas
Acute and chronic CV diseases; psychiatry; aging; intensive care; neurodegenerative disordes; hematooncology.
Position
from 2004 Member, Commission of Human Experimentation of the Italian Drug Agency (AIFA)
2001-2003 Member, Commissione Unica del Farmaco (CUF), Ministry of Health
from 2002 Director, Consorzio Mario Negri Sud, S. Maria Imbaro, Chieti.
1996-2002 Coordinator, Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche
"Mario Negri", Milano
from 1990 Co-Director, Scuola Superiore di Ricerca in Medicina Generale (CSeRMEG)
from 1976 Founding member of the International Society of Drug Bulletins (ISDB)
Coordinator, Commission of Human Experimentation, Regione Lombardia
from 1983 Founder and in the Editorial Board of the nursing research Journal Rivista
dell'Infermiere/Assistenza Infermieristica e Ricerca
from 1977 Consultant to WHO and other UN agencies for drug selection and policy; training in
methods of clinical and epidemiological research in developing countries mainly in Latin
America and Africa
1976-1999 Head, Laboratory of Clinical Pharmacology of the Istituto di Ricerche Farmacologiche
from 1975
"Mario Negri", Milano
Head, Regional Centre for Drug Information (CRIF), Regione Lombardia, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano
1969-1974 Research Assistant, Laboratory of Clinical Pharmacology, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano
Selected publications
• Landolfi R, Marchioli R, Kutti J, Gisslinger H, Tognoni G, Patrono C, Barbui T, for the European Collaboration on Low-
Dose Aspirin in Polycythemia Vera Investigators. Efficacy and safety of low-dose aspirin in polycythemia vera. N. Engl.
J. Med. 2004; 350: 114-124
• Eden T, Pui CH, Schrappe M, Tognoni G, Masera G, et al. All children have a right to full access to treatment for cancer.
Lancet 2004; 364: 1121-1122
• Marchioli R, Finazzi G, Landolfi R, Kutti J, Gisslinger H, Patrono C, Marilus R, Villegas A, Tognoni G, Barbui T.
Vascular and neoplastic risk in a large cohort of patients with polycythemia vera. J. Clin. Oncol. 2005; 23: 2224-2232
• Berger JS, Roncaglioni MC, Avanzini F, Pangrazzi I, Tognoni G, Brown DL. Aspirin for the primary prevention of
cardiovascular events in women and men: a sex-specific meta-analysis of randomized controlled trials. JAMA. 2006 Jan
18;295(3):306-13. Erratum in: JAMA. 2006 May 3;295(17):2002
• Strippoli GF, Tognoni G, Navaneethan SD, Nicolucci A, Craig JC. Haemoglobin targets: we were wrong, time to move
on. Lancet 2007;369:346-350
127
ANNUAL REPORT 2008

IRFMN
• GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini R,
Lucci D, Nicolosi GL, Porcu M, Tognoni G). Effect of n-3 polyunsaturated fatty acids in patients with chronic heart
failure (the GISSI-HF trial): a randomised, double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230
Giovanna Balconi got her degree at the School for Technicians of Biomedical Institutes of the
University of Milan, with a specialisation in Histology in the Pathological Anatomy Laboratory of the
same University (1968).
Main fields of interest
Isolation, culture and characterization of peripheral blood circulating progenitor cells of patients with
heart failure.
“In vitro” culture and characterization of stem cells for repair of myocardial infarction in experimental
animal models.
Positions
from July 2005 Head of Tissue Culture Unit, Cardiovascular Clinical Pharmacology Laboratory,
Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy
Oct 1995 - June 2005 Head of Tissue Culture Unit, Vascular Biology Laboratory, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
Dec 1983 - Oct 1995 Head of Tissue Culture Unit, Anticancer Chemotherapy Laboratory, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano, Italy
Oct 1968 - Nov 1983 Researcher, Anticancer Chemotherapy Laboratory, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
Selected publications
• Condorelli G, Borello U, De Angelis L, Latronico M, Sirabella D, Coletta M, Galli R, Balconi G, Follenzi A, Frati G,
Cusella De Angelis MG, Gioglio L, Amuchastegui CS, Adorini L, Naldini L, Vescovi A, Dejana E, Cossu G.
Cardiomyocytes induce endothelial cells to trans-differentiate into cardiac muscle: Implications for myocardium
regeneration. Proc Natl Acad Sci USA 2001; 98: 10733-10738
• Cattelino A, Liebner S, Gallini R, Zanetti A, Balconi G, Corsi A, Bianco P, Wolburg H, Moore R, Oreda B, Kemler R,
Dejana E. The conditional inactivation of the beta-catenin gene in endothelial cells causes a defective vascular pattern
and increased vascular fragility. J Cell Biol 2003; 162: 1111-1122
• Cusella De Angelis MG, Balconi G, Bernasconi S, Zanetta L, Boratto R, Galli D, Dejana E, Cossu G. Skeletal myogenic
progenitors in the endothelium of lung and yolk sac. Exp Cell Res 2003; 290: 207-216
• Galli D, Innocenzi A, Staszewsky L, Zanetta L, Sampaolesi M, Bai A, Martinoli E, Carlo E, Balconi G, Fiordaliso F,
Chimenti S, Cusella G, Dejana E, Cossu G, Latini R. Mesoangioblasts, vessel-associated multipotent stem cells, repair
the infarcted heart by multiple cellular mechanisms. A comparison with bone marrow progenitors, fibroblasts, and
endothelial cells. Arterioscler Thromb Vasc Biol 2005; 25: 692-697
• Sarto P, Balducci E, Balconi G, Fiordaliso F, Merlo L, Tuzzato G, Pappagallo GL, Frigato N, Zanocco A, Forestieri C,
Azzarello G, Mazzucco A, Valenti M T, Alborino F, Noventa D, Vinante O, Pascotto P, Sartore S, Dejana E, Latini R.
Effects of exercise training on endothelial progenitor cells in patients with chronic heart failure. J Card Fail 2007; 13:
701-708
• Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G, Cuccovillo
I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G. Cardiac mesoangioblasts are committed, self-renewable
progenitors, associated with small vessels of juvenile mouse ventricle. Cell Death Differ 2008; 15: 1417-1428.
Paola Di Giulio got her Nursing Diploma in the Nursing School of Istituto Nazionale dei Tumori in
Milano and her Master in Oncology Nursing at Guildford University (UK) in 1995.
Main fields of activity
Coordination of multicentre and observational studies studies in cardiology and palliative care.
Coordination of nursing networks.
Position
from March 2001 Associated professor at the Turin University.
from 1997 Responsible of the Nursing Research Unit
from 1995 Senior researcher of the Cardiovascular Research Department
from 1989 Consultant of the Clinical Phrmacology Laboratory
since 1998 Coordinator of the Editorial Board of Assistenza Infermieristica e Ricerca
Selected publications
• Laquintana D, Di Giulio P: Per un ruolo infermieristico nella farmacovigilanza. Assistenza Infermieristica e ricerca
2002; 21: 198-210
• Di Giulio P, Saiani L, Laquintana D, Palese A, Gruppo PARI-ETLD. Studio clinico randomizzato controllato in doppio
cieco sull’efficacia dei trattamenti delle lesioni da decubito. Assistenza Infermieristica e Ricerca 2004; 23: 201-218
128
ANNUAL REPORT 2008

IRFMN
• Toscani F, Di Giulio P, Brunelli C, Miccinesi G. Laquintana D. How people die in hospital general wards: a descriptive
study. J Pain Symptom Manage 2005; 30: 33-40
• Saiani L, Di Giulio P, Gruppo PARI-FV (Percorsi Assistenziali e Ricerca Infermieristica-Farmaco Vigilanza)
Epidemiologia dei problemi assistenziali legati a farmaci e presidi in RSA e distretto. Assistenza Infermieristica e
Ricerca 2007; 26: 123-164
• Lepore V, Cecchetto G, Di Giulio P, Saiani L, Samarelli V, Saugo M, Romero M, Scurti V, Tognoni G, Valerio M. Età
anziana-molto-anziana, e “aspettativa di vita” Assistenza Infermieristica e Ricerca 2007; 26: 234-242
• Di Giulio P, Toscani F, Villani D, Brunelli C, Gentile S, Spadin P. Dying with advanced dementia in long-term care
geriatric institutions: a retrospective study. J Palliat Med 2008; 11: 1023-1028.
Fabio Fiordaliso got his Biological Science degree in 1995 at the University of Milan.
Education
1998 Postdoctoral degree in Pharmacological Research, Istituto di Ricerche Farmacologiche
“Mario Negri”, Milan, Italy
1995 Doctoral degree in Biological Sciences, University of Milan, Italy
Main fields of activity
Therapeutical potential of stem cell and antioxidant treatments in experimental model of diabetic
cardiomyopathy and in primary myocyte cultures exposed to hyperglycemia.
Morphological and structrural analysis of cells and tissue by optical, confocal and electron microscopy.
Positions
from 2007
from 2006
from 2005
from 2005
from 2001
Head of Bio-imaging Unit, Department of Cardiovascular Research, Istituto di Ricerche
Farmacologiche “Mario Negri”, Milan
Member of the Heart Failure Association (HFA) of the European Society of Cardiology
Member of the Working group on myocardial function (WG 4) of the European Society of
Cardiology
Member of the steering committee of the Consorzio of Microscopy and Image Analysis
(MIA)
Senior Research Scientist, Laboratory of Cardiovascular Clinical Pharmacology
(Department of Cardiovascular Research), Istituto di Ricerche Farmacologiche “Mario
Negri”, Milan
1997-2001 Post-Doctoral Research Fellow at Cardiovascular Research Institute (Department of
Medicine), New York Medical College, Valhalla, New York
1994-1997 Research Fellow, Laboratory of Cardiovascular Clinical Pharmacology (Department of
Cardiovascular Research), Istituto di Ricerche Farmacologiche “Mario Negri”, Milan
1992-1994 Research training, Institute of General Pathology, University of Milan (Italy)
Selected publications
• Fiordaliso F, Bianchi R, Staszewsky L, Cuccovillo I, Doni M, Laragione T, Salio M, Savino C, Melucci S, Santangelo F,
Scanziani E, Masson S, Ghezzi P, Latini R. Antioxidant N-Acetyl-L-Cysteine attenuates hyperglycemia-induced
cardiomyocyte death in rats. J Mol Cell Cardiol 2004; 37: 959-968
• Fiordaliso F, Chimenti S, Staszewsky L, Bai A, Carlo E, Cuccovillo I, Doni M, Mengozzi M, Tonelli R, Ghezzi P,
Coleman T, Brines M, Cerami A, Latini R. A non-erythropoietic derivative of EPO effectively ameliorates experimental
cardiac ischemia with reperfusion. Proc Natl Acad Sci USA 2005; 102: 2046-2051
• Fiordaliso F, Cuccovillo I, Bianchi R, Bai A, Doni M, Salio M, De Angelis N, Ghezzi P, Latini R, Masson S.
Cardiovascular oxidative stress is reduced by an ACE inhibitor in a rat model of streptozotocin-induced diabetes . Life Sci
2006; 79: 121-129
• Fiordaliso F, De Angelis N, Cuccovillo I, Bai A, Salio M, Serra DM, Bianchi R, Razzetti R, Latini R, Masson S. Effect
of β-adrenergic and renin-angiotensin system blockade on myocyte apoptosis and oxidative stress in diabetic
hypertensive rats. Life Sci 2007; 81: 951-959
• Latini R, Brines M, Fiordaliso F. Do non-hemopoietic effects of erythropoietin play a beneficial role in heart failure
Heart Fail Rev 2008; 13: 415-423
• Dossena S, Imeri I, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M,
Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant
prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model.
Neuron 2008; 60: 598-609
129
ANNUAL REPORT 2008

IRFMN
Serge Masson obtained his doctorate (PhD) in Biochemistry and Cellular Biology in 1990 at the
University of Marseilles (France), followed by a postdoctoral stay at the Panum Institute in Copenhagen
(Denmark)
Education
1988-1990 Doctorate fellow, Faculty of Medicine, University of Aix-Marseilles, France
1990-1993 Post-doctoral Researcher, Panum Institute and Assistant Lecturer, University of
Copenhagen, Denmark
1993 Research Scientist, NMR Laboratory, Hospital “San Raffaele”, Milan, Italy
from 1994 Research Scientist, Department of Cardiovascular Research, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
Main fields of activity
Physiopathology, diagnostic and prognostic role of the activation of neuroendocrine systems in
cardiovascular disease
Position
from 2002
from 2002
from 2002
Head of the Cardiovascular Endocrine Unit, responsible for Quality Assurance for the
Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche "Mario
Negri", Milano, Italy
Tutor of fellows of the School of Specialists in Pharmacological Research, Istituto di
Ricerche Farmacologiche "Mario Negri", Milano, Italy
Fellows of the American Heart Association (Basic Council) and the Working Group on
Myocardial Function of the European Society of Cardiology
Selected publications
• Anand IS, Latini R, Florea VG, Kuskowski MA, Masson S, Signorini S, Mocarelli P, Hester A, Glazer R, Cohn JN, for
the Val-HeFT Investigators. C-reactive protein in heart failure: prognostic value and the effect of valsartan. Circulation
2005; 112: 1428-1434.
• Masson S, Latini R, Anand I S, Barlera S, Judd D, Salio M, Perticone F, Perini G, Tognoni G, Cohn JN, on behalf of the
Val-HeFT Investigators. The prognostic value of Big endothelin-1 in more than 2,300 patients with heart failure enrolled
the Valsartan in Heart Failure Trial (Val-HeFT). J Card Fail 2006; 12: 375-380.
• Latini R, Masson S, Anand I S, Missov E, Carlson M, Vago T, Angelici L, Barlera S, Parrinello G, Maggioni AP,
Tognoni G, Cohn J N, Val-HeFT Investigators. Prognostic value of very low plasma concentrations of troponin T in
patients with stable chronic heart failure. Circulation 2007; 116: 1242-1249.
• Masson S, Latini R, Anand IS, Vago T, Angelici L, Barlera S, Missov ED, Clerico A, Tognoni G, Cohn JN, on behalf of
the Val-HeFT Investigators. Direct comparison of B-type natriuretic peptide (BNP) and amino-terminal proBNP in a
large population of patients with chronic and symptomatic heart failure. The Valsartan Heart Failure (Val-HeFT) data.
Clin Chem 2006; 52: 1528-1538.
• Staszewsky L, Wong M, Masson S, Barlera S, Carretta E, Maggioni AP, Anand IS, Cohn JN, Tognoni G, Latini R,
Valsartan Heart Failure Trial Investigators. Clinical, neurohormonal, and inflammatory markers and overall prognostic
role of chronic obstructive pulmonary disease in patients with heart failure: data from the Val-HeFT Heart Failure Trial.
J Card Fail 2007; 13: 797-804.
• Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators.
Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure Trial).
J Am Coll Cardiol 2008; 52: 997-1003
Enrico Bjørn Nicolis has attended the courses in Computer Science at the University of Milan.
Education
1991-1999 “Research fellow”, Istituto di Ricerche Farmacologiche "Mario Negri", Milano, Italy
Main fields of activity
Data management and analysis of randomized clinical trials. Developing of database and tools for studies
of population genetics, particularly for linkage analysis.
Position
from 2001 Head of the Bioinformatics Unit, Istituto di Ricerche Farmacologiche "Mario Negri",
Milano, Italy
from 1999 Research fellow of the Laboratory of Clinical Drugs Evaluation
from 1997 System administrator at the EDP center, Istituto di Ricerche Farmacologiche "Mario Negri",
Milano, Italy
from 1991 Research fellow at the Medical Informatics and Applied Statistics Unit, Istituto di Ricerche
Farmacologiche "Mario Negri", Milano, Italy
130
ANNUAL REPORT 2008

IRFMN
Selected publications
• Nobili A, Gebru F, Rossetti A, Schettino F, Zahn R W, Nicolis E, Macario G, Celani L, Acik V O, Farina ML, Naldi L.
Doctorline: A private toll-free telephone medical information service. Five years of activity: Old problems and new
perspectives. Ann Pharmacother 1998; 32: 120-125
• Santoro E, Nicolis E, Franzosi MG.Telecommunication technology for the management of large scale clinical trials: The
GISSI experience. Comput Methods Programs Biomed 1999; 60: 215-223
• Santoro E, Nicolis E, Franzosi MG, Tognoni G. Internet for clinical trials: Past, present, and future. Control Clin
Trials 1999; 20: 194-201
• Tognoni G, Franzosi MG, Nicolis E, Barlera S, Specchia C, Chiodini B, Crociati L, Ferrario L, PROCARDIS
Consortium. A trio family study showing association of the lymphotoxin-alfa N26 (804A) allele with coronary artery
disease. Eur J Hum Genet 2004; 12: 770-774
• Barlera S, Specchia C, Farrall M, Chiodini BD, Franzosi MG, Rust S, Green F, Nicolis E, Peden J, Assmann G, Collins
R, Hamsten A, Tognoni G, PROCARDIS Consortium. Multiple QTL influence the serum Lp(a) concentration: a
genome-wide linkage screen in the PROCARDIS study. Eur J Hum Genet 2007; 15: 221-227
• Specchia C, Barlera S, Chiodini BD, Nicolis EB, Farrall M, Peden J, Collins R, Watkins H, Tognoni G, Franzosi MG,
PROCARDIS Consortium. Quantitative trait genetic linkage analysis of body-mass index in familial coronary artery
disease. Hum Hered 2008; 66: 19-24
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The areas of interest of the Department of Cardiovascular Research include the experimental,
clinical, genetic, epidemiological aspects of acute myocardial infarction, cardiac failure, cardiac
arrhythmias, as well as the clinical and epidemiological investigation of cardiovascular
prevention, hypertension and stroke. Following the successful experience of the GISSI-trials
(Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto), the activation of large
collaborative networks in the setting of the National Health Service hospitals and in general
practice has become a key characteristics of the Department, which can now rely on the
permanent collaboration of over 300 clinical groups and of several hundred general
practitioners. Over the years, firm links have also been established with international leading
research groups.
The experimental research activity concerns the physiopathology, the pharmacological
modulation and the prognostic role of the activation of the renin-angiotensin-aldosterone
system, as well as other neurohormonal systems, in myocardial infarction and heart failure, the
physiopathology, the pharmacological modulation and prognostic role of the activation of the
inflammatory processes in myocardial infarction and heart failure; a more recent research topic
is the cell therapy of myocardial infarction.
The activity in clinical research includes the clinical assessment of therapeutic strategies with
large scale clinical trials in the field of acute coronary syndromes, congestive heart failure and
atrial fibrillation. A recently developing area is the genetic epidemiology of myocardial
infarction and heart failure. Several studies have been conducted in the area of clinical
epidemiology and risk factors assessment of myocardial infarction.
The collaboration with a large network of General Practitioners in the area of cardiovascular
prevention allowed to test new hypotheses through large scale clinical trials and to evaluate the
actual transferability of evidence based interventions in the every day practice through
epidemiological or outcome research studies. Pharmacoepidemiological studies through the
analysis of a large sample of Local Health Units drug prescriptions were also performed. A
research network of nurses has been developed with the main focus on the assessment of healthrelated
quality of life of patients and on the epidemiology of nursing interventions and their
implications for patients' well being and outcomes.
131
ANNUAL REPORT 2008

IRFMN
FINDINGS/MAIN RESULTS
The GISSI-HF Study, a randomized multicenter trial conducted in nearby 7000 patients from
357 cardiology sites in Italy, showed that a simple, safe, one-a-day capsule of n-3
polyunsaturated fatty acids (PUFA) can reduce mortality and admission to hospital for
cardiovascular reasons in patients with heart failure.
Patients received either n-3 PUFA in a capsule once daily (3494 patients) or placebo (3481).
955 patients in the PUFA group (27%) died, compared with 1014 (29%) in the placebo group,
meaning a relative risk reduction of 9% in the PUFA group. A higher proportion of patients in
the placebo group (2053/59%) died or were admitted to hospital for cardiovascular reasons than
in the PUFA group (1981/57%) a relative reduction of 8% in the PUFA group. In absolute
terms, 56 patients needed to be treated with PUFA for just under four years to avoid one death,
or 44 patients to avoid one event of either death or admission to hospital for cardiovascular
causes. Statin treatment with rosuvastatin does not affect clinical outcomes in patients with
chronic heart failure.
In 2500 patients with chronic heart failure, enrolled to the GISSI-HF multicenter trial we
showed that microalbuminuria, a marker of vascular and renal injury, is one of the strongest
predictors of death and hospitalization for cardiovascular reasons. The relation with outcomes is
still strong after adjustment for major markers of risk, including creatinine, BNP, CRP. It's the
largest study showing the prognostic value of microalbuminuria in heart failure.
GISSI-AF, a randomized, prospective, parallel group, placebo-controlled, multicenter study on
the use of valsartan, an angiotensin II AT1-receptor blocker (ARBs) in the prevention of Atrial
Fibrillation (AF) recurrence, has recruited and treated for 12 months 1400 patients with a
history of recent AF associated with cardiovascular diseases/comorbidities. In GISSI-AF, the
largest trial ever conducted with ARBs in pts with AF, Valsartan did not reduce AF recurrence.
A trend favoring valsartan was observed only in the small group of pts with heart failure and/or
left ventricular dysfunction.
The PROCARDIS consortium was conceived to study the complex genetics of coronary artery
disease (CAD) susceptibility in Europeans and has assembled clinical resources to undertake
genetic association studies. Recently PROCARDIS has undertaken a case-control study in 4,251
CAD cases and 4,443 controls using previously reported and tagging SNPs (Single Nucleotide
Polymorphisms) of a region on chromosome 9p that is associated with CAD and with type 2
diabetes (T2D). PROCARDIS replicated the SNPs and showed that the strong consistent
association detected by these SNPs is a consequence of a “yin-yang” of markers spanning 53
kb. There was no evidence of additional CAD susceptibility alleles over the major risk
haplotype. CAD patients without myocardial infarction (MI) showed a trend towards stronger
association than MI patients. The CAD susceptibility conferred by this locus did not differ by
sex, age, smoking, obesity, hypertension or diabetes. A simultaneous test of CAD and diabetes
susceptibility with CAD and T2D-associated SNPs indicated that these associations were
independent of each other.
A genetic study conducted in the patients of the GISSI-Prevenzione (GISSI-P) trial showed that
Apolipoprotein E polymorphisms influence effect of pravastatin on survival after myocardial
infarction. We analyzed 3304 Italian patients with MI randomized to pravastatin or no treatment
+with a median follow-up time of 24 months and we found that epsilon-4 allele is a determinant
of the response to pravastatin in terms of survival. Though in the entire population investigated
we found a beneficial effect of pravastatin in terms of survival, only the epsilon-4 carriers
seemed to have gained a significant benefit from this treatment. We suggest that the effect of
132
ANNUAL REPORT 2008

IRFMN
statins is of particular interest in this fraction of the population and that genetic markers can
help in identifying patients that benefit more from statin treatment.
NATIONAL COLLABORATIONS
Angiogenesis and Tumor Targeting Research Unit & Telethon Institute for Gene Therapy,
Ospedale San Raffaele, Milano
ANMCO (Associazione Nazionale Medici Cardiologi Ospedalieri)
Centro Cardiologico Monzino IRCCS, Milano
CINECA (Consorzio Interuniversitario per il Calcolo Automatico dell'Italia Nord-Orientale)
CSeRMEG (Centro Studi e Ricerche in Medicina Generale)
Dipartimento di Cardiologia e UTIC, Istituto Clinico Humanitas IRCCS, Milano
Dipartimento Cardio-Vascolare ed Endocrino-Metabolico, Ospedale Casa Sollievo della
Sofferenza IRCCS, San Giovanni Rotondo
Ematologia, Ospedale Sant’Anna, Torino
Fondazione Don Gnocchi IRCCS, Milano
Gruppi organizzati di MMG (FIMMG, CoS, Ass.Cu.M.I., AMISI)
IEO (Istituto Europeo di Oncologia), Milano
IFOM-FIRC, Milano
Istituto di Anestesiologia e Rianimazione IRCCS, Ospedale Maggiore Policlinico, Mangiagalli,
Regina Elena, Milano
Istituto di Ricerca in Cure palliative Lino Maestroni, Cremona
Laboratorio di Endocrinologia, Ospedale Luigi Sacco, Milano
Regione Lombardia
SIBioC (Società Italiana di Biochimica Clinica e Biologia Molecolare)
SIFO (Società Italiana di Farmacia Ospedaliera)
Stem Cell Research Institute, Ospedale San Raffaele, Milano
Unità Operativa Semplice di Neuroanestesia e Neurorianimazione, Dipartimento di Medicina
Perioperatoria e Terapie Intensive, Ospedale San Gerardo, Monza
Università degli Studi di Milano, Dipartimento di Medicina Interna
Università degli Studi di Milano, Dipartimento di Scienze Farmacologiche
Università degli Studi di Torino, Dipartimento di Anatomia, Farmacologia e Medicina Forense
Università degli Studi di Torino, Dipartimento di Sanità Pubblica e Microbiologia
Università degli Studi di Verona, Dipartimento di Sanità Pubblica
Università degli Studi di Verona, Istituto di Anatomia Umana
INTERNATIONAL COLLABORATIONS
Cecomet (Centro de Epidemiologia comunitaria y Medicina tropical, Esmeraldas) Ecuador
Cochrane Collaboration, Oxford, UK
Clinical Trial Research Unit, Auckland University, New Zealand
CTSU (Clinical Trial Service Unit) /ISIS (International Studies on Infarct Survival), Oxford,
UK
Division of Genetics and Development, Guy's, King's and St Thomas' School of Medicine,
King's College, London, UK
DSAN SUPSI (Scuola Universitaria Professioni Sanitarie), Lugano CH
ECLA (Estudios Cardiologicos de Latino-America)
133
ANNUAL REPORT 2008

IRFMN
EVGN (European Vascular Genomics Network) VI Framework Program, European Union
JW Goethe University, Department of Cardiology, Frankfurt, Germany
Karolinska Institutet, Stockholm, Sweden
PHRI (Population Health Research Institute), McMaster University, Hamilton, Ontario, Canada
SIOP (International Society of Paediatric Oncology)
University of Cambridge, UK
University of Minnesota, Minneapolis, USA
University of Oslo, Norway
Wellcome Trust Centre for Human Genetics, University of Oxford, UK
WONCA (World Organization of Family Doctors)
EDITORIAL BOARD MEMBERSHIP
Circulation, International Journal of Health Services, European Heart Journal (Gianni Tognoni)
Journal of Cardiac Failure, Journal of Cardiovascular Medicine (Roberto Latini)
Assistenza Infermieristica e Ricerca, European Journal of Cancer Care, European Journal of
Oncology Nursing, International Nursing Perspectives (Paola Di Giulio)
PEER REVIEW ACTIVITIES
American Heart Journal, Atherosclerosis Thrombosis and Vascular Biology, Cardiovascular
Research, Circulation, Circulation Research, Clinical Pharmacology and Therapeutics,
European Heart Journal, European Journal of Cancer Care, European Journal of Cardiovascular
Nursing, European Journal of Oncology Nursing, International Journal of Cardiology,
International Journal of Obesity, Italian Journal of Cardiology, Journal of Cardiac Failure,
Journal of Internal Medicine, Journal of Cardiovascular Medicine, Life Sciences, The Lancet,
PharmacoEconomics.
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Ethical Committee of the ASL of Milan
Ethical Committee of Lombardy Region
Ethical Committee of the Provincia of Verona
Agenzia Italiana del Farmaco, Direzione Generale Farmaci e Dispositivi Medici
EVENT ORGANIZATION
Investigator's Meeting - Stato di avanzamento dello studio Prono/Supino II
16/01/08, Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
Investigator's Meeting – Presentazione dei risultati dello studio GISSI-AF
134
ANNUAL REPORT 2008

IRFMN
01/06/08, Fortezza da Basso, Firenze
Investigator's Meeting – GISSI-HF: un modello di ricerca collaborativa
06/09/08, Aula Magna Santa Lucia, Bologna
Investigator's Meeting – Stato di avanzamento dello studio "NeuroMorfeo"
08/10/08, Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
Investigator's Meeting - Stato di avanzamento dello studio Rischio & Prevenzione
18/10/08, Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
PROCARDIS Research Program – PROCARDIS Ethics Workshop
- Ethical and regulatory aspects of the Italian participation in the PROCARDIS program
- Differences in attitudes and common concerns on genetic research within Europe
17-3-08, Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
Master di I° Livello in Ricerca Clinica dell’Università degli Studi di Milano, Facoltà di
Medicina e Chirurgia, Dipartimento di Medicina Interna (Anno Accademico 2007-2008)
15/02/08 La ricerca clinica oggi: profit e no-profit. Il protocollo dello studio
18/02/08 Il disegno degli studi clinici
28/02/08 Legislazione sulla sperimentazione clinica e ruolo dei Comitati Etici
10/03/08 Monitoraggio degli studi clinici
20/03/08 Calcolo della dimensione campionaria
28/03/08 Dalla preclinica alla clinica: sviluppo di nuovi farmaci cardiovascolari
02/04/08 Studi di fase 2: Obiettivi, disegno e stima del campione in oncologia
09/04/08 Le revisioni sistematiche e metanalisi
08/05/08 Ricerca clinica nel campo dell'epilessia
14/05/08 Farmacovigilanza
15/05/08 Ricerca clinica nell'ictus
23/05/08 Interazione tra farmaci
13/06/08 Internet e le nuove tecnologie per l’aggiornamento medico
18/06/08 Regolamentazione dei farmaci in Europa
09/09/08 Studi di fase 3: obiettivi, disegno e indicatori
16/09/08 Scienza ed etica
17/09/08 Studi osservazionali: obiettivi, disegno ed aspetti regolatori
23/09/08 Studi osservazionali: obiettivi, disegno ed aspetti regolatori - II^ parte
Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
Master di I° Livello in Ricerca Clinica dell’Università degli Studi di Milano, Facoltà di
Medicina e Chirurgia, Dipartimento di Medicina Interna (Anno Accademico 2008-2009)
10/11/08 Esercitazioni di statistica descrittiva
Il disegno dello studio in epidemiologia
Il disegno degli studi clinici
11/11/08 Esercitazioni di inferenza statistica
Studi di fase 2: obiettivi, disegno e stima del campione in oncologia
12/11/08 Legislazione sulla sperimentazione clinica e ruolo dei Comitati Etici
Misure di rischio in epidemiologia
Internet e le nuove tecnologie per l'aggiornamento medico-scientifico
17/11/08 Analisi della sopravvivenza
Scienza ed etica
18/11/08 Farmacovigilanza
Monitoraggio degli studi clinici profit. Report delle Reazioni Avverse
135
ANNUAL REPORT 2008

IRFMN
19/11/08 Monitoraggio degli studi clinici no-profit
Report delle Reazioni Avverse negli studi clinici no profit
Dalla preclinica alla clinica: sviluppo di nuovi farmaci cardiovascolari
24/11/08 Studi di fase 3: obiettivi, disegno e indicatori in oncologia
Le revisioni sistematiche e metaanalisi
25/11/08 Ricerca clinica nel campo dell'epilessia. Ricerca clinica nell'ictus
Interazioni tra farmaci
La ricerca bibliografica oggi
26/11/08 Ricerca in medicina generale
Medicina di domani, etica di ieri
Istituto di Ricerche Farmacologiche “Mario Negri”, Milano
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT
WAS INVOLVED
European Society of Cardiology. Winter Meeting on Translational Cardiology - Heart Failure
and Regeneration, 23-26/01/08, Garmisch-Partenkirchen, Germany
- Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction
Università di Modena e Reggio Emilia – Master Gestione delle Sperimentazioni – Statistica
Medica II, 25-28/2/08, Dipartimento di Oncologia ed Ematologia, Università di Modena e
Ferrara, Italy
Il concetto di prognosi nei diversi tipi di studi in epidemiologia:
- Outcome negli studi osservazionali
- Outcome negli studi sperimentali
- Definizione di Endpoint surrogato
Metodi statistici (1) per l’analisi della prognosi:
- Modelli di regressione lineare
- Modello logistico
Guida alla lettura critica di un articolo scientifico
Metodi statistici (2) per l’analisi della prognosi:
- Modello a rischi proporzionali di Cox
- Analisi della sopravvivenza
Guida alla lettura critica di un articolo scientifico
Istituto Clinico Humanitas. IV Corso di specializzazione in emodinamica e cardiologia
interventistica, 04/03/08, Auditorium Istituto Clinico Humanitas, Rozzano (MI), Italy
- La cardiomiopatia postinfartuale: il futuro delle cellule staminali
American College of Cardiology. 57th Annual Scientific Session, 29/03-01/04/08, Chicago,
USA
- Multiple RAAS inhibition influences the changes of biohumoral markers in patients with heart
failure. Data from Aliskiren Observation of Heart Failure Treatment (ALOFT) Study
Istituto di Ricerche Farmacologiche “Mario Negri” – European Clinical Research
Infrastructures Network-ECRIN. Donne & Ricerca Clinica, Giornata Internazione della ricerca
clinica, 21/05/08, Istituto di Ricerche Farmacologiche “Mario Negri”, Milano, Italy
136
ANNUAL REPORT 2008

IRFMN
- Fatti, problemi e difficoltà nella ricerca clinica al femminile in cardiologia
Dipartimento di Malattie Cardiovascolari e A.S.O. Santa Croce e Carle di Cuneo. Luci ed
ombre nell’utilizzo dei sartani nelle malattie cardiovascolari: i dati della letteratura, 25/05//08,
Salone “Costantia” Castello Rosso, Costigliole-Saluzzo (CN)
- L’uso dei sartani nei pazienti con scompenso cardiaco
Associazione Nazionale Medici Cardiologi Ospedalieri. XXXIX° Congresso Nazionale di
Cardiologia, 30/05-02/06/08, Firenze, Italy
- Studi clinici
- Razionale e disegno dello Studio GISSI-AF
- Razionale per l’uso di un protocollo dinamico infusionale di insulina
- Valutazione dell’utilità di un intervento infermieristico di supporto telefonico per migliorare
gli stili di vita dopo un ricovero per sindrome coronarica acuta
- Valore prognostico delle variazioni di NT-proBNP in pazienti con insufficienza cardiaca
coronarica. Dati dallo studio Val-HeFT
- Determinanti di disfunzione endoteliale (microalbuminuria) e infiammazione vascolare
(Pentraxina-3) in pazienti con insufficienza cardiaca cronica. Dati dallo studio GISSI-HF
- La pentraxina lunga PTX3 è un marcatore di comorbidità nei pazienti con insufficienza
cardiaca. Dati dallo studio GISSI-HF
- Interazione fra un marcatore del turnover del collagene (PIIINP) e un antagonista
dell’aldosterone (Canrenone) sul rimodellamento ventricolare in pazienti con insufficienza
cardiaca lieve. Dati dallo studio AREA IN.CHF
- Trattamento dell’iperglicemia in corso di sindrome coronarica acuta: efficacia e sicurezza di
un protocollo di infusione endovenosa di insulina a gestione infermieristica
Roche Diagnostics. 6th International Symposium onNF-proBNP, 18-19/06/08, Baveno, Italy
- hs Troponin T: The Val-HeFT and GISSI Studies
- Serial measurements of NT-proBNP in chornic heart failure
European Society of Cardiology. ESC Congress 2008, 30/08-03/09/08, Munich, Germany
- Effectiveness and safety of a new nurse-implemented insulin infusion protocol (DDD) for
intensive glucose control in diabetic patients with acute coronary syndromes
- Clinical determinants of pentraxin-3 (PTX3), a novel marker of vascular inflammation, in
patients with chronic heart failure. Data from the GISSI-HF study
- Categorial changes of NT-proBNP concentrations predict outcome in ambulatory patients with
chronic stable heart failure. Data from the valsartan heart failure trial
WONCA EUROPE 2008. 14th Regional Conference. Overcoming the distance - Family doctor,
bringing the art of medicine to the patients, 04-07/09/08, Istanbul, Turkey
- The optimisation of cardiovascular prevention in everyday practice: preliminary results of the
Risk & Prevention Study
Clinical Forum S.r.l. Easy future. Nuove prospettive terapeutiche, 12-13/09/08, Istituto di
Ricerche Farmacologiche “Mario Negri”, Milano, Italy
- Revisione dell’ipotesi metabolico-infiammatoria nello scompenso. Implicazioni terapeutiche
Dipartimento Cardiovascolare dell’Azienda Ospedaliero Universitaria degli Ospedali Riuniti di
Trieste e Centro Cardiovascolare dell’Azienda per i Servizi Sanitari N. 1 Triestina, 10-11/10/08,
Trieste, Italy
- Citochine, infiammazione e BNP nello scompenso cardiaco: aiuteranno realmente a
comprendere e curare meglio la sindrome
137
ANNUAL REPORT 2008

IRFMN
Struttura Complessa di Cardiologia dell’Azienda Ospedaliera G Brotzu di Cagliari e Unità
Operativa di Cardiologia dell’Ospedale SS Annunziata-ASL N. 1 di Sassari. La Sardegna nel
cuore. 2° Congresso di cardiologia a carattere nazionale, 23-25/10/08, Cagliari, Italy
- Le terapie geniche e cellulari
American Society of Nephrology. ASN Congress, 4-9/11/08, Philadelphia, USA
- Safety and tolerability profile of aliskiren added to optimized therapy in patients with heart
failure and renal impairment
American Heart Association. AHA Scientific Session 2008, 8-12/11/08, New Orleans, USA
- Elevated albumin excretion is an independent risk factor in patients with chronic heart failure.
Data from the GISSI-Heart Failure Trial
- Circulating markers of myocyte injury predict first recurrence of atrial fibrillation. Data from
the GISSI-Atrial Fibrillation Trial
- Role of inflammatory markers in the prediction of first recurrence of atrial fibrillation. Data
from the GISSI-Atrial Fibrillation Trial
- Stable precursor fragments of vasoactive peptides predict outcome in patients with chronic
heart failure. Data from the GISSI-Heart Failure Trial
- The role of valsartan in the prevention of atrial fibrillation recurrence: the GISSI-AF results
Ospedale Luigi Sacco. La comorbidità scompenso cardiaco e fibrillazione atriale: quali approcci
terapeutici La gestione multidisciplinare, 29/11/08, Milano, Italy
- Luci ed ombre nel trattamento farmacologico: cosa ci suggeriscono le nuove linee guida
GRANTS AND CONTRACTS
AIFA (Agenzia Italiana del Farmaco), AstraZeneca, Azienda Ospedaliera San Gerardo Monza,
Chiesi Farmaceutici, Boehringer Ingelheim Italia SpA, BRAHAMS AG, Centro Cardiologico
Monzino IRCCS Milano, Cambridge University, CONGENIA, Collegio Interprovinciale
Milano-Lodi, Fondazione Don Gnocchi Milano, Fondazione San Raffaele Milano, Heart Care
Foundation, International Biomedical System SpA, Istituto Auxologico di Milano, Kinetic
Concepts Inc., Ministero della Salute, Novartis Pharma, Ospedale Casa Sollievo della
Sofferenza IRCCS San Giovanni Rotondo, Oxford University, Population Health Research
Institute-Mc Master University, Pfizer Italia, Regione Lombardia, Roche Diagnostics, Sanofi-
Aventis, Sigma Tau, SPA Società Prodotti Antibiotici S.p.A., Takeda Italia S.p.A., Università
degli Studi Torino
138
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Amigoni M, Bellani G, Scanziani M, Masson S, Bertoli E, Radaelli E, Patroniti N, Di Lelio A, Pesenti A, Latini R
Lung injury and recovery in a murine model of unilateral acid aspiration: functional, biochemical, and morphologic
characterization
Anesthesiology 2008; 108: 1037-1046
Anand SS, Islam S, Rosengren A, Franzosi MG, Steyn K, Yusufali AH, Keltai M, Diaz R, Rangarajan S, Yusuf S, on
behalf of the the INTERHEART Investigators
Risk factors for myocardial infarction in women and men: insights from the INTERHEART study
Eur Heart J 2008; 29: 932-940
Bertele' V, Angelici L, Barlera S, Garattini S
Thrombolysis or nothing for acute myocardial infarction It's all the same!
Br J Clin Pharmacol 2008; 65: 955-958
Broadbent HM, Peden JF, Lorkowski S, Goel A, Ongen H, Green F, Clarke R, Collins R, Franzosi MG, Tognoni G,
Seedorf U, Rust S, Hamsten A, Farrall M, Watkins H, for the PROCARDIS Consortium
Susceptibility to coronary artery disease and diabetes is encoded by distinct, tightly linked, SNPs in the ANRIL locus
on chromosome 9p
Hum Mol Genet 2008; 17: 806-814
Cholesterol Treatment Trialists' CTT Collaboration
Efficacy of cholesterol-lowering therapy in 18,686 people with diabetes in 14 randomised trials of statins: a metaanalysis
Lancet 2008 371: 117-125
Connolly SJ, Pogue J, Eikelboom J, Flaker G, Commerford P, Franzosi MG, Healey JS, Yusuf S, on behalf of the
ACTIVE W Investigators
Benefit of oral anticoagulant over antiplatelet therapy in atrial fibrillation depends on the quality of international
normalized ratio control achieved by centers and countries as measured by time in therapeutic range
Circulation 2008; 118: 2029-2037
CPR CHD Genetics Collaboration
Collaborative pooled analysis of data on C-reactive protein gene variants and coronary disease: judging causality by
Mendelian randomisation
Eur J Epidemiol 2008; 23: 531-540
Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M,
Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R
Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse
model
Neuron 2008; 60: 598-609
Galvez BG, Sampaolesi M, Barbuti A, Crespi A, Covarello D, Brunelli S, Dellavalle A, Crippa S, Balconi G,
Cuccovillo I, Molla F, Staszewsky L, Latini R, DiFrancesco D, Cossu G
Cardiac mesoangioblasts are committed, self-renewable progenitors, associated with small vessels of juvenile mouse
ventricle
Cell Death Differ 2008; 15: 1417-1428
Latini R, Brines M, Fiordaliso F
Do non-hemopoietic effects of erythropoietin play a beneficial role in heart failure
Heart Fail Rev 2008; 13: 415-423
Maggioni AP, Franzosi MG, Latini R
Beta-adrenoceptor antagonists and antianginal drugs
In: Side effects of drugs, Annual 30 Elsevier, Amsterdam, 2008; 223-230
Masson S, Latini R
Amino-terminal pro-B-type natriuretic peptides and prognosis in chronic heart failure
Am J Cardiol 2008; 101 Suppl: 56A-60A
139
ANNUAL REPORT 2008

IRFMN
Masson S, Latini R, Anand IS, Barlera S, Angelici L, Vago T, Tognoni G, Cohn J N, for the Val-HeFT Investigators
Prognostic value of changes in N-termianl pro-brain natriuretic peptide in the Val-HeFT (Valsartan Heart Failure
Trial)
J Am Coll Cardiol 2008; 52: 997-1003
McMurray JJV, Pitt B, Latini R, Maggioni AP, Solomon SD, Keefe DL, Ford J, Verma A, Lewsey J, for the
Aliskiren Observation of Heart Failure Treatment (ALOFT) Investigators
Effects of the oral direct renin inhibitor aliskiren in patients with symptomatic heart failure
Circulation Heart Failure 2008; 1: 17-24
Montalvo G, Avanzini F, Anselmi M, Prandi R, Ibarra S, Marquez M, Armani D, Moreira J M, Caicedo C,
Roncaglioni MC, Colombo Fabio, Camisasca P, Milani V, Quimi' S, Gonzabay F, Tognoni G
Diagnostic evaluation of people with hypertension in low income country: cohort study of "essential"
method of risk stratification
BMJ 2008; 337: a1387
Pedrazzini GB, Masson S, Latini R, Klersy C, Rossi M G, Pasotti E, Faletra FF, Siclari F, Minervini F, Moccetti T,
Auricchio A
Comparison of brain natriuretic peptide plasma levels versus logistic EuroSCORE in predicting in-hospital and late
postoperative mortality in patients undergoing aortic valve replacement for symptomatic aortic stenosis
Am J Cardiol 2008; 102: 749-754
Pedrazzini G, Santoro E, Latini R, Fromm L, Franzosi MG, Moccetti T, Staszewsky L, Barlera S, Tognoni G,
Maggioni A P, for the GISSI-3 Investigators
Causes of death in patients with acute myocardial infarction treated with angiotensin-converting enzyme inhibitors:
Findings from the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto (GISSI)-3 trial
Am Heart J 2008; 155: 388-394
Salio M, Chimenti S, De Angelis N, Molla F, Maina V, Nebuloni M, Pasqualini F, Latini R, Garlanda C, Mantovani
A
Cardioprotective function of the long pentraxin PTX3 in acute myocardial infarction
Circulation 2008; 117: 1055-1064
Specchia C, Barlera S, Chiodini BD, Nicolis EB, Farrall M, Peden J, Collins R, Watkins H, Tognoni G, Franzosi
MG, on behalf of the PROCARDIS Consortium
Quantitative trait genetic linkage analysis of body-mass index in familial coronary artery disease
Hum Hered 2008; 66: 19-24
GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini
R, Lucci D, Nicolosi GL, Porcu M, Tognoni G)
Effect of n-3 polyunsaturated fatty acids in patients with chronic heart failure (the GISSI-HF trial): a randomised,
double-blind, placebo-controlled trial. Lancet 2008; 372: 1223-1230
GISSI-HF Investigators (Writing Committee: Tavazzi L, Maggioni AP, Marchioli R, Barlera S, Franzosi MG, Latini
R, Lucci D, Nicolosi GL, Porcu M, Tognoni G)
Effect of rosuvastatin in patients with chronic heart failure (the GISSI-HF trial): a randomised, double-blind, placebocontrolled
trial
Lancet 2008; 372: 1231-1239
LAY PRESS SELECTION PUBLISHED IN 2008
Latini R
Cytokines and heart failure
Nova Acta Leopold 2008; n. 351: 27-31
Latini R, Masson S
Prognostic value of circulatory cardiac troponins in heart failure
Rev Esp Cardiol 2008; 61: 667-669
140
ANNUAL REPORT 2008

IRFMN
Latini R, Masson S
Il valore diagnostico della troponina T nell'insufficienza cardiaca
Recenti Prog Med 2008; 99: 505-508
Masson S, Latini R, Salio M
NT-proBNP and prognosis in chronic heart failure
In: NT-proBNP as a biomarker in cardiovascular diseases. Prous Science, Barcelona, 2008; 75-81
Tognoni G
Per una cultura-pratica cardiovascolare non-arrogante
Informazione sui Farmaci 2008; 32, n. 5: 118-123
Turazza FM, Masciocco G, Galvanin S, Macera F, Lenardon A, Iannì S, Foti G, Frigerio M
Introduzione alla cardiomiopatia ipertrofica: eziologia, presentazione e storia naturale
In: Cardiologia 2008. Atti del 42° convegno internazionale del dipartimento cardiologico A. De Gasperis. 15-19
settembre 2008, Milano. Fond. Centro Cardiologia e Cardiochirurgia A. De Gasperis, Milano, 2008; 189-193
RESEARCH ACTIVITIES
Laboratory of Cardiovascular Clinical Pharmacology
The effects of mesoangioblasts and of different progenitor cells on injury
after experimental myocardial infarction in the mouse
Many studies have demonstrated that autologous and homologous cells of various origins can
repair myocardium damaged due to an acute ischemic insult. Mesangioblasts are potentially
interesting when compared with bone marrow precursors because (a) they are easily expanded
and (b) obtainable by a biopsy of skeletal muscle in man. Mesoangioblasts isolated from human
heart biopsies migrate and home in the heart of immunodeficient mice after coronary ligation,
and they survive for at least 4 weeks. Studies are ongoing to establish their protective and
regenerative potential, as a basis for possible clinical studies. The same model of coronary
ligationin immunodeficient mice is being used for testing in vivo the angiogenic potential of
other progenitor cells such as mononuclear cells Tie-2+ (collaboration with Michele De Palma,
HSR, Milano), CD34+ (collaboration with Franca Fagioli, Osp S. Anna, Torino), CD133+
(collaboration with Giulio Pompilio, Centro Cardiologico Monzino, Milano). This research is
partly done within the sixth EU program, European Vascular Genomics Network, that ended in
December 2008 (EVGN, www.evgn.org).
Pulmonary injury by hydrochloric acid in the mouse: a model of
Aspiration pneumonitis to test protective interventions
Aspiration pneumonitis (AP) occurs when the acid content of the stomach makes his way
through the larynx in the lower respiratory tract. Patients with consciousness disturbance are at
risk for this event. Specifically, it has been shown that Pulmonary Aspiration can complicate
between 0.47-1.41% general anesthesia procedures.
The main injurious mechanism of AP is the chemical insult due to the low pH of gastric
secretions, which causes a chemical burn of the airway tree and of the alveolar structures. The
course of AP can be extremely variable, ranging from the “silent aspiration” characterized by a
modest desaturation to the dramatic sequelae of Acute Lung Injury (ALI) and Acute Respiratory
Distress Syndrome (ARDS), requiring prolonged mechanical ventilation and potentially leading
to death. In a murine model of monolateral acid instillation established in our laboratory, we
have shown the protective effect of exogenous pulmonary surfactant instillation. We are
currently working on a model of ventilation-induced lung injury (VILI) to assess the effect of
141
ANNUAL REPORT 2008

IRFMN
exogenous surfactant and its modulation in transgenic mice for the acute-phase protein
pentraxin-3.
Conditional transgenic model of cardiac HGF expression to promote
neovascularization and to recruit stem cells in the myocardial infarction
Growth factors such as hepatocyte growth factor (HGF) through angiogenic and anti-apoptotic
effects may promote cardiac repair after myocardial infarction and in heart failure. The cardiacspecific
α-Myosin Heavy Chain (MHC-α) transactivator was used to direct expression of HGF
to cardiomyocytes: by this way the effects of HGF can be tested under cardiac ischemia and
reperfusion, without the need for administration of exogenous HGF. The Heart-HGF transgenic
model is being used to verify the ability of HGF in vivo to promote neovascularisation, to
protect cardiomyocytes from apoptosis, to recruit and activate endogenous or transplanted stem
cells and to sustain their cellular replication and differentiation into cardiomyocytes after
ischemic damage and reperfusion.
Roles of macrophages in cardiac ischemia/reperfusion injury and in
cardiac repair
Macrophages either resident or from blood-borne monocytes play several key roles in the
response of the heart to ischemic injury. They may be useful in particular during cardiac repair,
when collagen deposition occurs and neonagiogenesis, stimulated by growth factors produced
by macrophages. The aim of the project is to assess the relevance of macrophages in myocardial
repair and scar formation after myocardial infarction, in the attempt to dissect the role of
inflammatory cells in myocardial injury vs repair.
The effects of coronary ligation (with/without reperfusion) is being tested in mice in which
machrophages can be ablated by administration of Dyphteria toxin in transgenic mice
expressing human diphteria toxin receptor (CD11b-DTR). We established a regimen of
administration of dyptheria toxin that allows the halving of monocytes level, and hence of
peritoneal macrophages. A preliminary experiment of cardiac ischemia/reperfusion has shown
that the number of macrophages was also halved in the infarct area in transgenic mice treated
with dyphteria toxin. We are now running experiments to evaluate the structural and functional
consequences of this reduction in macrophages number.
Effects of dipeptidyl peptidase-4 (DPP-4) inhibition on progenitor cells and
cardiac repair in type 2 diabetes
In patients and in experimental animal models with type 2 diabetes mellitus, inhibition of
plasma dipeptidyl peptidase IV (DPP-4) activity, increased plasma levels of the glucagonlike
peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) improving glucose
tolerance and islet cell function. Stromal cell-derived factor 1 (SDF-1α) is also cleaved by DPP-
4 and is crucially important in the mobilization and selective homing of endothelial progenitor
cells (EPCs) to ischemic tissues. EPCs have the potential to differentiate into functional mature
endothelial cells, which can substitute diseased endothelium and contribute to repair ischemic
tissues, including the heart. This regenerative action of EPCs is compromised in both type 1 and
type 2 diabetic patients.
The present study aimed at evaluating whether the administration of DPP-4 inhibitor to
obese/diabetic ob/ob mice improves mobilization/homing of EPCs by increasing levels of SDF-
1, and by this way contributing to the regeneration of myocardium and blood vessels in a model
of isoproterenol-induced cardiac infarction. To further increase cardiac injury a group of
animals were forced to swim for 5 minutes.
Hypoxic areas, detected by Hypoxyprobe-1, were found in the epicardium of mice treated
with isoproterenol and forced to swim. The phenomenon was exacerbated in the diabetic ob/ob
mice treated in the same way. The hypoxic myocardial damage determined a significative
142
ANNUAL REPORT 2008

IRFMN
mobilization of EPCs in control mice whereas this protective response was compromised in the
diabetic ob/ob mice.
Whether SDF-1 levels paralleled the EPC mobilization, the hypoxic damage determined a
decrease of capillary density and whether forced swim play a role in the cardiovascular damage
will be evaluated before beginning with the treatment of these animals with an DPP-4 inhibitor
GISSI-HF: biohumoral substudy and microalbuminuria
The clinical trial GISSI-HF (see related paragraph) was designed to assess whether two
treatments (a statin and n-3 polyunsaturated fatty acids or PUFA) can improve the prognosis of
patients with heart failure of any etiology, with preserved or compromised left ventricular
ejection fraction. Main results have recently been published (see references). The biomarker
substudy aims at exploring the pathophysiology of heart failure and mechanisms of action of the
treatments in study. Overall, 1237 patients have been enrolled in the substudy. Blood samples
were collected at enrolment and after three months to measure plasma PUFA, and markers of
ventricular myocyte stress (brain natriuretic peptides, BNP and N-terminal proBNP, MRproANP),
myocardial damage (cardiac troponin T, measured with the standard assay or with a
newly developed high sensitive method) and inflammation (C-reactive protein, CRP, pentraxin-
3, PTX3). Baseline fraction of n-3 PUFA averages 3.4 and increases by about 50% over 3
months. Baseline BNP concentration in 1223 patients is 141 pg/mL (median) and correlates
with the severity of heart failure. The number of circulating endothelial progenitor has been
measured in a smaller group of 68 patients, in collaboration with the laboratory of E. Dejana
(IFOM). Number of endothelial progenitor cells in patients with heart failure is 30-50% lower
than in healthy volunteers was inversely correlated to morbidity/mortality in a population of
patients from 5 Italian centers and a Cardiology center in Frankfurt (collaboration with Andreas
Zeiher). Microalbuminuria (defined as the ratio between urinary concentrations of albumin and
creatinine) is being measured in more than 2000 patients enrolled in the GISSI-HF trial as an
indicator of renal endothelial dysfunction. Microalbuminuria (albumin/creatinine = 30-299
mg/g) is present in 19% of the patients and is associated to inflammation, endothelial
dysfunction and neurohormonal activation. Novel and more stable circulating biomarkers, have
been successively assayed. They belong to the family of natriuretic peptides (mid-regional proatrial
natriuretici peptide, MR-proANP), endothelin (C-terminal pro-endothelin-1, CT-proET-1),
vasopressin (C-terminal pro-vasopressin, CT-proAVP) and adrenomedullin (mid-regional proadrenomedullin,
MR-proADM). Other markers related to infection and innate immunity are
currently measured (cromogranin A, mannose-binding lectin, osteoprotegerin, adiponectin and
alpha-defensin). First data on the biomarkers substudy have been presented to national and
international scientific meetings and original publications are under preparation.
PTX-3, a novel long pentraxin is a marker of severity of disease and of
outcome in cardiovascular diseases, independent of C-reactive protein
PTX-3 is a novel long pentraxin whose expression is induced by cytokines in endothelial and
mononuclear cells, mostly in striated muscle and heart, while C-reactive protein (CRP) is
mainly synthesized in the liver. PTX3 was shown to peak in plasma around 7 h after onset of
symptoms of MI and to be an independent predictor of 3-month mortality. PTX3 has been
assayed with a more accurate method in 1200 patients with symptomatic heart failure (GISSI-
HF) and in 380 patients with atrial fibrillation (GISSI-AF) to explore its role in other
cardiovascular diseases. First results indicate that PTX3 is associated with different clinical
characteristics in patients with heart failure, including advanced age, ventricular dysfunction,
functional class (NYHA class), and comorbidities such as atrial fibrillation and diabetes. PTX3
independently predicts mortality and morbidity in the patients with chronic heart failure.
143
ANNUAL REPORT 2008

IRFMN
Echocardiographic and biohumoral substudy – GISSI-AF trial
The GISSI Atrial Fibrillation trial (GISSI-AF) tests the efficacy of Valsartan, an angiotensin II
AT1-receptor blocker, in the prevention of atrial fibrillation recurrence in 1400 patients. A
substudy of the GISSI-AF has recently been concluded; it evaluated the potential role of
biohumoral factors and cardiac structural remodeling in the reoccurrence and severity of atrial
fibrillation. In approx. 400 patients three serial echocardiographic exams (at randomization, 6
months and 1 year) and contemporaneous blood collection were performed. Left ventricular and
atrial dimensions were determined by echocardiography, whereas plasma levels of natriuretic
peptides (BNP, NT-proBNP and MR-proANP), troponin T (high sensitive method), stable
vasopactive peptides (endothelin-1, Adrenomedullin and vasopressin), inflammatory markers
(C-reactive protein, interleukin-6 and pentraxin-3) and procalcitonin have been measured.
Besides giving clues on the pathophysiology of atrial fibrillation, the most common arrhythmia
in elderly, this substudy aims at providing mechanical insights of the potential benefits of the
study drug. The study was completed on January 31, 2008 and results being analyzed. First data
on circulating biomarkers have been presented to the scientific meeting of the American Heart
Association (November 2008).
CandHeart: effects of candesartan on BNP and left ventricular function in
patients with symptomatic heart failure
Candesartan, an antagonist of angiotensin II type 1 receptors, significantly reduces mortality
and morbidity in heart failure, as shown by the CHARM trials. The principal objective of the
trial is to assess the effects Candesartan on circulating levels of brain natriuretic peptide (BNP)
in patients suffering from CHF with depressed or preserved left ventricular (LV) systolic
function. The study enrolled 487 patients in 70 clinical centers with a follow-up of 1-year. Serial
circulating blood samples and echocardiographic examinations have been performed at baseline
and after 3 and 12 months (end of study). Besides BNP, other prognostic biomarkers such as
aldosterone and microalbuminuria have been assayed. A statistical plan of analyses has been
written and main results are expected for the beginning of 2009.
Trial Prone/Supine 2: effects of prolonged prone position on survival in
severe acute respiratory distress syndrome (ARDS)
This trials extends the findings of the previous Prone/Supine trial that showed in 304 patients
with ARDS that 6-hour pronation in the first 10 days improved blood gas parameters, but did
not reduce mortality (around 50% in the first 6 months). This was one of the rare examples of a
multicenter trial in Intensive Care performed entirely in Italy. The same group of Centers
(headed by the Istituto di Anestesia e Rianimazione from the Policlinico di Milano, L Gattinoni
with data management by Mario Negri) has recently concluded a new study that aims at
evaluating the effect of a prolonged pronation (20 hours a day) in patients with severe ARDS, a
subgroup that showed a trend towards improved survival on pronation in the previous study.
Three hundred and forty two patients have been enrolled in 20 centers and the main results of
this study have been submitted for publication.
DyDa: left ventricular dysfunction in diabetes. Prevalence and incidence
of left ventricular dysfunction in diabetics patients without clinical cardiac
disease
This is a prospective, multicentric, national and epidemiological trial aimed at evaluating the
prevalence of left ventricular dysfunction (systolic or diastolic) in 1000 patients with type 2
diabetes mellitus but no clinical cardiovascular disease at enrolment. The incidence of left
ventricular dysfunction is monitored during a 2-year follow-up using ECG and
echocardiography. The Biomarker Core Laboratory evaluated the biohumoral profile of these
144
ANNUAL REPORT 2008

IRFMN
patients at study entry, measuring the circulating levels of brain natriuretic peptide, C-reactive
protein, microalbuminuria and glycated hemoglobin. Enrolment started in July 2006 and ended
in March 2008 with 970 patients recruited. The assays of the biomarkers are concluded and first
data on the association between theses markers and baseline clinical characteristics are under
analysis. In a subgroup of patients from the study, levels of circulating progenitor cells and their
angiogenic potential in vitro will be measured to assess whether they are associated to risk of
developing cardiovascular disorders in diabetics.
Albumin Italian Outcome Sepsis Study. The ALBIOS Study (AIFA)
ALBIOS is a multicenter, controlled, randomized clinical trial that compares the efficacy of
human albumin and a crystalloid solution for volume replacement in patients with severe sepsis
or septic shock. The primary endpoint is survival at 28 and 90 days after enrolment. Secondary
endpoints include the number of organ dysfunctions, severity of organ dysfunction (SOFA
scale), and lengths of stay in (intensive care unit) ICU and in hospital. More than 150 ICU in
Italy are expected to participate to this large study, coordinated by the Ospedale Maggiore
Policlinico in Milan and the Consorzio Mario Negri Sud. A group of 48 ICUs participates to a
biomarkers substudy, coordinated by the laboratory of Clinical Cardiovascular Pharmacology,
with the aims of enrolling 800 patients. Serial blood samples are collected to measure the
possible effects of albumin on markers of inflammation, infection, cardiac function and
coagulation. 21 patients have been randomized by the end of 2008.
Clinical, biochemical and instrumental predictors of outcome in
rehabilitation after cardiac surgery (MIUR)
Due to the short length of hospital recovery in patients with cardiac disease, the period of
rehabilitation becomes crucial and is indicated for almost all the cardiomyopathies, including
myocardial revascularization and surgery of cardiac valves. The objective of this study is to
evaluate the prognostic role of circulating biomarkers in 250 patients enrolled in 5 Centers for
rehabilitation after cardiac surgery. A plasma bank will be collected under the responsibility of
the Laboratory of Cardiovascular Clinical Pharmacology from the Mario Negri Institute to assay
potential markers of interest. The project is coordinated by the Fondazione Don Gnocchi in
Milan. The first patient was enrolled in 2008.
Evaluation of different anesthesiological strategies for supratentorial
neurosurgery. The NeuroMorfeo Study (AIFA)
The aim of the study is to evaluate whether an anesthesia with volatile anesthetics is equivalent
to endovenous anesthetics for elective supratentorial surgery. This is a multicenter, randomized,
controlled and opened study, based on a design of equivalence for comparison of different
anesthesiological strategies. Patients to be included (n= 393) will be selected in 14
neurosurgical centers in Italy for elective intracranial surgery with supratentorial lesions without
signs of endocranial hypertension (range of age 18-75 years). The biohumoral response to
surgical stress will be measured as an indicator of homeostasis and neurovegetative status. The
urinary excretion of catecholamines and cortisol and the plasma concentration of cortisol will be
measured in a central laboratory. The study is coordinated by the San Gerardo Hospital in
Monza and by the Department of Cardiovascular Research from the Mario Negri Institute. The
first patient has been enrolled in December 2007, and since then 314 patients have been
randomized. Preliminary data show that the urinary excretion of catecholamines (normalized by
creatinine concentration) is within the expected range.
Prevalence of asymptomatic cardiac dysfunction and heart failure in a
population of elderly subjects from Lazio. The PREDICTOR study
This observational study aims at evaluating the prevalence of asymptomatic cardiac dysfunction
145
ANNUAL REPORT 2008

IRFMN
and heart failure in a random sample of elderly subjects from the Lazio area. The secondary
objective is to identify clinical, biohumoral (natriuretic peptides) and non-invasive instrumental
(echocardiography and ECG) markers of asymptomatic cardiac dysfunction and heart failure.
The population under observation is a randomly selected sample of elderly subjects (age ranging
from 65 and 84 years) resident in the area of 10 hospital cardiology centers. In a first phase,
1000 subjects have been recruited; a second phase has recently started with the objective of
enrolling another 2000 subjects. Blood samples are collected for each subject and stored in the
biobank in the Laboratory of Clinical Cardiovascular Pharmacology. Preliminary data obtained
in the first 600 subjects show a good pathophysiological agreement between the circulating
levels of NT-proBNP and indexes of left or right asymptomatic cardiac dysfunction or the
severity of heart failure.
Laboratory of Clinical Drug Evaluation
PROCARDIS: A genome-wide strategy to identify susceptibility loci in
precocious coronary artery disease
The PROCARDIS research programme, a genome-wide strategy to identify susceptibility loci
in precocious coronary artery disease (CAD) supported by the 5th Framework Programme of
the EC, was initiated as a collaboration between the universities of Oxford and Mƒnster, the
Karolinska Institute, the Mario Negri Institute with the support of the GISSI group, Oxagen, a
biotechnology company, and AstraZeneca. The objectives of the first stage of this programme
were to collect a minimum of 2000 affected sibling pairs (ASPs) and families with precocious
CAD and to apply genome-wide linkage mapping techniques, by typing anonymous highly
informative markers spaced throughout the genome, to identify chromosomal regions which are
linked to the susceptibility to early-onset CAD. The study design is based on family
ascertainment, to allow non-parametric linkage analyses (in ASPs). The PROCARDIS collected
2,036 CAD families from four European countries, in order to maximise the power of detecting
genes that confer modest risks. A genome-wide linkage scan identified three promising regions
for intensive study; one of the linked regions (Chromosome 17) was confined to families with
multiple cases of myocardial infarction and was replicated in a second independent series of
families. In addition the linkage scan confirmed a previously identified locus on Chromosome 2.
These results demonstrate that novel CAD susceptibility genes are tractable to positional
cloning which promises to lead to the identification of new molecular insights into this
condition, and hopefully, new treatments.
Additionally, the program has collected nuclear families (e.g. parent-offspring trios) for
transmission-based tests of association for fine mapping by linkage disequilibrium analysis and
testing of positional candidate genes.
Extensive clinical and biochemical intermediate phenotype data have also been collected and
assessed.
The second stage of PROCARDIS, supported by the EC 6th Framework Programme, is
conducting a large GWAS (genome wide association study), where the patients with myocardial
infarction enrolled in the first stage will be compared with control subjects to identify novel
candidate genes. The results will be replicated in different populations. The collaboration has
been extended to include complementary expertise, as proteomics, bioinformatics, functional
analysis.
GISSI-HF: A large scale clinical trial testing the effects of n-3 PUFA and
Rosuvastatin on mortality/morbidity of patients with symptomatic
Congestive Heart Failure
146
ANNUAL REPORT 2008

IRFMN
The GISSI (Gruppo Italiano per lo Studio della Sopravvivenza nell'Insufficienza cardiaca) is a
collaborative group endorsed by ANMCO (Associazione Nazionale Medici Cardiologi
Ospedalieri) and by the Istituto Mario Negri, active from 20 years in the cardiovascular research
field. The GISSI-HF was the fifth large scale clinical trial conducted by the Group and was a
prospective, multicenter, randomized, double blind, placebo controlled study, with randomized
allocation of patients with a clinical diagnosis of heart failure to:
Randomization 1 (R1): n-3 PUFA vs corresponding placebo;
Randomization 2 (R2): rosuvastatin vs corresponding placebo.
The primary objective of the GISI-HF was to demonstrate that, in patients with heart failure
treated at the best of recommended treatment, long-term administration of n-3 PUFA and/or
rosuvastatin is more effective than the corresponding placebo in the reduction of all-cause
mortality and all-cause mortality or cardiovascular hospitalizations.
The study started in September 2002 with the participation of 357 departments of cardiology
and nearby 7000 patients; 3494 patients received n-3 PUFA in a capsule once daily and 3481
received placebo; in a follow-up time of four years, 955 patients in the PUFA group (27%) died,
compared with 1014 (29%) in the placebo group, meaning a relative risk reduction of 9% in the
PUFA group. A higher proportion of patients in the placebo group (2053/59%) died or were
admitted to hospital for cardiovascular reasons than in the PUFA group (1981/57%) a relative
reduction of 8% in the PUFA group. In absolute terms, 56 patients needed to be treated with
PUFA for just under four years to avoid one death, or 44 patients to avoid one event of either
death or admission to hospital for cardiovascular causes. Statin treatment with rosuvastatin did
not affect clinical outcomes in patients with chronic heart failure.
Several substudies focus on possible mechanistic effects of the study treatments: ventricular
remodeling (echo); biohumoral; genetic; arrhythmic and autonomic pattern (Holter monitoring);
exercise capacity; cognitive function; burden of care. The analysis of these results are in
progress.
The project is conducted in collaboration with the Laboratory of Cardiovascular Clinical Pharmacology.
GISSI-HF Genetic Substudy
The role of genetic factors in causes, evolution, prognosis and treatment of heart failure is
largely unexplored, with the exception of heart failure originated by specific cardiomyopathies
(such as dilated, hypertrophic, arrhythmogenic right ventricular cardiomyopathies), for which
the role of heritable gene mutations is increasingly well understood. Heart failure (HF) is a
syndrome with different etiologies, and more than one half is caused by coronary heart disease
(CHD). A genetic substudy to GISSI-HF (see "ad hoc" section) offers the opportunity to
improve knowledge on the role of genetic factors involved in heart failure, through a collection
of blood samples of a large population of patients, involving cases of heart failure of different
etiologies, i.e. non-ischaemic and ischaemic heart disease.
The objective of the genetic substudy is 1) to assess the relationships between the
polymorphysms of various candidate genes and the clinical outcome in patients enrolled in
GISSI-HF study; 2) to assess whether these relationships are modified by the experimental
treatments.
The genetic markers to be studied will initially focus on polymorphisms of genes involved in
lipid metabolism and inflammation. However, as new genetic information is being accumulated
continually, at present it is not known if other polymorphisms/genes will be important at the
time of study completion. The possibility to assay other allelic variants is then foreseen. The
GISSI-HF genetic substudy is conducted by nearly 100 Centres that have included 2200
patients.
GISSI-Prevenzione-Genetic Study
Myocardial infarction is a multifactorial disease. While the role of known risk factors on
coronary heart disease susceptibility is well defined, the impact of the genetic components and
147
ANNUAL REPORT 2008

IRFMN
its interaction with environmental factors need investigation. The GISSI-Prevenzione trial
investigated the effects of pharmacological treatments with n-3 PUFA and pravastatin on
morbidity and mortality after myocardial infarction. During the study more than 8000 samples
of a large population of patients affected by this disease has been collected and stored with the
collaboration of SIBioC (Societê Italiana di Biochimica Clinica e Biologia Molecolare). The
GISSI-Prevenzione-Genetic Study investigates the role of genetic factors in ischaemic heart
disease. The objectives of the project are 1) to assess the relationships between the
polymorphysms of various candidate genes and the clinical outcome in patients enrolled in the
large clinical trial GISSI-Prevenzione study; 2) to assess whether these relationships are
modified by the pharmacological treatments. According to these objectives, we investigated the
relationship between APOE, mortality and the response to treatment in 3304 myocardial
infarction survivors randomized to pravastatin or no treatment. We found that epsilon 4 allele is
a determinant of pravastatin response in terms of survival. Though in the entire population
investigated we found a beneficial effect of pravastatin in terms of survival, only the epsilon 4
carriers seemed to have gained a significant benefit from this treatment. We suggest that the
effect of statins is of particular interest in this fraction of the population and that genetic markers
can help in identifying patients that benefit more from statin treatment.
Association studies in the same population on the adiponectin gene, the CRP (C-reactive
protein) gene variants, some genetic variants on Chromosome 9p21 are in progress.
GISSI-AF. Clinical trial testing the efficacy of an angiotensin II AT1-
receptor blocker in Atrial Fibrillation
The GISSI-AF is a randomized, prospective, parallel group, placebo-controlled, multicenter
study on the use of valsartan, an angiotensin II AT1-receptor blocker in the prevention of Atrial
Fibrillation (AF) recurrence.
Atrial fibrillation is the most frequent form of arrhythmia in clinical practice, affecting 6% of
people over 65 years old. The traditional therapies are often able to restore the sinus rhythm but
are subject to a very high percentage of relapses, above all when the AF has been present for a
long time and when there are structural changes at both atrial and ventricular level. The reninangiotensin-aldosterone
system (RAAS) plays a key role in the “remodeling” phenomenon,
which makes increasingly irreversible the electrical, mechanical and structural properties of the
atrial tissue. Existing experimental and clinical data do not allow any definite conclusion
regarding the efficacy of an angiotensin II AT1-receptor blocker in the prevention of AF. The
GISSI group decided to conduct a specific trial of adequate size versus placebo aimed to assess
if the addition of valsartan on top of established therapies can reduce the recurrence of atrial
fibrillation in patients with a history of recent AF associated with cardiovascular
diseases/comorbidities. The study has recruited and treated for 12 months 1400patients with a
history of recent AF associated with cardiovascular diseases/comorbidities. In GISSI-AF, the
largest trial ever conducted with ARBs in pts with AF, Valsartan did not reduce AF recurrence.
A trend favoring valsartan was observed only in the small group of pts with heart failure and/or
left ventricular dysfunction.
The project was conducted in collaboration with the Laboratory of Cardiovascular Clinical
Pharmacology.
NeuroMorfeo Study: see Laboratory of Cardiovascular Clinical Pharmacology
The Population Health Research Institute (PHRI), McMaster University, Hamilton, Ontario, is
the coordinating center of a multinational network of cardiology clinics that collaborate to the
conduction of multicenter large scale clinical trials (nearly 40 Countries and more than 600
cardiology clinics). The Laboratory of Clinical Drug Evaluation is responsible for the scientific
coordination in Italy of some of these trials (ACTIVE, RE-LY, CURRENT, FUTURA OASIS-
8).
148
ANNUAL REPORT 2008

IRFMN
ACTIVE study (Atrial Fibrillation Clopidogrel Trial with Irbesartan for
prevention of Vascular Events)
The aims of the study were to evaluate whether clopidogrel plus acetylsalicylic acid (ASA) is
superior to ASA alone (A study) and non-inferior to standard oral anticoagulant therapy (W
study) in preventing vascular events in patients with atrial fibrillation and to evaluate whether
blood pressure lowering with irbesartan is superior to placebo in preventing vascular events in
patients with atrial fibrillation (I study). The primary efficacy outcome is the first occurrence of
stroke, myocardial infarction, vascular death over the duration of follow-up, (a minimum of 2
and maximum of 4 years approximately). A sample size of 14000 patients was planned, 6500 in
the W study (testing the efficacy of warfarin vs ASA + clopidogrel) and 7500 in the A study
(testing the efficacy of ASA + clopidogrel vs ASA alone). The W study was stopped early by
the Data and Safety Monitoring Board in September 2005 after an interim analysis showing a
significant difference in favor of warfarin over the combination of ASA + clopidogrel. The
details of the ACTIVE W have been published (Lancet 2006; 367:1903-1912). The A and the I
studies are continuing and final results are expected for 2009.
RE-LY study (Randomized Evaluation of Long term anticoagulant therapY)
Non-valvular atrial fibrillation is implicated in nearly 15% of strokes. Dose-adjusted warfarin
decreases the risk of stroke by 62%. However, in practice, the risk of bleeding, the variability of
anticoagulation intensity and the need of frequent monitoring and dose adjustments limits
treatment with warfarin, leaving patients outside the therapeutic range almost half the time.
Underuse of warfarin in patients with atrial fibrillation at high risk of bleeding calls for safer,
more reliable alternatives. The direct thrombin inhibitor dabigatran could offer fixed oral dosing
without need for coagulation monitoring, rapid onset and offset of action, stable
pharmacokinetics with little potential for drug interactions, and no known food interactions. For
these reasons an international multicentre, randomized, active controlled, parallel group, noninferiority,
clinical trial (RE-LY study) was designed to evaluate the efficacy and safety of
dabigatran etexilate compared with open label adjusted warfarin for the prevention of stroke and
systemic embolism in patients with non-valvular atrial fibrillation. The recruitment of patients
was completed and the long-term follow-up is ongoing. Final results are expected for 2009.
CURRENT OASIS-7 study
OASIS 7 is a randomized, multinational, 2X2 factorial design, parallel-group, double-blind
study, comparing a high loading dose regimen of clopidogrel versus standard dose and high
dose regimen of aspirin versus standard dose, in patients with acute coronary syndrome
managed with an early invasive strategy. The study will involve approximately 25,000 patients
in 800 clinical centers worldwide. The primary objective of the study is to determine whether a
high dose regimen of clopidogrel is superior to a standard dose of clopidogrel in preventing CV
death, myocardial infarction or stroke and to determine if high dose of aspirin is as safe as low
dose in terms of TIMI major bleeding rate. Secondary objective is to evaluate the safety of the
clopidogrel high dose regimen compared to the standard dose regimen in terms of TIMI major
bleeding. Final results are expected for 2009.
INTER-HEART Study
INTER-HEART is an epidemiological study sponsored by the World Health Organization and
the World Heart Federation aimed to determine the association between risk factors and acute
myocardial infarction within populations defined by ethnicity and/or geographic region, and to
assess the relative importance of risk factors across these populations. The project is coordinated
149
ANNUAL REPORT 2008

IRFMN
by the PHRI, McMaster University, Hamilton, Canada. The Clinical Drug Evaluation
Laboratory serves as National Coordination Office for Italy.
INTER-HEART was conducted in 52 countries in Asia, Europe, Middle East, Africa, Australia,
and North and South America, utilizing a standardized protocol. The study evaluated the
importance of conventional and emerging risk factors within each geographic region, and
whether their impact varies by region in a population of nearly 28000 individuals (cases with
acute myocardial infarction and matched controls). The study collected data on demographic
factors (country of origin, first language), socioeconomic status (education, occupation,
income), lifestyle (tobacco use, physical activity, dietary patterns), and personal and family
history of cardiovascular disease and risk factors.
The INTER-HEART has documented that nine simple modifiable risk factors can account for
over 90% of the population attributable risk for heart disease globally. More importantly, these
risk factors appear to have similar predictability in all regions of the world, as well as in all
ethnic groups. A further analysis published in the 2005 redefined obesity: traditional definitions
have been based on Body Mass Index (BMI), but this paper very clearly showed that,
irrespective of BMI, the waist-to-hip ratio (WTHR) is a far better predictor of myocardial
infarction risk across diverse populations. The observations carry important implications for
people and populations hitherto considered to be low risk on the basis of their BMIs.
In a further analysis, the INTERHEART study established that all forms of tobacco exposure,
such as smoking, chewing tobacco or inhaling second hand smoke, increase the risk of heart
attack. Compared to people who had never smoked, smokers had a three-fold increased risk of a
heart attack. An ad hoc assessment of the gender influence, comparing risk factors for acute
myocardial infarction (MI) between women and men globally, the INTER-HEART showed that
women experience their first acute MI on average 9 years later than men. Nine modifiable risk
factors are significantly associated with acute MI in both men and women and explain greater
than 90% of the PAR. The difference in age of first MI is largely explained by the higher risk
factor levels at younger ages in men compared to women. The approach to prevention of MI in
men and women can be based on similar principles in all regions of the world.
Laboratory of General Practice Research
Risk and Prevention Study (R&P)
R&P is a study on the optimization of cardiovascular prevention of subjects at high risk
performed at national level by General Practitioners.
Study objective and design
- Controlled clinical trial, double-blind and randomised, of the efficacy of a n-3 PUFA treatment
in reducing the incidence of cardiovascular events, both fatal and non-fatal, in a population
defined as at high risk by participating GPs.
- Practicability and overall yield of the preventive interventions adopted (outcome study) The
epidemiological and care history of this population shall form the object of a specific evaluation
according to a plan of formal predefined analyses.
Study population
Inclusion criteria
Among the subjects deemed by GPs to be at high cardiovascular risk, patients are selected if
presenting:
- multiple risk factors (e.g. hypertension, hypercholesterolemia, diabetes, smoking, family
history of myocardial infarction, obesity, sex and old age)
- previous cardio-cerebrovascular events or clinical manifestations of atherosclerotic disease
(stroke, TIA, peripheral arteriopathy, previous arterial revascularisation procedures, angina
pectoris).
150
ANNUAL REPORT 2008

IRFMN
Exclusion criteria
- serious co morbidity with an unfavorable prognosis over the short term (e.g. cancer);
- expected non-compliance over a long period of time; contraindications (known allergies to n-
3PUFA)
- indications (previous MI) for treatment with n-3 PUFA.
Efficacy measures
The primary objective is to evaluate if a long-term administration of n-3 PUFA is more effective
than the corresponding placebo in reducing cardiovascular mortality and hospitalization for
cardiovascular causes. Randomisation is central, stratified by GP.
The experimental treatment consists of one capsule containing 1g of n-3 PUFA, or the
corresponding placebo, to be taken daily.
The duration of follow-up is 5 years.
In order to document with sufficient statistical reliability that the experimental treatment with n-
3 PUFA reduces of 15% the incidence of the events considered in the primary end-point, a total
of 12,000 patients is required.
Up-date of the study: From February 2004 to March 2007 12,521 patients have been enrolled
by a network of 860 GPs. The Local Health Authorities involved are 57 and in each one
investigator’s meeting has been organized. The characteristics of the population so far enrolled
are the following: mean age 65 years, males 62%, hypertension 79%, hypercholesterolaemia
62%, diabetes 56%, smokers 16%, obesity 35%, family history of premature myocardial
infarction 20%. Twenty five% of patients have a clinical manifestation of atherosclerotic
diseases, 50% have diabetes in association with another risk factor and 23% have multiple risk
factors. Currently the mean follow-up is 2.5 years. The observed incidence of cardiovascular
events (cardiovascular mortality and hospitalizations for cardiovascular causes) in this period
was 6%. Five % of patients prematurely withdrawn from the trial and 2.5% of patients
discontinued treatment because of side effects (mainly gastrointestinal) that disappeared after
discontinuation.
After 2 years of follow-up in patients with high blood pressure a statistically significant
reduction of blood pressure was observed , as well as significantly reduction of LDL levels in
patient with high cholesterol. During this time 20% of smokers stopped smoking. On the
contrary no changes were observed in glycated hemoglobin level in diabetic patients, and in
BMI in obese patients.
This data suggest that GPs’ interventions have a clinically relevant impact on risk factors
control and
life styles habits of their patients.
More information available on the website www.rischioeprevenzione.it.
The “attributability”of cardiovascular events in the GP’s perception
A cardiovascular event is the sudden manifestation - more or less expected - of clinical history,
life-style habits, social environment, patient and medical attitudes and health strategies adopted.
To carefully examine the reasons why a cardiovascular event occurred is part of a good care in
the context of general practice. To explicitate this process, for all the representative cases that
occurred during a defined period of care could transform a usual attitude of good clinical
practice into an “epidemiology of the attributability”.
The aim of this study is to evaluate the “epidemiology of the attributability” of cardiovascular
events through GPs’ perception. Clinical, socio-economic, psychological, risk factors and lifestyle
variables were investigated. The study started in October 2007and it has been concluded
in October 2008. 74 GPs were actively involved in study and have reported at least one event.
Overall 248 cases have been included during one year.
The events were expected by GPs in the 66% of the cases, while in 1/3 were not expected. The
majority of patients were known to GP and acute myocardial infarct was the event that occurred
151
ANNUAL REPORT 2008

IRFMN
more frequently. The relevant “area” most often reported by GPs were: clinical history, lifestyle
and the psychological factors.
The patients for whom the cardiovascular event was expected, were older, known by GP and
had more clinical disease in comparison with those for whom the event was not expected.
GPs have reported their consideration on the event in the 58% of the cases; particularly for the
younger patients with extra-clinical problems and when the cardiovascular event was not
expected.
It can be concluded that GPs, in their perception on the “attributability” of the events, taken
into account not only patient’s history (clinical and extra-clinical) but also their relationship
with the patient and often they feel that they “haven’t done in half” to avoid the event.
Epidemiological and clinical profile of diabetic patients in Lombardy
Region using administrative databases.
The study is part of an ongoing pharmacoepidemiological project in collaboration with the
Health Department of the Lombardy Region. Its main objective is the definition of a model to
assess and control the use of health resources of diabetic patients by means of integrated
administrative database.
Specific aims of the study are:
• To identify the diabetic population by specific drug consumption (ATC group = A10)
• To assess the co morbidities by co-administered cardiovascular and non-cardiovascular drugs
• To describe the diagnostic procedures by the assessment of laboratory, instrumental and
specialist examinations
• To estimate the rate and causes of hospitalization by the analyses of hospital admission data
Study population
Diabetic patients have been identified on the basis of the specific diabetic drugs. A first pilot
study has been carried out on the data of the ASL of Bergamo, testing also the feasibility of a
case-control study. Those resident that during year 2005 have received at least a prescription of
an antidiabetic drug (oral insulin or hypoglycaemic drugs, A10 group of ATC classification)
have been considered as diabetic. They then have been followed in the course of the year,
linking the registry office database with those of the prescription’s and the Hospital
Admission’s (HD) ones. The sample consisted of 1.043.411 inhabitants (10,9% of all the
Lombardy Region population). The prevalence of drug-treated diabetes is 4.1% (42,618
subjects); the number of patients treated with antidiabetic drugs increases with increasing of the
age, till to reach 15% in those > 75 years. The oral antidiabetics associations (OAD) are more
frequently prescribed followed by sulphanylureas, insulins alone and the biguanides. The
association between insulins and AOD was prescribed to 10,3% of diabetics. 80% of the
diabetic patients received at least a cardiovascular drug. Of the 42,618 diabetic ones, 12,057
(28,3%) have been hospitalized at least once during the same year.
Case-control study
Case-control methodology has been applied in order to estimate the use of health resources of
the cases compared to the control group. The cases are diabetic patients, having more than 50
years. The controls are persons without diabetes, matched to the cases for age, sex and for being
in charge to the same general practitioner. As expected, the cases have a greater pharmacologic
burden and a greater hospitalization rate. To represent the clinical complexity of the diabetic
patients opposed to the controls, co-morbidity diabetes-correlated hospitalizations were used as
indicators, in particular: acute myocardial infarction, coronary disease, heart failure,
cerebrovascular disease and chronic renal failure. The results show that all these events are
much more frequent in the diabetic patients than in the control group.
152
ANNUAL REPORT 2008

IRFMN
The stratification of global cardiovascular risk in hypertensive patients of
the district of Borbon – Ecuador
The Laboratory is involved in a collaborative project with the Cecomet (Centro de
Epidemiologia comunitaria y Medicina tropical) in Esmeralda, Ecuador, on the prevalence and
treatment of hypertension in the district of Borbon, a rural zone of Ecuador in the northern part
of the country.
In this area, 36% of the adult population is affected by hypertension and more than half of
hypertensive patients present blood pressure levels > 160/110 mmHg.
From 2001, in the District is ongoing an intensive follow-up of the hypertensive population with
the following aims: to evaluate the global cardiovascular risk of the population, to better control
blood pressure levels increasing the number of subjects treated with hypertensive therapy (in
particular those at high cardiovascular risk) and monitoring of the clinical complications.
Preliminary data show that:
• Patients treated with hypertensive therapy are increased from 39% to 59%
• Antihypertensive drugs are mainly prescribed to subjects with high blood pressure levels
(80% of those with systolic blood pressure >180mmHg are actually under treatment) or at
high cardiovascular risk (82%)
• Blood pressure control is improved (patients with systolic blood pressure levels
> 180mmHg decreased from 33% to 24% and those with levels

IRFMN
specific attention to drug-related problems. In fact an adverse drug reaction is likely to be the
cause of most unexpected problems that arise in elderly patients.
The project has been completed and more than 350 nurses have been involved in 95 Nursing
Homes and Districts. In the 6 observation days 1500 patients were observed. Overall 2224
problems were identified and 25% attributed to drugs and devices.
154
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF MOLECULAR
BIOCHEMISTRY AND
PHARMACOLOGY
STAFF
Head
Mario SALMONA, Sci.Prep.Alim.D., Ph.D.
Laboratory of Biochemistry and Protein Chemistry
Head
Mario SALMONA, Sci.Prep.Alim.D.,Ph.D.
Medical Biochemistry Unit
Head
Valentina BONETTO, Chem.Pharm.D.
Synaptic Transmission Unit
Head
Marco GOBBI, Pharm.D.
Laboratory of Molecular Biology
Head
Enrico GARATTINI, M.D.
Pharmacogenomics Unit
Head
Maddalena FRATELLI, Biol.Sci.D.
Gene Structure and Regulation Unit
Head
Mineko TERAO, Bioch.D., Ph.D.
Laboratory of Receptor Pharmacology
Head
Tiziana MENNINI, Pharm.D.
Laboratory of Neuroimmunology
Head
Pietro GHEZZI, Ph.D.
Pharmacology of Septic Shock Unit
Head
Pia VILLA, Pharm.D.
Metabolic Neuropathies Unit
Head
Roberto BIANCHI, Biol.Sci.D.
Laboratory of Molecular Pathology
Head
Lavinia CANTONI, Biol.Sci.D.
Laboratory of Systems Biology
Head
Gianfranco BAZZONI, M.D.
155
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Mario Salmona obtained his doctorate degree in Biochemistry and Food Technology at the University of
Milan in 1971. His background is in biochemistry, biophysics and pharmacology. His scientific interests
relate to problems of human and animal diseases originating from the aberrant folding of proteins. In this
context, a major portion of his studies was devoted to the etiopathogenesis and therapy of prion diseases.
He has published over 200 articles on peer reviewed scientific journals.
1971-1975 Ph.D in Pharmacology, Mario Negri Institute
1975 Visiting Fellow in the Department of Biology of the Weizmann Institute of Science, Rehovot, Israel
1976-1997 Head, Laboratory of Enzyme Research, Mario Negri Institute
1995 to date Dean of the School of Advanced Pharmacology, Mario Negri Institute
1997 to date Head, Department of Biochemistry and Molecular Pharmacology, Mario Negri Institute
2003 to date Member of the American Society of Biochemistry and Molecular Biology
Referee for international scientific journals
Selected publications
• Saracino GA, Villa A, Moro G, Cosentino U, Salmona M.
Spontaneous beta-helical fold in prion protein: The case of PrP(82-146).
Proteins. 2008 Oct 29. [Epub ahead of print]
• Massignan T, Casoni F, Basso M, Stefanazzi P, Biasini E, Tortarolo M, Salmona M, Gianazza E, Bendotti C, Bonetto V.
Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1 mouse.
Biochem Biophys Res Commun. 2007; 353:719-25
• De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Limido L, Forloni G, Tagliavini F,
Salmona M.
The efficacy of tetracyclines in peripheral and intracerebral prion infection.
PLoS ONE. 2008;3(3):e1888
• Cosentino U, Pitea D, Moro G, Saracino GA, Caria P, Varì RM, Colombo L, Forloni G, Tagliavini F, Salmona M.
The anti-fibrillogenic activity of tetracyclines on PrP 106-126: a 3D-QSAR study.
J Mol Model. 2008 14: 987-94
• Fioriti L, Angeretti N, Colombo L, De Luigi A, Colombo A, Manzoni C, Morbin M, Tagliavini F, Salmona M, Chiesa R,
Forloni G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-Sträussler-Scheinker
disease amyloid protein.
J Neurosci. 2007; 27:1576-83
• Gobbi M, Colombo L, Morbin M, Mazzoleni G, Accardo E, Vanoni M, Del Favero E, Cantù L, Kirschner DA, Manzoni
C, Beeg M, Ceci P, Ubezio P, Forloni G, Tagliavini F, Salmona M.
Gerstmann-Sträussler-Scheinker disease amyloid protein polymerizes according to the "dock-and-lock" model.
J Biol Chem. 2006; 281:843-9
Gianfranco Bazzoni got his Medicine and Surgery degree in 1988 (at the University of Milan) and the
specialisation in Pharmacological Research in 1992 (at the Mario Negri Institute, Milan). His area of
expertise is cell biology, with focus on the processes of cell adhesion and migration.
1988-2000 Research Fellow, Mario Negri Institute
1993-1997 Post-doctoral Fellow, Dana Farber Cancer Institute and Harvard Medical School, Boston, MA
2000-2002 Research Scientist, Mario Negri Institute
2003 Head, Unit of Cell Adhesion, Mario Negri Institute
2004 to date Head, Laboratory of Systems Biology, Mario Negri Institute
2004 Regular Member of The American Physiological Society, Bethesda, MD
Referee for international scientific journals
Selected publications
• Paris L, Bazzoni G The protein interaction network of the epithelial junctional complex: a system-level analysis Mol
Biol Cell 2008 19: 5409-5421
• Paris L, Tonutti L, Vannini C, Bazzoni G. Structural organization of the tight junction. Biochim Biophys Acta 2008
1778: 646-659
• Huang H, Cruz F, Bazzoni G. Junctional adhesion molecule-A regulates cell migration and resistance to shear stress. J.
Cell Physiol 2006; 209; 122-130.
• Martinez-Estrada OM, Manzi L, Tonetti P, Dejana E, Bazzoni G. Opposite effects of Tumor Necrosis Factor and soluble
fibronectin on Junctional Adhesion Molecule-A in endothelial cells. Am J Physiol (Lung Cell Mol Physiol) 2005; 288:
L1081-L1088.
• Bazzoni G, Tonetti P, Manzi L, Cera MR, Balconi G, Dejana E. Expression of Junction Adhesion Molecule-A prevents
spontaneous and random motility. J Cell Sci 2005; 118: 623-632.
156
ANNUAL REPORT 2008

IRFMN
• Bazzoni G, Dejana E. Endothelial cell-to-cell junctions: molecular organization and role in vascular homeostasis. Physiol
Rev 2004; 84(3): 869-901.
Lavinia Cantoni obtained her degree in Biological Sciences in 1973 at the University of Milan. Then
she specialized in pharmacological research at the Mario Negri Institute (1974-1977).
Research areas 1) biochemical-molecular mechanisms activated by oxidative stress 2) drug metabolism
3) porphyrias.
1977-1978 Post-doctoral Fellow, Medical Research Council, Toxicology Unit, Carshalton, UK
(Winner of a Welcome Trust Research Fellowship)
1979-1982 Research Scientist, Mario Negri Institute
1980-1990 Visiting Scientist, Toxicology Unit, Carshalton, UK, and Cornell Medical Center, New
York, NY (short periods)
1983-1998 Head, Unit of Heme and Hemoprotein Metabolism, Mario Negri Institute.
1998 to date, Head, Laboratory of Molecular Pathology, Mario Negri Institute.
Member of the National Roll of Biologists and of the Italian Toxicology Society.
Referee for international scientific journals, tutor for university degree thesis and teacher in the
Pharmacological Research Specialisation Course for graduates held at the Mario Negri Institute.
Selected publications
• Rizzardini M., Chiesa R., Angeretti N., Lucca E., Salmona M., Forloni G., Cantoni L. Prion protein fragment 106-126
differentially induces heme oxygenase-1 mRNA in cultured neurons and astroglial cells. J. Neurochem. 68: 715-720,
1997
• Rizzardini M., Zappone M., Villa P, Gnocchi P., Sironi M., Diomede L., Meazza C., Monshouwer M., Cantoni L.
Kupffer cell depletion partially prevents hepatic heme oxygenase 1 messenger RNA accumulation in systemic
inflammation in mice: role of interleukin 1 beta. Hepatology 27: 703-710, 1998
• Cantoni L.,Valaperta R., Ponsoda X., Castell, J.V., Barelli D., Rizzardini M., Mangolini A., Hauri L., Villa P. Induction
of hepatic heme oxygenase-1 by diclofenac in rodents: role of oxidative stress and cytochrome P-450 activity. J.
Hepatology 38: 776-783, 2003
• Babetto E., Mangolini A., Rizzardini M., Lupi M., Conforti L., Poletti A., Rusmini P., Cantoni L. Tetracycline-regulated
gene expression in the NSC-34-tTA cell line for investigation of motor neuron diseases. Mol. Brain Res. 140: 63-72,
2005
• Rizzardini M., Lupi M., Mangolini A., Babetto E., Ubezio P., Cantoni L. Neurodegeneration induced by complex I
inhibition in a cellular model of familial amyotrophic lateral sclerosis. Brain Res. Bull. 69: 465-474, 2006
• Raimondi A., Mangolini A., Rizzardini M., Tartari S., Massari S., Bendotti C., Francolini M., Borghese N., Cantoni L.,
Pietrini G. Cell culture models to investigate the selective vulnerability of motoneuronal mitochondria to familial ALSlinked
G93ASOD1. Eur. J. Neurosci. 24: 387-399, 2006
Enrico Garattini obtained his degree in Medicine and Surgery with full marks (110/110) in 1982 at the
University of Milan. His scientific interests relate to problems of Cellular Biology and Molecular
Biology.
1982-1990 Research Fellow of the National Research Council, Mario Negri Institute
1983-1987 Postdoctoral Researcher at the Roche Institute of Molecular Biology, Department of
Neurosciences Nutley, New Jersey, US
1991-1997 Senior Researcher Regione Lombardia and Head of the Molecular Biology Unit, Mario Negri
Institute
1997 to date Head, Laboratory of Molecular Biology, Mario Negri Institute
From 2005 Dean, Advanced School of Pharmacology (Philosophy Doctor), Mario Negri Institute
Member of the Editorial Board of the European Journal of Cancer and of Current Cancer Therapy
Reviews
Member of the American Society of Biochemistry and Molecular Biology (ASBMB)
Selected publications
• Gianni M, Boldetti A, Guarnaccia V, Rambaldi A, Parrella E, Raska I Jr, Rochette-Egly C, Del Sal G, Rustighi A, Terao
M, Garattini E
Inhibition of the peptidyl-propyl-isomerase Pin1 enhances the responses of acute myeloid leukemia cells to retinoic acid
via stabilization of RARα and PML-RARα. Cancer Res. 2008, in press
• Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A, Tiveron C,
Garattini E
Role of the molybdo-flavoenzyme, aldehyde oxidase homolog 2, in the biosynthesis of retinoic acid: generation and
characterization of a knock-out mouse, Mol Cell Biol 2008 [Epub ahead of print]
• Gianni M, Parrella E, Raska I Jr, Gaillard E, Nigro EA, Gaudon C, Garattini E, Rochette-Egly C. P38MAPK-dependent
phosphorylation and degradation of SRC-3/AIB1 and RARalpha-mediated transcription. EMBO J. 2006 Feb
22;25(4):739-51
157
ANNUAL REPORT 2008

IRFMN
• Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I,
Carminati P, Terao M, Pisano C. ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in
myeloid leukemia cells: Modulation of intracellular calcium homeostasis. Blood 2004; 103: 194-207
• Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene
cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with
selective expression in the olfactory mucosa. J Biol Chem 2004; 279: 50482-50498
• Pisano C, Kollar P, Gianni M, Kalac Y, Giordano V, Ferrara F F, Tancredi R, Devoto A, Rinaldi A, Rambaldi A, Penco
S, Marzi M, Moretti G, Vesci L, Tinti O, Carminati P, Terao M, Garattini E. Bis-indols a novel class of molecules
enhancing the cytodifferentianting properties of retinoids in myeloid leukemia cells. Blood 2002; 100: 3719-3730
Pietro Ghezzi
Research Areas: Cytokines and inflammation; redox regulation
1979-1990: Researcher, Mario Negri Institute
1991 to date: Head, Laboratory of Neuroimmunology, Mario Negri Institute
1998-2000: Research Associate at Stanford University School of Medicine, Department of Genetics
2000 to date: Member, Kenneth Warren Laboratory, Ossining, NY (USA)
Referee for international scientific journals
Selected publications
• Leist M, Ghezzi P, et al.. Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science. 2004 Jul
9;305(5681):239-42.
• Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi Gene expression profiling reveals a
signaling role of glutathione in redox regulation.Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13998-4003.
• Villa P, Bigini P, Mennini T, Agnello D, Laragione T, Cagnotto A, Viviani B, Marinovich M, Cerami A, Coleman TR,
Brines M, Ghezzi P. Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia
by targeting neuronal apoptosis. J Exp Med. 2003 Sep 15;198(6):971-5.
• Siren AL, Fratelli M, Brines M, Goemans C, Casagrande S, Lewczuk P, Keenan S, Gleiter C, Pasquali C, Capobianco A,
Mennini T, Heumann R, Cerami A, Ehrenreich H, Ghezzi P. Erythropoietin prevents neuronal apoptosis after cerebral
ischemia and metabolic stress. Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4044-9.
• Laragione T, Bonetto V, Casoni F, Massignan T, Bianchi G, Gianazza E, Ghezzi P. Redox regulation of surface protein
thiols: identification of integrin alpha-4 as a molecular target by using redox proteomics. Proc Natl Acad Sci U S A.
2003 Dec 9;100(25):14737-41.
Tiziana Mennini got her degree in Pharmacy at the University of Milano (1975). In the same year she
obtained a fellowship from the European Molecular Biology Organization, to learn sub-cellular
fractionation techniques and synaptosomes utilization in neurochemistry, at the laboratory of Prof. VP
Whittaker ( Stockholm, Sweden). In 1882 she spent a further period in Prof. Whittaker’s laboratories
(Max-Plank-Institut fur Biophysikalische Chemie, Abteilung Neurochemie Am Fassberg, Gottingen,
Germany). She spent all her scientific career at the Mario Negri Institute:
1967- 1975 Research Assistant in the Laboratory of Drug Metabolism
1975-1987 Chief of the Unit of Neurochemical Transmission,
1988 to date Chief of the Laboratory of Receptor Pharmacology
Speaker, chairman and organizer at many congresses and courses, author of more than 200
articles published in international journals in the area of receptor pharmacology and
neuropharmacology.
Selected publications
• Beghi E, Bendotti C, Mennini T. 2005. Merits of a new drug trial for ALS Science 308:632-633
• Gobbi M, Mennini T. 2001. Is St John's wort a 'Prozac-like' herbal antidepressant Trends Pharmacol Sci 22:557-559.
• The Italian ALSSG. 1996. Ceftriaxone in amyotrophic lateral sclerosis. Eur J Neurol 3:295-298.
• Mennini T, Mocaer E, Garattini S. 1987. Tianeptine, a selective enhancer of serotonin uptake in rat brain. Naunyn-
Schmiedebergs Arch Pharmacol 336:478-482.
• Mennini T, Garattini S. 1983. Benzodiazepines receptor binding in vivo: pharmacokinetic and pharmacological
significance. Advances Biochemical Psychopharmacol 38:189-199.
• Mennini T, Bernasconi S, Manara L, Samanin R, Serra G. 1977. The effect of intracerebral 6-hydroxy dopamine on 3Hreserpine
binding to different brain regions of the rat. Pharmacol Res Commun 9:857-862.
Roberto Bianchi got his degree in Biological Sciences at University of Milan, Italy in 1992. Since 1975
he served as student teacher and supervisor at Mario Negri Institute and from 1989 to 1997 at Houston
University. His main interests are diabetic complications (peripheral and autonomic neuropathies) and
neurodegenerative disorders (Multiple Sclerosis, drugs induced neuropathies).
1971 Technician in the Laboratory of Biochemical Pharmacology, Mario Negri Institute
158
ANNUAL REPORT 2008

IRFMN
1980-1981 Visiting Scientist in the Center for Neurosciences Behavioral Research, The Weizmann
Institute of Science, Rehovot, Israel
1981-1995 Research Assistant in the Laboratory of Biochemical Pharmacology, Mario Negri Institute
1988-1997 Research Fellow in the Dept. Biochemical Biophysical Sciences, University of Houston, US
(3 mo. a year)
1993-1995 Research Fellow in the Dept. Medicine, Case Western Reserve University, Cleveland, US (1
mo. a year)
Since 1996 Head, Unit of Metabolic Neuropathies, Mario Negri Institute
Selected publications
• Bianchi R., Berti-Mattera L.N., Fiori M.G., Eichberg J.:Correction of altered metabolic activities in sciatic nerves of
streptozotocin-induced diabetic rats. Effects of ganglioside treatment. Diabetes 39: 782-788 (1990).
• Scarpini E., Bianchi R., Moggio M., Sciacco M., Fiori M.G., Scarlato G.: Decrease of nerve Na+,K+-ATPase activity in
the pathogenesis of diabetic neuropathy. J. Neurol. Sci. 120: 159-167 (1993).
• Conti G., Scarpini E., Baron P.L., Livraghi S., Tiriticco M., Bianchi R, Vedeler C., Scarlato G.: Macrophage infiltration
and death in the nerve during the early phases of experimental diabetic neuropathy: a process concomitant with
endoneurial induction of IL-1 and p75NTR. J. Nuerol. Sci. 195: 35-40 (2002)
• Bianchi R., Buyukakilli B., Brines M., Savino C., Cavaletti G., Oggioni N., Lauria G., Borgna M., Lombardi R., Cimen
B., Comelekoglu U., Kanik A., Tataroglu C., Cerami A., Ghezzi P. Erythropoietin both protects from and reverses
experimental diabetic neuropathy. Proc Natl Acad Sci USA 101: 823-828 (2004)
• Leist M., Ghezzi P., Grasso G, Bianchi R., Villa P., Fratelli M., Savino C., Bianchi M., Nielsen J., Gerwien J., Kallunki
P., Larsen A.K., Helboe L., Christensen S., Pedersen L.O., Nielsen M., Troup L., Sager T., Sfacteria A., Erbayktar S,
Erbayktar Z., Gokmen N., Yilmaz O., Cerami-Hand C., Xie, Q-W., Coleman T., Cerami A., Brines M. Erythropoietinderived
tissue-protective cytokines that do not bind to the classical erythropoietin receptor. Science, 305(5681) 239-242,
2004.
• Savino C., Pedotti R., Baggi F., Furlan R., Ubiali F., Gallo B, Nava S., Bigini P., Barbera S., Fumagalli E., Mennini T.,
Vezzani A., Rizzi M., Coleman T., Cerami A.,Brines M., Ghezzi P., Bianchi R.Delayed administration of erythropoietin
and its non-erythropoietic derivatives ameliorates chronic murine autoimmune encephalomyelitis. Journal of
Neuroimmunology, 2005, E-pub December 6, 2005
Valentina Bonetto has got the degree in Pharmaceutical Chemistry and Technology at the University of
Padua, Italy in 1993. She has got the Ph.D. in Medical Biochemistry and Biophysics at Karolinska
Institutet, Stockholm, Sweden.
Her principal lines of research are: 1) Study of the pathogenetic mechanisms at the basis of amyotrophic
lateral sclerosis (ALS); 2) Identification of biomarkers of ALS; 3) Role of the oxidative modification in
neurological disorders. These issues are investigated by different experimental approaches, including
proteomics and mass spectrometry.
Since 2000 she is at the Mario Negri Institute in the Laboratory of Biochemistry and Protein Chemistry,
from 2002 is also Assistant Telethon Scientist at Dulbecco Telethon Institute. Since October 2007 she is
the Head of the Unit of Medical Biochemistry inside the Laboratory of Biochemistry and Protein
Chemistry. She is author of 26 publications from 1994 to 2007, in peer-reviewed journals. Among them
she is first author in 11 and last author in 4. She is also author of 2 reviews. She is reviewer for scientific
journals in the field of Proteomics and Neuroscience.
Selected publications
• Massignan, T., Casoni, F., Basso, M., Stefanazzi, P., Biasini, E., Tortarolo, M., Salmona, M., Gianazza, E., Bendotti, C.,
Bonetto V. (2007) Proteomic analysis of spinal cord of presymptomatic amyotrophic lateral sclerosis G93A SOD1
mouse. Biochem. Biophys. Res. Commun., 353:719-25.
• Basso, M., Massignan, T., Samengo, G., Cheroni, C., De Biasi, S., Salmona, M., Bendotti, C., Bonetto, V. (2006)
Insoluble mutant SOD1 is partly oligoubiquitinated in amyotrophic lateral sclerosis mice. J. Biol. Chem., 281:33325-
33335.
• Ghezzi, P., Casagrande, S., Massignan, T., Basso, M., Bellacchio, E., Mollica, L., Biasini, E., Tonelli, R., Eberini, I.,
Gianazza, E., Dai, W.W., Fratelli, M., Salmona, M., Sherry, B., Bonetto. V. (2006) Redox regulation of cyclophilin A by
glutathionylation. Proteomics, 6: 817-825.
• Casoni, F., Basso, M., Massignan, T., Gianazza, E., Cheroni, C., Salmona, M., Bendotti, C., Bonetto, V. (2005) Protein
nitration in a mouse model of familial amyotrophic lateral sclerosis: Possible multifunctional role in the pathogenesis. J.
Biol. Chem., 280: 16295-16304.
• Laragione, T., Bonetto, V., Casoni, F., Massignan, T., Bianchi, G., Gianazza, E., Ghezzi, P. (2003) Redox regulation of
surface protein thiols: identification of integrin alpha-4 as a molecular target by using redox proteomics. Proc. Natl.
Acad. Sci. U S A 100: 14737-14741.
• Casagrande, S.* , Bonetto, V.*, Fratelli, M., Gianazza, E., Eberini, I., Massignan, T., Salmona, M., Chang, G., Holmgren,
A., Ghezzi, P. (2002) Glutathionylation of human thioredoxin: a possible crosstalk between the glutathione and
thioredoxin systems. Proc. Natl. Acad. Sci. U S A 99, 9745-9749. *These authors contributed equally to the study.
159
ANNUAL REPORT 2008

IRFMN
Maddalena Fratelli got her degree in Biological Sciences at the University of Pisa and at the Scuola
Normale Superiore di Pisa in 1983. Then the specialization in Pharmacological Research at the Mario
Negri Institute in 1986.
Her main fields of interest are: 1. High throughput genomic systems for the study of drug action and
pharmacoresistance. 2. Redox regulation of protein function and gene expression: glutathionylation and
gene expression profiling of glutathione dependent responses to oxidant challenge.
1988-1989 Postdoctoral Research Fellow in the Medical Research Council, Neurobiology Unit,
Cambridge, UK.
Since 1995, Head, Unit of Mediators of inflammation, Laboratory of Neuroimmunology, Mario Negri
Institute
Since 2005, Head, Unit of Pharmacogenomics, Laboratory of Molecular Biology, Mario Negri Institute
Selected publications
• Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi P. Gene expression profiling reveals
a signaling role of glutathione in redox regulation. Proc Natl Acad Sci U S A. 2005;102:13998-4003.
• Brines M, Grasso G, Fiordaliso F, Sfacteria A, Ghezzi P, Fratelli M, Latini R, Xie QW, Smart J, Su-Rick CJ, Pobre E,
Diaz D, Gomez D, Hand C, Coleman T, Cerami A. Erythropoietin mediates tissue protection through an erythropoietin
and common beta-subunit heteroreceptor. Proc Natl Acad Sci U S A. 2004; 101:14907-12.
• Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P,
Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S,
Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M. Derivatives of
erythropoietin that are tissue protective but not erythropoietic. Science. 2004; 305:239-42
• Fratelli M, Minto M, Crespi A, Erba E, Vandenabeele P, Del Soldato P, Ghezzi P. Inhibition of nuclear factor-kappaB
by a nitro-derivative of flurbiprofen: a possible mechanism for antiinflammatory and antiproliferative effect. Antioxid
Redox Signal. 2003; 5:229-35
• Fratelli M, Demol H, Puype M, Casagrande S, Eberini I, Salmona M, Bonetto V, Mengozzi M, Duffieux F, Miclet E,
Bachi A, Vandekerckhove J, Gianazza E, Ghezzi P. Identification by redox proteomics of glutathionylated proteins in
oxidatively stressed human T lymphocytes. Proc Natl Acad Sci U S A. 2002; 99:3505-10
• Siren AL, Fratelli M, Brines M, Goemans C, Casagrande S, Lewczuk P, Keenan S, Gleiter C, Pasquali C, Capobianco A,
Mennini T, Heumann R, Cerami A, Ehrenreich H, Ghezzi P. Erythropoietin prevents neuronal apoptosis after cerebral
ischemia and metabolic stress. Proc Natl Acad Sci U S A. 2001; 98:4044-9
Marco Gobbi got his degree in Pharmacy at the University of Milan, Italy, in 1989.
Currently, his main fields of interest are: 1) the study of presynaptic mechanisms, such as
neurotransmitter release/reuptake with a particular focus on plasma membrane transporters; and 2) the
study of protein misfolding and aggregation and in particular the characterization of the prion protein
amyloid formation, investigated by different approaches including surface plasmone resonance.
Since 1981, Researcher in the Laboratory of Neuropharmacology and, from 1988, in the Laboratory of
Receptor Pharmacology, Mario Negri Institute
Starting from 1989 Chief, Unit of Synaptic Transmission, Mario Negri Institute
In Jan 2006, his group joined the Laboratory of Biochemistry and Protein Chemistry, Mario Negri Institute
Co-author in more than 70 scientific publications on peer-reviewed international journals. First or last
author in more than 40 of them. Reviewer for international scientific journals operating in the
Neuroscience/Neuropharmacology fields.
Selected publications
• Gerstmann-Straussler-Scheinker disease amyloid protein polymerizes according to the "dock-and-lock" model. Gobbi M,
Colombo L, Morbin M, Mazzoleni G, Accardo E, Vanoni M, Del Favero E, Cantù L, Kirschner D A, Ceci P, Ubezio P,
Manzoni C, Forloni G, Tagliavini F, Salmona M J Biol Chem. 2006; 281: 843-9
• Funicello M, Conti P, De Amici M, De Micheli C, Mennini T, Gobbi M. 2004. Dissociation of [3H]L-glutamate uptake
from L-glutamate-induced [3H]D-aspartate release by 3-hydroxy-4,5,6,6a-tetrahydro-3aH-pyrrolo[3,4-d]isoxazole-4-
carboxylic acid and 3-hydroxy-4,5,6,6a-tetrahydro-3aH-pyrrolo[3,4-d]isoxazole-6-carboxylic acid, two
conformationally constrained aspartate and glutamate analogs. Mol Pharmacol 66:522-529.
• Gobbi M, Moia M, Pirona L, Ceglia I, Reyes-Parada M, Scorza C, Mennini T. 2002. p-Methylthioamphetamine and 1-
(m-chlorophenyl)piperazine, two non-neurotoxic 5-HT releasers in vivo, differ from neurotoxic amphetamine
derivatives in their mode of action at 5-HT nerve endings in vitro. J Neurochem 82:1435-1443.
• Gobbi M, DallaValle F, Ciapparelli C, Diomede L, Morazzoni P, Verotta L, Caccia S, Cervo L, Mennini T. 1999.
Hypericum perforatum L. extract does not inhibit 5-HT transporter in rat brain cortex. Naunyn Schmiedebergs Arch
Pharmacol 360:262-269.
• Gobbi M, Gariboldi M, Piwko C, Hoyer D, Sperk G, Vezzani A. 1998. Distinct changes in peptide YY binding to, and
mRNA levels of, Y1 and Y2 receptors in the rat hippocampus associated with kindling epileptogenesis. J Neurochem
70:1615-1622.
160
ANNUAL REPORT 2008

IRFMN
• Crespi D, Mennini T, Gobbi M. 1997. Carrier-dependent and Ca(2+)-dependent 5-HT and dopamine release induced by
(+)-amphetamine, 3,4-methylendioxymethamphetamine, p-chloroamphetamine and (+)-fenfluramine. Br J Pharmacol
121:1735-1743.
Mineko Terao obtained her doctorate degree in Pharmaceutical Science from the Kobe Women’s
College of Pharmacy, Japan in 1978. Her scientific interests relate to problems of Cellular Biology and
Molecular Biology.
1983 Ph.D in Molecular Biology, Kyoto University, Japan
1982-1983 Research Fellow, Department of Medical Chemistry, Kyoto University Faculty of Medicine,
Japan
1983-1987 Postdoctoral Associate of the Institute for Cancer Research, Philadelphia, US
From 1987 Visiting Scientist of Mario Negri Institute
From 1998 Head of the Unit of Gene Structure and Regulation, Mario Negri Institute
Selected publications
• Terao M, Kurosaki M, Barzago MM, Varasano E, Boldetti A, Bastone A, Fratelli M, Garattini E.
Avian and canine aldehyde oxidases. Novel insights into the biology and evolution of molybdo-flavoenzymes.
J Biol Chem. 2006 Jul 14;281(28):19748-61
• Garattini E, Parrella E, Diomede L, Gianni M, Kalac Y, Merlini L, Simoni D, Zanier R, Ferrara F F, Chiarucci I,
Carminati P, Terao M, Pisano C. ST1926, a novel and orally active retinoid-related molecule inducing apoptosis in
myeloid leukemia cells: Modulation of intracellular calcium homeostasis. Blood 2004; 103: 194-207
• Vila R, Kurosaki M, Barzago M M, Kolek M, Bastone A, Colombo L, Salmona M, Terao M, Garattini E. Regulation and
biochemistry of mouse molybdo-flavoenzymes. The DBA/2 mouse is selectively deficient in the expression of aldehyde
oxidase homologues 1 and 2 and represents a unique source for the purification and characterization of aldehyde oxidase.
J Biol Chem 2004; 279: 8668-8683
• Kurosaki M, Terao M, Barzago M M, Bastone A, Bernardinello D, Salmona M, Garattini E. The aldehyde oxidase gene
cluster in mice and rats: Aldehyde oxidase homologue 3, a novel member of the molybdo-flavoenzyme family with
selective expression in the olfactory mucosa. J Biol Chem 2004; 279: 50482-50498
• Parrella E, Gianni’ M, Cecconi V, Nigro E, Barzago MM, Rambaldi A, Rochette-Egly C, Terao M and Garattini E.
Phosphodiesterase 4 inhibition by piclamilast potentiates the cyto-differentiating action of retinoids in myeloid leukemia
cells. J Biol Chem 2004; 279: 42026-42040
• Garattini E, Gianni’ M and Terao M. Retinoid related molecules an emerging class of apoptotic agents with promising
clinical potential in oncology: pharmacological activity and mechanisms of action. Curr Pharm Design 2004, 10: 433-
448
Pia Emilia Villa got her degree in Pharmaceutical Chemistry and Technology at the University of Pavia
in 1975 and the specialisation in Pharmacological Research at the Mario Negri Institute, Milan in 1978.
Her scientific interests are the physiopathologic factors of sepsis and their pharmacological modulation,
the cellular and molecular mechanisms of neurodegeneration and neuroprotection in experimental
cerebral ischemia.
1976-1992: Research fellow, laboratory of Perfusion of Isolated Organs and Toxicology, Mario Negri
Institute
1979-1980: Visiting fellow, laboratory of Cultured Hepatocytes, Toxicology Unit, MRC, Carshalton,
England
1983: Visiting fellow, Unité de Recherche Hépatologique, Rennes, France
1993-1995: Research Scientist, laboratory of Neuroimmunology, Mario Negri Institute
1995 to date: Head, Unit of Pharmacology of Septic Shock, Mario Negri Institute
1982 Regular member of the Italian Society of Toxicology
1992 Regular member of Celltox
1996 Regular member of the International Cytokine Society.
Selected publications
• Villa P, Shaklee CL, Meazza C, Agnello D, Ghezzi P, Senaldi G. Granulocyte colony-stimulating factor and antibiotics
in the prophylaxis of a murine model of polymicrobial peritonitis and sepsis. J Infect Dis. 1998; 178: 471-7.
• Villa P, Saccani A, Sica A, Ghezzi P. Glutathione protects mice from lethal sepsis by limiting inflammation
and potentiating host defense. J Infect Dis. 2002; 185: 1115-20.
• Erbayraktar S, Grasso G, Sfacteria A, Xie QW, Coleman T, Kreilgaard M, Torup L, Sager T, Erbayraktar Z,
Gokmen N, Yilmaz O, Ghezzi P, Villa P, Fratelli M, Casagrande S, Leist M, Helboe L, Gerwein J,
Christensen S, Geist MA, Pedersen LO, Cerami-Hand C, Wuerth JP, Cerami A, Brines M.
Asialoerythropoietin is a nonerythropoietic cytokine with broad neuroprotective activity in vivo. Proc Natl
Acad Sci U S A. 2003;100 (11):6741-6.
• Villa P, Bigini P, Mennini T, Agnello D, Laragione T, Cagnotto A, Viviani B, Marinovich M, Cerami A,
Coleman TR, Brines M, Ghezzi P. Erythropoietin selectively attenuates cytokine production and
inflammation in cerebral ischemia by targeting neuronal apoptosis. J Exp Med. 2003;198 (6):971-5.
161
ANNUAL REPORT 2008

IRFMN
• Leist M, Ghezzi P, Grasso G, Bianchi R, Villa P, Fratelli M, Savino C, Bianchi M, Nielsen J, Gerwien J, Kallunki P,
Larsen AK, Helboe L, Christensen S, Pedersen LO, Nielsen M, Torup L, Sager T, Sfacteria A, Erbayraktar S,
Erbayraktar Z, Gokmen N, Yilmaz O, Cerami-Hand C, Xie QW, Coleman T, Cerami A, Brines M. Derivatives of
erythropoietin that are tissue protective but not erythropoietic. Science. 2004;305: 239-42.
• Garau A, Bertini R, Colotta F, Casilli F, Bigini P, Cagnotto A, Mennini T, Ghezzi P, Villa P. Neuroprotection with the
CXCL8 inhibitor repertaxin in transient brain ischemia. Cytokine. 2005;30:125-31.
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department comprises six laboratories. Research is heterogeneous in terms of scientific
interests and aims, but it is unified by the structural and functional study of specific,
pharmacologically important gene products, using a common body of techniques. Classical
biochemistry and molecular biology methods are used to define proteins that might be targets
for the pharmacological activity of drugs. Potential direct interactions between drugs and
proteins are studied at the molecular level by a variety of approaches ranging from animal
studies to computer simulation.
FINDINGS/MAIN RESULTS
Identification of tetracylines as potential anti-prion drugs.
Synthesis and physicochemical and biological characterization of peptides deduced from the
primary sequence of prion protein.
Identification of a relationship between cholesterol synthesis and production of prion protein.
Protein identifications by mass spectrometry and data base searching using a combination of
techniques.
Characterization of the Pentraxin 3 role in the organization of the cumulus oophorus
extracellular matrix and in female fertility.
System-level analysis of protein interactions in the epithelial junctional complex.
Characterization of the role of Junctional Adhesion Molecule-A (JAM-A) in the control of cell
motility.
Characterization of the effect of inflammatory cytokines on JAM-A function.
Development of a constitutive and of an inducible motor neuron cellular model to unravel the
toxicity of mutant G93A superoxide dismutase 1 responsible for some forms of familial
amyotrophic lateral sclerosis.
Conditions of oxidative stress or the presence of compounds impairing the electron transport
chain are a risk factor to motor neurons of individuals carrying mutant forms of superoxide
dismutase 1.
Mitochondrial damage due to mutant G93A superoxide dismutase 1 occurs selectively in motor
neurons.
Mitochondrial damage by mutant G93A superoxide dismutase 1 in motor neurons is modulated
by the level of expression of the mutant protein.
Identification and characterization of a novel class of retinoids endowed with strong and
selective apoptogenic acivity on the neoplastic cell. Pre-clinical development of these agents for
the treatment of acute leukemia.
Identification and characterization of novel retinoid-based pharmacological combinations for
the treatment of acute myelogenous leukemia.
Molecular cloning and characterization of the cDNAs and genes of four novel members of the
mammalian molybdo-flavoprotein family. Definition of a novel gene cluster on human
chromosome 2 and mouse chromosome 1.
Development of knok-out animals for molybdo-flavoproteins: AOX1, AOH1, AOH2, AOH3.
162
ANNUAL REPORT 2008

IRFMN
Creation of integrated instruments for the rationalization of Microarray analysis
processes.
Identification of erythropoietin as a neuroprotective agent and of new molecules with
neuroprotective
activity.
Identification of the pharmacological action of erythropoietin against peripheral neuropathy of
diabetes.
Identification of erythropoietin derivatives that retain its neuroprotective actions but have lost
its hemopoietic ones.
Discovery of proteins that are regulated by the redox state through the formation of reversible
disulfide bonds with glutathione (protein glutathionylation).
Identification of exofacial proteins undergoing thiol redox regulation.
The use of antioxidant molecules in models of sepsis and inflammation diminishes the
inflammatory response while potentiates the innate immune response.
Identification of the gene expression profile regulated by thiols.
The treatment with a non haematopoietic derivate of Erythropoietin (CEPO) reduces motor
neuron loss and clinical progression in a mouse model of ALS related to alterations in vesicle
trafficking, the wobbler mouse.
Treatment with a soluble TNF receptor in the wobbler mouse, reduces motor neuron
degeneration and the phosphorylation of the two main stress kinases (p38 e JNK) activated by
TNF receptors.
Riluzole treatment reduces motor neuron loss and clinical progression of wobbler mouse by
increasing the endogenous BDNF expression .
Oxidative stress, glial activation and inflammation occur in the retinopathy as well as in
cerebral and spinal cord dysfunction in the mnd mouse, a model of progressive epilepsy with
mental retardation related to mutation in the CLN8 gene. These findings provide further
evidence for the implication of TNF death receptor signalling in the pathology of Neuronal
Ceroid Lipofuscinosis
The affinity of pergolide for human cloned 5-HT 2A and 5-HT 2B receptors is similar and higher
that that for the human cloned D 2L receptor. These findings, together with the fact that it acts as
agonist at both h5-HT 2A and h5-HT 2B receptors, could explain its potential toxicity mediated by
activation of cardio-pulmonary 5-HT 2B receptor.
New conformationally constrained aspartate and glutamate analogues dissociate glutamate
uptake inhibition and reverse transport-mediated release.
Dimethyl sulfoxide, a solvent commonly utilized to dissolve hydrophobic compound for in
vitro experiments, interferes with the 5-HT6 agonists activity when the scintillation proximity
assay is used for evaluating 35S-GTP-γ-S binding; but does not interfere with the europium
labelled GTP binding determined by “time-resolved fluorescence” .
NATIONAL COLLABORATIONS
Advanced Biology Center, Genoa
Dip. Anatomia, Farmacologia, Medicina Legale, University of Turin
Dip. Biotecnologie, Università degli Studi, Milan
Dip. Chimica Biochimica e Biotecnologie per la Medicina, Università degli Studi, Milan
Dip. Chimica Farmaceutica e Tossicologica, Università degli Studi, Milan
Dip. Farmaco-Chimico, Università degli Studi, Messina
Dip. Farmaco-Chimico-Tecnologico, University of Siena
Dip. Farmacologia Medica, Università degli Studi, Milan
Dip. Scienze Biochimiche, University of Florence
163
ANNUAL REPORT 2008

IRFMN
Dip. Scienze Farmaceutiche, University of Catania
Dip. Scienze Farmaceutiche, University of Genoa
Dip. Scienze Farmacologiche, Università degli Studi, Milan
Dip. Scienze Fisiologiche e Farmacologiche, University of Pavia
Dip. Scienze Molecolari, University of Milan
Dip. Studi pre-clinici, University of Milan
Facoltà di Biologia, Università degli Studi, Milan
Facoltà di Chimica, Università degli Studi, Milan
Facoltà di Chimica, University of Ferrara
Fondo Edo Tempia, Biella
GlaxoSmithkline, Verona
IFOM Fondazione Istituto FIRC di Oncologia Molecolare, Milan
IRCCS Fondazione "Istituto C. Mondino", Laboratorio di Neurobiologia Sperimentale, Pavia
Istituto di Biologia Molecolare Buzzati Traverso, Naples
Istituto di Biomedicina e Immunologia Molecolare CNR, Palermo
Istituto di Endocrinologia, Centro di Eccellenza per le Malattie Neurodegenerative, Università
degli Studi, Milan
Istituto di Clinica Neurologica, Ospedale Maggiore Policlinico, Milan
Istituto Clinico Humanitas, Milan
Istituto di Neuroscienze C.N.R., Pisa
Istituto Nazionale dei Tumori, Milan
Istituto Nazionale dei Tumori, Naples
Istituto Nazionale Neurologico "C. Besta", Milan
Istituto Oncologico Europeo, Milan
Istituto Regina Elena, Rome
Istituto Toscano Tumori, Florence
Newron Pharmaceuticals, Milan
Ospedale Maggiore Policlinico, Milan
Ospedale Pediatrico Bambino Gesu', Rome
Ospedale Pediatrico "Gaslini", Genoa
Ospedale S. Gerardo, Monza, Milan
Sigma-Tau, Pomezia, Rome
Zambon, Milan
INTERNATIONAL COLLABORATIONS
The Babraham Institute, Cambridge, UK
Boston College, Boston, MA, USA
Case Western Research University, Cleveland, OH, USA
Dept. of Neurology, Keio University, Tokyo, Japan
Dept. de Quimica-Fisica de Macromoleculas Biologicas, CSIC, Madrid, Spain
Faculdad de Ciencias Medicas, Universidad de Santiago de Chile, Chile
Dept. of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The
Hebrew University of Jerusalem, Jerusalem, Israel
Flanders Interuniversity Institute for Biotechnology (VIB) University of Gent, Belgium
FMP, Berlin, Germany
Giessen Polyclinic University, Giessen, Germany
Houston University, TX, USA
IBSN CNRS, Marseille, France
Indiana University, Indianapolis, IN, USA
164
ANNUAL REPORT 2008

IRFMN
Institut de Genetique et Biologie Moleculaire et Cellulaire, Strasbourg, France
Institut Pasteur, Paris, France
John Innes Centre, Norwich, UK
Kenneth S. Warren Institute, Ossining, NY, USA
Max-Planck-Institut für experimentelle Medizin, Göttingen, Germany
National Institute of Health, Bethesda, MD, USA
Nippon University, Tokyo, Japan
North Shore University Hospital, Manhasset, NY, USA
Pepscan System BV, Lelystad, Holland
Polichem S.A., Lugano, Switzerland
Stanford University School of Medicine, Stanford, CA, USA
Technical University Braunschweig, Germany
Trinity College, Dublin, Ireland
Universidad de La Laguna, Tenerife, Spain
Universidad Nova, Lisbon, Portugal
Universitat des Saarlandes, Hamburg, Germany
Universitat Freiburg, Germany
Université Paris, France
Université Victor Segalen Bordeaux 2, Bordeaux, France
University of Aberdeen, UK
University of Birmingham, UK
University of Cardiff, UK
University of Colorado, School of Medicine, Denver, CO, USA
University of Glasgow, UK
University of Gottingen, Germany
University of Muenster, Germany
University of Southampton, UK
University of Sussex, UK
University of Vienna, Austria
Waring-Webb Institute, University of Colorado, Denver CO, USA
Weizmann Institut, Rehovot, Israel
Westfaelische Wilhelms-Universitaet Muenster, Germany
EDITORIAL BOARD MEMBERSHIP
Neurobiology of Lipids (L. Diomede)
Neuroimmunomodulation (P. Ghezzi)
Newsletters of the International Cytokine Society (P. Ghezzi)
European Journal of Cancer (E. Garattini)
PEER REVIEW ACTIVITIES
American Journal Physiology, Biochemical Journal, Biochemical Pharmacology, Biochimica
Biophysica Acta, Brain Research, Cancer Research, Cell Death and Differentiation, Cell
Research, Circulation, Drug Investigation, European Journal of Cancer, European Journal of
Immunology, European Journal of Neuroscience, International Journal of Cancer, Journal of
Cell Biology, Journal of Hepatology, Journal of Immunology, Journal of Investigative
Dermatology, Journal of Lipid Mediators, Journal of Neurochemistry, Journal of
Neuroimmunology, Journal of Translational Medicine, Neuroscience Letters, Pharmacological
165
ANNUAL REPORT 2008

IRFMN
Research, Physiological Genomics, Proceedings of the National Academy of Sciences, Life
Sciences.
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
Meeting: “First European Synapse Meeting”, “SNAP-25 negatively modulates voltage-gated
calcium channels and network activity”, 26 March, Bordeaux, France
Meeting: “Associazione Italiana di Neuropatologia (AINP) e Associazione Italiana per la
Ricerca sull’Invecchiamento Cerebrale (AIRIC), “Analysis of C1-INH to MBL, using Surface
Plasmon Resonance”, “Synaptic dysfunction in a mouse model of a familial prion disease:
impairment of glutamate exocytosis”, “Synaptic dysfunction in a mouse model of a familial
prion disease: impairment of calcium dynamics”, 18 June, Milan, Italy
Conference: “ICAD Alzheimer’s Associations International Conference on Alzheimer’s Disease
and Related Disorders”, “Beta-amyloid interferences with cognitive functions in an aggregation
status-dependent manner”, 30 July, Chicago, USA
Meeting: “8th Siena Meeting, From Genome to Proteome: Integration of proteome completion”,
“Comparison of proteomic approaches for protein biomarker discovery in amyotrophic lateral
sclerosis”, 31 August, Siena, Italy
Meeting: “XIX National Meeting on Medicinal Chemistry”, “New thienopyrimidine derivatives
as potential 5-HT7 receptor ligands”, “Heteroaryloxy analogues of WA4101: synthesis and
biological evaluation at alfa1-adrenoreceptor subtypes”, 14 September, Verona, Italy
Simposium: “SFB – Symposium”, “Partial agonism for the induction of SERT-mediated 5-HT
release and currents in vitro”, “Glutamate uptake is dissociated from substrate-induced
glutamate release as evidence by conformationally constrained aspartate and glutamate
analogues”, 26 September Vienna, Austria
Congress: “Congresso annuale Società Italiana di Neurologia”, “Analgesic effect of
buprenorphine in an experimental model of painful diabetic peripheral neuropathy”, 18
October, Napoli, Italy
GRANTS AND CONTRACTS
Agenzia Italiana del Farmaco, Rome, Italy
Biotecnologies BT - Perugia, Italy
Dompè, L' Aquila, Italy
European Union, Bruxelles, Belgium
Istituto Auxologico Italiano, Milan, Italy
Istituto Nazionale Neurologico "C. Besta", Milan, Italy
Italian Association for Cancer Research (AIRC), Milan, Italy
Italian Ministry of University and Research (MIUR), Rome, Italy
Kenneth S. Warren Institute, NY, USA
166
ANNUAL REPORT 2008

IRFMN
Lundbeck A/S, Copenhagen, Denmark
Cariplo Foundation, Milan, Italy
Don Gnocchi Foundation, Milan, Italy
Mariani Foundation, Milan, Italy
Monzino Foundation, Milan, Italy
Ministry of Health, Rome, Italy
National Research Council (CNR), Palermo, Italy
North Shore University Hospital, NY, USA
Perfetti-Van Melle, Lainate (Mi), Italy
Sigma Tau, Pomezia (Rome), Italy
Telethon, Milan, Italy
University of Florence, Italy
University of Milan-Bicocca, Italy
University of Siena, Italy
Weizmann-Pasteur-Negri Foundation, Paris, France
Zambon Group, Bresso (Mi), Italy
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Bate C, Marshall V, Colombo L, Diomede L, Salmona M, Williams A
Docosahexaenoic and eicosapentaenoic acids increase neuronal death in response to HuPrP82-146 and Abeta(1-42
Neuropharmacology 2008 54 : 934-943
Bate C, Tayebi M, Diomede L, Salmona M, Williams A
Docosahexaenoic and eicosapentaenoic acids increase prion formation in neuronal cells
BMC Biol 2008 6 : 39
Bazzoni G
Signalling pathways and adhesion molecules as targets for antiangiogenesis therapy in tumors
Adv Exp Med Biol 2008 610 : 74-87
Bigini P, Repici M, Cantarella G, Fumagalli E, Barbera S, Cagnotto A, De Luigi A, Tonelli R, Bernardini R, Borsello
T, Mennini T
Recombinant human TNF-binding protein-1 (rhTBP-1) treatment delays both symptoms progression and motor
neuron loss in the wobbler mouse
Neurobiol Dis 2008 29 : 465-476
Brines M, Patel N S A, Villa P, Brines C, Mennini T, De Paola M, Erbayraktar Z, Erbayraktar S, Sepodes B,
Thiemermann C, Ghezzi P, Yamin M, Hand C C, Xie Q W, Coleman T, Cerami A
Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin
Proc Natl Acad Sci USA 2008 105 : 10925-10930
Colleoni S, Jensen A A, Landucci E, Fumagalli E, Conti P, Pinto A, De Amici M, Pellegrini-Giampietro D E, De
Micheli C, Mennini T, Gobbi M
Neuroprotective effects of the novel glutamate transporter inhibitor (-)-3-hydroxy-4,5,6,6a-tetrahydro-3aHpyrrolo[3,4-d]-isoxazole-4-carboxylic
acid, which preferentially inhibits reverse transport (glutamate release)
compared with glutamate reuptake
J Pharmacol Exp Ther 2008 326 : 646-656
Colombo Alessio, Bastone A, Ploia C, Sclip A, Salmona M, Forloni G, Borsello T
JNK regulates APP cleavage and degradation in a model of Alzheimer's disease
Neurobiol Dis 2008 [Epub ahead of print]
Cosentino U, Pitea D, Moro G, Saracino A A G, Caria P, Vari' M R, Colombo L, Forloni G, Tagliavini F, Salmona M
The anti-fibrillogenic activity of tetracyclines on PrP 106-126: a 3D-QSAR study
J Mol Model 2008 14 : 987-994
167
ANNUAL REPORT 2008

IRFMN
Deban L, Jarva H, Lehtinen M J, Bottazzi B, Bastone A, Doni A, Jokiranta T S, Mantovani A, Meri S
Binding of the long pentraxin PTX3 to factor H: interacting domains and Function in the regulation of complement
activation
J Immunol 2008 181 : 8433-8440
De Luigi A, Colombo L, Diomede L, Capobianco R, Mangieri M, Miccolo C, Mangeri M, Limido L, Forloni G,
Tagliavini F, Salmona M
The efficacy of tetracyclines in peripheral and intracerebral prion infection
PLoS One 2008 3 : e1888
De Paola M, Diana V, Bigini P, Mennini T
Morphological features and responses to AMPA receptor-mediated excitotoxicity of mouse motor neurons:
comparison in purified, mixed anterior horn or motor neuron/glia cocultures
J Neurosci Methods 2008 170 : 85-95
Fumagalli E, Funicello M, Rauen T, Gobbi M, Mennini T
Riluzole enhances the activity of glutamate transporters GLAST, GLT1 and EAAC1
Eur J Pharmacol 2008 578 : 171-176
Garattini E, Fratelli M, Terao M
Mammalian aldehyde oxidases: genetics, evolution and biochemistry
Cell Mol Life Sci 2008 65 : 1019-1048
Ghezzi P, Di Simplicio P
Glutathionylation pathways in drug response
Curr Opin Pharmacol 2008 7 : 398-403
Gianni M, Boldetti A, Guarnaccia V, Rambaldi A, Parrella E, Raska I Jr, Rochette-Egly C, Del Sal G, Rustighi A,
Terao M, Garattini E
Inhibition of the peptidyl-propyl-isomerase Pin1 enhances the responses of acute myeloid leukemia cells to retinoic
acid via stabilization of RARα and PML-RARα
Cancer Res. 2008, in press
Gobbi M, Funicello M, Gerstbrein K, Holy M, Moya P R, Sotomayor R, Forray M I, Gysling K, Paluzzi S, Bonanno
G, Reyes-Parada M, Sitte H H, Mennini T
N,N-dimethyl-thioamphetamine and methyl-thioamphetamine, two non-neurotoxic substrates of 5-HT transporters,
have scant in vitro efficacy for the induction of transporter-mediated 5-HT release and currents
J Neurochem 2008 105 : 1770-1780
Inforzato A, Rivieccio V, Morreale A P, Bastone A, Salustri A, Scarchilli L, Verdoliva A, Vincenti S, Gallo G,
Chiapparino C, Pacello L, Nucera E
Structural characterization of PTX3 disulfide bond network and its multimeric status in cumulus matrix organization
J Biol Chem 2008 283 : 10147-10161
Manzoni C, Colombo L, Messa M, Cagnotto A, Cantù L, Del Favero E, Salmona M
Overcoming synthetic Aβ peptide aging: a new approach to an age-old problem
Amyloid, 2008 in press
Mengozzi M, Cervellini I, Bigini P, Martone S, Biondi A, Pedotti R, Gallo B, Barbera S, Mennini T, Boraso M,
Marinovich M, Petit E, Bernaudin M, Bianchi R, Viviani B, Ghezzi P
Endogenous erythropoietin as part of the cytokine network in the pathogenesis of experimental autoimmune
encephalomyelitis
Mol Med 2008 14 : 682- 688
Micale N, Colleoni S, Postorino G, Pellicano' A, Zappala' M, Lazzaro J, Diana V, Cagnotto A, Mennini T, Grasso S
Structure-activity study of 2,3-benzodiazepin-4-ones noncompetitive AMPAR antagonists: identification of the 1-(4-
amino-3-methylphenyl)-3,5-dihydro-7,8-ethylenedioxy-4H-2,3-benzodiazepin-4-one as neuroprotective agent
Bioorg Med Chem 2008 16 : 2200-2211
Modica M N, Romeo G, Salerno L, Pittala' V, Siracusa M A, Mereghetti I, Cagnotto A, Mennini T, Gaspar R, Gal A,
Falkay G, Palko M, Maksay G, Fulop F
Synthesis and receptor binding of new thieno[2,3-d]-pyrimidines as selective ligands of 5-HT(3) receptors
168
ANNUAL REPORT 2008

IRFMN
Arch Pharm (Weinheim) 2008 341 : 333-343
Natalello A, Prokorov V V, Tagliavini F, Morbin M, Forloni G, Beeg M, Manzoni Claudia, Colombo L, Gobbi M,
Salmona M, Doglia S M
Conformational plasticity of the Gerstmann-Sträussler-Scheinker disease peptide as indicated by its multiple
aggregation pathways
J Mol Biol 2008 381 : 1349-1361
Paris L, Bazzoni G
The protein interaction network of the epithelial junctional complex: a system-level analysis
Mol Biol Cell 2008 19 : 5409-5421
Paris L, Bononi E, Bazzoni G
Network analysis of cell adhesion. Adhesomes as context-defined subnetworks
Communicative & Integrative Biology 2008, in press
Paris L, Tonutti L, Vannini C, Bazzoni G
Structural organization of the tight junctions
Biochim Biophys Acta 2008 1778 : 646-659
Pera M, Martinez-Otero A, Colombo L, Ruiz-Molina D, Badia A, Clos M V
Acetylcholinesterase as an amyloid enhancing factor in PrP82-146 aggregation process
Mol Cell Neurosci 2008 [Epub ahead of print]
Pinto A, Conti P, De Amici M, Tamborini L, Grazioso G, Colleoni S, Mennini T, Gobbi M, De Micheli C
Synthesis of enantiomerically pure HIP-A and HIP-B and investigation of their activity as inhibitors of excitatory
amino acid transporters
Tetrahedron Asymmetry 2008 19 : 867-875
Pittala' V, Modica M, Salerno L, Siracusa M, Guerrera F, Mereghetti I, Cagnotto A, Mennini T, Romeo G
Synthesis and endothelin receptor binding affinity of a novel class of 2-substituted-4-aryl-3-quinolinecarboxylic acid
derivatives
Med Chem 2008 4 : 129-137
Repici M, Mare L, Colombo A, Ploia C, Sclip A, Bonny C, Nicod P, Salmona M, Borsello T.
c-Jun N-terminal kinase binding domain-dependent phosphorylation of mitogen-activated protein kinase kinase 4 and
mitogen-activated protein kinase kinase 7 and balancing cross-talk between c-Jun N-terminal kinase and extracellular
signal-regulated kinase pathways in cortical neurons.
Neuroscience 2008 [Epub ahead of print]
Roglio I, Bianchi R, Camozzi F, Carozzi V, Cervellini I, Crippa D, Lauria G, Cavaletti G, Melcangi R C
Docetaxel-induced peripheral neuropathy: protective effects of dihydroprogesterone and progesterone in an
experimental model
J Peripher Nerv Syst 2008, in press
Roglio I, Bianchi R, Gotti S, Scurati S, Giatti S, Pesaresi M, Caruso D, Panzica G, Melcangi R C
Neuroprotective effects of dihydroprogesterone and progesterone in an experimental model of nerve crush injury
Neuroscience 2008 155 : 673-685
Roglio I, Giatti S, Pesaresi M, Bianchi R, Cavaletti G, Lauria G, Garcia-Segura L M, Melcangi R C
Neuroactive steroids and peripheral neuropathy
Brain Res Rev 2008 57 : 460-469
Saracino A A G, Villa A, Moro G, Cosentino U, Salmona M
Spontaneous β-helical fold in prion protein. The case of PrP(82-146)
Proteins 2008 [Epub ahead of print]
Siracusa M A, Salerno L, Modica M N, Pittala' V, Romeo G, Amato M E, Nowak M, Bojarski A J, Mereghetti I,
Cagnotto A, Mennini T
Synthesis of new arylpiperazinylalkylthiobenzimidazole, benzothiazole, or benzoxazole derivatives as potent and
selective 5-HT1A serotonin receptor ligands
J Med Chem 2008 51 : 4529-4538
169
ANNUAL REPORT 2008

IRFMN
Taoufik E, Petit E, Divoux D, Tseveleki V, Mengozzi M, Roberts M L, Valable S, Ghezzi P, Quackenbush J, Brines
M, Cerami A, Probert L
TNF receptor I sensitizes neurons to erythropoietin- and VEGF-mediated neuroprotection after ischemic and
excitotoxic injury
Proc Natl Acad Sci USA 2008 105 : 6185-6190
Terao M, Kurosaki M, Barzago M M, Fratelli M, Bagnati R, Bastone A, Giudice C, Scanziani E, Mancuso A,
Tiveron C, Garattini E
Role of the molybdo-flavoenzyme, aldehyde oxidase homolog 2, in the biosynthesis of retinoic acid: generation and
characterization of a knock-out mouse
Mol Cell Biol 2008 [Epub ahead of print]
Valli C, Paroni G, Di Francesco A M, Riccardi R, Tavecchio M, Erba E, Boldetti A, Gianni M, Fratelli M, Pisano C,
Merlini L, Antoccia A, Cenciarelli C, Terao M, Garattini E
Atypical retinoids ST1926 and CD437 are S-phase-specific agents causing DNA double-strand breaks: significance
for the cytotoxic and antiproliferative activity
Mol Cancer Ther 2008 7 : 2941-2954
RESEARCH ACTIVITIES
Laboratory of Receptor Pharmacology
Studies on the role of dysfunction of the endoplasmic reticulum (ER) or
the Golgi apparatus (GA) in neurodegenerative disease
The secretory pathway starts at the endoplasmic reticulum (ER) where proteins are synthesized
and folded and chaperone–mediated quality control prevents misfolded proteins to reach their
destination and interfere with normal metabolism. Protein transport by mean of vesicles
continues through the Golgi Apparatus (GA), that is most abundant in neurons, and finishes in
the plasma membrane, secretory vesicles or lysosomes. The endocytic pathway enables
internalized macromolecules to be delivered via endosomes to lysosomes where they are
enzymatically digested.
In mammalian cells, the Golgi-associated retrograde protein (GARP) complex is involved in
retrograde transport of endosomes to the trans GA network. Defective intracellular membrane
trafficking is common to several neurodegenerative diseases. Among the neuronal population,
motor neurons, due the their high energy requirement and long axons are, together with retinal
cells, the most sensitive ones.
The Laboratory of Receptor Pharmacology utilizes two mouse models of neurodegeneration
related to cellular transport disruption, carrying mutation in proteins resident in the ER (the
mnd mouse) or in the GARP complex (the wobbler mouse).
The neuronal ceroid lipofuscinosis (NCLs) are a group of autosomal recessive
neurodegenerative diseases and a significant cause of childhood progressive intellectual and
neurological deterioration, for which no curative or preventive treatment are available. Among
them, the progressive epilepsy with mental retardation is the newest form with mutation in the
CLN8 gene encoding a novel ER transmembrane protein with undefined function. An
orthologue of CNL8 is mutated in the motor neuron degeneration mouse (mnd) which shows
early retinopathy and delayed motor neuron dysfunction without degeneration. How CLN8
mutation leads to NCL defect is unknown. Our laboratory is studying mnd mice since many
years: we have already reported decreased spinal cord GLT-1 glial glutamate transporter and
increased plasma glutamate concentration already at presymptomatic stage in mnd mice, with
increased GluR2 and lowered GluR3 AMPA receptor subunits in the lumbar spinal cord. The
AMPA receptor antagonists ZK 187638 (non-competitive), like NBQX (competitive),
ameliorate motor behavior in mnd mice. We also found that TNF and TNFR1 is increased in
the spinal cord of mnd mice already at presymptomatic stage, when intensive astrocytes and
170
ANNUAL REPORT 2008

IRFMN
microglial proliferation occurs. The rate of oxygen consumption (QO2), and mitochondrial
functions were decreased in mnd spinal cord. The level of lipid peroxide derivatives reacting
with thiobarbituric acid (TBARS) were increased in mnd spinal cord and retina. L-carnitine
treatment delayed the onset of motor behaviour impairment, increased the mitochondrial
enzyme activities and was effective in enhancing QO 2 and decreasing TBARS levels.
At present we are testing the susceptibility to convulsions in mnd mice at pre-symptomatic
stage , in order to find a further link to the human pathology. In these studies new non-invasive
approaches, like Optical Imaging , MRI, MicroCT and Confocal Angiography are applied, in
order to allow a better translation of the results to the human disease.
Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder involving
primarily motor neurons in the spinal cord, brainstem and cerebral motor cortex and leading to
denervation, muscular atrophy and paralysis. The disease is sporadic in approximately 90% of
cases, and the mechanism(s) responsible for the selective motor neuron degeneration are far
from being elucidated.
A missense mutation in Vps54 has been described in the wobbler mouse, which share many
pathological features with ALS patients. In mammalian cells Vps54 forms heterotrimeric
complexes with Vps52 and Vps53 to form the GARP complex, involved in retrograde transport
of endosomes to the trans GA network. Thus we use the wobbler mouse as a reliable tool to
understand the interplay between endosomal dynamics and the selective loss of motor neurons.
A series of experiments are in progress in cultured neural cells obtained from wobbler and
healthy homozygous mice to investigate the possible effect of Vps54 mutation on intracellular
trafficking, mitochondrial activity, and lysosome accumulation.
We have already reported that wobbler mice are sensitive to riluzole treatment, without
marked changes in AMPA/NMDA receptor subunits expression in motor neurons of early
symptomatic mice. In addition we found increased levels of TNF and TNFR1 in the cervical
spinal cord and a significant effect of chronic treatment with a soluble h-TNF binding protein,
resulting in slower clinical symptoms progression, reduced motor neuron loss and selective
inhibition of the two main stress-kinases (p38 and JNK) associated to TNF receptors activation.
Finally we investigated two different approaches of cell therapy. Undifferentiated adult neural
stem cells (in collaboration with Dr. Parati, Istituto Besta) produced a weak and transient
protective effect in clinical progression but significantly reduced motor neuron loss occurring in
the wobbler mouse. Transplantation of mononucleate cells from human cord blood (in
collaboration with Dott. Lazzari, Policlinico), although did not replace degenerating motor
neurons, produced a marked neuroprotective effect by slowing the clinical progression and
reducing motor neuron loss, biceps atrophy and neuroinflammation (reactive gliosis).
Neuropharmacology of the glutammatergic and serotonergic systems
Glutamate is the major excitatory amino acid in the central nervous system; the extracellular
glutamate concentration has to be maintained at physiological level by active uptake mediated
by specific transporters (Excitatory Amino Acid Transporters, EAATs) located on the plasma
membranes of neurons and glia. Alterations in this process might lead to a relevant increase in
extracellular glutamate concentrations, that is highly toxic for neurons in the central nervous
system. A research project is in progress in order to better characterize the involvement and the
role of the neuronal compartment in the general process of glutamate homeostasis. The
functional properties are evaluated using biochemical assays and the quantitative characteristics
are evaluated by western blot for specific neuronal and glial proteins and flow cytometry
analysis (in collaboration with Dr. Bernasconi, Department of Oncology, IRFMN). These
evaluations are done on purified preparations from mouse spinal cord. The same characteristics
are evaluated on two different animal models of motor neuron degeneration, the wobbler mouse
and the transgenic SOD1G93A mouse (in collaboration with Dott. Bendotti, Department of
171
ANNUAL REPORT 2008

IRFMN
Neuroscience, IRFMN), in order to understand the possible involvement in neurodegenerative
diseases.
Results obtained to date indicate that there is an active neuronal component in the glutamate
uptake. The comparison between the two animal models of neurodegeneration suggest that this
component has a different importance in the motor neuron death processes. In the wobbler
mouse the excitotoxicity related to alterations of glutamate uptake is likely to be not involved,
while in the transgenic SOD1G93A mouse we found a significant reduction of glutamate
uptake in the neuronal fraction. This alteration probably contribute to the exacerbation of
processes leading to motor neuronal death in this animal model.
A research project is in progress in collaboration with professor Bonanno (University of
Genova) to characterize mechanisms of glutamate release from synaptosomes obtained from
spinal cord of wobbler mouse, in order to evaluate if the motor neuronal death might depend on
altered processes of neurotransmitter release in the nerve endings.
We recently concluded a project focused on the study of the interaction between glutamate
transporters and riluzole. Riluzole is a drug with a complex mechanism of action, and is the
only drug approved for the treatment of amyotrophic lateral sclerosis (ALS). We used as
experimental model cell cultures that stably express the main three glutamate transporters
(GLT1, GLAST and EAAC1) and we demonstrated for the first time that riluzole acts on the
glutamate uptake mediated by the three EAAT subtypes, significantly increasing the efficiency
of the process. This mechanism might be relevant in pathological conditions characterized by
glutamate concentrations over the physiological threshold, as occurs in ALS.
AMPA receptor-mediated excitotoxicity is one of the main events involved in motor neuron
degeneration in ALS pathogenesis. We established a new model of primary cocultures of
purified motor neurons over a glial layer. This in vitro model allows to obtain healthier motor
neurons maintained in more physiological conditions and was used to demonstrate that AMPA
receptor agonists can induce both apoptotic or non-apoptotic death pathways depending on their
concentrations. We studied the interaction between excitotoxicity and other potentially
neurotoxic factors, such as the inflammatory mediators, and we demonstrated that the proinflammatory
chemokine IL-8 induces motor neuron death through the CXCR2 receptor.
Preliminary studies on the effect of TNF-α, whose levels were found increased in the spinal
cords of animal models of motor neuron degeneration, indicated the pivotal role of glial cells in
mediating neurotoxicity of this cytokine. At present, we are studying the main intracellular
biochemical pathways involved in neurodegenerative mechanisms (such as calcium influx) and
testing new potential treatments to selectively interfere with each of them. We have also started
a study on the neurotoxic effect of serum from professional soccer players aimed at identifying
potential risk factors present in the blood which could support the high incidence of ALS in this
group of athletes.
Studies on primary spinal cord or hippocampal neuronal cultures from wild type or mnd mice
are in progress in order to investigate their sensitivity to AMPA receptor agonists and
antagonists, and to evaluate the role of astrocytes obtained from mnd mice in affecting motor
neuron viability. Studies on astrocytes derived from neural stem cells of wobbler or control
mice are in progress in order to evaluate whether typical biochemical alterations of wobbler
mice are already evident in precursor-derived cells : we study the role of glutamate transporters
and possible effects of this kind of cells on healthy motor neurons. Finally, studies on the role
of Vsp54 mutation on intracellular trafficking are in progress in primary cultures of astrocytes
obtained from the spinal cord of adult wobbler or control mice.
The Laboratory of Receptor Pharmacology is maintaining, since many years, collaborations
with medicinal chemistry laboratories to characterize the affinity and selectivity of newly
synthesized compounds on neurotransmitter receptors using in vitro binding methods. The
results are utilized for molecular modelling (QSAR) studies and/or for further
pharmacological evaluations. Particularly, a collaboration is ongoing with Prof. S. Grasso
172
ANNUAL REPORT 2008

IRFMN
(University of Messina) and Prof. C. De Micheli (University of Milan), for the development of
new non-competitive AMPA receptor antagonists.
An useful application of this technique is also the evaluation of the agonist/antagonist activity
of compounds acting on G-protein coupled receptors (GPCR), measuring their effects on 35S-
GTP-γ-S binding. New non-radioactive methods based on “time-resolved fluorescence” are
under characterization as functional assays to monitor GPCR activity on cell membranes.
Recently we have verified the effect of dimethyl sulfoxide, a solvent commonly utilized to
dissolve hydrophobic compound for in vitro experiments, on different in vitro assays utilizing
HEK 293 cells expressing the 5-HT6 serotonin receptors. Our results indicate that dimethyl
sulfoxide interferes with the agonist activity on 5-HT6 receptor when the scintillation
proximity assay 35S-GTP-γ-S binding is used. On the contrary, it does not interferes with
europium labelled GTP binding determined by “time-resolved fluorescence”.
In addition, the laboratory perform autoradiography binding studies in order to evaluate ex vivo
drug-receptor occupancy.
Finally, we have ongoing collaborative studies with Dr. Gobbi (Lab. biochemistry and protein
chemistry, IRFMN), to characterize the role of the allosteric site at the serotonin transporter on
the mechanism of action of SSRI (selective serotonin reuptake inhibitors) antidepressant, like
escitalopram.
Laboratory of Neuroimmunology
Redox regulation
The study of the so-called oxidative stress has led to the identification of biochemical events
that are modified by antioxidant molecules even in the absence of oxidative stress intended as
overproduction of toxic oxygen intermediates (free radicals). We use the term redox regulation
to define the pattern of cell functions (gene expression, activity of enzymes or transcription
factors) that are in some way modified by the redox state of the cell, defined as the ratio
between oxidizing and reducing species (usually: the oxidized glutathione / reduced glutathione
ratio). Our work focuses on the molecular mechanisms by which small changes in the redox
state can affect proteins, with particular attention to the reversible modification of cysteines to
form disulfide bonds (with proteins or with small molecular weight thiols such as glutathione).
We recently focused our attention on the identification of the redox state of proteins present on
the outside of the plama membrane since these often have key functions (e.g.: transporters,
receptors) and are the closest target of extracellular oxidants. We also apply proteomics
techniques and gene expression profiling using DNA microarrays to identify the pathways
susceptible of redox regulation.
Neuroprotection and erythropoietin
The pathologies of the central or peripheral nervous systems studied in the lab are: cerebral
ischemia, experimental autoimmune encephalomyelitis, and diabetic neuropathy). Using animal
models and cell culture, we try to clarify the relationships between neuronal death and
inflammation, and to intervene with protective agents. Among the latter, we are studying
endogenous molecules that have shown an unexpected anti-apoptotic action on neuronal cells,
particularly erythropoietin and anti-inflammatory drugs.
Laboratory of Molecular Biology
Novel retinoids for the treatment of acute myeloid leukemia
The synthetic and natural derivatives of retinoic acid (retinoids) have shown promising activity
in the treatment of leukemia and solid cancer. Retinoids exert their therapeutic activity through
173
ANNUAL REPORT 2008

IRFMN
three distinct types of effects: cyto-differentiation, growth inhibition and apoptosis. The three
effects can be dissociated, albeit partially, as retinoids endowed primarily with cytodifferentiating
or anti-proliferative activities are known. Recently, we identified and
characterized a novel class of retinoids with strong and selective apoptotic activity towards the
neoplastic cell. These compounds (RRMs, retinoid related molecules), which were originally
developed as selective agonists of the gamma-types of the nuclear receptors of retinoic acid
(RARy), induce apoptosis in different types of leukemia and solid cancer cells through a largely
unknown mechanism. The process of apoptosis set in motion by RRMs is different from that of
other known chemotherapeutic agents and does not require the activation of the nuclear retinoic
acid receptors. RRMs are active not only in vitro but also in vivo on a number of pre-clinical
models of acute myeloid leukemia. Currently, some of these innovative molecules are in an
advanced phase of pre-clinical development.
Novel retinoic-acid-based pharmacological combinations for the treatment
of acute leukemia
The clinical use of retinoic acid for the treatment of acute promyelocytic leukemia (APL) is
based on the ability of this compound to induce the maturation of the leukemic blast along the
normal myelocytic/granulocytic pathway. At present, the clinical use of retinoic acid for the
treatment of patients suffering from APL is the sole example of differentiation therapy.
Differentiation therapy is worth pursuing as it is theoretically associated with a lower level of
toxicity relative to what observed following treatment with the classical cyto-toxic agents.
However, the clinical use of retinoic acid is still burdened by a number of problems including
natural and induced resistance as well as systemic and local toxicity. One of the possible ways
to increase the therapeutic index of retinoic acid is based on the identification of compounds or
drugs that potentiate the pharmacological activity of the retinoid. We have recently
demonstrated that a series of agents, such as G-CSF, interferons, cAMP analogs,
phosphodiesterase IV inhibitors and a number of other novel compounds sensitize the leukemic
blast to the pharmacological activity of retinoic acid. More recently, Pin1, a peptidyl-prolylisomerase,
was identified as an important molecular target for the sensitization of leukemia and
cancer cells to the affects of retinoic acid and derivatives. In the long run, it is our objective to
develop novel combinations and bring some of the above mentioned retinoic-acid-based
combinations to the clinic. In addition we intend to use some of the combinations as
pharmacological tools to dissect the intricacies of the cyto-differentiation process set in motion
by retinoic acid in the leukemic blast. We are widening the scope of our scientific activity to
the study of solid tumors with particular reference to breast carcinoma. In this last type of
tumor we are investigating the cross-talk between estrogens and retinoids with the development
of appropriate cellular models genetically engineered for the expression or silencing of estrogen
receptors.
The family of molybdo-flavoproteins
Molybdo-flavoenzymes are proteins of medical and industrial interest. They are the sole
enzymes that require molybdenum, in the form of the molybdenum cofactor, for their catalytic
activity. The laboratory has a long-standing and specific interest in the biochemistry and
biology of mammalian molybdo-flavoproteins. In the past, the laboratory contributed to the
elucidation and characterization of the primary structure of the two mammalian molybdoflavoproteins,
aldehyde oxidase (AOX1) and xanthine oxidoreductase (XOR). In the last few
years, the group identified and cloned the cDNAs and the genes coding for three novel mouse
molybdo-enzymes (AOH1, AOH2 and AOH3) belonging to the sub-family of molybdoflavoproteins.
The long-term goal of our studies is to define the structure, the substrate
specificity, the mechanisms of catalysis as well as the physo-pathological function of the three
new proteins. We will also continue the biochemical and functional studies on mammalian
174
ANNUAL REPORT 2008

IRFMN
AOX1 and XOR. To achieve our aims, we have recently developed cell lines over-expressing
XOR in a tetracycline inducible fashion. In addition, we have generated knock-out mice for the
genes encoding AOH2 and AOH3. The first knock-out model suggests that AOH2 is involved
in the synthesis of retinoic acid from retinaldehyde in the skin and the Harderian gland, which is
the main intra-orbital exocrine gland present in rodents.
Creation of integrated instruments for the rationalization of Microarray
analysis processes
An interdisciplinary group has been created in the Institute, with several external
collaborations, that deals, at different levels and with different professional
backgrounds, with microarray data analysis.
The procedures adopted by the different groups have been discussed, compared and
formally described with the aim of establishing a single workflow (that is a software
that provides end users with an easier way to orchestrate and describe complex
processing of data in a visual form).
The advantages of this approach are manifold. In fact, the highest possible automation
of the processes makes them more reliable and facilitates documentation and
reproducibility of the entire analysis process. Moreover it allows an easy comparison
among the different methodological choices, with the aim of reaching a shared
procedure among the collaborative groups.
The activity has required the development of a software for the analysis the preparation
of a hardware platform (Beowulf cluster), to support the required computational power.
Laboratory of Biochemistry and Protein Chemistry
Chimico-physical and molecular-biochemical studies on the prion protein
and on the peptides deduced from its aminoacid sequence
Prion protein fibril formation is associated with neuronal cytotoxicity and astrogliosis observed
in prion diseases. The formation of fibrils is the consequence of a conformational switch
between the structure of the native and the pathological proteins and it is considered of pivotal
importance for the appearance and progression of prion protein diseases.
Identifying the molecular determinants responsible for the conformational transition is likely to
give insight into the pathogenetic process leading to prion diseases. A reductionist appraoch to
the problem calls for the generation of simple experimental models in which the dynamics of
the conformational transition can be studied in detail. In our laboratory we developed synthetic
peptides that mimic the fibrillogenic properties of pathological prion protein. With the use of
various types of biochemical and biophysical techniques we studied the conformation of these
peptides through the evaluation of their secondary structure, the resistance to protease digestion
as well as the aggregating and amyloidogenic properties. Our approach gave detailed
informations on the conformational plasticity of various types of prion protein fragments.
To understand the correlation between the chemico-physical properties of prion protein-derived
peptides and their biological effects we used appropriate cellular models. These experiments,
performed in collaboration with the Laboratory of the Biology of Neurodegenerative Disorders,
gave informations on the sub-cellular distribution of the peptides to identify the intracellular
biological targets.
Development of a therapeutical strategy against prion-related diseases
Currently, no therapeutic options for the cure of prion-related diseases are available and the
identification of new molecules for the prevention and treatment of the infettivity is of great
interest. Althought some compounds gave positive results in prion cellular models, in human
175
ANNUAL REPORT 2008

IRFMN
they exhert low efficacy for toxicity and for their inability to pass the blood brain barrier. A first
approach for the development of anti-prion candidates involved molecules capable of interfering
with the fibrils formation, directly interacting with the β-sheet conformation of PrPSc causing
its disaggregation. In collaboration with the Laboratory of the Biology of the Neurodegenerative
Disorders we aimed to identify compounds endowed with anti-fibrillogenic activity and test
their efficacy in different in vitro and in vivo pre-clinical models of prion diseases.
Our studied indicated that tetracyclines are new good candidates as anti-prion drugs since they
act as anti-fibrillogenic compunds increasing the prion sensitivity to protease K digestion.
Moreover, they inhibited neuronal cell death and astrogliosis caused by prion peptides and
prolonged the survival of prion-infected animals.
Another therapeutic approach is based on the development of molecules that inhibited the PrP
formation by interacting with the lipid metabolism and destabilizing specific cellular membrane
domains. In collaboration with the Department of Infective Pathology and Diseases of the
Royal Veterinary School (Hawkshead, UK) we reported that statins, inhibiting cholesterol
synthesis, reduced the in vitro prion production. On the other hand, compounds such as
polyunsaturated fatty acids, known for their reducing effects on cholesterol levels, increased the
PrP production. Our future studies are aimed at understanding the relationship between
cholesterol cellular membrane distribution, lipid domain stability and the conversion of cellular
prion protein in its pathologic form.
Oxidative stress and protein aggregation in amyotrophic lateral sclerosis:
a proteomic approach
The molecular mechanisms at the basis of neurodegenerative diseases including the geneticallylinked
ones, such as amyotrophic lateral sclerosis (ALS), are still unknown. However, there is
evidence that oxidative stress and protein aggregation play central roles in the pathogenesis of
such diseases. The Unit of Medical Biochemistry, conducted proteome analysis of an animal
model of familial ALS. In collaboration with the laboratory of Molecular Neurobiology, we
focused the attention on the analysis of protein expression changes and protein modifications,
such as tyrosine nitration and ubiquitination, in a transgenic mouse, which over-express human
mutated (G93A) superoxide dismutase (SOD1). We analyzed, by proteomic tools, spinal cord of
presymptomatic G93A SOD1 mice, we identified nitrated proteins and quantified the level of
nitration for each protein in comparison with healthy controls. We have revealed that there is a
substantial increase of the nitration level in at least five proteins: actin, alpha and gamma
enolase, ATP synthase and a chaperone protein, HSC71. The alteration of the function of these
proteins may have important consequences on the cellular metabolism and catabolism, and
therefore may be at the basis of the molecular mechanisms leading to neurodegeneration. In
addition, by mass spectrometry, we have identified the specific nitrated tyrosines for a number
of proteins. We have observed that al least enolase and glyceraldehyde 3-phosphate
dehydrogenase are nitrated at the same tyrosine site known to be phosphorylated. This is an
important finding, which is worthwhile further studying. In fact, it may indicate a possible
involvement of nitration in signaling pathways and phosphorylation cascades. Regarding the
aggregation studies, we have isolated detergent insoluble-protein fractions from spinal cord of
G93A SOD1 mice and we have completed the comprehensive characterization of all the
proteins contained. With this study we have identified the protein constituents of aggregates,
still unknown, and therefore have contributed to the comprehension of the role of protein
inclusions in ALS pathogenesis.
Laboratory of Molecular Pathology
In vitro models for investigating motor neuron pathologies
176
ANNUAL REPORT 2008

IRFMN
Presence of mutant forms of specific proteins plays a key role in many neurodegenerative
diseases.
Experimental models in vivo and in vitro are sorely needed to study the effects of these toxic
proteins. We have applied the pTet-Off system to control gene expression through the level of
tetracyclines to a murine motor neuron-like cell line (NSC-34) establishing new cell lines
(called NSC-34 tTA) that stably express the transactivating protein tTA. These cell lines are
suitable to study the pathogenic mechanisms of motor neuron diseases after transient/stable
transfection with genes of interest for these pathologies.
We used this approach to establish NSC-34 tTA cell lines that express in a doxycycline
inducible fashion the human G93A mutant superoxide dismutase 1. Mutant forms of superoxide
dismutase 1 are responsible for some of the familial forms of amyotrophic lateral sclerosis.
These conditional cell lines as well as others previously obtained that constitutively express
mutant G93A superoxide dismutase 1 are used to study the pathogenic mechanisms of
amyotrophic lateral sclerosis.
Novel intracellular targets in the selective degeneration of motor neurons
in amyotrophic lateral sclerosis
Amyotrophic lateral sclerosis is a rapidly fatal neurodegenerative disease characterized by loss
of motor neurons. The management of this disease remains essentially supportive and
symptomatic. Understanding the mechanisms underlying the disease is a way to favor more
efficient therapeutic strategies. Alterations of mitochondria morphology were observed in the
early stages of the disease in the motor nerve terminals of ALS patients and in murine
experimental models. For these reasons we addressed our studies to determine biochemicalmolecular
alterations involved in the mitochondrial damage utilizing our cellular models. We
have studied the toxicity of mutant forms of superoxide dismutase 1, responsible for some
familial forms of amyotrophic lateral sclerosis. We showed that mutant superoxide dismutase 1
(G93ASOD1) altered the mitochondrial morphology and that motor neurons are selectively
susceptible to this damage. Mitochondrial damage was modulated by the extent of expression of
G93ASOD1 protein. Furthermore the expression of G93ASOD1 protein increased the
susceptibility of motor neurons to inhibitors of the electron transport chain (ETC) and to
oxidants. Exposure to drugs or exogenous compounds impairing the ETC could thus be a risk to
motor neurons of individuals carrying mutant superoxide dismutase 1.
Cytochrome P-450 superfamily
Cytochrome(s) P-450 have evolved into a large superfamily that varies enormously in substrate
affinity and product formation. This system plays a major role in the metabolism of drugs and
other chemicals. The majority of existing drugs depends on the P-450 system for terminating
their biological effects or for side effects or adverse reaction. The laboratory has a long-standing
interest in the induction/degradation mechanisms of specific cytochrome P-450 families due to
drug administration or to disease states. Our recent research focused on cytochrome P-450
induction by herbal remedies such as Hypericum perforatum extracts (St. John’s Wort), which
have an alleged activity in mild to moderate depression but interfere with the effect of several
drugs.
Activation of enzymes of the heme metabolic pathway (heme oxygenase
system, biliverdin reductase) as a protective response to stress
The enzymatic system of heme oxygenase (HO) is devoted to cellular degradation of heme
containing molecules, like cytochromes and hemoglobin, and to recycling of iron. Products
formed by the catalytic activity of HO - carbon monoxide and bile pigments - are important
regulating factors in the cell. An increase of HO activity (which is usually sustained by
activation of the inducible form HO-1) is now considered a protective mechanism against
177
ANNUAL REPORT 2008

IRFMN
untoward stimuli particularly when oxidative stress is involved. In the past, the laboratory of
Molecular Pathology identified cytokines as inducers of HO activity and as transcriptional
activators of the HO-1 gene. We are currently investigating the functional significance of HO-1
activation in neurodegeneration.
Laboratory for the Study of Biological Systems
System-level analysis of protein interactions in the epithelial junctional
complex
Inter-cellular junctions form the apical junctional complex and mediate adhesion between
adjacent cells, thus representing the cellular basis for tissue cohesion (for instance, the epithelial
lining of the intestine). In order to acquire system-level understanding of the apical junctional
complex, we have studied (using a methodological approach of ‘network analysis’) all the
protein interactions that have been described at the junctions in epithelial cells of human origin.
We also found that proper ‘hubs’ (i.e., very rare proteins with an exceedingly high number of
interactions with other proteins) were absent from the junctional network. Nevertheless, we
observed that the most connected (albeit non-hub) proteins were also essential proteins. In
addition, we have detected modules within the junctional networks (i.e., densely inter-connected
groups of proteins). Analysis of the modules has highlighted general organizing principles of
the junctional complex, thus confirming the usefulness of network analysis for studying the
components and the interactions of the cell.
Novel regulators of cell motility
Cell motility plays a central role in several biological processes, under both normal (e.g.
embryonic development) and pathological conditions (e.g. tumor cell dissemination). Thus, it is
important to identify the molecular mechanisms that regulate cell motility. In recent years, we
have characterized Junctional Adhesion Molecule-A (JAM-A), a membrane molecule that
localizes to the intercellular tight junctions and binds PDZ-type intracellular proteins. In the
course of these studies, we have discovered that JAM-A expression reduces cell motility. In
addition, we have found that JAM-A enhances microtubule stability and focal adhesion
formation, which are the adhesive points of contact between cells and extracellular matrix. All
these functional changes require amino acid residues that mediate binding to PDZ-type
intracellular proteins. These findings have highlighted a novel mechanism of motility inhibition
that requires the interaction between a membrane protein and PDZ-type intracellular proteins.
Effect of inflammatory cytokines on Junctional Adhesion Molecule-A
(JAM-A)
In the course of inflammatory responses, JAM-A contributes to the leakage of plasma proteins
and the transmigration of circulating leukocytes. Although it has been reported that the
inflammatory cytokine Tumor Necrosis Factor (TNF) causes the disassembly of JAM-A from
the intercellular junctions, the mechanism has not been elucidated fully. Recently, we found that
TNF enhances the solubility of JAM-A in non-ionic detergents and increases the amount of
detergent-soluble JAM-A at the cell surface. In addition, we found that, upon cell treatment with
TNF, higher levels of JAM-A become detectable at the cell surface (by FACS analysis). As
these higher levels of JAM-A derive from the intercellular junctions (and not from intracellular
stores), we propose that TNF causes not only the disassembly of JAM-A from the junctions and
its subsequent redistribution to the cell surface, but also its dispersal in such a way that JAM-A
becomes more easily accessible to the antibodies used for FACS analysis. These findings are
important to highlight potential mechanisms of permeability regulation during inflammation
that might be modulated by inflammatory interventions.
178
ANNUAL REPORT 2008

IRFMN
179
ANNUAL REPORT 2008

IRFMN
180
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF EPIDEMIOLOGY
STAFF
Head
Carlo LA VECCHIA, M.D.
LABORATORY OF GENERAL EPIDEMIOLOGY
Head
Carlo LA VECCHIA, M.D.
Cancer Epidemiology Unit
Head
Cristina BOSETTI, Mat.Sci.D.
Lifestyle Habits and Prevention Unit
Head
Liliane CHATENOUD, Biol.Sci.D.
Epidemiology for Clinical Research Unit
Head
Silvano GALLUS, Comp.Sci.D.
LABORATORY OF EPIDEMIOLOGICAL METHODS
Head
Eva NEGRI, Mat.Sci.D.
LABORATORY OF EPIDEMIOLOGY OF CHRONIC DISEASES
Head
Alessandra TAVANI, Biol.Sci.D.
LABORATORY OF MEDICAL INFORMATICS
Head
Eugenio SANTORO, Comp.Sci.D.
181
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Carlo La Vecchia holds a Doctor of Medicine from the University of Milan, a Master of Science in
Clinical Medicine (epidemiology) from Oxford University and a Diploma from the Post-Graduate School
of Pharmacological Research at the “Mario Negri” Institute for Pharmacological Research in Milan.
Work experiences: He is Head of the Department of Epidemiology at the Mario Negri Institute for
Pharmacological Research in Milan, Italy. He is also Associate Professor of Epidemiology at the
University of Milan, Adjunct Professor of Epidemiology, University of Lausanne, Switzerland (since
2002) and Adjunct Professor of Medicine, School of Medicine, Vanderbilt University, Nashville, TN
(2002-2009).
Dr. La Vecchia is a temporary advisor at the International Agency for Research on Cancer IARC/WHO in
Lyon and at the WHO in Geneva, and a registered journalist in Milan. He was an Honorary Senior
Lecturer in Oral Medicine at the Eastman Dental Institute at the University College of London (1996-
2003) and Adjunct Associate Professor of Epidemiology at the Harvard School of Public Health (1996-
2001).
He is Associate Editor to: European J. of Cancer Prevention and presently serves on the editorial boards
of the Jornals: Alimentazione e Prevenzione; Archives of Medical Science; Asian Pacific Journal of
Cancer Prevention; Current Cancer Therapy Reviews; Dermatology Research and Practice; Digestive and
Liver Disease; Economia Politica del Farmaco; European Journal of Cancer Prevention; European Journal
of Nutrition; In Scope Oncology & Haematology; Journal of Nephrology; Maturitas; Nutrition and
Cancer; Oncology; Open Cancer Journal; Oral Oncology; Revisiones en Ginecologia y Obstetricia;
Revista Española de Nutrición Comunitaria; Revue d'Epidémiologie et de Santé Publique; The Lancet,
italian edition; Tumori. In 1993, he received the Glaxo Prize for medical publication.
He has authored or co-authored over 1,800 publications in peer reviewed journals (1420 included in
Pubmed), with over 52,000 quotations.
Eva Negri got a degree in Mathematics in 1985 at the University of Milan, School of Mathematics.
Awards: EEC scholarship for postgraduate training in Epidemiology (1988).
Areas of interest: Design, conduction and analysis of epidemiologic studies on chronic diseases (e.g.
cancer and myocardial infarction) and injuries, analysis of mortality of cohorts of workers, analysis of
temporal trends and geographic distribution of mortality from cancer, cardiovascular disease, injuries and
other selected conditions, analysis of national health surveys, application of linear modeling techniques to
the analysis of epidemiological data, collaborative re-analyses and meta-analyses of epidemiological
studies.
Work experiences: Since 2007: Laboratory Chief, Unit of Epidemiologic Methods, Department of
Epidemiology; 1992-2006: Unit Chief, Unit of Epidemiologic Methods, Laboratory Epidemiology; since
1990-1992: Researcher at the Laboratory of Epidemiology; 1984-1990: Collaborator of the Laboratory of
Epidemiology.
Selected publications
• Negri E, La Vecchia C, Pelucchi C, Tavani A The risk of acute myocardial infarction after stopping drinking Prev Med
2005; 40: 725-728
• Negri E, Pelucchi C, Talamini R, Montella M, Gallus S, Bosetti C, Franceschi S, La Vecchia C Family history of cancer
and the risk of prostate cancer and benign prostatic hyperplasia Int J Cancer 2005; 114: 648-652
• Negri E, Little D, Boiocchi M, La Vecchia C, Franceschi S. B-cell non-Hodgkin’s lymphoma and hepatitis C virus
infection: A systematic review Int J Cancer 2004; 111: 1-8
• Negri E, Ron E, Franceschi S, La Vecchia C, Preston-Martin S, Kolonel L, et al. Risk factors for medullary thyroid
carcinoma: A pooled analysis Cancer Causes Control 2002; 13: 365-372
• Levi F, La Vecchia C, Boyle P, Lucchini F, Negri E Western and eastern European trends in testicular cancer mortality
Lancet 2001; 357: 1853-1854
182
ANNUAL REPORT 2008

IRFMN
Alessandra Tavani degree in Biological Sciences, University of Milan, Italy (July 1977);
Pharmacological Research Specialist, “Mario Negri” Institute for Pharmacological Research, Milan,
Italy (July 1979).
Work experiences: 1979-81: Researcher at the laboratory of Drug Metabolism, “Mario Negri” Institute
for Pharmacological Research. 1981: Researcher at the Unit for Research on Addictive Drugs (director
prof. H.W. Kosterlitz), University of Aberdeen, Scotland, U.K. 1982-1990: Head of the Unit of Opioid
Neuropharmacology, “Mario Negri” Institute for Pharmacological Research. 1990: Researcher at the
Unit of Clinical Perinatal Pharmacology, “Mario Negri” Institute for Pharmacological Research. From
1991-2006: Head of the Unit of Epidemiology of Chronic Diseases of the Laboratory of Epidemiology,
“Mario Negri” Institute for Pharmacological Research. 2007-: Head of the Laboratory of Epidemiology
of Chronic Diseases of the Department of Epidemiology, “Mario Negri” Institute for Pharmacological
Research.
Awards: "Rafaelsen Scholar Award" from the Collegium Internationale Neuro-Psychopharmacologicum
(CINP), 16th Meeting, Munich (F.R.G.), 1988.
Areas of interest: Epidemiology of cancer and coronary heart disease. Organization of case-control
studies and color studies on cancer and coronary heart disease, including biological sample collection.
Analyses of risk factors related to genetic factors and lifestyles, particularly coffee, diet, physical activity.
Selected publications
• Tavani A, Zucchetto A, Dal Maso L, Montella M, Ramazzotti V, Talamini R, Franceschi S, La Vecchia C. Lifetime
physical activity and the risk of renal cell cancer. Int J Cancer 2007; 120: 1977-1980
• Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C. Tobacco smoking, smoking
cessation, and cumulative risk of upper aerodigestive tract cancers Am J Epidemiol 2008; 167: 468-473
• Tavani A, Pelucchi C, Parpinel M T, Negri E, Franceschi S, Levi F, La Vecchia C. n-3 Polyunsaturated fatty acid intake
and cancer risk in Italy and Switzerland. Int J Cancer 2003; 105: 113-116
• Herrero R, Castellsague X, Pawlita M, Lissowska J, Kee F, Balaram P, Rajkumar T, Sridhar H, Rose B, Pintos
J, Fernandez L, Idris A, Sanchez M J, Nieto A, Talamini R, Tavani A, et al. Human papillomavirus and oral cancer: The
International Agency for Research on Cancer Multicenter Study. J Natl Cancer Inst 2003; 95: 1772-1783
• Tavani A, Pelucchi C, Negri E, Bertuzzi M, La Vecchia C. n-3 polyunsaturated fatty acids, fish, and nonfatal acute
myocardial infarction. Circulation 2001; 104: 2269-2272
Eugenio Santoro got his degree in Computer Science in 1990 at the Milan University. He started to
work at the “Mario Negri” Institute in 1985 as a research fellow. He was Head of the Applied Statistics
and Informatics Unit and of the Applied Statistics and Informatics laboratory, which was part of the
Department of Cardiovascular Research. Since 2001 he is Head of the Laboratory of Medical
Informatics that is currently part of the Department of Epidemiology. His main areas of interest have
been biostatistics and clinical informatics with the development of software for data management and
data analyses of large scale clinical trials in cardiology, such as the GISSI studies (Gruppo Italiano per
lo Studio della Sopravvivenza nell’Infarto miocardico). His main current area of interest is the Internet,
and more recently the web 2.0, and their application in the medical field, in clinical research, and in
medical education through the development of health related websites. He is author or co-author of more
than 160 scientific papers published in peer reviewed journals, and of more than 70 scientific abstracts
submitted to the main international meetings in the cardiology and in the computer science fields. He is
also author of three books (available in Italian) about the use of the Internet in medicine (“Web 2.0 and
medicine”, “Guida alla medicina in rete” and “Internet in medicina. Guida all’uso e applicazioni
pratiche”, published by the Pensiero Scientifico Editore, Rome) and of one section about Internet and
medicine, included in one of the most important italian medical encyclopedia (“Enciclopedia Medica
Italiana”, UTET 2007). He also collaborates to the publication of the Italian National Bioethics
Committee’s guidelines about ethics, health, and the new information technologies.
Selected publications
• Santoro E. Podcast, wiki e blog: il web 2.0 al servizio della formazione e dell’aggiornamento del medico. Recenti Prog
Med 2007;98:484-494.
• Santoro E, Rossi Valentina, Pandolfini C, Bonati M. DEC-NET: The development of the European register of clinical
trials on medicines for children. Clin Trials 2006; 3: 366-375
• Clivio L, Tinazzi A, Mangano S, Santoro E. The contribution of information technology: Towards a better clinical data
management. Drug Dev Res 2006; 67: 245-250
• Santoro E. Internet and information on breast cancer: an overview. Breast 2003; 12: 424-431
• Santoro E, Nicolis E, Franzosi M G, Tognoni G. Internet for clinical trials: Past, present, and future. Control Clin Trials
1999; 20: 194-201
183
ANNUAL REPORT 2008

IRFMN
• Franzosi M G, Santoro E, Zuanetti G, Latini R, Maggioni A P, Tognoni G, GISSI. Indications for ACE inhibitors in the
early treatment of acute myocardial infarction. Systematic overview of individual data from 100.000 patients in
randomized trial. Circulation 1998; 97: 2202-2212
• Franzosi M G, Santoro E, De Vita C, Geraci E, Lotto A, Maggioni A P, Mauri F, Rovelli F, Santoro L, Tavazzi
L, Tognoni G, GISSI.
Ten-year follow-up of the first megatrial testing thrombolytic therapy in patients with acute myocardial infarction.
Results of Gruppo Italiano per lo Studio della Sopravvivenza nell’Infarto-1 study. Circulation 1998; 98: 2659-2665
• Franzosi M G, Latini R, Maggioni A P, Barlera S, Negri E, Nicolis E, Santoro E, Santoro L, Tognoni G, Garattini
S, GISSI-3.
GISSI-3: Effects of lisinopril and transdermal glyceryl trinitrate singly and together on six-week mortality and
ventricular function after acute myocardial infarction. Lancet 1994; 343: 1115-1122
• Maggioni A P, Maseri A, Fresco C, Franzosi M G, Mauri F, Santoro E, Tognoni G, GISSI-2. Age-related increase in
mortality among patients with first myocardial infarctions treated with thrombolysis. N Engl J Med 1993; 11: 1442-1448
Cristina Bosetti got her degree in Mathematics in 1994 at the University of Milan, School of
Mathematics, and the Post-Graduate Diploma in Pharmacological Research in 1999 at the “Mario Negri”
Institute for Pharmacological Research in Milan.
Areas of interest: Epidemiology of cancer, cardiovascular diseases and other chronic conditions. In
particular case-control studies on cancers of the upper respiratory and digestive sites, thyroid, breast,
hormone-related cancers, and on ischemic heart disease. Analysis of risk related to diet, alcohol, tobacco,
reproductive and hormonal factors, occupational and environmental exposure to toxic substances, through
the application of generalized linear models.
She is author/coauthor of more than 130 publications on these issues on peer-reviewed scientific journals.
Work experiences: Unit Head, Unit of Cancer Epidemiology, Department of Epidemiology, Istituto di
Ricerche Farmacologiche “Mario Negri”, Milan (since Sept. 2005); Collaboration with the “International
Epidemiology Institute”, Rockville, MD, USA (since 2002- ); Visiting scientist at the Unit of “Field and
intervention studies”, International Agency for Research on Cancer (IARC), Lyon, France (sett-2000-giu.
2001); Visiting scientist at the Department of Epidemiology, Harvard School of Public Health, Boston,
MA (sett-nov. 1998); Researcher at the Laboratory of Epidemiology, Istituto di Ricerche Farmacologiche
“Mario Negri”, Milan (1998-2005); Researcher at the Laboratory of Mother and Child Health, Istituto di
Ricerche Farmacologiche “Mario Negri”, Milan (1996-1997).
Selected publications:
• Bosetti C, Bertuccio P, Levi F, Lucchini F, Negri E, La Vecchia C. Cancer mortality in the European Union, 1970-2003,
with a joinpoint analysis. Ann Oncol. 2008;19:631-40.
• Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C.Tobacco Smoking, Smoking
Cessation, and Cumulative Risk of Upper Aerodigestive Tract Cancers.Am J Epidemiol. 2007; 167:468-73.
• Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, 1970-
2000. Ann Oncol 2005; 16: 489-511.
• Bosetti C, Spertini L, Parpinel M T, Gnagnarella P, Lagiou P, Negri E, et al. Flavonoids and breast cancer risk in Italy.
Cancer Epidemiol Biomarkers Prev 2005; 14: 805-808.
• Bosetti C, Micelotta S, Dal Maso L, Talamini R, Montella M, Negri E, et al. Food groups and risk of prostate cancer in
Italy. Int J Cancer 2004; 110: 424-428.
• Smith J S, Herrero R, Bosetti C, Munoz N, Bosch F X, Eluf-Neto J, et al. IARC Multicentric Cervical Cancer Study
Group Herpes simplex virus-2 as a human papillomavirus cofactor in the etiology of invasive cervical cancer. J Natl
Cancer Inst 2002; 94: 1604-1613.
Liliane Chatenoud, Doctor in Science Biology, University of Milan (1987); Postgraduate Doctor in
Health Statistics University of Milan (1995).
Areas of interest: Epidemiological studies on obstetric diseases. Dermato-epidemiology. Cancer
epidemiology (case-control studies on cancers of the breast, female genital tract). Analysis of temporal
trends and geographical distribution of perinatal, infant mortality, cancer and other selected conditions
(over 100 publications on these topics, 1993-2005).
Work experiences: Since Sept. 2005: Unit Head, “Lifestyle and Prevention”, Department of
Epidemiology. 1993-2005: Researcher at the Laboratory of Epidemiology; 1988-1990: Staff Statistician
Bracco S.p.A., Milan.
Selected publications
• Naldi L, Chatenoud L, Belloni A, Peserico A, Balato N, Virgili A R, Bruni P L, Ingordo V, Lo Scocco G, Solaroli C,
Schena D, Di Landro A, Pezzarossa E, Arcangeli F, Gianni C, Betti R, Carli P, Farris A, Barabino G F, La Vecchia C,
Parazzini F Medical history, drug exposure and the risk of psoriasis. Evidence from an Italian case-control study
Dermatology 2008 216 : 125-132
184
ANNUAL REPORT 2008

IRFMN
• Valentini L G, Casali C, Chatenoud L, Chiaffarino F, Uberti Foppa C, Broggi G Surgical site infections after elective
neurosurgery: a survey of 1747 patients. Neurosurgery 2008 62 : 88-95
• Chatenoud L, Mosconi P, Malvezzi M, Colombo P, La Vecchia C, Apolone G. Impact of a major thermoelectric plant on
self-perceived health status. Prev Med. 2005;41:328-33.
• Naldi L, Chatenoud L, Linder D, Belloni Fortina A, Peserico A, Virgili AR, et al. Cigarette smoking, body mass index,
and stressful life events as risk factors for psoriasis: results from an Italian case-control study. J Invest Dermatol.
2005;125:61-7.
• Bosetti C, Malvezzi M, Chatenoud L, Negri E, Levi F, La Vecchia C. Trends in cancer mortality in the Americas, 1970-
2000. Ann Oncol 2005; 16: 489-511.
• Chatenoud L, Tavani A, La Vecchia C, Jacobs D R, Negri E, Levi F, Franceschi S. Whole grain food intake and cancer
risk. Int J Cancer 1998; 77: 24-28.
Silvano Gallus got his degree in Computer Science in 1999 at the Milan University.
Areas of interest: Design, data managing, and statistical analyses of case-control studies on the
associations between several risk factors (including in particular tobacco smoking, alcohol drinking and
Mediterranean diet) and risk of cancer, coronary heart disease and several other conditions. Analyses of
occupational cohort studies. Monitoring of prevalence and trends of smoking habit and obesity in Italy.
Author/coauthor of over 130 publication in peer-reviewed journals since 1998.
Work experiences: Chief of the Unit of Epidemiology for Clinical Research of the Department of
Epidemiology (since 2006); computer analyst, graphic designer, and statistical and epidemiological
consultant, Milan and Bergamo (since 2002); researcher at the Laboratory of Epidemiology (since 1997);
creator, designer and webmaster of the website of one of the major Italian public hospital, Milano (1999-
2002).
Since 2008, Dr Gallus is Associate Editor of the journal BMC Public Health, and is member of the
editorial board of The Open Obesity Journal
Selected publications
• Gallus S, Foschi R, Negri E, Talamini R, Franceschi S, Montella M, Ramazzotti V, Tavani A, Dal Maso L, La Vecchia
C. Dietary zinc and prostate cancer risk: a case-control study from Italy. Eur Urol. 2007;52:1052-6.
• Gallus S, Naldi L, Carli P, La Vecchia C; Italian Group for Epidemiologic Research in Dermatology (GISED). Nevus
count on specific anatomic sites as a predictor of total body count: a survey of 3,406 children from Italy. Am J
Epidemiol. 2007;166:472-8.
• Gallus S, Scotti L, Negri E, Talamini R, Franceschi S, Montella M, Giacosa A, Dal Maso L, La Vecchia C. Artificial
sweeteners and cancer risk in a network of case-control studies. Ann Oncol. 2007;18:40-4.
• Gallus S, Schiaffino A, La Vecchia C, Townsend J, Fernandez E. Price and cigarette consumption in Europe. Tob
Control. 2006;15:114-9.
• Gallus S, Zuccaro P, Colombo P, Apolone G, Pacifici R, Garattini S, La Vecchia C. Effects of new smoking regulations
in Italy. Ann Oncol. 2006;17:346-7.
• Clifford GM, Gallus S, Herrero R, Muñoz N, Snijders PJ, Vaccarella S, Anh PT, Ferreccio C, Hieu NT, Matos E,
Molano M, Rajkumar R, Ronco G, de Sanjosé S, Shin HR, Sukvirach S, Thomas JO, Tunsakul S, Meijer CJ, Franceschi
S; IARC HPV Prevalence Surveys Study Group. Worldwide distribution of human papillomavirus types in cytologically
normal women in the International Agency for Research on Cancer HPV prevalence surveys: a pooled analysis.Lancet.
2005;366:991-8.
185
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT’S ACTIVITIES
The Department of Epidemiology is involved in the epidemiology of several common cancers
(including cancers of the breast, female genital tract, respiratory and digestive sites, prostate and
urinary organs, lymphoid malignancies, melanoma, etc.) and of cardiovascular diseases, both
through a descriptive and an analytical approach. Among the activities of descriptive
epidemiology are the analysis of temporal trends and geographical distribution of mortality
from cancer, cardiovascular diseases, and other selected conditions, in Italy and Europe; the
analysis of trends in tobacco consumption in the Italian population, and the corresponding
effects on incidence and mortality from lung and other tobacco-related neoplasms. The analytic
epidemiology activities include the conduction and analysis of case-control studies, aimed at
identifying and better quantifying the association between genetic factors (family history),
selected lifestyle habits (diet, tobacco, alcohol, etc.), use of exogenous hormones and exposure
to various substances and the development of various forms of cancers and cardiovascular
diseases. In particular, the Department works on the analysis of dietary correlates of cancer and
cardiovascular disease risk; quantification of health effects of tobacco smoking, alcohol
consumption and implications for prevention; epidemiological studies on the risk related to oral
contraceptive and hormone replacement therapy use; evaluation of the impact of screening in
the early diagnosis and prevention of cancer. Other activities include: the conduction of
quantitative reviews and meta-analysis of published data; the re-analysis of original data from
epidemiological studies of cancers of the oral cavity and pharynx, pancreas, thyroid, breast,
ovary, and cervix; epidemiological and clinical studies in dermatology in collaboration with the
“Gruppo Italiano per gli Studi Epidemiologici in Dermatologia” (GISED); the analysis of
historical cohort studies of occupational exposures to aromatic amines, asbestos, herbicides and
other known or potential carcinogens; monitoring and prevention of injuries. Other
Department’s activities are related to the development of medical websites, the study of the
quality of medical information available on the Internet, and the training and research on issues
related to medical informatics and those concerning the use in the medical field of the Internet
and web 2.0 applications.
FINDINGS/MAIN RESULTS
The risk of cancers of the pharynx, larynx and oesophagus is significantly increased for smokers
of 2 cigarettes/day as compared to nonsmokers. This highlights the absence of any harmless
level for cigarette smoking in relation to upper aero-digestive tract cancers.
There is a significant inverse association between diet diversity and risk of cancers of the oral
cavity, pharynx and esophagus. Diversity within consumption of vegetables and fruits has also a
protective effect on upper aero-digestive tract cancers, including laryngeal cancer.
According to the type of epidemiological study, the risk of oral and pharyngeal cancer is
decreased by 35% to 48% for high vegetable consumption and by 22% to 45% for high fruit
consumption.
Subjects with a family history of gastric cancer have a 2.5-fold increased risk of gastric cancer.
The risk might be higher when the affected relative is a sibling rather than a parent.
186
ANNUAL REPORT 2008

IRFMN
A diet rich in cereals and potatoes and poor in vegetables and fruit has an unfavorable role on
gastric cancer. Among micronutrients, vitamin E, alpha-carotene and beta-carotene intake are
inversely associated with gastric cancer, while sodium is directly associated with risk.
Vitamin D intake might decrease the risk of colon cancer, particularly of the proximal colon,
while it is unrelated to rectal cancer in the Italian population investigated.
Intake of vitamin D over 3.57 μg/day or 143 IU appears to have a protective effect against
breast cancer.
Our analyses of data from a study conducted in Harbin, Northeast China, give further support to
a positive association between indoor air pollution, history of selected non-malignant lung
diseases, and lung cancer risk in both sexes.
Extended information on family history of cancer, including data on second-degree relatives,
could be useful to improve the discriminatory power of the Gail model risk factors for breast
cancer.
Consistently with previous epidemiological data, in our Italian study alcohol is more strongly
related to estrogen receptors positive than to estrogen receptors negative breast tumors.
In a case-control study on endometrial cancer, we found a significant direct association with
consumption of red meat and a moderate increase in risk for consumption of sweets and poultry.
On the other hand, a high consumption of cereals and vegetables decreased endometrial cancer
risk by 44% and 41%, respectively.
Frequent use of allium vegetables, including onions and garlic, might reduce the risk of
endometrial as well as of various other cancers.
A selection of carotenoids, particularly beta-carotene, beta-cryptoxanthin and lutein/zeaxanthin,
is inversely related with endometrial cancer. Thus, all our findings indicate that diet might have
an important role on endometrial cancer risk.
In our meta-analysis of published studies on coffee and endometrial cancer, the risks are
decreased by 13% for low-to-moderate coffee drinkers and by 36% for heavy drinkers, as
compared to nondrinkers.
Our results rule out a strong relation between physical activity (both occupational and
recreational) and endometrial cancer.
Women with at least one first-degree relative diagnosed with endometrial cancer have a more
than doubled risk of developing the same neoplasm. Also, a family history of uterine and
colorectal, but not breast, cancers increases the risk of endometrial cancer in the Italian
population.
Isoflavones and flavonols may have a distinct role in explaining the effect of fruit and vegetable
against ovarian cancer.
Using data from a collaborative reanalysis of 45 epidemiological studies from 21 countries, we
estimate that oral contraceptives already prevented some 200,000 ovarian cancers and 100,000
deaths from the disease, and that over the next few decades the number of cancers prevented
will rise to at least 30,000 per year.
187
ANNUAL REPORT 2008

IRFMN
Our study of renal cell cancer, based on a large dataset, provides further evidence that coffee,
decaffeinated coffee and tea consumption are not related to renal cell cancer risk.
When 6 major macronutrients are included in the same model of renal cell cancer, the adverse
effect of high intake of starch and the protective effect of polyunsaturated fatty acids remain
statistically significant.
In a meta-analysis of thirty-nine published studies on glycemic index and load, we found a
stronger direct association with colorectal and endometrial cancer than with breast cancer. No
association was found for pancreatic cancer.
Mortality from cancer in the European Union (EU) shows favorable patterns over recent years
in both sexes.
Mortality from cancer in Italy showed a favorable trend over recent years.
The epidemic of non Hodgkin lymphoma observed during the second half of the 20th century
has now started to level off in Europe as in other developed areas of the world.
Mortality from hepatocellular carcinoma remains largely variable across Europe. Favorable
trends were observed in several European countries (including Italy) mainly over the last
decade, particularly in women and in young adults.
For men in the EU, mortality rates from esophageal cancer were stable around 6/100,000
between the early 1980’s and the early 1990’s, and slightly declined in the last decade
(5.4/100,000 in the early 2000s). Oesophageal cancer mortality rates remained comparatively
low in European women.
Over recent years, most countries showed decreasing trends in bladder cancer mortality.
In men in the EU, mortality rates from kidney cancer peaked at 4.8 per 100,000 in 1990-1994,
and declined to 4.1 in 2000-2004. In women in the EU, the corresponding values were 2.1 in
1990-1994 and 1.8 in 2000-2004.
The Mediterranean diet is rich in fat and starch, and hence may be related to overweight.
However, in our study adherence to the major characteristics of the Mediterranean diet is
unrelated to body mass index (BMI) and waist-to-hip ratio, confirming previous data from
Greece and Spain.
Body mass index influences the response to treatment with biological drugs in patients with
moderate to severe psoriasis.
In a survey conducted in 2008 on a sample of 3,035 individuals, representative of the Italian
population aged 15 years or over, 22.0% of Italians described themselves as current cigarette
smokers. Subjects with less privileged socio-economic characteristics should be considered
target populations for tobacco control.
In a sample of 20,800 immigrants, those who are illegal (16,033 subjects), have a higher
frequency (10%) of acute infections of the upper respiratory tract, esophagus, stomach and
duodenum diseases when compared to those immigrants with a regular residence card. No other
major differences emerged between these two subgroups of subjects.
188
ANNUAL REPORT 2008

IRFMN
In a survey on 3406 Italian schoolchildren aged 12-17 years, 17.4% of children had one or more
congenital melanocytic naevi or congenital naevus-like naevi. A higher number of common
melanocytic naevi, family history of malignant melanoma and personal history of diabetes were
directly associated with congenital nevi.
A systematic review of risk factors for falls in community-dwelling elderly people, including
relevant studies published up to 2006, showed that the strongest associations were found for
history of falls, vertigo, gait problems, use of walking aids, and use of antiepileptics.
NATIONAL COLLABORATIONS
Agenzia giornalismo scientifico Zadig, Milano
Associazione Nazionale dei Medici Cardiologi Ospedalieri (ANMCO)
Centro di Riferimento Oncologico, Servizio di Epidemiologia, Aviano (PN)
Comune di Milano, Direzione centrale salute, Settore politiche per la salute
Fondazione LuVI
Fondazione SmithKline, Milano
Gruppo Italiano per lo Studio della Sopravivenza nell’Infarto miocardico (GISSI)
Gruppo Italiano Studi Epidemiologici in Dermatologia GISED, Bergamo
International Centre for Pesticides and Health Risk Prevention, Milano
Istituto Auxologico Italiano, Divisione Malattie Metaboliche III, IRCCS, Piancavallo (VB)
Istituto Auxologico Italiano, Laboratorio Sperimentale di Ricerche Endocrinologiche (LSRE),
IRCCS, Milano
Istituto Europeo di Oncologia, Divisione di Epidemiologia e Biostatistica, Milano
Istituto Europeo di Oncologia, Divisione di Chirurgia Cervico Facciale, Milano
Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione (INRAN), Roma
Istituto Nazionale Neurologico “Carlo Besta”, Milano
Istituto Nazionale per lo Studio e la Cura dei Tumori, Oncologia Sperimentale, Unità di Eredità
Poligenica, Milano
Istituto Tumori “Fondazione Pascale”, Servizio di Epidemiologia, Napoli
Novartis Vaccines SpA, Siena
Ordine dei Medici della Provincia di Bari
Ospedale Casa Sollievo della Sofferenza San Giovanni Rotondo
Ospedale Niguarda Ca’ Granda, Dipartimento Trapianti di Fegato, Milano
Ospedale Niguarda Ca’ Granda, Istituto di Fisiologia Clinica CNR, Sezione di Milano, Milano
Ospedali Riuniti di Bergamo
Policlinico di Monza, Unità Operativa di Endoscopia I, Monza (MI)
Prima Clinica Ostetrico Ginecologica, Mangiagalli, Milano
Società Italiana Attività Regolatorie
Università Bocconi di Milano, Dipartimento di Analisi Istituzionale e Management Pubblico
Università degli Studi di Milano-Bicocca, Dipartimento di Statistica, Milano
Università degli Studi di Milano-Bicocca, I Clinica Otorinolaringoiatria, DNTB, Monza
Università di Milano, Clinica Pediatrica De Marchi, Milano
Università di Milano, Istituto di Statistica Medica e Biometria “G.A. Maccacaro”, Milano
Università di Milano, Prima Clinica Ostetrico Ginecologica, Milano
Università di Pavia, Azienda di Servizi alla Persona, Pavia
Università di Torino, Istituto di Medicina del Lavoro, CTO, Torino
Università di Verona, Clinica Ostetrico Ginecologica, Verona
189
ANNUAL REPORT 2008

IRFMN
INTERNATIONAL COLLABORATIONS
Catalan Institute of Oncology, Institut d’Investigaciò Biomédica de Bellvitge (IDIBELL),
Cancer Prevention and Control Unit, L’Hospitalet de Llobregat, Spagna
Center of Oncology, Dept. of Epidemiology and Cancer Prevention, Varsavia, Polonia
Centre for Research in Environmental Epidemiology (CREAL) and Municipal Institute of
Medical Research (IMIM), Barcellona, Spagna
Harvard School of Public Health, Department of Epidemiology, Boston, USA
Hôpital Necker - Enfants Malades, Centre of the Association Claude Bernard on Auto-immunes
diseases, Parigi, Francia
International Agency for Research on Cancer, Lione, Francia
International Epidemiology Institute (IEI), Rockville, USA
International Life Science Institute (ILSI), Bruxelles, Belgio
National Cancer Institute, Environmental Studies Section, Bethesda, USA
National Cancer Institute, Radiation Epidemiology Branch, Bethesda, USA
National School of Public Health, WHO, Atene, Grecia
Registre Vaudois des Tumeurs, Institut Universitaire de Médecine Sociale et Préventive,
Losanna, Svizzera
Senologic International Society
Society for Internet in Medicine
UNDP/UNFPA/WHO/WORLD Bank special program of research development and research
training in human reproduction, Ginevra, Svizzera.
Universitat Pompeu Fabra, Department of Experimental and Health Sciences, Barcellona,
Spagna
University of Athens Medical School, Department of Hygiene and Epidemiology, Atene, Grecia
University of Las Palmas de Gran Canaria, Department of Clinical Sciences, Las Palmas de
Gran Canaria, Spagna
Vanderbilt University, Department of Medicine, School of Medicine, Nashville, USA
EDITORIAL BOARD MEMBERSHIP
Advances in Therapy (Eva Negri)
Alimentazione e Prevenzione (Carlo La Vecchia)
Archives of Medical Science (Carlo La Vecchia)
BMC Public Health (Silvano Gallus, Associate Editor)
Cancer Causes and Control (Carlo La Vecchia)
Cancer Letter (Carlo La Vecchia, Associate Editor)
Current Cancer Therapy Reviews (Carlo La Vecchia)
Dermatology Research and Practice (Carlo La Vecchia)
Digestive and Liver Disease (Carlo La Vecchia)
Economia Politica del Farmaco (Carlo La Vecchia)
European Journal of Cancer Prevention (Carlo La Vecchia, Associate Editor)
European Journal of Clinical Nutrition (Carlo La Vecchia)
European Journal of Nutrition (Carlo La Vecchia)
Evidence Based Dermatology (Carlo La Vecchia, Liliane Chatenoud)
In Scope Oncology & Haematology (Carlo La Vecchia)
190
ANNUAL REPORT 2008

IRFMN
International Journal of Cancer (Carlo La Vecchia)
Maturitas (Carlo La Vecchia)
Nutrition and Cancer (Carlo La Vecchia)
Open Cancer Journal (Carlo La Vecchia)
Oral Oncology (Carlo La Vecchia)
Portale Partecipasalute.it – http://www.partecipasalute.it (Eugenio Santoro)
Revisiones en Ginecologìa y Obstetricia (Carlo La Vecchia)
Revista Española de Nutriciò Comunitaria (Carlo La Vecchia)
Revue d’Epidémiologie et de Santé Publique (Carlo La Vecchia)
Società Italiana Attività Regolatorie News, SIARNews (Eugenio Santoro)
The Lancet, edizione italiana (Carlo La Vecchia)
The Open Obesity Journal (Silvano Gallus)
Tumori (Carlo La Vecchia)
PEER REVIEW ACTIVITIES
Acta Psychiatrica Scandinavica, American Journal of Clinical Nutrition, American Journal of
Epidemiology, Annals of Epidemiology, Annals of Oncology, Archives of Internal Medicine,
BMC Public Health, British Journal of Cancer, British Journal of Nutrition, British Medical
Journal, Canadian Journal of Physiology and Pharmacology, Cancer, Cancer Causes and
Control, Cancer Detection and Prevention, Cancer Epidemiology Biomarkers and Prevention,
Computer Methods and Programs in Biomedicine, Diabetes/Metabolism Research and Reviews,
Digestive Liver Disease, Epidemiologia & Prevenzione, Epidemiology, Epidemiology &
Biostatistic, European Heart Journal, European Journal of Cancer, European Journal of Cancer
Prevention, European Journal of Clinical Nutrition, European Journal of Epidemiology,
European Journal of Public Health, Evidence-Based Healthcare and Public Health,
Gynecological Endocrinology, Gut, Hepatology, Human Reproduction, International Journal of
Cancer, International Journal of Epidemiology, International Journal of Obesity, JAMA, Journal
of American College of Nutrition, Journal of Clinical Endocrinology and Metabolism, Journal
of Clinical Epidemiology, Journal of Epidemiology and Community Health, Journal of
Investigative Dermatology, Journal of Medical Internet Research , Journal of the National
Cancer Institute, Lancet Oncology, Maturitas, Melanoma Research, Nicotine & Tobacco
Research, Nutrition and Cancer, Obstetric and Gynecology, Oncology, Preventive Medicine,
Public Health Nutrition, Radiation Research, Revue d’Epidèmiologie et de Santé Publique, The
Breast, The Cancer Journal, The Lancet, Tobacco Control, Tumori.
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Advisory Committee of the Oxford Collaborative group on Aetiological Factors in Cancers of
the Female Genital Tract
Comitato Scientifico del Gruppo Italiano Studi Epidemiologici in Dermatologia
Comitato Scientifico della Società Italiana di Colposcopia e Patologia Cervico Vaginale
Data and Safety Monitoring Board of the “Phase II therapeutic trial with a humanized
nonmitogenic CD3 (ChAgly CD3) monoclonal antibody in recently diagnosed type I diabetic
patients”
IARC/OMS di Lione e OMS di Ginevra
Ministero della Salute, Sottocomitato fumo.
191
ANNUAL REPORT 2008

IRFMN
Ministero della Salute, Commissione Oncologica Nazionale.
Scientific Review Committee del UND/WHO/World Bank Human Reproduction Programme
Società Italiana della Riproduzione
EVENT ORGANIZATION
I° edizione del corso ECM “"La ricerca bibliografica su database biomedici", Istituto di
Ricerche Farmacologiche “Mario Negri”, Milano, 13 maggio 2008
I° edizione del corso ECM "Internet e l’aggiornamento professionale in ambito medico", Istituto
di Ricerche Farmacologiche “Mario Negri”, Milano, 28 maggio 2008
I° edizione del corso ECM "Il web 2.0 al servizio della formazione e dell’aggiornamento del
medico", Istituto di Ricerche Farmacologiche “Mario Negri”, Milano, 11 giugno 2008
HiWATE Mid-term review meeting (Project EC FP6), Institute of Pharmacological Research
“Mario Negri”, Milan, 3-5 June 2008
II° edizione del corso ECM “"La ricerca bibliografica su database biomedici", Istituto di
Ricerche Farmacologiche “Mario Negri”, Milano, 24 novembre 2008
II° edizione del corso ECM "Internet e l’aggiornamento professionale in ambito medico",
Istituto di Ricerche Farmacologiche “Mario Negri”, Milano, 25 novembre 2008
II° edizione del corso ECM "Il web 2.0 al servizio della formazione e dell’aggiornamento del
medico", Istituto di Ricerche Farmacologiche “Mario Negri”, Milano, 26 novembre
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
Meeting. FP6 Network of Excellence CCPRB. “Assessor”. Malmo, Svezia. 21-23 Gennaio
2008.
Meeting. Pancreatic Cancer Case-control Consortium.”New Members and Studies”. Rockville,
MD. 23 Gennaio 2008.
International Workshop. Screening for lung cancer and management of early stage disease.
Istituto Clinico Humanitas. “An update in lung cancer epidemiology. Prevention perspectives”.
Rozzano (MI). 25 Gennaio 2008.
Riunione Commissione Droghe Istituto Superiore Sanità. Roma. 30 Gennaio 2008
Seminario a Nerviano Medical Sciences. Cancer mortality and control in the European Union.
Nerviano (MI). 1 Febbraio 2008
192
ANNUAL REPORT 2008

IRFMN
5th International Conference. Cancer Prevention 2008. “Aspirin and NSAID’s in Cancer
Prevention: Attempts of an International Consensus. Consensus finding roundtable”. St. Gallen,
Switzerland. 6-8 Marzo 2008.
Meeting. IARC Working group on burden of cancer caused by asbestos. “Estimates of asbestosrelated
cancer burden: Europe, Australia and New Zealand, Asia and North America”. Lyon,
France. 6-7 Marzo 2008.
XIV Congresso Nazionale delle Malattie Digestive (FIMAD) “Caffe’ e rischio di tumori con
particolare riferimento al tumore colorettale ed epatico” Palacongressi della Riviera di Rimini,
Rimini. 8-12 marzo 2008
VII Barcelona International Congress on the Mediterranean Diet. “2008 Grande Covian
Awardees Conference”. Barcelona, Spagna. 11-12 Marzo 2008.
Convegno Lega Italiana per la Lotta contro i Tumori. I sapori della salute. “Dieta e prevenzione
del cancro: dati italiani”. Roma. 17 Marzo 2008.
Epidemiology Specialist Panel Meeting. WHO headquarters. A cluster randomized controller
trial to evaluate the effectiveness of the clinically integrated RHL-Evidence-based Medicine
Course (New proposal). Ginevra, Svizzera. 31 Marzo 2008.
Commissione Lega Italiana per la Lotta Contro i Tumori. Alimentazione-Oncologia geriatrica.
Roma. 2 Aprile 2008
20° Congresso Nazionale ANDID. “Food nutrition, physical activity and the prevention of
cancer: a global perspective”. Firenze 9-12 Aprile 2008.
15° Congresso nazionale del collegio dei docenti di odontoiatria. “Epidemiologia e fattori di
rischio del carcinoma orale”. Roma. 16-19 Aprile 2008
3rd International meeting methodological issues in oral health research Palazzo Feltrinelli.
“Survival analysis of orthodonic brackets with clustered observations”. Gargnano del Garda. 16-
18 Aprile 2008
X Convegno nazionale tabagismo e servizio sanitario nazionale. ISS. “L’andamento dei tumori
tabacco correlati nell’ultimo decennio”. Roma. 30 Maggio 2008
Meeting – Bladder cancer from pathogenesis to prevention. “Coffee and alcohol consumption
and bladder cancer”. Stockholm, Sweden. 24-25 Aprile 2008
10 th Congress of the European Society of Contraception. “Epidemiology”. Praga, Repubblica
Ceca. 30 Aprile – 3 Maggio 2008
Corso di epidemiologia descrittiva. Porto, Portogallo. 7-8 Maggio 2008
First International Congress on nutrition and cancer. “Diet and cancer with a focus on
Mediterranean diet”. Antalya, Turchia 19-23 Maggio 2008
Toxicology course Vienna. “Nutrition and disease in epidemiological studies”
,”Epidemiological studies on cancer risk factors”. Vienna, Austria 29 Maggio 2008
193
ANNUAL REPORT 2008

IRFMN
X Convegno nazionale tabagismo e servizio sanitario nazionale. “L’andamento dei tumori
tobacco correlate nell’ultimo decennio”. Roma 30 Maggio 2008
Convegno. Prevenzione del carcinoma della cervice uterina: incontro-confronto su conoscenze e
prospettive. “Epidemiologia”. Milano 17 Giugno 2008
1 a Conferenza italiana di oncologia toracica. “The changing in the Epidemiology of Lung
Cancer: the epidemiological evidence”. Napoli 26-28 Giugno 2008
Quinquennial Review of the Cancer Research UK Centre for Epidemiology. Mathematics &
Statistics, directed by Professor Jack Cuzick at the Wolfson Institute of Preventative Medicine,
Barts & the London School of Medicine & Dentistry. Londra, UK, 25 Giugno 2008
Review on Man-made fibers and cancer risk. Visit to the International Epidemiology Institute.
Rockville, MD. 27 Luglio-2 Agosto 2008.
Meeting. Contraception over 40. “Hormonal contraception in aged women: cardiovascular and
cancer risk. Capri 31 Agosto – 1 Settembre 2008
Corso Partecipasalute: Divieto di fumare nei locali pubblici: prime valutazioni dell’effetto della
legge. Istituto di Ricerche Farmacologiche Mario Negri, Milano, 15 Settembre 2008
Membro del working group. Atlas of cancer mortality in Europe. Lione, Francia 5 Ottobre 2008
2nd Congress of the International Society on Nutrigenetics and Nutrigenomics. "Fish
consumption, omega-3 FA and the risk of cancer - cohort and case-control studies" Geneva,
Svizzera 6-8 ottobre 2008
UNDP/UNFPA/WHO/World bank special programme of research, development and research
training in human reproduction. Membro del working group. The safety of quinacrine in
humans when administered as intrauterine doses for non-surgical sterilization. Geneva, Svizzera
8-10 Ottobre 2008.
2 nd European Conference on injury prevention and safety promotion. Membro del working
group. Parigi, Francia. 9-10 Ottobre 2008
6th APOLLO WP Leaders meeting. “WP4: Falls among elderly”. Parigi, Francia. 11 Ottobre
2008
X Congresso Nazionale di Oncologia Medica. “Controllo del cancro in Europa: attuali
andamenti”. Verona 11-14 Ottobre 2008
Meeting Europa Donna. The European breast cancer coalition. “Prevention: Lifestyle factors
and their impact on breast cancer. Milano. 15 Ottobre 2008
XXXII Congresso annuale. Epidemiologia per la Prevenzione. “Consumo di aglio e cipolla e
rischio di tumore”,”Profili dietetici e rischio di tumore dell’alto apparato digerente e respiratorio
in Italia”,”Mortalità per tumori in Italia, dal 1970 al 2002”,”Nutrizione, dieta e attività fisica
nell’epidemiologia e Prevenzione del cancro”. Milano 15-17 Ottobre 2008
194
ANNUAL REPORT 2008

IRFMN
7 th International Symposium on Multiple risk factors in cardiovascular diseases. Prevention and
Intervention – Health Policy. “Smoking and other risk factors for cardiovascular disease: a
quantification”. Venezia (Lido), 22-25 ottobre 2008
Convegno Extra Virgin Olive Oil International week. “Olio d’oliva e tumori: i dati italiani”.
Bari, 4 novembre 2008
Corso di formazione psicologi disassuefazione progetto tabagismo e scuola. “Fumo e tumori: I
rischi del fumo e i benefici della cessazione”. Como, 8 Novembre 2008
II International Conference on olive and health. “Olive oil and cancer risk”. Jaen & Cordoba,
20-22 Novembre 2008
Giornata Nazionale sulla Prevenzione primaria e secondaria del cancro del colon-retto. “Dieta”.
Roma, 4 Dicembre 2008
Convegno Nazionale. Cancerogenesi Professionale: della valutazione del rischio e sorveglianza
sanitaria, alla diagnosi per il riconoscimento della malattia professionale. “Risultati falsopositivi
negli studi epidemiologici sulla cancerogenesi occupazionale ed ambientale”. Torino,
16 dicembre 2008
11° European Conference of Medical and Health Libraries, Helsinki 23-28 giugno 2008. Titolo
relazione: “Supporting the evidence: Clinical Trials, Where to Find and How to Search”
Corso, “Corso di Formazione per volontari dei punti di Accoglienza e Informazione in
oncologia” promosso dall’Istituto Superiore di Sanità e da Alleanza Contro il Cancro, Roma 11-
13 Dicembre 2008. Titolo relazione: “La valutazione di qualità tecnico-formale del materiale
cartaceo e dei siti web”
Corso, “PubMed e ClinicalTrials.gov per l’aggiornamento professionale del medico” promosso
dall’Ordine dei Medici della Provincia di Bari, Bari 21 novembre 2008.
III Edizione del corso di formazione per rappresentanti di associazioni di cittadini & pazienti
“Orientarsi in salute e sanità per fare scelte consapevoli”, promosso da Partecipasalute, Milano
18 novembre 2008. Titolo della relazione: “Come navigare in Internet alla ricerca di
informazioni di qualità. Siti utili per le associazioni di pazienti”
Master Universitario di I° livello in Ricerca Clinica, Università di Milano, anno accademico
2008-2009. Ruolo di docenze nel modulo “Internet e le nuove tecnologie per l’aggiornamento
medico-scientifico”, Milano 12 novembre 2008
Corso "Epidemiologia clinica e metodologia della ricerca" promosso dal Centro per la ricerca e
lo studio del dolore dell’Istituto Mario Negri, Rimini 27-28 ottobre 2008. Titolo relazione:
“Internet in medicina”
Corso “Internet e l’uso dei nuovi strumenti del web 2.0 in ambito medico: potenzialità e
applicazioni”, Istituto Nazionale dei Tumori, Milano 8 ottobre 2008
Corso, “Medicina basata sulle prove di efficacia” promosso dall’Azienda Unità Sanitaria
Locale n.1 di Sassari, Sassari 23 maggio 2008. Titolo della lezione “Siti e applicazioni
Internet in ambito medico: tipologia di informazione e modalità di reperimento”.
195
ANNUAL REPORT 2008

IRFMN
Corso, “Medicina basata sulle prove di efficacia. Corso avanzato” promosso dall’Azienda
Unità Sanitaria Locale n.1 di Sassari, Sassari 12 settembre 2008. Titolo della lezione “Siti e
applicazioni web 2.0 in ambito medico: tipologia di informazioni e metodologia d’uso”.
AIFA
Associazione Italiana per la Ricerca sul Cancro
Comune di Milano
Lega Italiana Lotta contro i Tumori
European Commission (FP7)
GISED
Grunenthal-Formenti
Ministero della Salute
Regione Lombardia
Weber Shandwich
Consorzio Melinda
ISA
GRANTS AND CONTRACTS
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Bosetti C, Gallus S, Peto R, Negri E, Talamini R, Tavani A, Franceschi S, La Vecchia C.
Tobacco smoking, smoking cessation, and cumulative risk of upper aerodigestive tract cancers.
Am. J. Epidemiol., 167: 468-473 (2008).
Bosetti C, Levi F. Boffetta P, Lucchini L, Negri E, La Vecchia C.
Trends in mortality from hepatocellular carcinoma in Europe, 1980-2004.
Hepatology, 48: 137-145 (2008).
Garavello W, Foschi R, Talamini R, La Vecchia C, Rossi M, Dal Maso L, Tavani A,
Levi F, Barzan L, Ramazzotti V, Franceschi S, Negri E.
Family history and the risk of oral and pharyngeal cancer.
Int. J. Cancer, 122: 1827-1831 (2008)
La Vecchia C.
Coffee and liver cancer prevention: is it a fact or just a potential
Hepatology, 48: 7-9 (2008).
Pelucchi C, Dal Maso L, Montella M, Parpinel M, Negri E, Talamini R, Giudice A,
Franceschi S, La Vecchia C.
Dietary intake of carotenoids and retinol and endometrial cancer risk in an Italian
case-control study.
Cancer Causes Control., 19: 1209-1215 (2008).
Smedby KE, Vajdic CM, Falster M, Engels EA, Martinez-Maza O, Turner J,
Hjalgrim H, Vineis P, Seniori Costantini A, Bracci PM, Holly EA, Willett E, Spinelli JJ,
La Vecchia C, Zheng T, Becker N, De Sanjosé S, Chiu BC-H, Dal Maso L, Cocco P,
Maynadié M, Foretova L, Staines A, Brennan P, Davis S, Severson R, Cerhan JR,
Breen EC, Birmann B, Grulich AE, Cozen W.
Autoimmune disorders and risk of non-Hodgkin lymphoma subtypes: a pooled analysis within the
InterLymph Consortium.
Blood, 111: 4029-4038 (2008).
196
ANNUAL REPORT 2008

IRFMN
Fustinoni S, Campo L, Liesivuori J, Pennanen S, Vergieva T, van Amelsvoort LGPM,
Bosetti C, Vam Loveren H, Colosio C.
Biological monitoring and questionnaire for assessing exposure to ethylenebisdithiocarbamates
In a multicenter European field study.
Human Exp. Toxicol., 27: 681-691 (2008).
Rossi M, Negri E, Bosetti C, Dal Maso L, Talamini R, Giacosa A, Montella M,
Franceschi S, La Vecchia C.
Mediterranean diet in relation to body mass index and waist-to-hip ratio.
Publ. Health Nutr., 11: 214-217 (2008).
Galeone C, Pelucchi C, La Vecchia C, Negri E, Bosetti C, Hu J.
Indoor air pollution from solid fuel use, chronic lung diseases and lung cancer in Harbin
and Northeast China.
Eur. J. Cancer Prev., 17: 473-478 (2008).
Bosetti C, McLaughlin JK, Tarone RE, Pira E, La Vecchia C.
Formaldehyde and cancer risk: a quantitative review through 2006.
Ann. Oncol., 19: 29-43 (2008).
Pelucchi C, Naldi L, Di Landro A, La Vecchia A, on behalf of the Oncology Study Group
of the Italian Group for Epidemiologic Research in Dermatology (GISED).
Anthropometric measures, medical history and risk of basal cell carcinoma in an Italian
case-control study.
Dermatology, 216: 271-276 (2008)
Naldi L., Chatenoud L., Belloni A., Peserico A., Balato N., Virgili A.R., Bruni P.L.,Ingordo V.,
Lo Scocco G., Solaroli C., Schena D., Di Landro A., Pezzarossa E., Arcangeli F., Gianni C.,
Betti R., Carli P., Farris A., Barabino G.F., La Vecchia C., Parazzini F.
Medical history, drug exposure and the risk of psoriasis. Evidence from an Italian
case-control study.
Dermatology 216: 125-132, 2008.
Lucenteforte E, Talamini R, Montella M, Dal Maso L, Tavani A, Deandrea S, Pelucchi C,
Greggi S, Zucchetto A, Barbone F, Parpinel M, Franceschi S, La Vecchia C, Negri E.
Macronutrients, fatty acids and cholesterol intake and endometrial cancer.
Ann. Oncol., 19: 168-172 (2008).
Gallus S, Negri E, Boffetta P, McLaughlin JK, Bosetti C, La Vecchia C,
European studies on long-term exposure to ambient particulate matter and
lung cancer
Eur. J. Cancer Prev., 17: 191-194 (2008).
La Vecchia C, Zhang ZF, Altieri A.
Alcohol and laryngeal cancer: an update.
Eur. J. Cancer Prev., 17: 116-124 (2008).
Levi F, Boffetta P, La Vecchia C.
High constant incidence rates of second primary neoplasms.
Eur. J. Cancer Prev., 17: 385-388 (2008).
Bagnardi V., Zatonski W., Scotti L., La Vecchia C., Corrao G:
Does drinking pattern modify the effect of alcohol on the risk of coronary heart disease
Evidence from a meta-analysis.
J. Epidemiol. Community Health, 62: 615-619 (2008).
Pelucchi C., Galeone C., Montella M., Polesel J., Crispo A., Talamini R., Negri E.,
Ramazzotti V., Grimaldi M., Franceschi S., La Vecchia C.
Alcohol consumption and renal cell cancer risk in two Italian case-control studies.
Ann. Oncol., 19: 1003-1008 (2008).
Randi G, Ferraroni M, Talamini R, Garavello W, Deandrea S, Decarli A, Franceschi S,
La Vecchia C.
197
ANNUAL REPORT 2008

IRFMN
Glycemic index, glycemic load and thyroid cancer risk.
Ann. Oncol., 19: 380-383 (2008).
Pandolfini C, Bonati M, RossiV, Santoro E, Choonara I, Naylor C, Sammons H,
Jacqz-Aigrain E, Zarrabian S, Arnau JM, Castel JM, Danés I, Fuentes I.
The DEC-net European register of paediatric drug therapy trials: its content and their context.
Eur.J. Clin. Pharmacol., 64: 611-617 (2008).
Pelucchi C, Gallus S, Garavello W, Bosetti C, La Vecchia C.
Alcohol and tobacco use, and cancer risk for upper aerodigestive tract and liver.
Eur. J. Cancer Prev., 17: 340-344 (2008).
Bosetti C, Levi F, Ferlay J, Garavello W, Lucchini F, Bertuccio P, Negri E, La Vecchia C.
Trends in oesophageal cancer incidence and mortality in Europe.
Int. J. Cancer, 122: 1118-1129 (2008) Num. Reg.: 10978
Pedrazzini G, Santoro E, Latini R, Fromm L, Franzosi M G, Mocetti T, Staszewsky L,
Barlera S, Tognoni G, Maggioni A P, GISSI-3
Causes of death in patients with acute myocardial infarction treated with angiotensinconverting
enzyme inhibitors: Findings from the Gruppo Italiano per lo Studio della
Sopravvivenza nell'Infarto (GISSI)-3 trial
Am. Heart J., 155 : 388-394 (2008).
Garavello W., Giordano L., Bosetti C., Talamini R., Negri E., Franceschi S., La Vecchia C.
Diet diversity and risk of oral and pharyngeal cancer risk.
Eur. J. Nutrition, 47: 280-284 (2008).
Hu J, La Vecchia C, DesMeules M, Negri E, Mery L, and the Canadian Cancer Registries
Epidemiology Research Group.
Meat and fish consumption and cancer in Canada.
Nutr. Cancer, 60: 313-324 (2008).
Bidoli E, Talamini R, Zucchetto A, Polesel J, Bosetti C, Negri E, Maruzzi D, Montella M,
Franceschi S, La Vecchia C.
Macronutrients, fatty acids, cholesterol and renal cell cancer risk.
Int. J. Cancer, 122: 2586-2589 (2008).
Willett EV, Morton LM, Hartge P, Becker N, Bernstein L, Boffetta P,Bracci P, Cerhan J,
Chiu BCH, Cocco PL, Dal Maso L, Davis S, De Sanjose S, Smedby KE, Ennas MG, Foretova L,
Holly EA, La Vecchia C, Matsuo K, Maynadie M, Melbye M, Negri E, Nieters A, Severson R,
Slager SL, Spinelli JJ, Staines A, Talamini R, Vornanen M, Weisenburger DD, Roman E for the
Interlymph Consortium.
Non-Hodgkin lymphoma and obesity: a pooled analysis from the InterLymph consortium.
Int. J. Cancer, 122: 2062-2070 (2008).
Iacobelli N, Gallus S, Petridou E, Zuccaro P, Colombo P, Pacifici R, La Vecchia C, Negri E.
Smoking behaviors and perceived risk of injuries in Italy, 2007.
Prev. Med., 47: 123-126 (2008).
Petridou ET, Pourtsidis A, Dessypris N, Katsiardanis K, Baka M, Moschovi M,
Polychronopoulou S, Koliouskas D, Sidi V, Athanasiadou-Piperopoulou F, Kalmanti M,
Belechri M, La Vecchia C, Curado MP, Skalkidis I.
Childhood leukaemias and lymphomas in Greece (1996-2006): a nationwide registration study.
Arch. Dis. Childhood., 93: 1027-1032 (2008).
Polesel J, Talamini R, La Vecchia C, Levi F, Barzan L, Serraino D, Franceschi S, Dal Maso L.
Tobacco smoking and ther risk of upper aero-digestive tract cancers: A re-analysis of
case-control studies using spline models.
Int. J. Cancer, 122: 2398-2402 (2008).
Bosetti C, Bertuccio P, Levi F, Lucchini F, Negri E, La Vecchia C.
Cancer mortality in the European Union, 1970-2003, with a jointpoint analysis.
Ann. Oncol., 19: 631-640 (2008).
198
ANNUAL REPORT 2008

IRFMN
Lagiou P, Rossi M, Lagiou A, Tzonou A, La Vecchia C., Trichopoulos D.
Flavonoid intake and liver cancer: a case-control study in Greece.
Cancer Causes Control, 19: 813-818 (2008).
Lucenteforte E, Scita V, Bosetti C, Bertuccio P, Negri E, La Vecchia C.
Food groups and alcoholic beverages and the risk of stomach cancer: a case-control
study in Italy.
Nutr. Cancer, 60: 577-584 (2008).
Ferketich AK, Gallus S, Colombo P, Fossati R, Apolone G, Zuccaro P, La Vecchia C.
Physician-delivered advice to quit smoking among Italian smokers
Am. J. Prev. Med., 35: 60-63 (2008).
Gnagnarella P, Gandini S, La Vecchia C, Maisonneuve P.
Glycemic index, glycemic load and cancer risk: a meta-analysis.
Am. J. Clin. Nutr., 87: 1793-801 (2008)
Edefonti V, Decarli A, La Vecchia C, Bosetti C, Randi G. Franceschi S, Dal Maso L, Ferraroni M.
Nutrient dietary patterns and the risk of breast and ovarian cancers.
Int. J. Cancer, 122: 609-613 (2008)
Levi F, Ferlay J, Galeone C, Lucchini F, Negri E, Boyle P, La Vecchia C.
The changing pattern of kidney cancer incidence and mortality in Europe.
BJU Int., 101: 949-958 (2008).
Gallus S, Naldi L and the Oncology Study Group of the Italian Group
for Epidemiologic Research in Dermatology (GISED).
Distribution of congenital melanocytic naevi and congenital naevus-like naevi in a survey
of 3406 Italian schoolchildren
Br. J. Dermatol., 159: 433-438 (2008).
Corrao G, Zambon A, Conti V, Nicotra F, La Vecchia C, Fornari C, Cesana G, Contiero P,
Tagliabue G, Nappi RE, Merlino L.
Menopause hormone replacement therapy and cancer risk: an Italian record linkage investigation.
Ann. Oncol., 19: 150-155 (2008).
Deandrea S, Talamini R, Foschi R, Montella M, Dal Maso L, Falcini F, La Vecchia C,
Franceschi S, Negri E.
Alcohol and breast cancer risk defined by estrogen and progesterone receptor status:
A case-control study.
Cancer Epidemiol. Biomarkers Prev., 17: 2025-2028 (2008).
Foschi R, Lucenteforte E, Bosetti C, Bertuccio P, Tavani A, La Vecchia C, Negri E.
Family history of cancer and stomach cancer risk.
Int. J. Cancer, 123: 1429-1432 (2008).
Pelucchi C, Tavani A, La Vecchia C.
Coffee and alcohol consumption and bladder cancer.
Scand. J. Urol. Nephrol., 42 (Suppl. 218): 37-44
Negri E, La Vecchia C, Collaborative Group on Epidemiological Studies of Ovarian Cancer.
Ovarian cancer and oral contraceptives: collaborative reanalysis of data from 45
epidemiological studies including 23 257 women with ovarian cancer and 87 303 controls.
Lancet 371: 3003-314 (2008).
Rossi M, Negri E, Lagiou P, Talamini R, Dal Maso L, Montella M, Franceschi S, La Vecchia C.
Flavonoids and ovarian cancer risk: A case-control study in Italy.
Int. J. Cancer, 123: 895-898 (2008).
Levi F., Randimbison L., Te VC, Maspoli Conconi M., La Vecchia C.
Risk of prostate, breast and colorectal cancer after skin cancer diagnosis.
Int. J. Cancer, 123: 2899-2901 (2008).
199
ANNUAL REPORT 2008

IRFMN
Boffetta P, McLaughlin JK, La Vecchia C, Tarone RE, Lipworth L, Blot WJ
False-positive results in cancer epidemiology: a plea for epistemological modesty.
JNCI, 100: 988-995 (2008).
Bosetti C, Levi F, Ferlay J, Lucchini F, Negri E, La Vecchia C.
Incidence and mortality from non-Hodgkin lymphoma in Europe: the end of an epidemic
Int. J. Cancer, 123: 1917-1923 (2008). Num. Reg.: 11294
Malvezzi M, Bosetti C, Negri E, La Vecchia C, Decarli A.
Cancer mortality in Italy, 1970-2002.
Tumori, 94: 640-657 (2008).
Poli A, Marangoni F, Paoletti R, MannarinoE, Lupattelli G, Notarbartolo A, Aureli P,
Bernini F, Cicero A, Gaddi A, Catapano A, Cricelli C, Gattone M, Marrocco W,
Porrini M, Stella R, Vanotti A, Volpe M, Volpe R, Cannella C, Pinto a, del Toma E,
La Vecchia C, Tavani A, Manzato E, Riccardi G, Sirtori C, Zambon A.
Non-pharmacological control of plasma cholesterol levels.
Nutr. Metab. & Cardiovasc. Dis. 18: S1-S16 (2008).
Bosis S, Esposito S, Niesters HGM, Zuccotti GV, Marseglia G, Lanari M, Zuin G,
Pelucchi C, Osterhaus ADME, Principi N.
Role of respiratory pathogens in infants hospitalized for a first episode of
wheezing and their impact on subsequent recurrences.
Clin. Microbiol. Infect., 14: 677-684 (2008).
Zucchetto A, Talamini R, Dal Maso L, Negri E, Polesel J, Ramazzotti V, Montella M,
Canzonieri V, Serraino D, La Vecchia C, Franceschi S.
Reproductive, menstrual, and other hormone-related factors and risk of renal cell cancer.
Int. J. Cancer, 123: 2213-2216 (2008).
Ju J, La Vecchia C, DesMeules M, Negri E, L. Mery and the Canadian Cancer Registries
Epidemiology Research Group.,
Nutrient and fiber intake and risk of renal cell carcinoma.
Int. J. Cancer, 60: 720-728 (2008).
Lucenteforte E, Garavello W, Bosetti C, Talamini R, Zambon P, Franceschi S,
Negri E, La Vecchia C.
Diet diversity and the risk of squamous cell esophageal cancer.
Int. J. Cancer, 123: 2397-2400 (2008).
Crispo A, D’Aiuto G, De Marco MR, Rinaldo M, Grimaldi M, Capasso I, Amore A,
Bosetti C, La Vecchia C, Montella M.
Gail model risk factors: impact of adding an estended family history for breast cancer.
Breast J., 14: 221-227 (2008).
Valentini LG, Casali C, Chatenoud L, Chiaffarino F, Uberti-Foppa C, Broggi G.
Surgical site infections after elective neurosurgery: a survey of 1747 patients.
Neurosurgery, 61: 88-95 (2008).
Boffetta P, McLaughlin JK, La Vecchia C.
Reply: “Environment” in cancer causation and aetiological fraction: limitations and ambiguities.
(by Boffetta, P et al. (2007) Carcinogenesis, 28, 913-915).
Carcinogenesis, 29: 1850 (2008).
Dal Maso L, Zucchetto A, Talamini R, Serraino D, Stocco CF, Vercelli M, Falcini F,
Franceschi S, for Prospective Analysis of Case-control studies on Environmental factors and health
(PACE) study group, La Vecchia C, Negri E, Gallus S, Galeone C, Pelucchi C,
Effect of obesity and other lifestyle factors on mortality in women
with breast cancer.
Int. J. Cancer 123: 2188-2194 (2008)
200
ANNUAL REPORT 2008

IRFMN
Pelucchi C, Negri E.
Bladder cancer.
In: Heggenhougen IH, Quah S, eds., International Encyclopedia of Public Health,
First Edition, San Diego, Academic Press, 1: 311-318 (2008).
Parazzini F, Cipriani S, Bianchi S, Gotsch F, Zanconato G, Fedele L.
Risk factors for deep endometriosis: a comparison with pelvic and ovarian endometriosis.
Fertil. Steril., 90:174-179 (2008).
Naldi L, Addis S, Chimenti S, Giannetti A, Picardo M, Tomino C, Maccarone M,
Chatenoud L, Bertuccio P, Caggese E, Cuscito R and the Psocare Study Centres.
Impact of body mass index and obesity on clinical response to sistemic treatment for psoriasis.
Dermatology,, 217: 365-373 (2008).
Boers D, van Amelsvoort L, Colosio C, Corsini E, Fustinoni S, Campo L, Bosetti C,
La Vecchia C, Vergieva T, Tarkowski M, Liesivuori J, Steerenberg P, van Loveren H.
Asthmatic symptoms after exposure to ethylenebisdithiocarbamates and other
pesticides in the Europit field studies.
Human Exp. Toxicol., 27: 721-727 (2008).
Van Almelsvoort LGPM, Mohren DCL, Slangen JJ, Swaen G, Corsini E, Fustinoni S,
Vergieva T, Bosetti C, Liesivuori J, Tarkoeski M, Colosio C, van Loveren H.
Immune effects and exposure to ethylenebisdithiocarbamate pesticides in re-entry workers
in the Netherlands.
Human Exp. Toxicol., 27: 693-699 (2008).
Swaen GMH, van Amelsvoort LGPM, Boers D, Corsini E, Fustinoni S, Vergieva T,
Bosetti C, Pennanen S, Liesivuori J, Colosio C, van Loveren H.
Occupational exposure to ethylenebisdithiocarbamates in agriculture and allergy: results
From the EUROPIT field studies.
Human Exp. Toxicol., 27: 715-720 (2008).
Negri E.
Message 3: Prevent falls.
Arch. Ellenic med., 25 (S 1): 19-26 (2008).
201
ANNUAL REPORT 2008

IRFMN
LAY PRESS SELECTION PUBLISHED IN 2008
Santoro E. I podcast al servizio della formazione e dell'aggiornamento del cardiologo. Recenti Prog Med 2008 99:
163-170
Santoro E. Health-e-child e il grid computing in ambito sanitario: un nuovo supporto alla dignostica e alla clinica.
SIAR News 2008 52: 65-66
Santoro E. Tecnologia RSS e aggregatori di notizie. Ricerca & Pratica 2008 n.139: 23-26
Santoro E. I podcast nell'aggiornamento professionale del medico (parte I)
Ricerca & Pratica 2008 n.140: 68-70
Santoro E. I podcast nell'aggiornamento professionale del medico (parte II)
Ricerca & Pratica 2008 n.141: 110-111
Santoro E. Le cartelle cliniche personali in rete. Un nuovo modo di controllare le proprie informazioni sanitarie
Ricerca & Pratica 2008 n.142: 157-159
Santoro E. Social network e ricercatori biomedici. Ricerca & Pratica 2008 n.143: 199-200
Santoro E. I blog e l’aggiornamento medico. Ricerca & Pratica 2008 n.144: 255-257
Santoro E. Web 3.0 e medicina. CARE 2008 2: 6-7
Santoro E. Web 3.0 e medicina: un nuovo web all'orizzonte Partecipasalute 2008;
http://www.partecipasalute.it/cms_2/node/790
Santoro E. Cartelle cliniche informatizzate: un aiuto alla pratica clinica e alla ricerca medica. Partecipasalute 2008;
http://www.partecipasalute.it/cms_2/node/902
Santoro E. Le cartelle cliniche personali. Partecipasalute 2008; http://www.partecipasalute.it/cms_2/node/875
Santoro E. Google health e le cartelle sanitarie on line: una nuova sfida per Google. Partecipasalute 2008;
http://www.partecipasalute.it/cms_2/node/919
Santoro E. Cartelle cliniche informatizzate, standard e interoperabilità: un aiuto alla pratica clinica e alla ricerca
medica. SIAR News 2008 53: 51-52
Santoro E. Google Flu Trends e la previsione di epidemie. Partecipasalute 2008;
http://www.partecipasalute.it/cms_2/node/983
OTHER PRODUCTS PUBLISHED IN 2008
Pistotti V, Santoro E. “Navigare sulla rete alla ricerca di informazioni di salute”. In: La dispensa di Partecipasalute:
orientarsi in salute e sanità per fare scelte consapevoli. IRFMN, Milano, 2008; 71-81
202
ANNUAL REPORT 2008

IRFMN
RESEARCH ACTIVITIES
LABORATORY OF GENERAL EPIDEMIOLOGY
Case-control studies on cancer
The collection and analysis of data from case-control studies on cancer have continued in 2008.
This is an integrated research program, designed to identify and quantify the role of
environmental, genetic factors, and their interaction on several common neoplasms, with
specific reference to the Italian population. The program started in the early 1980's and has
generated, during the last five years, over 200 publications. The project has evolved by
continuously updating the datasets, by refining and validating the data collection instruments,
by developing and integrating a detailed food composition database and introducing a larger
number of relevant covariates. Further developments are on-going, including follow-up of
subjects through a record-linkage approach and collection and analysis of biological samples.
The dataset includes now about: 6,500 cases of colorectal and breast cancers; over 1,000 of oral
and pharyngeal, stomach, endometrial, ovarian, prostate and kidney cancers; several hundred
cases of other common neoplasms, and over 19,000 controls. Collection of biological samples
has been pursued for new studies on several cancer sites, including laryngeal, bladder and
colorectal cancer, which will allow to analyze selected genetic polymorphisms, insulin-like
growth factors (IGF), adiponectin and other biomarkers.
The results of the main analyses conducted within 2008 are summarized below.
Tobacco smoking and upper aero-digestive tract cancers: a reanalysis
Although tobacco smoking has long been recognized as a major risk factor for cancer of the
upper aero-digestive tract (UADT, i.e., oral cavity, pharynx, larynx, and oesophagus), only a
few studies provided estimates of the effect of very low tobacco consumption. We evaluated the
dose-response relationship between UADT cancers and tobacco smoking using logistic
regression spline models. We included 1,241 UADT male cases and 2,835 male controls pooled
from a large series of case-control studies conducted in northern Italy and in the Swiss Canton
of Vaud during the last 2 decades. For cancers of the pharynx, larynx and oesophagus the risk
steadily increased with number of cigarettes/day and was significantly higher already for
smokers of 2 cigarettes/day as compared to nonsmokers. The effect of tobacco smoking at low
levels seemed less evident for laryngeal cancer since the excess risk begun with 6
cigarettes/day. In conclusion, for all the examined UADT sites, a dose-response relationship
between cancer risk and cigarette smoking emerged. The excess risk among people smoking 2
cigarettes/day highlights the absence of any harmless level for cigarette smoking.
Diet diversity and upper aero-digestive tract cancers
We analyzed the role of diet diversity (the variety of foods consumed) on the risk of UADT
cancers, including oral and pharyngeal, oesophageal and laryngeal cancer. The study of oral and
pharyngeal cancer included 805 cases and 2,081 controls. A significant inverse association was
observed with total diet diversity: the multivariate odds ratio (OR) was 0.78 for subjects in the
highest tertile of diversity. Inverse relations were found also for diversity within vegetables (OR
= 0.62) and fruits (OR = 0.67). The study of squamous cell esophageal cancer included 304
cases and 743 controls. ORs in the highest quartile of intake were 0.42 for total diversity, 0.34
for vegetable diversity and 0.51 for fruit diversity. The study of laryngeal cancer included 527
cases and 1297 controls. A significant inverse association was observed for vegetable diversity
203
ANNUAL REPORT 2008

IRFMN
(OR=0.41 for the highest versus the lowest quartile) as well as for fruit diversity (OR=0.40).
Conversely, a direct association was found for meat diversity (OR=1.67).
Dietary factors and oral and pharyngeal cancer
We reviewed data from six cohort studies and approximately 30 case-control studies on the
relation between selected aspects of diet and the risk of oral and pharyngeal cancer. The pooled
relative risk (RR) for high vegetable consumption were 0.65 from three cohort studies on
UADT cancer, and 0.52 from 18 case-control studies of oral and pharyngeal cancer.
Corresponding RRs for fruit consumption were 0.78, and 0.55.
Family history and gastric cancer
The relation between family history of cancer in first-degree relatives and the risk of stomach
cancer has been considered in our study including 230 cases and 547 matched controls. Relative
to subjects with no history, those with a family history of gastric cancer had an OR of gastric
cancer of 2.5. The OR was higher when the affected relative was a sibling (OR = 5.1) rather
than a parent (OR = 2.2), although the heterogeneity test was not significant.
Food groups, micronutrients and gastric cancer
In the same study of gastric cancer, we examined the relation with selected dietary factors. A
direct association was observed for cereals (OR=2.07 for the highest compared to the lowest
quintile of intake), soups (OR=1.94), and potatoes (OR=2.04). Conversely, inverse relations
were observed with vegetables (OR=0.47) and fruit intake (OR=0.53). No specific constituent
of plant foods has been consistently identified to explain the inverse association between nonstarchy
vegetables and fruit and gastric cancer. In our study, we estimated micronutrient intake
using information from a validated and reproducible food frequency questionnaire, through an
Italian food composition database. We found decreased ORs for the highest vs. lowest quartile
of vitamin E (OR = 0.50), alpha-carotene (OR = 0.52) and beta-carotene (OR = 0.42) intake.
Gastric cancer was directly associated with sodium, with ORs of 2.22 for the second, 2.56 for
the third and 2.46 for the fourth quartile of intake. No significant relation emerged with iron,
calcium, potassium, zinc, vitamin C, thiamin, riboflavin, niacin, vitamin B6, folate, vitamin D,
retinol, beta-cryptoxanthin, lycopene and lutein plus zeaxanthin.
Vitamin D and colorectal cancer
Epidemiologic evidence indicates that vitamin D is inversely associated with risk of colon or
rectal cancer or both. In our analyses on 1953 cases and 4154 controls, we found that adjusted
ORs for colon cancer decreased after the fifth decile of vitamin D intake, reaching 0.69 for the
ninth and tenth deciles. The inverse association appeared to be somewhat more pronounced for
the proximal than the distal colon, and was similar among strata of geographic region and
calcium intake. Rectal cancer was unrelated to vitamin D intake in this population.
Vitamin D and breast cancer risk
We investigated whether vitamin D was associated with risk of breast cancer in our study
including 2569 cases of breast cancer and 2588 hospital controls. After allowance for major risk
factors for breast cancer and dietary covariates, including calcium and energy intake, there was
no association up to the seventh decile between vitamin D intake and breast cancer. Thereafter,
the OR declined, so that the overall trend was statistically significantly inverse. The OR for
subjects in the three highest deciles of consumption of vitamin D compared with those in the
lowest ones combined was 0.79 (95% CI, 0.70-0.90). Only intake of vitamin D over 3.57 μg or
143 IU appeared to have a protective effect against breast cancer.
204
ANNUAL REPORT 2008

IRFMN
Indoor air pollution from solid fuel use and lung cancer
In some areas of China, indoor air pollution (IAP) originating principally from the combustion
of solid fuels has a relevant role in lung cancer. We analyzed the relationship between IAP from
solid fuel use and selected chronic lung diseases and lung cancer risk in Harbin, Northeast
China, an area with a very high baseline risk of lung cancer for both sexes. Data included 218
patients with incident, histologically confirmed lung cancer and 436 hospital controls. We
calculated an index of IAP from solid fuel use exposure using data on heating type, cooking fuel
used, and house measurements. Cases reported more frequently than controls an exposure to
coal fuel for house heating and/or cooking, and the OR for ever vs. never exposed was 2.19. The
ORs of lung cancer according to subsequent tertiles of IAP exposure index were 1.82 and 1.99
as compared with the lowest tertile. The ORs of lung cancer for participants with a history of
chronic bronchitis and tuberculosis were 3.79 and 3.82, respectively. This study gave further
support and quantification of the positive association between IAP, history of selected nonmalignant
lung diseases, and lung cancer risk for both sexes.
Gail model risk factors for breast cancer
We evaluated the feasibility and impact of adding an extended family history as a new breast
cancer risk factor into the Gail model using data from a hospital-based case-control study
conducted by the National Cancer Institute of Naples (southern Italy) between 1997 and 2000.
We compared the model with a first-degree relative (FDR) (standard Gail model) with a model
with information on second-degree relative (SDR); and the FDR with a model with the
combination of FDR and SDR. We computed the C-statistic by comparing the risks found in our
population to those in the Gail-US population. The concordance for the model with FDR was
0.55; the model with SDR showed a modest but significant discriminatory accuracy (0.56), and
the model with the combination of FDR and SDR gave a concordance statistic of 0.57,
indicating a good comparison between the two models. The results of the study showed that
extended family history information could be useful to improve the discriminatory power of the
Gail model risk factors.
Alcohol and breast cancer, according to estrogen receptor status
Studies suggested that the association between alcohol consumption and breast cancer risk is
stronger or limited to tumors expressing estrogen receptors (ER). We considered the issue using
data from a subset of subjects (989 cases and 1350 controls) from our breast cancer study, for
which biological information was available. Alcohol drinking was significantly associated with
ER+ tumors (OR = 2.16 for an intake of ≥13.8 g/day as compared with nondrinkers), but not
with ER- tumors (OR = 1.36). When breast cancers were further classified according to
progesterone receptors (PR), the findings for ER+PR+ cancers were similar to those for all ER+
ones, with an OR of 2.34 for an intake of ≥13.8 g/d. No significant association emerged for ER-
PR- tumors (OR = 1.25). Consistently with previous data, in this study alcohol was more
strongly related to ER+ than to ER- breast tumors.
Diet and endometrial cancer
We analyzed data from a case-control study on endometrial cancer, conducted during 1992-
2006 in three areas of Italy, including 454 cases and 908 controls, to investigate the relationship
between various aspects of diet and risk of endometrial cancer. With reference to main food
groups, we found a significant direct association with consumption of red meat, and a moderate
increase in risk for consumption of sweets and poultry. On the other hand, a high consumption
of cereals and vegetables decreased the risk of endometrial cancer by 44% and 41%,
respectively, while other food groups investigated were not associated with the risk of this
cancer. Another study considered the role of allium vegetables, which were found to be
inversely related with risk of various other cancers in an earlier Italian study. For endometrial
205
ANNUAL REPORT 2008

IRFMN
cancer too, we found significant inverse associations with consumption of both onions (OR =
0.40 for the highest vs. lowest level of consumption) and garlic (OR = 0.62). With reference to
micronutrients, a selection of carotenoids was inversely related with risk of endometrial cancer.
In particular, we found a beneficial effect for high levels of beta-carotene (OR = 0.69), betacryptoxanthin
(OR = 0.65) and lutein/zeaxanthin (OR = 0.59), while other carotenoids (such as
alpha-carotene and lycopene) and micronutrients were unrelated with endometrial cancer. In
conclusion, this study suggested that diet might have an important role on endometrial cancer,
and in particular that a diet rich in meat and poor in vegetables and related micronutrients could
have an adverse effect.
Family history of cancer and endometrial cancer risk
With reference to family history of cancer, our questionnaires had detailed information on
history of all cancers in first-degree relatives and thus allowed a thoroughly investigation of the
relation with endometrial cancer risk. The OR of endometrial cancer for women with at least
one first-degree relative diagnosed the same neoplasm was 2.13, while those who had relatives
with a diagnosis of uterine and colorectal cancer had OR of endometrial cancer of 1.76 and
1.62, respectively. Risks were also increased in women younger than 55 years at diagnosis (OR
= 2.60 for family history of uterine cancer, OR = 2.79 for family history of colorectal cancer).
We found direct associations with some other cancer sites, while there was no association with
the family history of breast cancer. Thus, our study confirmed that a family history of
endometrial, uterine and colorectal cancer increases the risk of endometrial cancer.
Flavonoids and ovarian cancer
Flavonoids intake has been associated with a decreased risk of various cancers in a number of
epidemiological studies. We considered the issue in relation to ovarian cancer, using data from
an Italian hospital-based case-control study including a total of 1031 cases and 2411 controls.
We found an inverse relation between flavonols (OR = 0.63 for the highest vs. lowest quintile)
and isoflavones (OR = 0.51) and ovarian cancer, with significant trend in risk. Adjustment for
fruit and vegetable intake did not modify these associations, suggesting that isoflavones and
flavonols may have a distinct role in explaining the effect of fruit and vegetable against ovarian
cancer.
Oral contraceptives and ovarian cancer
Data of our case-control study of ovarian cancer were included in a collaborative reanalysis of
45 epidemiological studies from 21 countries, for a total of 23,257 women with ovarian cancer
(cases) and 87,303 without ovarian cancer (controls). These data were analysed to assess the
relation between oral contraceptives (OC) use and ovarian cancer. Though an inverse
association is well known, the public-health effects of this reduction in risk will depend on how
long the protection lasts after use ceases. Overall 7308 (31%) cases and 32,717 (37%) controls
had ever used OC, for average durations among users of 4.4 and 5.0 years, respectively. The
longer that women had used OC, the greater the reduction in ovarian cancer risk (p

IRFMN
etiology of renal cell cancer (RCC). We found a direct association with starch intake (OR = 1.9
for highest vs. lowest quintile of intake), and an inverse association with fats from vegetable
sources (OR = 0.6), unsaturated fatty acids (OR = 0.5) and polyunsaturated fatty acids (OR =
0.5), particularly linoleic acid (OR = 0.5) and linolenic acid (OR = 0.7). When 6 major
macronutrients were included in the same model, the adverse effect of high intake of starch
remained statistically significant, together with the protective effect of polyunsaturated fatty
acids.
Reproductive and hormone-related factors and renal cell cancer
Using information from our study of RCC, including 273 female cases and 546 female controls,
we evaluated the effect of reproductive, menstrual and other gender-specific variables among
women. RCC risk was inversely related to age at first birth (OR = 0.7, for ≥ 25 vs.

IRFMN
HI-WATE project on colorectal cancer and drinking water by-products
In 2008, the Department started the collection of data for a case-control study on colorectal
cancer within a project of the 6-FP on the Health Impact of long term exposure to disinfection
by-products in drinking water (DBPs) (HI-Wate study). The study is conducted in the greater
Milan area and in the Provinces of Pordenone and Udine, and includes incident, histologically
confirmed cases enrolled in the major general and teaching hospital of the study area, and
frequency-matched controls, admitted to the same hospital as cases for acute, nonneoplastic
conditions. Cases and controls are interviewed using an extensive questionnaire. Detailed
information on water use and water-related habits for etiologically relevant time periods is
collected, including not only water consumption, but also water related activities, such as
showering, bathing and swimming, that may influence exposure assessment. Moreover, in order
to assess the subjects’ exposure to DBPs and THMs in water, current and historical THM levels,
water source and year of starting chlorination in the study area will be collected from local
companies, authorities and municipalities. About 250 colorectal cancer cases and 250
corresponding controls have been collected, i.e., about half of the total number of cases and
controls which should be enrolled at the end of the study. Giving the large number of people
exposed to chlorinated drinking-water and its DBPs, the study has potentially important
implications in terms of public health. Even modest excess risk in relation to DBP exposure
may in fact have a relevant impact on a population level, and may be responsible of a
considerable number of colorectal cancer cases.
Health status of a sample of illegal immigrants in the municipality of Milan
from 2005 to 2008
We conducted an analysis on a sample of 20,800 immigrants (of which 85% did not have a
regular VISA) collected by a “Consultorio” from Milan over the period 2005-2008. From this
analysis we observed that the diseases most frequently recorded in this sample were various
symptoms, such as asthenia, paresis, headache (ICD-9: 780-789), followed by arthritis and
backpain (found in 10-11% of subjects). It was also noted that the subjects who do not have a
regular VISA tend to have a higher frequency of acute infections of the upper respiratory tract
and diseases of the esophagus, stomach and duodenum (about 10% of the 16,033 irregular
immigrants, compared to around 6% of those in possession of a regular VISA). There were no
other important differences in the frequencies of disease between these two groups of
immigrants.
Impact of body mass index and obesity on clinical response to systemic
treatment for Psoriasis. Evidence from the PSOCARE project
To assess the role of BMI on early clinical response to systemic treatment for psoriasis, we
analyzed data from a nationwide registry and cohort study involving compelling registration of
patients receiving for the first time in their life a new systemic treatment for psoriasis at
reference centres in Italy. Information was gathered by treating physicians with the aid of a webbased
electronic form. Patients with a clinical diagnosis of chronic plaque psoriasis and with 8 and 16
weeks of completed follow-up by March 31 th , 2007 were eligible. A relative reduction of Psoriasis
Area Severity Index (PASI) of at least 75% at follow-up compared to baseline was evaluated as
clinical endpoint (PASI-75). A total of 2,368 patients were analysed at 8 weeks and 2,042 at 16
weeks. PASI-75 was achieved by 819 (34.5%) patients at 8 weeks and 1,034 (50.6%) patients at 16
weeks. The proportion steadily decreased with increased values of BMI, from 41.7% in patients
with BMI < 20 to 29.1% in patients with BMI ≥30 at 8 weeks, and from 59% in patients with
BMI

IRFMN
Distribution of congenital melanocytic naevi and congenital naevus-like
naevi in a survey of 3406 Italian children
To estimate the prevalence of congenital melanocytic naevi (CMN) and congenital naevus-like
naevi (CNLN) in Italian schoolchildren, and to assess variations according to potential risk
factors for melanoma, we conducted a survey in 13 Italian areas on 3406 schoolchildren aged
12-17 years. Children were examined by dermatologists who made a count of small (6-15 mm
in diameter) and medium/large (> 15 mm) CMN/CNLN on 19 anatomical areas. Overall, 592
children (17.4%) had one or more CMN/CNLN. Prevalence of small CMN/CNLN was 16.1%,
and that of medium/large CMN/CNLN was 1.8%. There was no difference between age groups
and sexes. CMN/CNLN were more frequent in children with a higher number of common
melanocytic naevi (multivariate odds ratio, OR = 7.1 for the highest vs. the lowest quartile).
Family history of malignant melanoma (OR = 1.4) and personal history of diabetes (OR = 4.4)
appeared to be directly, and sun exposure inversely associated with CMN/CNLN. No relation
was evident between CMN/CNLN and pigmentary traits, anthropometric characteristics,
dietary habits, freckles, sunburns, sunscreen use or history of selected diseases.
Public health prevention and information
The major products of our activity have also been published in the lay press, in order to increase
the project impact on prevention and public health.
LABORATORY OF EPIDEMIOLOGICAL METHODS
Strategies and best practices for the reduction of injuries
Since 2006, our Department is involved in the Apollo project, an European project aiming at
identifying strategies and best practices for the reduction of injuries. Within this project, our
Department is coordinator of the Workpackage 4 on the development and implementation of
recommendations for the prevention of falls among elderly people in the European Union (EU).
The majority of falls in elderly people are due to a combination of several interacting factors.
Many studies have investigated the role several factors on the risk of falling. Thus we conducted
a systematic review and a quantitative meta-analysis of risk factors for falls in communitydwelling
elderly people, including relevant studies published up to 2006. Fifty-nine studies were
included and 28 risk factors were analyzed, including socio-demographic characteristics,
mobility, sensory, psychological and medical factors, and medication use. The strongest
associations were found for history of falls, vertigo, gait problems, use of walking aids, and use
of antiepileptics.
Moreover, we investigated the amount of information on the participation rates reported from
studies investigating the effectiveness of selected intervention for the prevention of falls among
community-dwelling elderly people. We identified 32 studies implementing interventions based
on an exercise program, an exercise program in combination with other measures, or other types
of interventions. Fourteen studies did not report information on the refusal rate; most studies
reporting the information had a refusal rate between 25 and 50%. We also conducted a survey to
investigate the attitudes of elderly people towards selected intervention for the prevention of
falls. The study was conducted in Italy, Poland, Greece, Hungary, and Slovenia, countries for
which limited data is available on this issue. A total of 1497 subjects aged 65 to 89 years were
interviewed to evaluate their beliefs and attitudes towards two evidence-based interventions, i.e.
a social activity aimed at improving muscle strength and balance (such as exercise class or
dancing), and a home safety assessment and modification program. Among the respondents
about 47% would accept to participate to a social activity, and about 38% would accept a
209
ANNUAL REPORT 2008

IRFMN
program of home hazard assessment and modification. However, we found marked differences
between countries in the acceptability of the two proposed interventions.
Monitoring of cancer mortality in Europe
In 2008, we conducted various analyses within a project launched in 1992 for the monitoring of
cancer mortality in Europe and other areas of the world. The main objective of the project is to
maintain and improve an integrated system for analysis, modeling and interpretation of death
statistics in Europe, created by our group. The project is based on mortality data of the WHO,
and includes the number of cancer deaths in many countries, sex, cause, period, and age of death
in Europe and some other countries outside Europe, together with the estimates of the resident
population. The project is not only descriptive, as a specific effort is devoted to the
interpretation of the observed data on the basis epidemiological evidence. It offers a unique
opportunity for the continued exploitation of mortality data in Europe, with the primary aim of
improving the monitoring and prevention of cancer by optimizing the allocation of economic
resources in health.
Cancer mortality in the European Union
We analyzed cancer mortality in the EU over the period 1970-2003. Overall, cancer mortality
leveled off in men since 1988 and declined in 1993-2003 (annual percent change, APC = -
1.3%). In women, a steady decline has been observed since the early 1970s. The decline in male
cancer mortality has been driven by lung cancer, which leveled off since the late 1980s and
declined thereafter (APC = 2.7% in 1997-2003). Recent decreases were also observed for other
tobacco-related cancers, as oral cavity/pharynx, esophagus, larynx and bladder, as well as for
colorectal and prostate cancers. In women, breast cancer mortality leveled off since the early
1990s and declined thereafter (APC = -1.0% in 1998-2003). Female mortality declined through
the period 1970-2003 for colorectal and uterine cancer, while it increased over the last three
decades for lung cancer (APC = 4.6% in 2001-2003). In both sexes, mortality declined in 1970-
2003 for stomach cancer and for a few cancers amenable to treatment. This update analysis of
the mortality from cancer in the EU shows favorable patterns over recent years in both sexes.
Cancer mortality in Italy
We updated a previous work on Italian cancer mortality with data for 2002 and analyzed trends
over the 1970-2002 period. Total cancer deaths for 2002 in Italy were 163,070 (93,398 men,
69,672 women). Male cancer mortality rose until 1988 and since then has had a 1.4% yearly
fall. The first cause of cancer death in males was lung cancer, accounting for 28% of deaths.
The decrease in mortality from male lung cancer started in the late 1980’s and was the main
reason for the favorable trends in total male cancer mortality, reflecting the change in smoking
prevalence in Italian males. Female total cancer mortality trends have also been favorable, with
an overall yearly drop of 1.1% since 1992. The most frequent causes of cancer deaths in females
were breast and colorectal cancers, accounting for 16% and 14% of cancer deaths, and both
showed declining trends. Female lung cancer rose over the last decades because of the increase
in cigarette smoking since the 1970's in Italian women. Thus, mortality from the most common
cancers in Italy showed a favorable trend over recent years.
Incidence and mortality from non-Hodgkin lymphoma
We updated trends in mortality from non-Hodgkin lymphomas (NHL) considering data up to
2004 in several European countries, and for comparative purpose in the USA and Japan. We
also analyzed patterns in incidence for selected European countries providing national data. In
most European countries, NHL mortality rose up to the mid 1990s, and started to level off or
decline in the following decade. The rates were, however, still increasing in eastern Europe.
Overall, in the EU, mortality from NHL declined from 4.3/100,000 to 4.1 in men and from 2.7
210
ANNUAL REPORT 2008

IRFMN
to 2.5 in women between the late 1990s and the early 2000s. Similarly, NHL mortality rates
declined from 6.5/100,000 to 5.5 in US men and from 4.2 to 3.5 in US women. In most
countries considered, NHL incidence rates rose up to 1995-99, while they tended to level off or
decline thereafter, with particular favorable patterns in countries from northern Europe. Thus,
the epidemic of NHL observed during the second half of the 20th century has now started to
level off in Europe as in other developed areas of the world.
Trends in mortality from hepatocellular carcinoma in Europe
Age-standardized (world standard) mortality rates from hepatocellular carcinoma (HCC) were
computed for 23 European countries over the period 1980-2004. Male overall mortality from
HCC increased in Austria, Germany, Switzerland, and other western countries, while it
significantly decreased over recent years in countries such as France and Italy, which had large
upward trends until the mid-1990s. In the early 2000s, among countries allowing distinction
between HCC and other liver cancers, the highest HCC rates in men were in France
(6.8/100,000), Italy (6.7), and Switzerland (5.9), whereas the lowest ones were in Norway (1.0)
and Ireland (0.8). In women, a slight increase in overall HCC mortality was observed in Spain
and Switzerland, while mortality decreased in several other European countries, particularly
since the mid-1990s. In the early 2000s, female HCC mortality rates were highest in Italy
(1.9/100,000), Switzerland (1.8), and Spain (1.5) and lowest in Greece and Ireland (0.3). HCC
mortality remains largely variable across Europe. Favorable trends were observed in several
European countries mainly over the last decade, particularly in women and in young adults.
Trends in oesophageal cancer incidence and mortality in Europe
We analyzed recent trends in mortality from oesophageal cancer in 33 European countries. For
selected European cancer registration areas, we also analyzed incidence rates for different
histological types. For men in the EU, mortality rates were stable around 6/100,000 between the
early 1980’s and the early 1990’s, and slightly declined in the last decade (5.4/100,000 in the
early 2000s, annual percent change, APC = -1.1%). In several western European countries, male
rates have started to level off or decline during the last decade (APC = -3.4% in France, and -
3.0% in Italy). Also in Spain and the UK, which showed upward trends in the 1990 s, the rates
tended to level off in most recent years. A leveling of rates was observed only more recently in
countries of central and eastern Europe, which had had substantial rises up to the late 1990’s.
Oesophageal cancer mortality rates remained comparatively low in European women, and
overall EU female rates were stable around 1.1-1.2/100,000 over the last 2 decades (APC = -
0.1%). In northern Europe a clear upward trend was observed in the incidence of oesophageal
adenocarcinoma, and in Denmark and Scotland incidence of adenocarcinoma in men is now
higher than that of squamous-cell carcinoma. Squamous-cell carcinoma remained the prevalent
histological type in southern Europe. Changes in smoking habits and alcohol drinking for men,
and perhaps nutrition, diet and physical activity for both sexes, can partly or largely explain
these trends.
Declining mortality from bladder cancer
We updated trends in bladder cancer mortality in 32 European countries and the EU as a whole,
over the period 1970-2004. In the EU overall, bladder cancer mortality rates (age-standardized,
world standard population) were stable up to the early 1990’s at approximately 7/100,000 men
and 1.5/100,000 women, and declined thereafter by approximately 16% in men and 12% in
women, to reach values of 6 and 1.3/100,000, respectively, in the early years of the present
decade. Over recent years, most countries showed decreasing trends. The favorable trends in
men are partly or largely due to the recent declines in the prevalence of smoking in European
men, together with reduced occupational exposure to occupational carcinogens. The decreases
in women are more difficult to explain. Better control of urinary tract infections has probably
211
ANNUAL REPORT 2008

IRFMN
played a role, while the role of diet and other potential urinary tract carcinogens remains
undefined.
The changing pattern of kidney cancer incidence and mortality in Europe
We updated trends in kidney cancer mortality in 32 European countries and the EU as a whole,
over the period 1980-2004. In men in the EU, mortality rates from kidney cancer peaked at 4.8
per 100,000 in 1990-1994, and declined to 4.1 (-13%) in 2000-2004. In women in the EU, the
corresponding values were 2.1 in 1990-1994 and 1.8 (-17%) in 2000-2004. The main decreases
were in Scandinavian countries, and other western European countries. In most eastern
European countries kidney cancer mortality rates tended to stabilize, even if values remained
high, especially in the Czech Republic and Baltic countries. For kidney cancer incidence, there
were decreases in rates for both sexes in Sweden throughout the 25-year calendar period
considered. In the last 10 years considered, incidence rates decreased or tended to stabilize also
in other northern European countries in both sexes, except in the UK.
Record-linkage for cohort analyses
With reference to our project to create an Italian cohort study based on record-linkage, during
2008 we prepared the archives that will allow to link data between our network of case-control
studies (conducted since 1982) and those from the historical archives of the Local Health Unit
(ASL) of Milan, that include several health information such as vital status and date of death of
the subjects. Several stages of the project have been undertaken and completed during the year.
In particular, we finished the identification of subjects recruited in the case-control studies
among the historical archives of the ASL, and a database with univocal information on subjects
from the two sources is now available, thus allowing a linkage of the archives; at the same time,
we completed data collection from all case-control studies in a unique database (using original
data and through the preparation of a codebook that allowed to re-codify studies conducted in
various periods and on several diseases) that includes information from interviews to
approximately 27,000 subjects. During 2008, we searched in various other historical databases
individuals that were not linked in historical archives of the ASL. On 18,257 residents in
Lombardy, 15,860 (86.9%) subjects, including 7,680 cases (82.2%) and 8,911 controls (91.8%)
were linked.
LABORATORY OF EPIDEMIOLOGY OF CHRONIC DISEASES
Organization for data and biological sample collection for case-control
studies
Data collection of epidemiological data is going on and it includes: 1) interview and interviewer
management and training activity for new interviewers; 2) contacts with hospital department
and ethical committee for study approval and conduction; 3) check and codification of patient
questionnaires; 4) diagnosis and histological exam check; 5) organization and management of
biological sample collection; 6) data input management.
Ongoing case-control studies include: cancer of the oral cavity, pharynx, larynx, esophaguscardias,
biliary tract, colorectum, lymphomas, myelomas and sarcomas.
The overall updated dataset include about: 1.250 cases of cancers of oral cavity and pharynx,
700 of the esophagus, 1.100 of the stomach, 6.500 of the colorectum, 600 of the liver, 120 of the
biliary tract, 600 of the pancreas, 850 of the larynx, 500 cutaneous malignant melanoma, 7.000
of the breast, 1.000 of the cervix, 1.000 of the endometrium, 200 of trophoblastic gestational
disease, 200 of the vulva, 2.000 of the ovary, 1.300 of the prostate, 700 of the bladder, 800 of
the kidney and renal pelvis, 600 of the tyroid, 200 of Hodgkin disease, 500 of non-Hodgkin
212
ANNUAL REPORT 2008

IRFMN
disease, 150 of sarcomas, 300 of myelomas and about 18.000 controls. Biological sample
collection includes cancers of the oral cavity, pharynx, larynx, bladder and colorectum aimed to
study genetic polymorphisms.
Definition and management of a cohort of subjects living at Asti for
cardiovascular risk assessment
This study is conducted in collaboration with the ALMA association and the “Ordine dei
Medici di Asti” and includes data collection and first analyses of follow-up.
Case-control studies
Physical activity and endometrial cancer
In the case-control study of endometrial cancer, we investigated the relation with physical
activity at different ages. Taking into account various potential confounding factors, including
age, body mass index, diabetes, hormonal/reproductive factors and energy intake, the OR of
endometrial cancer in women at high level of occupational physical activity, as compared with
those at the lowest level, were 1.69 at 12 years of age, 1.33 between 15 and 19 years, 1.17
between 30 and 39 years and 0.82 between 50 and 59 years. No significant trend in risk with
level of occupational physical activity was found at any age. Similarly, no significant
association was detected with recreational physical activity in different periods of life, since the
OR for the highest vs. lowest level of physical activity were 0.82 at 12 years of age, 0.78
between 15 and 19 years, 1.12 between 30 and 39 years and 0.97 between 50 and 59 years.
Therefore, our results ruled out a strong relation between physical activity and endometrial
cancer.
Coffee consumption and endometrial cancer
Some studies found an inverse relation between coffee consumption and endometrial cancer
risk. We conducted a meta-analysis of published studies on this issue, including 2 cohort (201
cases) and 7 case-control studies (2409 cases). We observed a summary RR of 0.80 (95% CI:
0.68-0.94) for coffee drinkers as compared to nondrinkers. Compared to nondrinkers, the
summary RR were 0.87 (95% CI: 0.78-0.97) for low-to-moderate coffee drinkers and 0.64 (95%
CI: 0.48-0.86) for heavy drinkers.
Coffee, decaffeinated coffee, tea intake and risk of renal cell cancer
The relation between coffee, decaffeinated coffee and tea intake and RCC risk was analyzed in
our case-control study on 767 cases with RCC and 1,534 controls. Coffee intake (mostly
espresso and mocha) was not associated with RCC (OR = 1.02 in drinkers of 4 or more
cups/day compared with drinkers of less than 1 cup/day). No relation was observed with
decaffeinated coffee and tea intake, the ORs being 1.38 and 0.78 for drinkers compared with
nondrinkers respectively, although these two estimates were based on a low number of drinkers.
This study, based on a large dataset, provided further evidence that coffee, decaffeinated coffee
and tea consumption is not related to RCC risk.
Fish consumption and cancer risk
The relation between consumption of fish and n-3 polyunsaturated fatty acid (PUFA) and the
risk of selected neoplasms has been analysed in two series of integrated case-control studies
conducted in northern Italy during 1983-1990 and 1991-2008. In the first series of studies, the
OR for an increase in fish intake of 1 portion/week were 0.7 for rectal cancer; 0.8 for
oral/pharyngeal, esophageal, gastric, colon and laryngeal cancer; 0.90 for pancreatic,
endometrial and ovarian cancer and for multiple myelomas. No consistent relation was found
for liver, gallbladder, breast, prostate, bladder, kidney and thyroid cancers, Hodgkin disease and
213
ANNUAL REPORT 2008

IRFMN
non-Hodgkin lymphomas, although most OR were below unity for these neoplasms, too. In the
second series of studies, the OR for the highest quintile of n-3 PUFAs compared with the lowest
one were 0.5 for oral/pharyngeal, 0.5 for esophageal, 1.4 for gastric, 0.7 for colon, 0.8 for rectal,
1.5 for laryngeal, 0.8 for breast, 1.0 for endometrial, 0.6 for ovarian and 1.0 for renal-cell
cancers; the estimates and the trends in risk were significant for oral/pharyngeal, esophageal,
colon and ovarian cancer. Thus, although prospective studies overall do not provide evidence of
a significant association between n-3 PUFA and cancer incidence, these Italian studies suggest
that consumption of even relatively small amounts of fish decreases the risk of several cancers,
especially of cancers of some parts of the digestive tract.
LABORATORY OF MEDICAL INFORMATICS
Development of the BPCO.CARE website
This web based medical index has been developed by the Laboratory of Medical Informatics in
order to collect, classify, evaluate, and describe the most useful medical information on the web
related to the chronic obstructive pulmonary disease (COPD). The description associated to
each web site included in the BPCO.CARE index illustrates the main contents, the services, and
the tools offered to the users, including those related to the web 2.0 technology such as
podcasts, RSS feeds, blogs, webcasts, and webinars. A section of the BPCO.CARE website also
includes a classified collection of the podcast services available on the net in the COPD and
pulmonology area and information on how to subscribe to. The BPCO.CARE website (available
at http://www.bpcocare.it) has been developed in collaboration with the Department of
Cardiovascular Research.
Maintenance of the CARDIO.CARE, ONCO.CARE, GASTRO.CARE,
NEURO.CARE, PNEUMO.CARE, PAIN.CARE and DERMA.CARE websites
These indexes have been developed by the Laboratory of Medical Informatics in order to
collect, classify, evaluate, and describe the most useful medical information on the web, and to
provide Internet users with an easy means to surf the net. Several medical areas are covered
including oncology (http://www.oncocare.it), neurology (http://www.neurocare.it),
gastroenterology (http://www.gastrocare.it), cardiology (http://www.cardiocare.it),
pulmonology (http://www.pneumocare.it), the pain care and management
(http://www.paincare.it), and dermatology (http://www.dermacare.it). The project is in
collaboration with intramural departments (Department of Oncology, Laboratory of
Neurological Disorders and Department of Cardiovascular Research, Laboratory of General
Practice Research, Laboratory of Translational and Outcome Research in Oncology) and
extramural research groups (Italian Group for Epidemiologic Research in Dermatology,
GISED).
Studies on the typology of the web 2.0 applications in medicine
The Laboratory of Medical Informatics is involved in studies and surveys which aim is to
describe the typology of the web 2.0 applications and tools (including social networks, podcasts,
feed RSS, blogs, and wikis) in medicine available on the net, and how these are perceived by
the medical community.
Internet as a research and formative tool on the chronic pain in cancer
patient
This research project was born in the framework of the project "Pain in the patient with cancer",
in collaboration with the Laboratory of Medical Research and Consumer Involvement and the
Laboratory of Translational and Outcome Research in Oncology. Its aim is to make available
214
ANNUAL REPORT 2008

IRFMN
for doctors, patients and families correct information about therapies for cancer pain and to
produce greater tests on the effectiveness of therapies based on the use of analgesic drugs. This
project is articulated in two main activities:
- Paincare, set-up a catalogue of selected web pages dedicated to the cancer pain and relative
periodical up-to-date, http://www.paincare.it;
- adaptation of the methods and instruments of Evidence Based Medicine to the resources
available on the Internet about chronic pain in patients with cancer. This is done through a
specific questionnaire. We are also considering the opportunity to set up a new Italian cancer
pain website.
Studies on the quality of health information available on the Internet
We are conducting an analysis on the quality and reliability of the information available on the
Internet related to the management of cancer pain. Through an “ad hoc” form using worldwide
applied criteria and principles for the evaluation of health websites and the evidence based
medicine tools, an interdisciplinary team of reviewers is involved in the evaluation of the
quality of information offered by the websites included in the PAIN.CARE medical index. The
same study will also allow to compare the level of the quality of health related information
among different classes of websites. The study, in collaboration with the Laboratory of Medical
Research and Consumer Involvement, is ongoing and the results will be available in 2009.
Training activities
In 2008, the Laboratory of Medical Informatics continued its training activity on issues related
to the use of the Internet in medicine, and extended it to the use of the recent web 2.0
technologies and tools in the medicine area. The members of the laboratory staff activated (or
attended as invited teachers) a number of training courses, workshops, and master courses.
Onsite CME courses for the Italian physicians have also been organized using the
training/educational facilities and equipment available at the Mario Negri Institute.
215
ANNUAL REPORT 2008

IRFMN
216
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF PUBLIC HEALTH
STAFF
Head Department
Maurizio BONATI, MD.
"Angelo & Angela Valenti" Centre for Health Economics (CESAV)
Head of Laboratory
Livio GARATTINI, Econ.D.
Laboratory of Clinical Epidemiology
Head of Laboratory
Guido BERTOLINI, MD.
Clinical Knowledge Engineering Unit
Head Unit
Davide LUCIANI, MD.
Laboratory for Mother and Child Health
Head Laboratory
Maurizio BONATI, MD.
217
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Maurizio Bonati has a Medical School degree at the University of Milan.
Areas of interest: Monitoring and epidemiological evaluation of drug utilisation and effects of drugs and
vaccines in motherhood and childhood. Research methodology in general hospital and paediatric
community practice. Transfer of information to the community. Epidemiology of paediatric and perinatal
care.
Past and present roles both at the Mario Negri Institute and in other institutions: 1973-77 Research Fellow
at the IRFMN, within the Neurochemistry Lab.; 1977-85 Research Assistant at the IRFMN, within the
Clinical Pharmacology Lab.; 1986-93 Chief of the Perinatal Clinical Pharmacology Unit at the IRFMN;
Advisor to WHO for the Drug Utilization Research Group (pregnancy, paediatrics and breastfeeding);
1987-92 coordinator of the International Cooperative Study of Drug Use in Pregnancy, under the auspices
of WHO and the support of EEC; 1992-93 co-editor of The Kangaroo; 2000-05 coordinator of the
European Cooperative Study: “Development of the European register of clinical trials on medicines for
children” (DEC-net), under the 5 th Framework Programme’s Quality of life and Management of Living
Resources; since 1989 he has been director of the Centre for Drug Information; since 1993 head of the
Lab. for Mother and Child Health; since 1997 teacher for the Lombardy region’s professional training
courses; since 2000 teacher for the Lombardy region’s professional training courses; since 2002 Editor of
the Ricerca & Pratica scientific journal; since 2003 professor of the School of Specialisation in
Paediatrics - University of Milan Bicocca; teacher at the annual European course “Evaluation of
Medicinal Products in Children” (promoted by ESDPPP and Eudipharm); from May 2008 Head of
Department Public Health"Mario Negri" Institute for Pharmacology Research.
Selected publications
• Bonati M, Campi R. What Can We Do to Improve Child Health in Southern Italy PLos Medicine 2005;2(9):e250.
• Pandolfini C, Bonati M. A literature review on off-label drug use in children. Eur J Ped 2005;164: 552-558.
• Santoro E, Rossi V, Pandolfini C, Bonati M. DEC-net: the development of the European Register of Clinical Trials on
Medicines for Children. Clinical Trials 2006;3:366-375.
• Clavenna A, Rossi E, De Rosa M, Bonati M. Use of Psychotropic Medications in Italian Children and
Adolescents. Eur J Pediatr 2007;166:339-47.
• Maschi S, Clavenna A, Schiavetti B, Campi R, Bernat M, Bonati M. Neonatal outcome following pregnancy exposure to
antidepressants: a prospective controlled cohort study. BJOG 2008;115:283-289.
• Usala T, Clavenna A, Zuddas A, Bonati M. Randomised controlled trials of Selective Serotonin Reuptake Inhibitors in
treating depression in children and adolescents: a systematic review and meta-analysis. European
Neuropsychopharmacology 2008;18:62-73.
Livio Garattini: got his degree in Economics in March 1983 at the Bocconi University in Milan.
Educational activities: “King’s Fund College”, London: courses of health care management; “Centre for
Health Economics”, York: review of publications on the English NHS; “Ecole Nationale de la Santé
Publique”, Rennes: courses of health policy.
Areas of interest: Health Economics and Health Policy Analysis.
At present he is the Director of CESAV (Centre of Health Economics A. e A. Valenti - M. Negri
Institute); 1981-1983: researcher at M. Negri Institute; 1983-1984: clerk at Banca Commerciale Italiana
in Milan; 1984- 1985: junior consultant at “Sogess srl” in Milan; 1985-1990: researcher at Bocconi
University in Milan.
Selected publications:
• Garattini L, Ghislandi S (2006) “Off-patent drugs in Italy- A short-sighted view” The European Journal of Health
Economics 7(1): 79-83.
• Garattini L, Ghislandi S (2007) “Should we really worry about “launch delays” of new drugs in OECD countries”
(Editoriale) The European Journal of Health Economics 8(1): 1-3.
• Cornago D, Li Bassi L, De Compadri P, Garattini L (2007) “Pharmacoeconomic studies in Italy: a critical review of the
literature” The European Journal of Health Economics 8(2):89-95.
• Koleva D, Motterlini N, Banfi P, Garattini L (2007) “Healthcare costs of COPD in Italian referral centres: A prospective
study” Respiratory Medicine 101:2312-2320.
• Garattini L, Motterlini N, Cornago D (2008) “Prices and distribution margins of in-patent drugs in pharmacy: A
comparison in seven European countries” Health Policy 85(3):305-313.
• Garattini L, De Compadri P, Koleva D, Pasina L, Nobili A (2008) “A critical review of the full economic evaluations of
pharmacological treatments for colorectal cancer” JME 11(1):177-197.
218
ANNUAL REPORT 2008

IRFMN
Guido Bertolini got his Medical degree in 1989 at the University of Bologna, and the specialization in
Pharmacological Research in 1993 at the “Mario Negri” Institute and in Gastroenterology in 1994 at the
University of Pavia.
He founded and chaired from 1997 to 2000 the School of Clinical Methodology and Quality of Care
Improvement at the Ospedali Riuniti di Bergamo and the Istituto di Ricerche Farmacologiche Mario
Negri. From 1999 to 2003 he has been contract professor at the post-doctoral schools in Anaesthesia and
Intensive Care, University of Brescia and Milano; from 2002 to 2005 he has been contract professor of
Educational Science at the Faculty of Lettere e Filosofia, University of Bergamo.
Current research interests: Clinical Research Methodology, Continuous Quality of Care Assessment and
Improvement, Health services research and outcome, Medical decision making, Medical Education.
These interests are mainly developed within the fields of Intensive Care Medicine and Rare Diseases.
Since 1997 he chairs the GiViTI Coordinating Center for research in intensive care medicine. He has been
Head of the Unit of Epidemiology and Education for Clinical Practice at the “Mario Negri” Institute and
since 2001 he is the Head of the Laboratory of Clinical Epidemiology. From 2001 to 2005 he has been
Vice-chairman of the Research Group on Cost-effectiveness, Section on Health Services Research and
Outcomes – European Society of Intensive Care Medicine and, from 2001 to 2005, he has been President
of the Scientific Committee of the “Ospedale maggiore” in Crema.
Selected publications
• Malacarne P, Langer M, Nascimben E, Moro ML, Giudici Daniela, Lampati L, Bertolini G, GiViTI. Building a
continuous multicenter infection surveillance system in the intensive care unit: Findings from the initial data set of 9,493
patients from 71 Italian intensive care units. Crit Care Med 2008; 36: 1105-1113.
• Bertolini G, Rossi C, Anghileri A, Livigni S, Addis A, Poole D. Use of drotrecogin alfa (activated) in Italian intensive
care units: the results of a nationwide survey. Intensive Care Med 2007; 33: 426-434.
• Rossi C, Simini B, Brazzi L, Rossi G, Radrizzani D, Iapichino G, Bertolini G. Variable costs of ICU patients: a
multicenter prospective study. Intensive Care Med. 2006;32:545-52
• Vanoli M, Daina E, Salvarani C, Sabbadini MG, Rossi C, Bacchiani G, Schieppati A, Baldissera E, Bertolini G.
Takayasu's arteritis: A study of 104 Italian patients. Arthritis Rheum 2005; 15;53: 100-107. IF: 7.421.
• Galli M, Luciani D, Bertolini G, Barbui T. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of
thrombosis in the antiphospholipid syndrome. Blood 2003;102:2717-2723.
• Simini B, Bertolini G. Should same anaethesist do preoperative anaesthetic visit and give subsequent anaeshetic
Questionnaire survey of anaesthetists. BMJ 2003; 327: 79-80.
Davide Luciani got his Medical Degree at the University of Bologna in 1995, and the Diploma in
"Tropical Medicine and Hygiene" at the University of Liverpool in 1997. In 2001, he spent one year at
the Department of Statistical Science (University College London). Bayesian probabilistic applications,
decision theory and the graphical approach to pathophysiological modeling represent his main interests.
Within his research activity, these skills are meant as the main methodological ingredients in the
formalization of clinical reasoning, in order to improve its effectiveness and to exploit its educational
value.
Since 2005 he is responsible of the Unit of Clinical Knowledge Engineering.
Selected publications
• Luciani D, Cavuto S, Antiga L, Miniati M, Monti S, Pistolesi M, Bertolini G Bayes pulmonary embolism assisted
diagnosis: a new expert system for clinical use Emerg Med J 2007 ; 24 : 157-164
• M.Cesana, R.Cerutti, E.Grossi, E.Fagiuoli, M.Stabilini, F.Stella, D Luciani.Bayesian Data Mining Techniques: The
Evidence Provided by Signals Detected in Single-Company Spontaneous Reports Databases. Drug Information Journal,
vol. 41, pp. 11-21, 2007
• Latronico N, Bertolini G, Guarneri B, Botteri M, Peli E, Andreoletti S, Bera P, Luciani D, Nardella A, Vittorielli E,
Simini B, Candiani A Simplified electrophysiological evaluation of peripheral nerves in critically ill patients: the Italian
multi-centre CRIMYNE study Crit Care. 2007;11(1):R11.
• Bertolini G, Luciani D, Biolo G Immunonutrition in septic patients: A philosophical view of the current situation Clin
Nutr 2007 ; 26 : 25-29
• Luciani D, Marchesi M, Bertolini G. The role of Bayesian Network in the diagnosis of pulmonary embolism. J Thromb
Haemost 2003; 1: 698-707
• Galli M, Luciani D, Bertolini G, Barbui T. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of
thrombosis in the antiphospholipid syndrome. Blood ,2003 Oct 15;102(8):2717-23Haemostasis, 2003 Apr;1(4):698-707
219
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DIPARTMENT’S ACTIVITIES
The main aim of the Public Health Department is to understand which factors affect the health
of individuals or entire populations and to define effective interventions for responding to their
health needs. Special emphasis is therefore placed on prevention, so that the risks of contracting
illness are lowered, and on the dissemination of independent, evidence-based information.
The department’s effort cannot disregard the National Health System, however, which must
guarantee access to, and quality of, care that is based on principles of equity and appropriateness
and must guarantee it especially to the more vulnerable patient groups. It is in this context that
the Public Health Department carries out its activities.
In addition to its formal research activity, the department participates in, and organises,
initiatives involving information dissemination, training, and debate aimed at healthcare
operators and social care workers, but also at the general population. These activities are also
supported by the publication of the department’s two journals: Ricerca&Pratica and Quaderni
di Farmaco Economia.
"A. and A. Valenti" Centre for Health Economics (CESAV)
The "Angelo e Angela Valenti" Centre for Health Economics (CESAV) was established in 1992
at the "M. Negri Institute" and based at Villa Camozzi- Ranica (Bergamo)-Italy. CESAV is
primarily a research centre, but also does educational work. The centre is involved in health
economics and health policy research. The main areas of research are: Economic Evaluation of
Health Care Programs (i.e. assessment of costs and benefits of alternative health care treatments
and services) and Comparative Health Policy Analysis (i.e. study of domestic and foreign health
care systems, in particular aimed at identifying possible innovations for European countries).
The general aim of the Laboratory of Clinical Epidemiology is to contribute to the improvement
of health care in different medical fields. The guiding principles are mainly two: to help
physicians use the available knowledge and resources at their best; to play a role in the growth
of useful knowledge for clinical practice. The Laboratory operates particularly in the field of
Intensive Care Medicine and Rare Diseases.
Within the Laboratory, the Unit of Clinical Knowledge Engineering aims to bring the value of
clinical reasoning out, through the implementation of probabilistic models for its formalization,
thus favoring the evaluation and the continuous improvement of complex clinical activities.
Laboratory of Clinical Epidemiology
Research, as a multidimensional approach to producing knowledge, characterises the
Laboratory’s activity.
Research provides the basis for planning and carrying out the Laboratory’s activity in a critical
way and involves the participation of health professionals, social workers, mothers, children,
and parents.
Special attention is given to activities involving countries in the north and south of the world.
Laboratory for Mother and Child Health
The main objective of the Laboratory for Mother and Child Health is to ensure a better mother
and child well-being by undertaking interdisciplinary and collaborative work in the field.
Four broad areas, or spheres, of research have been selected:
- monitoring and epidemiological evaluation of utilisation and effects of drugs and vaccines;
- research methodology in general hospital and paediatric community practice;
220
ANNUAL REPORT 2008

IRFMN
- public health determinants of children’s well-being;
- transfer of health information to the community.
Special attention is given to activities involving countries in the north and south of the world.
In addition to the formal research activities, the Laboratory promotes initiatives in the public
health field, in particular those involving mother and child health care.
The initiatives involve the participation in, and the organisation of, educational, training, and
information-dissemination activities.
The critical and active transfer of scientific knowledge is a continuous, daily stimulus to the
Laboratory’s activity.
NATIONAL COLLABORATIONS
"A. and A. Valenti" Centre for Health Economics (CESAV)
Public and private institutions, other health care organizations (Ministry of Health, Regional and
Local Health Authorities, Hospital Trusts).
Laboratory of Clinical Epidemiology
−
−
−
−
−
−
Dipartimento di Neurologia, Ospedale Regionale S. Maria dei Battuti Cà Foncello,
Treviso
Dipartimento di Specialità Chirurgiche, Scienze Radiologiche e Medico Forensi,
Cattedra di Anestesia dell’Università degli Studi di Brescia
Servizio Anestesia e Rianimazione, Osp. San Giovanni Bosco, Torino
Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo
U.O. Anestesia e Rianimazione 1, Ospedale A. Manzoni, Lecco
Cattedra di Fisica e Informatica Medica, Facoltà di Medicina e Chirurgia, Firenze
Laboratory for Mother and Child Health
−
−
−
−
−
−
−
−
−
−
−
−
−
−
Centre for Child Health, (CSB)
Cultural Paediatric Association, (ACP)
Federfarma Lombardia
Hospital of Bergamo “Ospedali Riuniti”, Poison Control Center, Unit Clinical
Toxicology
Italian Drug Agency, (AIFA)
Italian Global Health Watch (OISG)
Italian National Institute of Health (ISS)
Italian Society of Preparers Pharmacists (SIFAP)
Italian Society of Clinical Pharmacy (SIFO)
Il Pensiero Scientifico Editore
Operating unity of Neuropsychiatry of the childhood and of the adolescence, Fondation
“Policlinico di Milano”, (UONPIA)
University of Cagliari, Department of Neuroscience, Clinic of Child and Adolescent
Neuropsychiatry
University of Milan-Bicocca, Faculty Medicine, Paediatric Clinic
University of Milan, Faculty of Political Science
221
ANNUAL REPORT 2008

IRFMN
INTERNATIONAL COLLABORATIONS
"A. and A. Valenti" Centre for Health Economics (CESAV)
− CES (Collège des Economistes de la Santé) of Paris
− Corvinus University of Budapest
− Global Fund of Geneva
− WidO of Bonn
− University Carlos III of Madrid
− University of Birmingham
− University of Hannover
− University of York
− University Pompeu Fabra of Barcelona
− University Erasmus of Rotterdam
Laboratory of Clinical Epidemiology
−
−
−
−
−
−
−
−
−
−
Bloomsbury Institute of Intensive Care Medicine, Institute of Biomedical Research,
University College London, UK
Department of Statistical Science, University College London, UK
Klink fur Anaesthesiologie und Intensivtherapie, Friedrich-Schiller-Universitat, Jena,
Germany
Machine Intelligence Group, University of Aalborg, Denmark
American Board of Family Medicine, Kentucky, US
Institute of Anaesthesia and Intensive Care, Semmelweis University, Budapest,
Hungary
Department of Anaesthesiology and Intensive Care, Warsaw Medical University,
Poland
Department of Intensive Care, General Hospital Novo Mesto, Slovenia
Department of Pulmonary and Intensive Care Medicine, Nicosia General Hospital,
Cyprus
Hospital Das Clínicas, Federal University of Minas Gerais, Belo Horizonte, Brazil
Laboratory for Mother and Child Health
− Catalan Institute of Pharmacology, Barcellona, Spagna
− Centre for Tropical Diseases (CECOMET), Ecuador
− Clínica Infantil Colsubsidio, Bogotà, Colombia
− European Medicines Agency (EMEA)
− European Society for Developmental, Perinatal and Paediatric Pharmacology.
(ESDPPP)
− European Union (EU)
− International Society of Drug Bulletins (ISDB)
− Hospital Robert Debré, Paris, France
− World Health Organization (WHO)
− University of Nottingham, Derbyshire Children’s Hospital, Derby, United Kingdom
222
ANNUAL REPORT 2008

IRFMN
EDITORIAL BOARD MEMBERSHIP
"A. and A. Valenti" Centre for Health Economics (CESAV)
INTERNATIONAL:
Acta Bio Medica; Applied Health Economics and Health Policy; Biomedical Statistics and
Clinical Epidemiology; Health Policy; Journal of Medical Economics; The European Journal of
Health Economics.
NATIONAL:
Economia e Politica del Farmaco; FarmacoEconomia News; Farmeconomia e Percorsi
Terapeutici; L'Internista; PharmacoEconomics Italian Research Articles; Quaderni di
FarmacoEconomia.
Laboratory of Clinical Epidemiology
Ricerca & Pratica;
Dedalo. Gestire i sistemi complessi in sanità.
Laboratory for Mother and Child Health
INTERNATIONAL:
European Journal Clinical Pharmacology; Journal of Clinical Pharmacology &
Pharmacoepidemiology; Paediatric & Perinatal Drug Therapy; Pediatria (São Paulo); Saludarte;
NATIONAL:
Dialogo sui Farmaci; Disturbi d’Attenzione e Iperattività; Quaderni ACP; Quaderni di
Farmacoeconomia; Ricerca & Pratica.
PEER REVIEW ACTIVITIES
"A. and A. Valenti" Centre for Health Economics (CESAV)
Applied Health Economics and Health Policy; Health Policy; PharmacoEconomics; The
European Journal of Health Economics.
Laboratory of Clinical Epidemiology
INTERNATIONAL:
British Medical Journal; Intensive Care Medicine; Critical Care Medicine; Clinical Journal of
the American Society of Nephrology.
NATIONAL:
Ricerca & Pratica
Laboratory for Mother and Child Health
INTERNATIONAL:
Archives of Diseases Childhood; British Medical Journal; BMC Pregnancy and Childbirth;
BMC Public Health; British Medical Journal; Current Drug Metabolism; European Journal of
Clinical Pharmacology; Future Medicine; Expert Review of Clinical Pharmacology; Health
223
ANNUAL REPORT 2008

IRFMN
Education Research; Journal of Antimicrobial Chemotherapy; Journal of Clinical Pharmacology
& Pharmacoepidemiology; Pharmacoepidemiology and Drug Safety; Prescrire.
NATIONAL:
Assistenza Infermieristica e Ricerca; Bollettino SIFO; Dialogo sui Farmaci; Epidemiologia e
Prevenzione; Giornale Italiano di Farmacia Clinica; Medico e Bambino; Quaderni ACP.
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
"A. and A. Valenti" Centre for Health Economics (CESAV)
− Commissione “Accordi di Programma”, AIFA, Roma
− Comitato di Bioetica, Provincia Autonoma di Trento
Laboratory of Clinical Epidemiology
−
−
−
Comitato Etico Azienda Ospedaliera “Bolognini” di Seriate (BG)
Commissione per la Ricerca sanitaria finalizzata, Provincia autonoma di Trento
Commissione di valutazione per il Bando Ricerca Finalizzata, Ministero della Salute
Laboratory for Mother and Child Health
− Comitato Etico dell'Azienda Ospedaliera "Ospedale Maggiore" di Crema
− Comitato scientifico ADHD, ISS
− Commissione Nazionale per le Vaccinazioni, Ministero della Salute
− Commissione tecnico-scientifica per la programmazione e verifica delle
vaccinazioni, Regione Lombardia
− Comitato Scientifico del Gruppo di Lavoro “Farmaci e Bambini”, AIFA
− Commissione tecnica per l'elaborazione, gestione e aggiornamento del
Prontuario Terapeutico Regionale (P.T.R.), Regione Autonoma Valle d'Aosta
− Gruppo di Lavoro Pediatrico, AIFA
− Gruppo di Lavoro "Promozione allattamento al seno", Regione Lombardia
− Paediatric Expert Group (P.E.G.), EMEA
EVENT ORGANIZATION
"A. and A. Valenti" Centre for Health Economics (CESAV)
Congress: Convegno Nazionale di Farmacoeconomia: “Economia del farmaco: fra soluzioni
tecniche e decisioni politiche”. 7-8 May, Ranica (BG)
Laboratory of Clinical Epidemiology
Congress, GiViTI Annual Meeting, October 29-31, Pesaro.
Workshop, Meeting Compact, April15, Ranica (BG).
224
ANNUAL REPORT 2008

IRFMN
Laboratory for Mother and Child Health
Stage in Cooperation and Public Health part of the Master’s Course in Analysis and
Management of Projects for Development. From 25 March at 4 april, Milan.
Course “La medicina basata sull’evidenza (EBM) dalla teoria alla pratica nella
neuropsichiatria dell’infanzia e dell’adolescenza. 11 June, Milano.
Seminary “Prevenzione della Cervice Uterina: incontro-confronto su conoscenze e
prospettive”. 17 June, Milan, Italy.
Course “La sperimentazione clinica in pediatria di famiglia”. 19-21 June, Santa Maria Imbaro
(CH), Italy.
PARTICIPATION IN EVENTS IN WHICH THE DEPARMENT WAS
INVOLVED
"A. and A. Valenti" Centre for Health Economics (CESAV)
May
Congress: “I Vaccini HPV: Le Esperienze Cliniche e le Strategie di Sanità Pubblica”.
“Le strategie di sanità pubblica”. “I costi e la sostenibilità”. Milan.
Course: “Le maculopatie e la degenerazione maculare legata all’età: gestione clinicoterapeutica
ed economia”. “L’impatto socioeconomico delle maculopatie: Farmacoeconomia
e Qualità di Vita”. Baveno (VB).
June
Congress: “Prevenzione del carcinoma della cervice uterina: incontro-confronto su
conoscenze e prospettive”. “Valutazione costi-benefici”. Milan.
Seminar: “Health Economics Ad Board for Almorexant” (panel of experts). Frankfort.
Workshop: “Strumenti e metodi di trasparenza per il processo decisionale”.
“Le possibili prospettive della prevenzione in funzione della qualità e della sostenibilità”.
“Introduzione alle principali tecniche adottate nelle valutazioni economiche in sanità”.
“Ruolo e sviluppo delle strategie di prevenzione vaccinale nel soggetto adulto”. Ragusa.
July
Congress: “7th European Conference on Health Economics”. Rome.
November
Course: “Corso di farmacoeconomia ed economia sanitaria”. Rome.
Laboratory of Clinical Epidemiology
February
Congress, Trauma e Subintensive, Bologna
Congress, "Riusciremo a morire in pace", Milano
Congress, Emorragia intracerebrale (ICH): quale paziente si giova di un trattamento
intensivo in ambiente neurochirurgico Torino
225
ANNUAL REPORT 2008

IRFMN
Course, Statistica di Base, Torino
March
Congress, Fine Vita, Padova
Course, Statistica di Base, Torino
Course, Statistica di Base, Torino
Course, Excel e MargheritaDue: applicazione pratica, Torino
Congress, GiViTI Bergamo, Zingogna (BG)
Congress, La sorveglianza delle infezioni in TI, Zingonia
April
Course, Statistica di Base, Torino
Congress, Le infezioni in TI: Presentazione dei dati regionali del gruppo GiViTI, Bologna
Course, Statistica di Base, Torino
Course, Statistica di Base, Torino
Course, Margherita Due (Course, avanzato), Torino
May
Congress, Analgesia, anestesia, terapia intensiva in ostetricia, Torino
Workshop, Perfezionamento e aggiornamento in medicina intensiva , Mendrisio
Course, Statistica di Base, Torino
Congress, Appropriatezza dei ricoveri in terapia intensiva, Torino
June
Congress, Valutazione della qualità della Assistenza sulla base degli indicatori di gravità del
paziente, Avezzano
September
Congress, ESPEN Congress, Firenze
Congress, Sepsi nel trauma - Trauma Update, Roma
Course, MargheritaTre: stato dell'arte e pianificazione dei lavori, Ivrea
October
Congress, Incontri GiViTI-Piemonte, Torino
Course, Migliorare la compilazione di Margherita Due ed imparare a leggere il report,
Torino
Course, Migliorare la compilazione di Margherita Due ed imparare a leggere il report,
Torino
Workshop, Accademia della cura, Rho (MI)
Congress, Meeting GiViTI, Pesaro
November
Congress, Trauma Update, Cesena
Course, MargheritaTre: presentazione dei nuovi moduli, Torino
December
Workshop, PNS: Euronetwork Sindromi Paraneoplastiche, Treviso
Congress, Sorveglianza delle infezioni in ti con margherita due: Presentazione dei dati 2007,
Torino
226
ANNUAL REPORT 2008

IRFMN
Laboratory for Mother and Child Health
January
Congress: “In Toscana, dai dati alle scelte: l’ospedalizzazione pediatrica e neonatale”.
Regione Toscana; Ospedale Meyer, Agenzia Regionale Sanità Toscana e Servizio Sanitario
della Toscana; Firenze.
Regional Congress: “Seminare e Raccogliere. Guadagnare Salute”. Associazione Culturali
Pediatri Campagna (ACP) e Università degli Studi di Napoli Federico II, Dipartimento di
Pediatria; Napoli.
February
Course of Formation: “Bioetica e Sperimentazione”. Centro di bioetica e biodiritto
dell’Università di Siena e Comitato Etico - AOU Senese; Siena.
Meetings in the library. Azienda U.L.SS. n. 6 Vicenza, International Society of Drug Bulletins
(ISDB), Biblioteca Biomedica Ospedale S. Bortolo; Vicenza.
March
III Regional Seminary: “Attualità in farmacovigilanza”. Università degli Studi di Siena,
Sistema di farmacovigilanza Regione Toscana; Siena.
IV National Congress: Pediatria On Line. Sirmione (BS).
April
Congress: “Un’esperienza sanitaria locale per un confronto globale”. Fondazione Ivo de
Carneri Onlus e Fondazione Policlinico San Matteo; Pavia.
Course: “Allestimento dei medicinali in pediatria”. Università degli Studi ¨ di Milano, Facoltà
di Farmacologia; Milano.
SIII 2008 Integrated interfaculty, interdisciplinary seminar: “Alimentazione: sicurezza,
accesso, qualità, culturaUniversità degli studi di Milano, Facoltà di Scienze Politiche; Milano.
May
Forum ISDB Italia: “Il ruolo dell’informazione indipendente”. Dialogo sui farmaci; Verona.
Seminary: “IX Giornata dell’allattamento al seno”. La Leche League Italia; Imola (BO).
4th Updating Course: “Novità e criticità nell’attività regolatoria di farmaci e dispositivi
medici”. Società Italiana di Farmacia Ospedaliera (SIFO) e Società Italiana Attività
Regolatorie (SIAR); Verona.
First Conference: “Anticipating changes in drug development for children building on
paediatric rheumatology”. Marie Curie ActionsTRiPR, Istituto Gaslini e Scuola Internazionale
di Scienze Pediatriche (SISP), Genova.
SEFAP Master (Servizio di Epidemiologia e Farmacologia Preventiva) :“Corso di
perfezionamento in Farmacovigilanza”. Università degli Studi di Milano, Dipartimento di
Scienze Farmacologiche; Milano.
June
Course: “2007 l’anno dei farmaci per i bambini”. Associazione Laureati della Facoltà di
Farmacia (a.l.far.). Istituto di Ricerche Farmacologiche “Mario Negri; Milano.
Congress: “Prevenzione del carcinoma della cervice uterine: incontro-confronto su conoscenze
e prospettive”. Regione Lombardia e Istituto Mario Negri; Milano.
September
227
ANNUAL REPORT 2008

IRFMN
XII Regional Congress Cure Primarie: “Il farmaco tra appropriatezza e sicurezza”. Regionale
¨ Friuli Venezia Giulia; Grado (GO).
First Conference: “Un disturbo conosciuto in tutto il mondo!”. Regionale Lombardia A.I.F.A.
onlus; Milano.
Festivaletteratura 2008: “Il corpo oltre. Come superare i limiti biologici: la vita dei farmaci”.
Mantova.
Congress: “Nuovi orizzonti per l’influenza”. ASL Provincia di Lodi Servizio Medicina
Preventiva nella città; Lodi.
October
XX National Congress ACP: “Venti di ACO”. Associazione Culturale Pediatri; Cagliari.
XXIX National Congress SIFO: “SIFO e istruzioni. Funzioni e competenze del farmacista per
un paese ed un SSN in evoluzione”. Società Italiana di Farmacia Ospedaliera; Napoli.
Seminary: “Star bene a scuola. Migliorare la qualità di vita per favorire l’apprendimento”.
Municipality of Milan; Milano.
Course: III edizione del percorso di formazione per rappresentanti di associazioni di cittadini
& pazienti “Orientarsi in salute & sanità per fare scelte consapevoli”. Partecipasalute, Istituto
di Ricerche Farmacologiche “Mario Negri”; Milano.
Meetings Beyond a look: “Salute: bene per pochi o diritto per tutti”. Coordinamento
Comasco per la Pace, Comune di Lurate Caccivio, Encuentro, Biblioteca Comunale e Pro
Loco; Lurate Caccivio (CO).
Meeting: “Premio Ercole Pisello”. Regione Umbria, Associazione Giuseppe Corradi, Comune
di Bevagna; Bevagna (PG).
November
Course “Formazione alla ricerca: inquinamento ambientale e funzionalità respiratoria in
bambini di un’area urbana”. ASL TA/1, Federazione Italiana Medici Pediatri (FIMP); Taranto.
Continuing education “Disagio scolastico e patologie neuropsichiatriche”. ASL Provincia di
Milano 3; Monza.
Bioetica Congress 1978-2008: “La 1978-2008 trent’anni di sanità tra bioetica e prassi
quotidiana”. Commissione Regionale di Bioetica, Regione Toscana, Servizio Sanitario della
Toscana; Firenze.
1th International Congress of UENPS: “Global Neonatology & Perinatology”. Union of
European Neonatal and Perinatal Societies (UENPD), Società Italiana di Neonatologia (SIN) e
Società Italiana di Medicina Perinatale (SIMP); Roma.
Course ECM for Paediatricians and Parents: “Dall’infanzia all’età avanzata: la
farmacovigilanza nelle età a maggior rischio”. Servizio Farmaceutico AUSL Ferrara; Ferrara.
Course: “Farmaci generici e note AIFA in pediatria: analisi dei dati prescrittivi”. Dipartimento
Cure Primarie ASL di Milano; Milano.
December
XXI National Congress Confronti in Pediatria: “Pediatria 2008: attraverso le immagini”.
IRCCS Burlo Garofolo, Università di Trieste; Trieste.
Formation Course: “In tema di antibioticoterapia, infezioni e malattie riemergenti”. Centro per
la Formazione Permanente e l'Aggiornamento del Personale del Servizio Sanitario (CEFPAS);
Associazione Culturale Pediatri Centro-Sicilia; Caltanisetta.
228
ANNUAL REPORT 2008

IRFMN
Seminary: “Prevenire il carcinoma della cervice uterina: serve il vaccino A che età”.
Consorzio Mario Negri Sud; Santa Maria Imbaro (CH)
GRANTS AND CONTRACTS
"A. and A. Valenti" Centre for Health Economics (CESAV)
− Abbott
− EG
− Grunenthal-Prodotti Formenti
− Novartis Farma SpA
− Ratiopharm
− Sanofi Aventis
− Sanofi Pasteur MSD
− Vivisol
Laboratory of Clinical Epidemiology
− ARESS Piemonte
− ASL TO-2 Piemonte
− AstraZeneca
− Azienda ULSS 16, Padova – Italia
− Bellco SpA
− Draeger Italia
− Regione Lombardia
− Regione Toscana
− Regione Piemonte
− Sanofi-Aventis Italia
Laboratory for Mother and Child Health
−
−
−
−
−
−
−
Boehringer Ingelheim
European Union
Il Pensiero Scientifico Editore
Italian Drug Agency, (AIFA)
Provinve of Milan
Regional Health Ministry - Lombardy Region
Regional Health Ministry - Valle d’Aosta Region
229
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
"A. and A. Valenti" Centre for Health Economics (CESAV)
Garattini L, Motterlini N, Cornago D. Prices and distribution margins of in-patent drugs in pharmacy: A comparison in seven
European countries. Health Policy 2008;85(3):305-313.
De Compadri P, Koleva D, Mangia A, Motterlini N, Garattini L. Cost minimisation analysis of 12 or 24 weeks of
peginterferon alfa-2b + ribavirin for hepatitis C virus. JME 2008;11(1):151-163.
Garattini L, De Compadri P, Koleva D, Pasina L, Nobili A. A critical review of the full economic evaluations of
pharmacological treatments for colorectal cancer. JME 2008;11(1):177-197.
Garattini L, Casadei G. Health technology assessment: for whom the bell tolls The European Journal of Health Economics
2008; http://springerlink.metapress.com/content/u361333hw68552w8/fulltext.pdf
Beghi E, Atzeni L, Garattini L. Economic Analysis of Newer Antiepilectic Drugs. CNS Drugs 2008;22(10):861-875.
Laboratory of Clinical Epidemiology
Bertolini G. From national to global outcome research in the intensive care unit: A challenge to win. Crit Care Med
2008;36:336-337.
Guarneri B, Bertolini G, Latronico N. Long-term outcome in patients with critical illness myopathy or neuropathy.
The Italian multi-centre CRIMYNE study. J Neurol Neurosurg Psychiatry 2008 79:838-41.
Malacarne P, Langer M, Nascimben E, Moro ML, Giudici Daniela, Lampati L, Bertolini G, GiViTI. Building a
continuous multicenter infection surveillance system in the intensive care unit: Findings from the initial data set of
9,493 patients from 71 Italian intensive care units. Crit Care Med 2008; 36: 1105-1113.
Di Bartolomeo S, Valent F, Rossi C, Beltrame F, Anghileri A, Barbone F. Geographical differences in mortality of
severely injured patients in Italy. Eur J Epidemiol. 2008; 23: 289-94.
Laboratory for Mother and Child Health
Bonati M. An independent contribution to the growth of paediatric clinical pharmacology in Italy. Eur J Clin
Pharmacol 2008;64:343-346.
Clavenna A, Bonati M. A missed opportunity. BMJ
http://www.bmj.com/cgi/eletters/337/nov24_2/a2245#205527:2008.
Font M, Miselli M, Conforti A, Bonati M. An Italian Story with wider implications BMJ
http://www.bmj.com/cgi/eletters/336/7655/1208-b#200313:2008.
Maschi S, Clavenna A, Schiavetti B, Campi R, Bernat M, Bonati M. Neonatal outcome following pregnancy
exposure to antidepressants: a prospective controlled cohort study. BJOG 2008;115:283-289.
Pandolfini C, Bonati M. European paediatric research and children’s therapeutic needs. A trial review. Acta
Paediatrica 2008;97:1232-1237.
Pandolfini C, Bonati M. Something is moving in European drug research for children, but a more focused effort
concerning all therapeutic needs is mandatory. Arch Dis Child 2008;93:715.
Pandolfini C, Bonati M, Rossi V, Santoro E, Choonara I, Naylor C, Sammons E, Jacqz-Aigrain E, Zarrabian S, Arnau
JM, Castel JM, Danés I, Fuentes I. The DEC-net European register of paediatric drug therapy trials: contents and
context. Eur J Clin Pharmacol 2008;64:611-617.
Santos DB, Clavenna A, Bonati M, Coelho HL. Off-label and unlicensed drug utilization in hospitalized children in
Fortaleza, Brazil. Eur J Clin Pharm 2008;64:1111-18.
Usala T, Clavenna A, Zuddas A, Bonati M. Randomised controlled trials of Selective Serotonin Reuptake Inhibitors
in treating depression in children and adolescents: a systematic review and meta-analysis. European
Neuropsychopharmacology 2008;18:62-73.
230
ANNUAL REPORT 2008

IRFMN
LAY PRESS SELECTION PUBLISHED IN 2008
"A. and A. Valenti" Centre for Health Economics (CESAV)
Garattini L, Gritti S. Confronto di prezzi e margini alla distribuzione coperti da brevetto in sette Paesi Europei. Focus
On 18 Febbraio 2008.
Garattini L, Gritti S. Prezzi dei farmaci coperti da brevetto. Ricerca & Pratica 2008;24:117-118.
Garattini L, Cornago D, De Compadri P, Gritti S. Confronto di prezzi e margini alla distribuzione di farmaci coperti
da brevetto in sette Paesi Europei. Quaderni di Farmaco Economia 2008;5:15-21 (1a parte).
Gritti S, Casadei G, De Compadri P, Garattini L. ECM: analisi comparativa in sei diversi Paesi europei. Quaderni di
Farmaco Economia 2008;6:17-28.
De Compadri P, Koleva D. I costi della psoriasis vulgaris nei pazienti sottoposti a terapia sistemica: una rassegna della
letteratura e una stima preliminare di costo in Italia. Quaderni di Farmaco Economia 2008;6:7-15.
Garattini L, Motterlini N, Cornago D. Confronto di prezzi e margini alla distribuzione di farmaci coperti da brevetto in sette
Paesi Europei. Quaderni di Farmaco Economia 2008;7:7-16 (2a parte).
Garattini L. Finanziaria 2008. Dialogo sui farmaci 2008;1:23-24.
Garattini L. Finanziaria e farmaci. Editoriale Ricerca & Pratica 2008;24(2):47-49.
Laboratory for Mother and Child Health
Biasini G, Bonati M, Corchia C, Marchetti F, Napoleone E, Panei P, Rossi Paolo, Rossi Rossella, Toffol G. Lettera
aperta di nove componenti esterni del Gruppo Italiano sui farmaci pediatrici dell'Agenzia Italiana del Farmaco. R&P
2008;24:163-164.
Bonati M. Farmaci essenziali per i bambini. Reazioni 2008;6:1.
Bonati M. Farmaci essenziali per i bambini. BIF 2008;XV:45-48.
Bonati M. Il Pap-test Certamente! La vaccinazione anti-HPV Sì, eventualmente. Medico e Bambino 2008;4:251-
53.
Bonati M. I determinanti della salute e le disuguaglianze sociali. Quaderni ACP 2008; 15:138.
Bonati M. Indipendenti da chi, da cosa Dialogo sui Farmaci 2008;3:154.
Bonati M. L’AIFA al servizio dei bambini. Medico e Bambino 2008;27:551-52.
Bonati M. L’AIFA dalla parte dei bambini. CARE 2008;4:16-17.
Bonati M. La vicenda dell’Agenzia Italiana del Farmaco (AIFA). Quaderni acp 2008;15: 224.
Bonati M. Prevenzione non è solo vaccinazione: il caso del carcinoma della cervice uterina. Quaderni di
farmacoeconomia 2008;5:5-6.
Bonati M. Redattori per passione, indipendenti per scelta. R&P 2008;24:141-43.
Clavenna A, Fortinguerra F. Contro la tosse è meglio il miele. Quaderni acp 2008;15:85.
Clavenna A, Fortinguerra F. Informazione sui farmaci: novità in Europa e in America. Quaderni ACP 2008;15:181.
Clavenna A, Fortinguerra F. Molte novità sui farmaci per bambini. Quaderni ACP 2008;15:131.
Clavenna A, Fortinguerra F. Paracetamolo e felilefrina: aggiornamenti sulla sicurezza di impiego. Quaderni acp
2008;15:259.
Font M, Miselli M, Conforti A, Bonati M. An Italian story with wider implications R&P 2008;24:209-220.
Gaillard P, Bullio P, Martinod D, Fortinguerra F, Bonati M. Valutazione dell’attività della Commissione tecnicoscientifica
della Valle d’Aosta. R&P 2008;24:187-98.
Maschi S, Bonati M. Additivi nei farmaci per bambini, effetti indesiderati e appropriatezza. Dialogo sui farmaci
2008;6:278-281.
231
ANNUAL REPORT 2008

IRFMN
Pandolfini C, Clavenna A, Bonati M. La qualità dell’informazione sulla fibrosi cistica nei siti internet italiani. R&P
2008;24:92-100.
Pirola ME, Bettinelli ME, Fortinguerra F. Prevalenza, esclusività e durata dell’allattamento al seno in Lombardia.
R&P 2008;24:31-33.
Taddia A, Zeni B, Campomori A, Campi R. Mifepristone e misoprostolo: efficacia e sicurezza nell'aborto medico.
R&P 2008;24:3-16.
OTHER PRODUCTS PUBLISHED IN 2008
"A. and A. Valenti" Centre for Health Economics (CESAV)
Boffelli S, Rossi C, Bertolini G. Progetto Margherita. Promuovere la ricerca e la valutazione in Terapia Intensiva.
Rapporto 2007. Bergamo: Ed. Sestante, 2008. [Report]
Laboratory for Mother and Child Health
Bonati M, Zuddas A. Metilfenidato, atomoxetina. In: Enciclopedia Medica Italiana. II tomo; III aggiornamento.
2008;2137-2150. [Chapter Book]
Bonati M, Campi R e Gruppo di Lavoro per la Convenzione sui Diritti dell’Infanzia e dell’Adolescenza. I diritti
dell’infanzia e dell’adolescenza in Italia. 4° rapporto di aggiornamento sul monitoraggio della Convenzione sui diritti
dell’infanzia e dell’adolescenza in Italia 2007-2008. Gruppo di lavoro per la CRC c/o Save the Children, Roma.
2008;6-150. [Chapter Book]
Bonati M. Cuba. In: Salute globale e aiuti allo sviluppo. Diritti, ideologie, inganni. 3° Rapporto dell’Osservatorio
Italiano sulla Salute Globale. Edizioni ETS, Pisa. 2008;354-365. [Chapter Book]
Bonati M. Neonatal Clinical Pharmacology 2008. In: 1st International Congress of UENPS. Rome, Italy, November
17-19, 2008. [abstract]
Clavenna A, Sequi M, Bortolotti A, Merlino L, Foretino I, Bonati M. Drug utilisation profile in the paediatric
population of Lombardy Region, Italy. In: Safe and Effective Medicines for Children. The 11th biennal ESDP
Congress. Rotterdam 2008;105. [abstract]
Fortinguerra F, Clavenna A, Bonati M. Drug related problems in children: the role of a drug information centre. In:
Safe and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008; 106. [abstract]
Fortinguerra F, Clavenna A, Labate L, Maschi S, Bonati M, Advertisement Reviewer Group. Pharmaceutical
advertisements in Italian paediatric general practitioner journals. In: Safe and Effective Medicines for Children. The
11th biennal ESDP Congress. Rotterdam 2008;107. [abstract]
Pandolfini C, Bonati M. Lesson from DEC-net. Development and application of the European register of clinical
trials on medicines for children. In: First Conference of the Marie Curie Actions TRiPR. Genoa, Italy, May 29 th -June
1 st , 2008. [abstract]
Racca F, Bonati M, Berta G, Testa R, Benini F, Benedetti M, Bignamini E, Maspoli M, Salvo I, Ranieri MV. Long
term ventilation of children in Italy: preliminary data from questionnaire survey. In: 21st Annual Congress European
Society of Intensive Care Medicine; Lisbon, Portugal. 21-24 September 2008. [abstract]
Racca F, Bonati M, Ottonello G, Berta G, Pavone M, Morandi F, Wolfler A, Tardivo I, Testa R, Sequi M, Maspoli
M, Bignamini E, Salvo I, Ranieri VM. Home discharge and monitoring program for home mechanically ventilated
children in Italy. In: 2nd Congress of the European Academy of Paediatrics. Nice, France, October 24-28, 2008.
[abstract]
Usala T, Rocchi F, Knellwolf AL, Bonati M, Masi G, Zuddas A, Arcieri R, Panei P. Long term safety of psychotropic
drugs used for Attention-deficit/hyperactivity disorder (ADHD) in Italian children and adolescent population. In: Safe
and Effective Medicines for Children. The 11th biennal ESDP Congress. Rotterdam 2008;119. [abstract]
During 2008 the laboratory’s activities were covered by the national media 96 times
232
ANNUAL REPORT 2008

IRFMN
RESEARCH ACTIVITIES
"A. and A. Valenti" Centre for Health Economics (CESAV)
Educational activity
Educational activities are developed only if related to research studies, in order to offer
original contributions which naturally reinforce the research aims.
Economic Evaluation of Health Care Programs
The aim of this research area is to assess the costs of pathologies and the cost-effectiveness
ratios of the diagnostic/therapeutic existing alternatives. In general, analyses can be
classified into two groups: partial economic evaluations (e.g. cost of illness analysis) and full
economic evaluations (e.g. cost-effectiveness analyses).
Comparative Health Policy Analysis
The aim of this research area is to study the organization of health care systems, in order to
draw lessons from international comparisons. This is particularly important in a "market"
like health care where economic competition lacks by definition and therefore public
regulation plays a crucial role.
Laboratory of Clinical Epidemiology
Appropriateness in Intensive Care Units
The main purpose of these research activities is the assessment and improvement of the
quality of care in Italian Intensive Care Units (ICUs). It is a multi-annual project promoted
on behalf of GiViTI, a collaborative network composed by half of the Italian ICUs and
coordinated by the Laboratory. The main focus is the “Margherita” project. Its aim is the
continuous evaluation of the quality of care and it is based on a free software developed by
the Laboratory and distributed to all the ICUs adhering to the GiViTI group. The software
has been realized on a modular structure, which enables to easily integrate the basic data
collection (the “core” of Margherita) with the data collection of specific research projects
(the “petals” of Margherita).
Additionally, in the current year, a software based on a probabilistic model (bayesian
network) covering a large set of clinical variables, has been further implemented. Such an
activity, lead by the Unit of Clinical Knowledge Engineering, and also involving the
Department of Statistical Science (University College London) and the Machine Intelligence
Group of the University of Aalborg (Denmark), pursues the realization of tools for the
retrospective evaluation of specific treatments in ICU. By means of this model, it is meant to
overcome the current limits of an overall evaluation of ICU, likewise it happens when the
traditional models for mortality prediction are adopted. Within this activity, a bayesian
system to monitor the Standardised Mortality Ratio of the single ICU. This system, which
resembles the bayesian pharamcovigilance system currently in use at the WHO and FDA,
will identify periods during which some problems occurred, allowing the physicians of the
Center to promptly and efficacy react.
Studies on Multiple Organ Failure pathological processes
The Laboratory of Clinical Epidemiology has lead several investigations to clarify which
pathophysiological mechanisms induce multiple organ failure, a condition still burdened
with high mortality. Among these, the investigation on the neuromuscular impairment in
critical patients (the observational study 'Crimyne'), the study on the impact of enteral
233
ANNUAL REPORT 2008

IRFMN
feeding, and the new treatments proposed for severe sepsis (Xigris and the removal of
inflammation mediators through specific filter applied to circuit of plasma-filtration).
The value of a strict glycemic control of critical patients has been recently emphasized, given
its connection to a drug like insulin that, in spite of its large availability and low cost,
induces a relevant reduction of mortality in ICU. The Unit of Clinical Knowledge
Engineering has developed a model based on a differential equations system whose aim is to
support the physician in dosing both insulin and glucose infusions, in order to extend the
possibility of a strict control even to patients with a high risk of hypoglycaemia. This model
represents a precious opportunity to investigate the pathophysiological mechanisms behind
the benefits of insulin already demonstrated at an empirical level, allowing the explanation
of the dynamic behaviour of glycemic fluctuations on the basis of the patient's metabolic
profile.
The reconstruction of clinical reasoning in the medical practice and
education
This area represents the main concern of the Unit of Clinical Knowledge Engineering, whose
objective is the valorization of clinical reasoning in solving complex clinical problems.
The diagnosis of pulmonary embolism still represents a relevant clinical challenge, due to
the complexity of the patient's clinical presentation and the variability of diagnostic
resources among Centres. In this regards, we are conducting an Italian multicenter study,
involving mainly Emergency Units, with the aim of prospectively validating the diagnostic
software BayPAD (Bayes Pulmonary embolism Assisted Diagnosis). Such a tool, relying on
a probabilistic model covering 72 clinical variables and doing without the need to input all
the contemplated observations, would overcome the main reasons which prevented ordinary
clinical guidelines to be largely accepted. Moreover, the results of the retrospective
validation of the system have been obtained.
The Unit started a project for the realization of a software assisting the physician in tracing
back the basis of his clinical decisions before the description provided by clinical reports,
among those that are typical of particular medical specialty. The software has the double
target to create specific applications based on probabilistic models representing complex
clinical decision problems, and to involve physicians in their construction. The last target is
achievable given the strong analogy between the causal structure of the exploited models
(bayesian networks) and the pathophysiological structure of medical knowledge. By this, it
will be given the chance to adopt this system within medical training projects, with a special
attention to e-learning programs.
Clinical Epidemiology of rare diseases and orphan medicine
Our purpose is to find out and describe clinical and research problems related either to rare
diseases or to neglected aspects of well-known diseases. We also focus on the needs of the
patients with rare diseases. A specific project is connected with this research activity: “PNS
– Euronetwork”. It is a European project on paraneoplastic neurological syndromes that is
financed by the Fifth and Sixth Framework Program of the European Community. Its
purposes are various: to develop a network of reference centers for these pathologies all
sharing a common database; to organize a sample bank of biological fluids and cerebrospinal
liquid to point out the best antibody indicators for the diagnosis and prognosis of these
patients; to realize some research projects on the treatment of this syndrome.
Laboratory for Mother and Child Health
Pharmacoepidemiology in the Lombardy Region
During 2005, 747,790 children and adolescents < 18 years (48% of the population) received
at least one drug prescription. A total of 2,177,469 prescriptions were dispensed,
corresponding to 3,122,745 medication packages. Each treated child received an average of 3
prescriptions and 4 packages (median 2). The highest prevalence was observed in the 1-5
234
ANNUAL REPORT 2008

IRFMN
year old age range (average value 65%), and then decreased to 38% in the 12-17 year range.
The prevalence was slightly higher in boys than girls (62% vs 59%). Antibiotics were the
most prescribed therapeutic class (41% of the population), followed by anti-asthmatics
(15%) and anti-histamines (5%). Altogether, these three therapeutic classes comprised 81%
of the prescribed packages. In all, 757 drugs were prescribed. Amoxicillin+clavulanic acid
was the most prescribed drug (18% of children), followed by amoxicillin (13%) and inhaled
beclometasone (9%). The 10 most prescribed drugs, 7 of which were antibiotics, represented
64% of the prescribed packages. Large differences were found in prevalence rates between
different LHUs; these ranged between 38 in Milan and 55% in Brescia. No correlation was
found between rank distribution at LHU level of the prevalence rate and hospitalisation rates
in the paediatric population. The age and residence of the child were the main determinants
of drug exposure. In particular, being 1-5 years old (OR 4.51; 95%CI 4.43 - 4.58) and living
in Brescia (OR 2.08; 95% CI 2.06 - 2.11) were the factors associated with the highest risk of
drug exposure. The general prescribing profile is similar to that observed in other Italian
studies, in particular in Northern Italy. Despite the fact that the Lombardy region is a quite
homogeneous context, quantitative and qualitative geographical differences were found. The
differences observed between local health units suggest that educational interventions for
health care professionals and parents could be effective in improving the rational drug use.
However, more efforts are needed also in certain contexts characterised by a low prevalence
rate, but also by a not always appropriate drug use.
Prescription, efficacy, and safety of psychotropic drugs in the Italian
paediatric population
The safety and effectiveness of psychotropic drug use in the paediatric population is widely
debite, in particolar because of the lack of data concerning the long term effects.
In Italy the prevalence of psychotropic drug prescriptions increased in the 2000-2002 period
and decreased afterwards. In such a context, the systematic continuous monitoring of
psychopharmacological treatments is essential. The Laboratory coordinates an independent
research project supported by the Italian Drug Agency (AIFA) entitled “Prescription,
efficacy, and safety of psychotropic drugs in the Italian paediatric population”, aimed to:
estimate the incidence and prevalence of psychotropic drug prescriptions in the paediatric
population, to evaluate the prescribing patterns, and to monitor the safety of these drugs. The
study population was composed by more than 76,000 children and adolescent < 18 years old
living in the local health unit of Verona. Between 1 January 2005 and 31 December 2006,
111 youths (0.8 per 1,000 of the study population) received at least one psychotropic drug
prescription. Only 29 patients were in charge to the child and adolescent outpatient
psychiatric services. Information concerning diagnostic and therapeutic approaches, and care
strategies were collected with the support of the treating physicians for 52 patients (47%).
Anxiety-depression syndrome and attention disorders were the most commonly diseases for
which psychotropic drugs were prescribed. In all, 20% of youths were chronically treated;
85% received also psychological support; child psychiatrists performed the first diagnosis in
50% of the cases; in 1/3 of the cases parents attended 2 or more different specialists.
Workgroup on the “Convention for child and adolescent rights”
The Laboratory for Mother and Child Health is part of the Workgroup on the “Convention
for child and adolescent rights” (CRC) in Italy.
The 4 th update report was published this year and, as with the previous reports, came out on
May 27 th , anniversary of the CRC’s ratification in Italy. The report testifies not only the
perseverance and effort of the participating associations in monitoring and keeping a
spotlight on child and adolescent rights throughout the years, but also the progress and
strengthening of the CRC group.
The 4 th CRC report offers an update of the issues addressed in the previous reports, adds
more in depth analyses, and enriches the analysis with the inclusion of new topics thanks to
the active contribution of an ever increasing number of associations.
235
ANNUAL REPORT 2008

IRFMN
The objective is to expand the workgroup’s area of observation so that it addresses all eight
groupings into which the United Nations Committee has divided the CRC’s articles,
corresponding to the report’s chapters, in the next United Nations Supplementary Report
expected in 2009 (year in which the United Nation’s Convention’s 20 th anniversary will be
celebrated.
The 4 th report provides an updated view of the putting into effect, or the violation, of Italian
children and adolescent’s rights. The CRC Group’s associations ask the government to
ponder this issue with the hope that such an effort, and the recommendations made, may
serve as a useful tool for those who in the new government will be responsible for the
welfare of children and adolescents in Italy.
In 2009 the Italian government will have to stand before the United Nations Committee and
describe the efforts made in the last few years to improve children’s rights and to put into
place the UN Committee’s conclusive observations concerning the extent of the CRC’s
fulfilment in Italy and of the two CRC Optional Protocols (from June 2006).
All this is carried out with the hope that it will stimulate, and contribute to, the development
of standard procedures and legislative reforms that will bring about a tangible improvement
in the condition of all youths in Italy.
National ADHD Register
The marketing authorisation for methylphenidate and atomoxetine in Italy has made
monitoring the use of these drugs in children with attention deficit hyperactivity disorder
(ADHD) necessary. In order to meet this need, a national registry coordinated by the Istituto
Superiore di Sanità’s drug department, in collaboration with the Italian Drug Agency and the
Conferenza permanente degli Assessori alla Sanità delle Regioni and the Province autonome
di Trento e Bolzano, was set up and activated on June 18, 2007. The Laboratory for Mother
and Child Health is part of the scientific committee and participated in writing the protocol,
which defines the structure and activities of the national ADHD register and its monitoring.
The registry’s aims are : to monitor the use of methylphenidate and atomoxetine; to evaluate
the safety and compliance of therapies with methylphenidate and atomoxetine, alone or in
combination with other therapeutic interventions (pharmacological or not) in the medium
and long term; to define the probability of developing ADHD requiring pharmacological
treatment in the school-aged population; to define the optimal management strategy for
ADHD through the standardisation of the most appropriate diagnostic and therapeutic
strategies; to evaluate the effectiveness of psychotropic drugs and/or behavioural therapy in
the evolution of ADHD in school-aged children; to calculate the long term risk of persistence
of the disorder, of being left behind in school, and of the development of psychosis or other
psychiatric disorders.
During the register’s first year of activity, a total of 851 patients were registered by 83
referral centres, 369 of whom were receiving methylphenidate and 482 atomoxetine.
Adverse reactions were reported in 23 patients, 7 of which were classified as severe. In all,
52% of patients concluded the follow-up after 6 months of treatment and 3% did so one year
after registration.
The Laboratory set up ADHDNews, a newsletter aimed at providing interested health care
operators a monthly bibliographic update of the recent scientific literature via email. This
initiative is part of the Italian Drug Agency’s (AIFA) independent research project
implemented in collaboration with the Istituto Superiore di Sanità and called “Long term
safety of drugs used to treat school-aged children with Attention Deficit Hyperactivity
Disorder and epidemiology of the disorder in the Italian population”. A total of 12 issues of
ADHDNews have been produced so far, identifying 440 scientific articles. Over 200 people
have registered to receive the update. The newsletter is also viewable on the Mario Negri’s
website under the section “The Mario Negri Institute for the physician” (L’Istituto Mario
Negri per il Medico): http://www.marionegri.it/mn/it/index.html
236
ANNUAL REPORT 2008

IRFMN
Ricerca & Pratica
Ricerca & Pratica was born in January, 1985, as a manifestation of the “Mario Negri”
Institute for Pharmacological Research. Today, the journal is part of the International Society
of Drug Bulletins (ISDB), which represents independent journals.
For more than twenty years, the journal has represented an arena for all those professionals
who collect data and carry out studies in general practice with the aim to increase their
knowledge and to improve their practice. Ricerca & Pratica is also appreciated for its ability
to go beyond the merely clinical aspect of medicine, without, however, forgetting that it is to
this aspect that the readers dedicate most of their time and effort.
Through its activity, Ricerca & Pratica can therefore represent an exclusive, independent
observation point. It is also an area that promotes contemplation, evaluation, and
information by applying “tools” such as data trustworthiness and importance, the balance
between benefits and risks and between benefits and costs, independence from conflicts of
interest, and the realistic objective to contribute to a progressive, equally distributed
improvement in the population’s health.
VII Master’s Course
An inter-university course aimed at specialising students in the planning and management of
co-operation projects for development. The seventh edition of a 10-day “Stage” in
Cooperation and Public Health, part of the Master’s Course in Analysis and Management of
Projects for Development (www.eco-dip.unimi.it), was organised by the Laboratory and held
at the “Mario Negri” Institute in Milan. The Stage involved a in-depth examination of the
problems associated with public health in developing countries, taking into consideration
their primary importance to the entire world’s population in this new millennium. The course
was based on both theory and practice. The lessons involved issues such as coping with
emergency situations, providing assistance for extreme needs, addressing the problem of
access to drugs, health and well-being indexes, analyses of living conditions, and
intervention programs, and were complemented by hands-on practice sessions that also
involved the basics of epidemiology.
OPINING of the Stage: health is a universal right and an essential need for all. Whether a
society is rich or poor, its development is judged on the basis of how justly distributed its
quality of care is to the population and to what extent it is guaranteed.
However, the right to health is often, and in many countries, not reached because economic
and social inequalities lead to wide health inequalities. This was the topic that the WHO’s
Commission on Social Determinants of Health addressed. The final report was sent to all
health and finance ministers and to healthcare and social work operators, and was presented
by a member of the international commission, Prof. Giovanni Berlinguer, on the stage’s
opening day.
CLOSING Stage: Gherardo Colombo, an ex magistrate (1974-2007) and author of numerous
publications who has been involved for years in explaining the meaning of justice to the
young, met with the students of the Master’s Course in Analysis and Management of
Development Projects during the Stage in Cooperation and Public Health. The topic of the
discussion was the “rules” in contemporary Italy and in the globalised world, sixty years
after the Universal Declaration on Human Rights (whose principles remain largely unmet
worldwide).
Social, economic, cultural, and health-related inequalities exist in all countries and
characterise the conditions and life expectancies of many populations. The justice that should
control and guarantee the distribution of rights and duties is often non-existent or distracted.
The rules (of justice) and adherence to them imply the participation in, and respect of, the
inalienable rights of mankind on the part of all: a utopia
237
ANNUAL REPORT 2008

IRFMN
“FARMACI E BAMBINI” workgroup
The huge ordeal with the Italian Drug Agency (AIFA) and the rapid reorganisation of its
structure caused the workgroup to suspend its work on drugs for children, after two years of
activity. The laboratory was one of the group’s members. The group’s activity in those two
years represented a new methodological approach that increased the AIFA’s attention on the
paediatric population and that included the organisation of educational and training
interventions aimed at health care operators. Given the group’s vast experience, the
technical-scientific, methodological, and managerial heritage that the AIFA has acquired
should not go lost, and the same is true for the approval and credibility gained by the
national regulatory agencies in Europe.
“NASCERE IN CASA”
The Associazione Nazionale Culturale Ostetriche Parto a Domicilio (National Cultural
Association of Home Birth Obstetricians) aims to increase home births, guaranteeing their
quality and safety, verifying and comparing the national and international experiences of
care outside the hospital, and defining professional selection and assistance criteria based on
scientific evidence from the World Health Organization’s recommendations and on women’s
wishes. The Laboratory for Mother and Child Health collaborates with the association,
analysing data collected by the obstetricians in order to improve the quality of care offered to
women and children through experience, but also through continuous study, continuing
professional training, and debate. These improvements will help protect out-of-hospital
births and safeguard the obstetricians’ specific professional role.
The prevention of cervical carcinoma
The Regional Council approved, with decree n.6683 dated 27 th February 2008, the regional
programme on the prevention of cervical carcinoma. The programme involves actions aimed
at incrementing adhesion to pap-test screening, especially in disadvantaged populations, and
to HPV vaccination in girls born in 2007.The complexity of the interventions to put into
place require the specific training of health professionals and the organisation of encounters
during which health professionals can discuss the different aspects of the disease, from its
epidemiology to its prevention and management. In this context, the laboratory, along with
the Lombardy Region, organised the first regional meeting entitled “Prevention of cervical
cancer: meeting and debate on knowledge and perspectives” at the Mario Negri Institute.
Co-operation with countries with limited resources
As an expression, test, and original method of expression of the choice to make the
Laboratory’s research transferable and accessible to all populations, the Laboratory
promoted and provided assistance to projects in, and for, the “South of the world”, also in
collaboration with the World Health Organization. The technical and organisational support
for local groups and international non-governmental organisations for carrying out sociosanitary
projects in countries with limited resources, especially Colombia, Ecuador, Brazil,
Bolivia, Cuba, Vietnam e the Balkans, continues.
ECUADOR - The laboratory supported the campaign “Adopt a mom”, promoted by the
MLAL World-Project, and aimed at the population of Borbón, a town in northern Ecuador.
About 25000 people, from both the black and indigenous races, live in this health district,
which is made up of 170 small communities dispersed along the rivers and from which it can
take up to 15 hours by motor-powered canoe to reach the Borbón hospital.
Objectives:
− improve maternal health conditions
− identify risk factors and obstetric complications in pregnancy, birth, and the perinatal
period early on and provide adequate treatment
− reduce cases of maternal death
− fight risks of foetal and neonatal death
238
ANNUAL REPORT 2008

IRFMN
− train local personnel in the mother and child health field: the communities’ midwives
(parteras), health promoters, and nurses along the rivers that normally assist with 70%
of births in the area.
COLOMBIA - The Colsubsidio Paediatric Investigation in the Americans 2008 Prize was
initiated in Colombia in 1992 with the aim to develop and promote paediatric research. The
prize, which has social and community connotations, is aimed at health care workers in
Central and South America and was designed to form a network though which knowledge,
analysis, study, and equity involving child health can be exchanged.
This network joins ethical solidarity with social and professional responsibility. The prize is
set up by Colsubsidio (Caja Colombiana Subsidio Familiar), a private, mutual society with
more than 850.000 members.
Colsubsidio is a non-profit organisation that has been working in the social, health, and
economic areas in different sectors of society since 1957 and is known for its high quality,
professional work. Over 90 projects from 10 countries were submitted for the 2004 edition.
The projects represented over 240 authors and were evaluated by an international jury
(Maurizio Bonati, Italy; Imti Choonara, United Kingdom; Celia Beatriz Gianotti, Brazil;
Raúl Mercer, Argentina; Carlos Bernal Parra, Gloria Arias Nieto, Jorge Mauricio Palau
Colombia).
The abstracts and integrated texts of the projects that received an award are published on the
SALUDARTE journal Vol. 6 n.2 November 2008 - February 2009.
239
ANNUAL REPORT 2008

IRFMN
240
ANNUAL REPORT 2008

IRFMN
LABORATORY OF REGULATORY
POLICIES
STAFF
Head
Vittorio BERTELE’, M.D.
241
ANNUAL REPORT 2008

IRFMN
CURRICULUM VITAE
Vittorio Bertele’ is a clinical pharmacologist. He got his MD degree in 1977 and the specialization in
Internal Medicine in 1982, both at the Milan University Medical School. He was research fellow at the
Harvard Medical School and then worked at the Milan University and the “Mario Negri” Institute.
His main areas of interest have been clinical pharmacology of drugs active on the haemostatic and
vascular system 1,2 , epidemiology of interventions in the cardiovascular area, and clinical trials and drug
utilization studies in the cardiovascular area 3,4 . He was CPMP expert at the EMEA, and member of the
Committee for Drug Price Negotiation at the Italian Ministry of Health 5,6 .
At present he is Head of the Regulatory Policies Laboratory at the "Mario Negri" Institute, and member
of the Technical-Scientific Committee at the Italian Drug Agency.
Selected publications
• Bertele' V., Falanga A., Tomasiak M., Dejana E., Cerletti C., De Gaetano G. Platelet thromboxane synthetase inhibitors
with low doses of aspirin: Possible resolution of the "aspirin dilemma". Science 1983; 220: 517-519
• Bertele' V., Falanga A., Tomasiak M., Chiabrando C., Cerletti C., De Gaetano G. Pharmacological inhibition of
thromboxane synthetase and platelet aggregation: Modulatory role of cyclooxygenase products. Blood 1984; 63: 1460-
1466 (1984).
• Bertele' V, Mussoni L, Pintucci G, Del Rosso G, Romano G, de Gaetano G, Libretti A. The inhibitory effect of aspirin on
fibrinolysis is reversed by iloprost, a prostacyclin analogue. Thromb Haemost 1989; 61: 286-288
• The i.c.a.i. Group (Gruppo di studio dell'Ischemia cronica Critica degli Arti Inferiori). Prostanoids for chronic critical leg
ischemia: A randomized, controlled, open-label trial with prostaglandin E 1 . Ann Int Med 1999; 130: 412-421
• Collaborative Group of the Primary Prevention Project (PPP). Low-dose aspirin and vitamin E in people at
cardiovascular risk: a randomised trial in general practice. Lancet 2001; 357: 89-95
• Garattini S, Bertele’ V. Adjusting regulatory rules to public health needs. Lancet 2001; 358: 64-67
• Garattini S, Bertele' V. Efficacy, safety, and cost of new anticancer drugs. BMJ 2002; 325: 269-271
• Garattini S, Bertele’ V, Li Bassi L. How can research ethics committees protect patients better BMJ 2003; 326:1199–
201
• Joppi R, Bertele' V, Garattini S. Disappointing biotech. BMJ 2005; 331: 895-897
• Garattini S, Bertele' V. Non-inferiority trials are unethical because they disregard patients' interests. Lancet 2007; 370 :
1875-1877
INTRODUCTION TO THE LABORATORY'S ACTIVITIES
Critical appraisal of clinical methodology
Evaluation of the appropriateness of drug legislation, institutions, and regulatory procedures
with respect to public health needs.
Cooperation to the design and conduct of pharmacovigilance and pharmacoepidemiology
studies in Europe
Cooperation to the development and functioning of the pan-European Infrastructure for clinical
trials (ECRIN, European Clinical Research Infrastructure Network)
Optimisation of drug use and healthcare fund stewardship including potential reforms and
initiatives to achieve this
Critical appraisal and recommendations for European Pricing and Reimbursement systems
including generics, interchangeable products within a class and new innovative medicines
Assessment of emerging technologies
Evaluation of marketing authorization applications submitted to the European regulatory agency
(EMEA) and of subsequent variations
242
ANNUAL REPORT 2008

IRFMN
Cooperation to the development and solution of regulatory issues in developing countries
FINDINGS/MAIN RESULTS
Critical appraisal of clinical research methodological aspects as the adoption of non-inferiority
design in clinical trials, the choice of inadequate comparators or outcome measures, etc
Raising awareness among interested parties about the deficiencies of the present EU
pharmaceutical legislation and about our proposals to improve it in the public health interest
Evaluation of pan-European clinical pilot studies to be run with the support of ECRIN
(European Clinical Research Infrastructure Network) to assess the validity of the overall
organisation and of the quality assurance system, and to refine cost evaluation
Participation in a consortium of public and private European pharmacovigilance and
pharmacoepidemiology centres aimed to plan and conduct a pharmacovigilance project in
Europe under the coordination of the European Medicines Agency (EMEA)
Development of Pan-European strategies for the rational use of new and existing drugs:
establishment of the Piperska Group
Recommendations for Pan-European pricing policies for generics as well as interchangeable
brands in a class once generics are available
Assessment of emerging technologies in the frame of the Italian Horizon Scanning Project
which provides decision makers with timely information on the potential clinical impact and
cost-effectiveness of new health technologies
Critical review of drug documentation at the basis of marketing authorizations
Critical review of the criteria to assess pharmaceutical innovation and include new drugs in the
national reimbursement schemes.
NATIONAL COLLABORATIONS
Italian Drug Agency (AIFA)
Istituto Superiore di Sanità
Department of Health Lombardy Region
Italian Horizon Scanning Project
243
ANNUAL REPORT 2008

IRFMN
European Medicine Agency (EMEA)
INTERNATIONAL COLLABORATIONS
European Clinical Research Infrastructures Network (ECRIN)
European Network of Centres in Pharmacoepidemiology and Pharmacovigilance (ENCePP)
Karolinska Institutet, Division of Clinical Pharmacology, Department of Laboratory Medicine,
and Department of Drug Management and Informatics, SE
University of Liverpool Management School, Prescribing Research Group, UK
International Information Network on New and Emerging Health Technologies (EuroScan)
World Health Organisation (Department of Essential Drugs and Medicines Policy)
Association of South East Asian Nations (ASEAN)
EDITORIAL BOARD MEMBERSHIP
Ricerca & Pratica
Dialogo sui Farmaci
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Technical Scientific Committee at the Italian Drug Agency (AIFA)
Subcommittee of the Centralised Procedure at the Italian Drug Agency (AIFA)
Scientific Committee of the Italian Horizon Scanning Project
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
Araszkiewicz AA, Szabert K, Godman B, Wladysiuk M, Barbui C, Haycox A. Generic olanzapine: health authority
opportunity or nightmare Expert Rev Pharmacoeconomics Outcomes Res 2008; 8:549-555
Bertele' V, Angelici L, Barlera S, Garattini S. Thrombolysis or nothing for acute myocardial infarction It's all the
same! Br J Clin Pharmacol 2008; 65: 955-958
Garattini S, Bertele' V. Do we learn the right things from clinical trials Eur J Clin Pharmacol 2008; 64: 115-125
Garattini S, Bertele' V. The mandate of the European Medicines Evaluation Agency. In : Pharmacoepidemiology and
therapeutic risk management Harvey Whitney Books, Cincinnati, 2008; 95-112
Garattini S, Bertele' V, Godman B, Haycox A, Wettermark B, Gustafsson L L, Piperska Group. Enhancing the
rational use of new medicines across European healthcare systems. Eur J Clin Pharmacol 2008; 64: 1137-1138
244
ANNUAL REPORT 2008

IRFMN
Godman B, Bucsics A, Burkhardt T, et al. Insight into recent reforms and initiatives in Austria; implications for key
stakeholders. Expert Rev. Pharmacoeconomcis Outcomes Res 2008; 8: 357-71
Godman B, Haycox A, Schwabe U, Joppi R, Garattini S. Having your cake and eating it: Office of Fair Trading
proposal for funding new drugs to benefit patients and innovative companies. Pharmacoeconomics 2008; 26: 91-8
Vitry A, Lexchin J, Sasich L, Dupin-Spriet T, Reed T, Bertele' V, Garattini S, Toop L, Hurley E. Provision of information on regulatory
authorities’ websites. Intern Med J 2008; 38: 559-567
Wettermark B, Godman B, Andersson K, Gustaffson L L, Haycox A, Bertele' V. Recent national and regional drug
reforms in Sweden. Implications for pharmaceutical companies in Europe. Pharmacoeconomics 2008 26 : 537-550
Wettermark B, Godman B, Andersson K, Gustafsson L, et al. Recent national and regional drug reforms in Sweden –
implications for pharmaceutical companies in Europe. Pharmacoeconomics 2008; 26: 537-50
RESEARCH ACTIVITIES
Critical appraisal of clinical methodology
Raising awareness about potential biases in clinical research
Critical evaluation of the EU pharmaceutical legislation
Raising awareness among interested parties about the deficiencies of the present EU
pharmaceutical legislation and about our proposals to improve it in the public health interest.
Critical appraisal of ongoing reforms including pricing reforms in major
European countries
Evaluation of ongoing reforms across Europe to enhance generic prescribing rates, drive down
generic prices and corresponding originator brands, as well as potential prices of
interchangeable brands once standards become available as generics, and the potential for cross
cultural learning to release valuable resources to fund increased volumes and new innovative
drugs in the future without prohibitive increases in general taxation or health insurances to
continue to provide equitable and comprehensive healthcare in Europe.
Development of a Pan-European Infrastructure for clinical trials
Participation in a distributed infrastructure linking national networks of clinical research centres
and clinical trials units (ECRIN, European Clinical Research Infrastructure Network) which
provides integrated ‘one-stop shop’ services to investigators and sponsors in multinational
studies.
Development of Pan-European strategies for pharmacovigilance
Developing and testing innovative methods to integrate and present information on benefits and
risks in order to provide all stakeholders (patients, prescribers, regulators and pharmaceutical
companies) with accurate and useful information on drug-related risks and benefits.
Development of Pan-European strategies for the rational use of drugs
Enhancing rational use in line with an approach that has become known as the ‘five Es’,
namely: evaluation; economics; enforcement; education and engineering to further fund
increased volumes and new valuable innovative drugs.
245
ANNUAL REPORT 2008

IRFMN
Assessment of emerging technologies
Collecting information on emerging medicines with respect to their potential clinical impact and
their cost effectiveness and ranking the new products according to their possible marketing
authorization date, their potential innovation grade, therapeutic and economic impact, possible
price and NHS sustainability with the aim to provide decision makers with timely information
on the potential clinical impact and cost effectiveness of new health technologies.
Assessment of drug dossiers for regulatory approvals
Expert support to the Rapporteurship for marketing authorisation applications and variations to
the conditions of marketing authorisation
Activities for the Technical Scientific Committee at the AIFA
Consultative activities for the Italian Drug Agency regarding regulatory duties with respect to
drug quality, safety, efficacy, and cost.
Activities for the sub-committee for the European Procedures
Assessment of the dossiers for marketing authorisation applications through centralized,
decentralized or mutual recognition procedures.
246
ANNUAL REPORT 2008

IRFMN
−
CENTRE OF COMPUTER SCIENCE
ENGINEERING
−
STAFF
Research and Communication Informatics
Head of Division
ROSSI Lorenzo Marco
Division of I.C.T. Services and Management
Head of Division
BAZZI Davide
247
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Lorenzo Marco Rossi graduated in Biomedical Engineering with specialization in Hospital Clinical
Instrumentation at Politecnico of Milan. He has been working with the Institute Mario Negri since 1998.
Main areas of interest:
1. Planning and realization of software for clinical research
2. Planning and realization of software system for in-plant automatization
3. Planning and realization of products for multimedial divulgation
Davide Bazzi graduated in Informatics with specialization in ABACUS at Istituto Tecnico Industriale
Statale of Corsico. He has been working with the Institute of Mario Negro since 1997.
Main areas of interest:
1. Planning, realization and management of communication Network and Data Center
2. Definement and management of quality levels in ICT services distribution
3. Planning and realization of technological innovation for ICT systems
4. Definement and application of organization’s methodologies and processes for the informatics security
management
INTRODUCTION TO THE CENTER'S ACTIVITIES
In order to fulfill even more specialization needs in informatics development, the Centre of
Computer Science Engineering during 2008 has been reorganized itself considering the acquired
skills and created three distinct division bound each other by a strong collaborative relationship.
The Centre of Computer Science Engineering gathering informatics multidisciplinary aspects
promotes and propose itself to coordinate and harmonize the development of the tools for the
management information, improving the integration between informative procedures making
more efficacious communication process and management of scientific and administrative
datas, in order to support and fasten decisional, management, clinical trials and scientific
processes.
RESEARCH ACTIVITIES
Implementation e of Clinical Trials’ gathering forms (E-CRF)
• Lab. Clinical Trials
o Trial COMETS
o Trial TAILOR
o Trial HEAD & NECK
• Outer
o Trial CIPOMO
Maintenance and management of datas’ gathering forms for the following
clinical trials
−
Lab. Neurological Disorders (Dep. Neuroscience):
o Estensione Registro Europeo SLA
o Trial L-ACETYLCARNITINE
o Trial ANTIEPILETTICI
o Trial EPILESSIA E STROKE
248
ANNUAL REPORT 2008

IRFMN
o
o
Trial EPO VS MP IN SPINAL SHOCK
Trial VALPROATO
−
−
−
−
Lab. Clinical Trials (Dep. Oncology)
o Trial FOLFOX
o Trial TOP
Lab. New Drug Development Strategies (Dep. Oncology)
o Trial MAPS
o Trial STARPAN
Dep. Epidemiology
o Trial CADASIL
Lab. Quality Assessment of Geriatric Therapies and Services (Dep. Neuroscience)
o Trial GISAS
o Patients registry for Polipathologies and Politherapies – SIMI web
Web based applications connected to the projects
- Utility system for the current trials gathering data program (realized)
- Analysis software for interaction between drugs and integrations between interaction
database and drugs reference book (in collaboration with Lab. Quality Assessment of
Geriatric Therapies and Services) (developed).
- Reusable electronic poll system for the Mango group
- Registration form to the INSILICO workshop (in collaboration with the Lab.
Environmental Chemistry and Toxicology) (developed)
- Forum "Under 40" (created)
- Electronic board "Negri Community" (developed and realized)
- Database concerning hospitalizations, recipes and medical visitation provided from
Regione Lombardia for covenant data analysis
Other projects
Web based application for in-plant automation
• New database for the Institute Orders
• New database for the Institute Distributed Publications
• Management of the database of the Institute Staff
Multimedial communication
- Planning videoconferences and web connection between quarters
- Presentation of the new Institute: interviews and videos
- DVD video editing and realization of new Institute presentation (in collaboration with
Photographer Office)
249
ANNUAL REPORT 2008

IRFMN
Websites
- Management of the Institute website
- "Adamant" project website
- "Fondazione Mattioli" website
Informatics infrastructures
- Realization of the MPLS communication data infrastructures among the Milan,
Bergamo and Ranica locations
- Planning and realization of the accesses and management and supporting processes
control system
250
ANNUAL REPORT 2008

IRFMN
ITALIAN COCHRANE CENTRE
STAFF
Head
Alessandro LIBERATI, M.D.
251
ANNUAL REPORT 2008

IRFMN
CURRICULUM VITAE
Alessandro Liberati obtained his MD Degree in 1978 at the University of Milano and his post doctoral
degree in Hygiene and Preventive Medicine in 1981 at the same university.
Teaching activities: Primary responsibility of several academic and non academic training courses on the
methodology of clinical research and systematic reviews/metanalyses. He is Director of the advanced
Master “Evidence based medicine e metodologia della ricerca sanitaria”, at the Università degli Studi di
Modena e Reggio Emilia.
Areas of scientific expertise: methodology of clinical research with particular reference to controlled
clinical trials, epidemiological methods for research synthesis (systematic reviews and metanalyses);
methods of practice guidelines production and implementation, evaluation of ethical implications of
clinical research.
Past and current roles at the Mario Negri Institute: 1980-1986 Junior researcher at the Laboratory of
Clinical pharmacology; 1987-1989 Head, Unit of Clinical Epidemiology and Health Services Research;
1990-1998 Head Laboratory of Clinical Epidemiology; since 1994 he is Director of the Italian Cochrane
Centre; since 1998 he is Associate Professor of Clinical Biostatistics and Epidemiology at the University
of Modena and Reggio Emilia; since 1997 he is President of the Associazione per la Ricerca sulla
Efficacia dell’Assistenza Sanitaria - Centro Cochrane Italiano (AREAS-CCI); since 2005 he is President
of the Local Ethics Committee of the Local Health Unit of Bologna; since 2004 he is Member of the
Commissione Nazionale Ricerca Sanitaria; since 2005 he is Member of the Commissione Ricerca e
Sviluppo dell’Agenzia Italiana del Farmaco (AIFA).
Selected publications
• Filippini G, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. When drug companies select what they want to
publish patients are denied relevant therapeutic information. Intern Emerg Med. 2008 Sep;3(3):255-7
• Moja L, Moschetti I, Cinquini M, Sala V, Compagnoni A, Duca P, Deligant C, Manfrini R, Clivio L, Satolli R, Addis A,
Grimshaw JM, Dri P, Liberati A. Clinical evidence continuous medical education: a randomised educational trial of an
open access e-learning program for transferring evidence-based information - ICEKUBE (Italian Clinical Evidence
Knowledge Utilization Behaviour Evaluation) - study protocol. Implement Sci. 2008 Jul 17;3:37
• Iorio A, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. Selecting references that match constructs: the difficult
job of citing the parachute hyperbole. Intern Emerg Med. 2008 Jun;3(2):151-4.
• Guyatt GH, Oxman AD, Kunz R, Jaeschke R, Helfand M, Liberati A, Vist GE, Schünemann HJ; GRADE Working
Group. Incorporating considerations of resources use into grading recommendations. BMJ. 2008 May
24;336(7654):1170-3.
• Guyatt GH, Oxman AD, Kunz R, Falck-Ytter Y, Vist GE, Liberati A, Schünemann HJ; GRADE Working Group. Going
from evidence to recommendations. BMJ. 2008 May 10;336(7652):1049-51.
• Banzi R, Moja L, Moschetti I, Liberati A, Gensini GF, Gusinu R, Conti AA. Rimonabant for overweight and "metabolic
syndrome": the attempt to supersize disease and risk by pharmaceutical marketing. Intern Emerg Med. 2008
Mar;3(1):53-6.
• De Palma R, Liberati A, Ciccone G, Bandieri E, Belfiglio M, Ceccarelli M, Leoni M, Longo G, Magrini N, Marangolo
M, Roila F; Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. Developing clinical
recommendations for breast, colorectal, and lung cancer adjuvant treatments using the GRADE system: a study from the
Programma Ricerca e Innovazione Emilia Romagna Oncology Research Group. J Clin Oncol. 2008 Mar 1;26(7):1033-9.
252
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE CENTRE'S ACTIVITIES
The Italian Cochrane Centre (ICC) (http://www.cochrane.it) was founded in 1994 and is
affiliated to the Cochrane Collaboration (CC). The CC is an international non profit
organization that prepares, maintains and promotes systematic reviews of the effects of health
care interventions. The main product of the Cochrane Collaboration is the Cochrane Library, a
quarterly publication containing Cochrane systematic reviews and other relevant databases of
other siblings international organizations.
The objectives of the ICC are centered around supporting various activities of the Cochrane
Collaboration within Italy. In particular:
a) to disseminate the knowledge of CC and CC activities throughout Italy;
b) to provide methodological and practical support to all individuals and groups who are
interested in collaborating with the CC;
c) to contribute to the adoption and dissemination of Evidence-based Medicine in Italy.
FINDINGS/MAIN RESULTS
The ICC has created a national network with researchers and health care providers who are
producing systematic reviews for the Cochrane Collaboration and are actively involved in other
activities related to the dissemination of evidence based medicine.
NATIONAL COLLABORATIONS
Agenzia Italiana del Farmaco (AIFA), Roma
Istituto Superiore di Sanità, Roma
Ministero della Salute, Roma
Centro Valutazione Efficacia Assistenza Sanitaria (CeVEAS), Modena
Dipartimento di Epidemiologia della ASL Roma E
Istituto Neurologico "Carlo Besta", Milano
Agenzia Sanitaria Regionale, Regione Emilia Romagna, Bologna
Università degli Studi di Milano
Università di Modena e Reggio Emilia
Azienda Sanitaria Locale 20, Alessandria
INTERNATIONAL COLLABORATIONS
The Cochrane Collaboration, Oxford, UK
Centre for Reviews and Dissemination, University of York, York, UK
British Medical Journal Publishing Group, London, UK
Centre for Statistics in Medicine, Oxford, UK
Thomas Chalmers Centre for Systematic Reviews, Ottawa, Canada
The Campbell Collaboration , Philadelphia, USA
253
ANNUAL REPORT 2008

IRFMN
EDITORIAL BOARD MEMBERSHIP
Clinical Evidence (Alessandro Liberati)
Evidence Based Health Policy and Research (Alessandro Liberati)
Journal of Clinical Epidemiology (Alessandro Liberati)
Journal of Health Services Research (Alessandro Liberati)
PEER REVIEW ACTIVITIES
Annals of Internal Medicine (Alessandro Liberati)
British Medical Journal (Alessandro Liberati)
JAMA (Alessandro Liberati)
Evidence Based Health Policy and Research (Alessandro Liberati)
Canadian Medical Association Journal (Alessandro Liberati)
EVENT ORGANIZATION
V Edition Master on “Evidence Based Medicine e metodologia della ricerca sanitaria”, March
2007 - April 2008, Università degli Studi di Modena e Reggio Emilia
1-day Cochrane Workshop in “Evidence for health care decision-making”
Focus in: Orthopedy and physiatry - 16 May 2008, Milan
Focus in: General Medicine - 12 June 2008, Milan
Workshop for physiotherapists - 26 June 2008, Milan
Focus in: Neurology and Internal Medicine - 10 July 2008, Milan
Annual Continental European Cochrane Entities Meeting (CECEM), 8-9 May 2008, Milan
Istituto Ortopedico Galeazzi “Raccomandazioni cliniche : come la ricerca potrebbe orientare la
pratica clinica” - 16 May 2008, Milan
Workshop on the 4th November 2008, Naples:
-The EBM in rehabilitation: the contribution of the Cochrane Collaboration
-The method GRADE to produce recommendations: how the research can guide clinical
practice
-Meeting of the Working Group on "New biotech drugs to treat breast cancer”
XIII Annual Meeting of the Italian Cochrane “Le conoscenze e le innovazioni in medicina: il
ruolo delle revisioni sistematiche” 3 November 2008, Naples
254
ANNUAL REPORT 2008

IRFMN
PARTICIPATION IN EVENTS IN WHICH THE CENTRE WAS
INVOLVED
V Edition Master on “Evidence Based Medicine e metodologia della ricerca sanitaria”, March
2007 - April 2008, Università degli Studi di Modena e Reggio Emilia
“Prove di efficacia e pratica terapeutica: EBM e Cochrane Collaboration”, Master per
Coordinatori di Unità Operativa e/o di Dipartimento per le professioni infermieristiche e
ostetriche, Università degli Studi di Milano, anno 2007-2008 (October 2008).
Annual Continental European Cochrane Entities Meeting (CECEM) 8 – 9 May 2008, Milan
XVI Cochrane Colloquium, 3-7 October 2008, Fribug, Germany
GRANTS AND CONTRACTS
Istituto di Ricerche Farmacologiche Mario Negri, Milano
AIFA - Agenzia Italiana del Farmaco, Roma
Ministero della Salute
255
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
De Andrea S, Montanari M, Moja L, Apolone G. Prevalence of undertreatment in cancer pain. A review
of published literature. Ann Oncol. 2008 Dec;19(12):1985-91.
Filippini G, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. When drug companies select what
they want to publish patients are denied relevant therapeutic information. Intern Emerg Med. 2008
Sep;3(3):255-7.
Moja L, Moschetti I, Cinquini M, Sala V, Compagnoni A, Duca P, Deligant C, Manfrini R, Clivio L,
Satolli R, Addis A, Grimshaw JM, Dri P, Liberati A. Clinical evidence continuous medical education: a
randomised educational trial of an open access e-learning program for transferring evidence-based
information - ICEKUBE (Italian Clinical Evidence Knowledge Utilization Behaviour Evaluation) -
study protocol. Implement Sci. 2008 Jul 17;3:37.
Iorio A, Moja L, Liberati A, Gensini GF, Gusinu R, Conti AA. Selecting references that match constructs:
the difficult job of citing the parachute hyperbole. Intern Emerg Med. 2008 Jun;3(2):151-4.
Guyatt GH, Oxman AD, Kunz R, Jaeschke R, Helfand M, Liberati A, Vist GE, Schünemann HJ; GRADE
Working Group. Incorporating considerations of resources use into grading recommendations. BMJ.
2008 May 24;336(7654):1170-3.
Guyatt GH, Oxman AD, Kunz R, Falck-Ytter Y, Vist GE, Liberati A, Schünemann HJ; GRADE Working
Group. Going from evidence to recommendations. BMJ. 2008 May 10;336(7652):1049-51.
Banzi R, Moja L, Moschetti I, Liberati A, Gensini GF, Gusinu R, Conti AA. Rimonabant for overweight
and "metabolic syndrome": the attempt to supersize disease and risk by pharmaceutical marketing. Intern
Emerg Med. 2008 Mar;3(1):53-6.
De Palma R, Liberati A, Ciccone G, Bandieri E, Belfiglio M, Ceccarelli M, Leoni M, Longo G, Magrini
N, Marangolo M, Roila F; Programma Ricerca e Innovazione Emilia Romagna Oncology Research
Group. Developing clinical recommendations for breast, colorectal, and lung cancer adjuvant treatments
using the GRADE system: a study from the Programma Ricerca e Innovazione Emilia Romagna
Oncology Research Group. J Clin Oncol. 2008 Mar 1;26(7):1033-9.
256
ANNUAL REPORT 2008

IRFMN
RESEARCH ACTIVITIES
Educational and dissemination activities
In 2008 the Italian Cochrane Centre (ICC) participated in the organization of fifth year of the
Master’s program in Evidence Based Medicine (EBM) and health research methodology in
collaboration with the University of Modena and Reggio Emilia in Italy. ICC organized and
conducted a series of workshops about basic EBM concepts for healthcare decision-making
directed toward general practitioners, specialists and doctors in training. These workshops were
held at the Mario Negri Institute in Milan.
In collaboration with AIFA and Zadig (a scientific publishing group), ICC has made a
significant contributions toward the ECCE project (continuing education centred on evidence
based medicine). The ECCE project was established in 2005 and promoted by AIFA as a part of
a large educational project for continuing medical education (CME) of all Italian doctors.
Within the first year of the project, ECCE attracted 17,000 doctors (2005); by the end of 2007
that number rose to 33000.
AIFA and Zadig have also been partners of the ICC in the translation, adaptation, distribution
and evaluation of Clinical Evidence - an international source of the best available evidence for
effective healthcare (published by BMJ publishing group). This past year we have been working
on the 6 th Italian edition of Clinical Evidence which corresponds to the 17th English edition.
In collaboration with PartecipaSalute project, ICC began to translate and disseminate the
Cochrane Collaborations' press releases (plain language summaries of Cochrane Systematic
Reviews) to scientific journalists, doctors and consumers
Research activities
ICC researchers have collaborated in the production of 7 Cochrane Systematic Reviews and 7
protocols currently available in the Cochrane Library.
http://www3.interscience.wiley.com/cgibin/mrwhome/106568753/HOMECRETRY=1&SRETRY=0
ICC researchers are involved in methodological projects focused on the study of the quality of
Systematic Reviews.
In 2008 the ICC has collaborated with PartecipaSalute project. PartecipaSalute aims to involve
consumers and their disease-related associations with follow objectives:
PartecipaSalute ("Participate in Health Care") is a project initiated at September 2003 to foster
a strategic alliance between patients’ groups and professional societies with the common goal
of promoting better health and shared decision-making. The project’s main aims are:
- to increase patients’ associations’ involvement in healthcare assistance and in decision
making processes;
- to promote a partnership between patients’ associations and medical societies soliciting
medical societies’ attention to patients’ need and perspectives
- to develop a partnership between citizens, patients and healthcare system.
The project developed a website - http://www.partecipasalute.it/cms/ - in order to offer critical
tools to read and understand medical and healthcare information, for better healthcare
decisions.
257
ANNUAL REPORT 2008

IRFMN
258
ANNUAL REPORT 2008

IRFMN
THE CATULLO AND DANIELA
BORGOMAINERIO CENTER
One of the buildings on the Mario Negri Institute campus is The Catullo and Daniela
Borgomainerio Center built in 1987 thanks to a donation from Mrs. Angela Marchegiano
Borgomainerio. This is a Center for the study of rare childhood diseases and even today some of
the laboratories housed in the building still conduct this research. For example, the study of new
therapies used to treat a very rare form of acute myeloid leukemia, know as acute promyelocytic
leukemia. A number of new studies are being done to identify new drugs having different
mechanisms able to synergize with trans retinoic acid.
Research on epidemiological childhood leukemia is also done at the Borgomainerio and a
similar line of research involves testicular cancer in adolescents and young adults.
We also do research aimed at finding evidence based therapies for children.
Paediatric research activities done at the Borgomainerio Center are also performed in
collaboration with groups located at other Institute locations including, The Aldo and Cele
Daccò Center for Clinical Research on Rare Diseases at Ranica in Bergamo, the Regional
Centre for Drug Information (CRIF) and the Laboratory for Mother and Child Health
(Department of Public Health) which are both located in Milan.
259
ANNUAL REPORT 2008

IRFMN
260
ANNUAL REPORT 2008

IRFMN
THE LIBRARY
STAFF
Head Librarian
Vanna Pistotti
261
ANNUAL REPORT 2008

IRFMN
The Library, specialised in pharmacology and clinical epidemiology, was founded in 1963
thanks to a generous donation from the Gustavus and Louise Pfeiffer Research Foundation, in
Denville, New Jersey, USA.
Numerous public and private organisations help keep it operative, through donations in money
or books, and subscriptions to periodicals.
STAFF
One Head and two Assistants plus a clerical worker
WHAT THE LIBRARY OFFERS
The library has a collection of about 5000 textbooks, monographs and congressional
proceedings, and 200 periodicals of which a major part are in an electronic format. The books
are classified according to the US National Library of Medicine Classification and the Medical
Subject headings of Medline (MeSH). Besides the internal collection, the Library has access to
the National Periodical Catalogue and to other Library systems (SBBL, GIDIF-RBM).
DATABESES AND ELECTRONIC JOURNALS
From every computer in the Institute it is now possible to have access to more than 2000
electronic journals and to three of the most important databases, PubMed, the Cochrane Library
and Embase.
SPECIAL PROJECTS
The Library cooperates to the realisation of the Italian Information Specialists’ (GIDIF,
RBM) journal catalog which is updated annually and to the catalog of the Lombardy
Biomedical Library Consortium, a network that serves, through Internet, the scientific
community in this District.
It collaborates to the maintenance of the Institute web site, particularly taking care of the
Publications section, both scientific and lay press.
TRAINING
Every year courses on the use of the database and electronic journals are organised. These
courses are designed for use by those working at the Institute but outsiders who are interested
may attend.
262
ANNUAL REPORT 2008

IRFMN
Pistotti V
Conoscere e usare Google
Ricerca & Pratica 2008 n.142 : 161
2008 PUBLICATIONS
Pistotti V, Santoro E
Navigare sulla rete alla ricerca di informazioni di salute
In :La dispensa di Partecipasalute: orientarsi in salute e sanità per fare scelte consapevoli
IRFMN, Milano, 2008; 71-81
CONTRACTS
Since 1994 the library has been part of the Lombard Biomedical Library System. 14 university
and research organisation libraries in Lombardy take part in this project, which allows easy, free
access to scientific information to over 140 centres and institutions the Lombardy Region.
263
ANNUAL REPORT 2008

IRFMN
264
ANNUAL REPORT 2008

IRFMN
Negri Bergamo Laboratories
ANNUAL
REPORT 2008
departments and laboratories
265
ANNUAL REPORT 2008

IRFMN
266
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF MOLECULAR
MEDICINE
STAFF
Head
Ariela BENIGNI, Biol.Sci.D., Ph.D.
Unit of Gene Therapy
Head
Susanna TOMASONI, Biol.Sci.D., Ph.D.
Laboratory of Cell Biology and Xenotransplantation
Head
Marina MORIGI, Biol.Sci.D., Ph.D.
Unit of Platelet-Endothelial Cell Interaction
Head
Miriam GALBUSERA, Biol.Sci.D.
Laboratory of Immunology and Genetics of Organ Transplantation and
Rare Diseases
Head
Marina NORIS, Chem.Farm.D., Ph.D.
Unit of Cellular biology of Autoimmunity and Transplant Rejection
Head
Sistiana AIELLO, Biol.Sci.D
Unit of Cellular and Molecular Biology of Transplantation Tolerance
Head
Federica CASIRAGHI, Chemist
Laboratory of Experimental Models of Kidney Diseases
Head
Carla ZOJA, Biol.Sci.D., Ph.D.
Unit of Pathology and Immunophatology
Head
Mauro ABBATE, M.D.
267
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Ariela Benigni got the Biol.Sci. degree in 1979 at the University of Milano, Italy, and the Ph.D. at
Maastricht University, Netherlands, in 2001.
Educational training: in 1979 Post Doctoral Fellow, Istituto di Ricerche Farmacologiche Mario Negri
(IRFMN), Laboratory of Cancer Chemotherapy, Milan, Italy; in 1980-1981 Post Doctoral Fellow,
Associazione Bergamasca per lo Studio delle Malattie Renali, Laboratory of the Division of Nephrology
and Dialysis, Ospedali Riuniti di Bergamo, Italy; in 1982 Post Doctoral Fellow, Centre Regional de
Transfusion Sanguigne de Strasbourg, France.
Areas of interest: vasoactive and inflammatory mediators of progressive renal injury with a particular
emphasis on endothelin-1; combined treatment of antipertensive and renoprotective drugs to halt
progressive renal injury; use of stem cells for tissue regeneration in acute renal failure in vivo e in vitro
gene transfer; prevention of acute graft rejection through gene therapy; induction of kidney transplant
tolerance by gene therapy; correction of genetic deficiency in rare diseases.
Employement: in 1983 Scientist, IRFMN, Laboratory of Kidney Disease, Bergamo, Italy; in 1990-
1994 Head Laboratory of Prostaglandin and Leukotriene Metabolism, IRFMN, Bergamo, Italy;
from January 1991 Scientific Secretary, IRFMN, Bergamo, Italy; in 1994-1999 Head Laboratory of
Vasoactive and Inflammatory Mediators of Tissue damage, IRFMN, Bergamo, Italy; from January
2000 Head, Department of Molecular Medicine, IRFMN, Bergamo, Italy; from March 2007
Consultant World Health Organization (WHO) for the multicentre observational study “Screening
for Pre-eclampsia: evaluation of the predictive ability of angiogenic factors for Pre-eclampsia”;
during 2007 Senior Fellow at the University of Oxford, Nuffield Department of Obstetrics &
Gynaecology.
Selected publications:
• Macconi D, Chiabrando C, Schiarea S, Aiello S, Cassis L, Gagliardini E, Noris M, Buelli S, Zoja C, Corna D, Mele
C, Fanelli R, Remuzzi G, Benigni A. Proteasomal processing of albumin by renal dendritic cells generates antigenic
peptides. J Am Soc Nephrol. 2009 Jan;20(1):123-30. Epub 2008 Dec 17.
• Vlahou A, Charonis A, Benigni A. Report on the first combined working group and management committee
meeting of EuroKUP (Urine and Kidney Proteomics cost action). J Proteomics. 2009 Jan 30;71(6):682-4. Epub
2008 Nov 17.
• Perico N, Benigni A, Remuzzi G. Present and future drug treatments for chronic kidney diseases: evolving targets
in renoprotection. Nat Rev Drug Discov. 2008 Nov;7(11):936-53. Epub 2008 Oct 10. Review.
• Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A,
Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury
and prolong survival in mice. Stem Cells. 2008 Aug;26(8):2075-82. Epub 2008 May 22.
• Pezzotta A, Mister M, Monteferrante G, Cassis L, Azzollini N, Aiello S, Satta M, Benigni A, Remuzzi G, Noris M.
Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in
rat. Transplantation. 2008 May 27;85(10):1476-82.
• de Borst MH, Benigni A, Remuzzi G. Primer: strategies for identifying genes involved in renal disease. Nat Clin
Pract Nephrol. 2008 May;4(5):265-76. Epub 2008 Mar 25. Review.
Marina Morigi got her Biol.Sci. degree in 1987 at the University of Milano, Milano, Italy and the Ph.D.
at Maastricht University, Netherlands, in 2005.
Educational training: in 1984-1987 Research training, IRFMN, Bergamo, Italy; in 1987 1995 Post
Doctoral Fellow, IRFMN, Bergamo, Italy; in 1991 Stage at Brigham and Women’s Hospital, Laboratory
of Dr. P. Marsden, Boston, USA.
Employement: since 1995 Scientist, IRFMN, Bergamo, Italy; in 1996-1999 Head, Unit of Renal and
Endothelial Cell Biology; since 2000 Head, Laboratory of Cell Biology and Xenotransplantation,
IRFMN, Bergamo, Italy.
Areas of interest: role of Shigatoxin in the pathogenesis of endothelial dysfunction and microvascular
thrombosis in Hemolytic Uremic Syndrome; in vitro model of hyperacute xenograft rejection (porcine
endothelium exposed to human serum as a source of xenoreactive natural antibodies and complement);
renal toxicity of the proteins filtered through the capillary barrier. In vitro model to study intracellular
signals, gene expression and production of inflammatory mediators in cultured proximal tubular cells
and glomerular epithelial cells; cell therapy and tissue regeneration: Capability of adult stem cells to
differentiate and to regenerate renal tissue in acute and chronic experimental models of renal disease.
268
ANNUAL REPORT 2008

IRFMN
Murine embryonic stem cell therapy to correct the genetic defect characteristic of Fabry disease in an
experimental mouse model.
Selected publications
• Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart transplant through the
generation of regulatory T cells. Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA,
Todeschini M, Solini S, Sonzogni A, Perico N, Remuzzi G, Noris M. J Immunol. 2008 Sep 15;181(6):3933-46.
• Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice.
Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A,
Remuzzi A, Remuzzi G. Stem Cells. 2008 Aug;26(8):2075-82. Epub 2008 May 22.
• The regenerative potential of stem cells in acute renal failure. Morigi M, Benigni A, Remuzzi G, Imberti B. Cell
Transplant. 2006;15 Suppl 1:S111-7. Review.
• M. Morigi, B. Imberti, C. Zoja, D. Corna, S. Tomasoni, M. Abbate, D. Rottoli, S. Angioletti, A. Benigni, N. Perico, M.
Alison, G. Remuzzi. Mesenchymal Stem Cells Are Renotropic, Helping to Repair the Kidney and Improve Function in
Acute Renal Failure. J Am Soc Nephrol 2004;15:1794-1804.
• Morigi M, Buelli S, Zanchi C, Longaretti L, Macconi D, Benigni A, Moioli D, Remuzzi G, Zoja C. Shigatoxin-induced
endothelin-1 expression in cultured podocytes autocrinally mediates actin remodeling. Am J Pathol. 2006 Dec;
169(6):1965-75.
• Imberti B, Morigi M, Tomasoni S, Rota C, Corna D, Longaretti L, Rottoli D, Valsecchi F, Benigni A, Wang J, Abbate
M, Zoja C, Remuzzi G. Insulin-like growth factor-1 sustains stem cell mediated renal repair. J Am Soc Nephrol. 2007
Nov;18(11):2921-8.
Marina Noris got her degree in Pharmaceutical Chemistry and Technologies in 1986 at the University
of Rome “La Sapienza) and the Ph.D. at Maastricht University, Netherlands, in 2005.
Educational training: in 1984-1986 Fellow, Istituto di Chimica Farmaceutica e Tossicologica, University
of Rome, Italy; in 1986-1987 Post Doctoral Fellow, Istituto di Chimica Farmaceutica e Tossicologica,
University of Rome, Italy; in September 1987-March 1994 Post Doctoral Fellow, IRFMN, Unit of
Mediators of Inflammation and Tissue Damage, Laboratory of Kidney Disease, Bergamo, Italy.
Areas of interest: immunology of transplantation, tolerance induction; genetics of hemolytic uremic
syndrome, thrombotic thrombocytopenic purpura, focal segmental glomerulosclerosis, diabetic
nephropathy, role of nitric oxide and arginine dysfunctions in uremia and in pre-eclampsia.
Employment: in 1994-1996 Head, Unit of Endothelial Cell Pathophysiology, IRFMN, Bergamo, Italy;
1996-1999 Head, Laboratory of Cellular and Molecular Biology of the immune response and
autoimmunity, IRFMN, Italy; from January 2000: Head, Laboratory of Immunology and Genetics of Rare
Diseases and Organ Transplantation, Department of Molecular Medicine, IRFMN, Bergamo, Italy.
Selected publications
• Castelletti F, Donadelli R, Banterla F, Hildebrandt F, Zipfel PF, Bresin E, Otto E, Skerka C, Renieri A, Todeschini M, Caprioli
J, Caruso RM, Artuso R, Remuzzi G, Noris M. Mutations in FN1 cause glomerulopathy with fibronectin deposits. Proc Natl
Acad Sci U S A. 2008;105(7):2538-43.
• Fang CJ, Fremeaux-Bacchi V, Liszewski MK, Pianetti G, Noris M, Goodship TH, Atkinson JP. Membrane cofactor protein
mutations in atypical hemolytic uremic syndrome (aHUS), fatal Stx-HUS, C3 glomerulonephritis, and the HELLP syndrome.
Blood;111(2):624-32.
• Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A, Perico
N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a semiallogeneic heart
transplant through the generation of regulatory T cells. J Immunol. 2008;181(6):3933-46.
• Caprioli J, Noris M, Brioschi S, Pianetti G, Castelletti F, Bettinaglio P, Mele C, Bresin E, Cassis L, Gamba S, Porrati F,
Bucchioni S, Monteferrante G, Fang CJ, Liszewski MK, Kavanagh D, Atkinson JP, Remuzzi G. Genetics of HUS: the impact
of MCP, CFH and IF mutations on clinical presentation, response to treatment, and outcome. Blood, 2006; 108(4):1267-79.
• Noris M, Casiraghi F, Todeschini M, Cravedi P, Cugini D, Monteferrante G, Aiello S, Cassis L, Gotti E, Gaspari F, Cattaneo
D, Perico N, Remuzzi G. Regulatory T Cells and T Cell Depletion: Role of Immunosuppressive Drugs. J Am Soc Nephrol.
2007; 18 (3):1007-1018.
• Noris M, Bucchioni s, Galbusera M, Donadelli R, Bresin E, Castelletti F, Caprioli J, Brioschi S, Scheiflinger F, Remuzzi G
and the International Registry of Recurrent and Familial HUS/TTP. Complement factor H mutation in familial thrombotic
thrombocytopenic purpura with ADAMTS13 deficency and renal involvement. J Am Soc Nephrol 2005; 16:1177-1183
• Noris M, Brioschi S, Caprioli J, Todeschini M, Bresin E, Porrati F, Gamba S, Remuzzi G, on behalf of the International
Registry of Familial and Recurrent HUS/TTP. Familial haemolytic uraemic syndrome and an MCP mutation. Lancet 2003;
362:1542-1547.
Carlamaria Zoja got her Biol.Sci. degree at the University of Milano, Italy, in 1979 and the Ph.D. at
the University of Maastricht, The Netherlands in 2001.
Educational Training: in 1979-1981 Post Doctoral Fellow, ‘Associazione Bergamasca per lo studio delle
Malattie Renali’, Laboratory of the Division of Nephrology and Dialysis, Ospedali Riuniti di Bergamo,
269
ANNUAL REPORT 2008

IRFMN
Italy; in 1981-1983 Post Doctoral Fellow, Center for Thrombosis and Vascular Research, Department of
Research Katholieke Universiteit, Leuven, Belgium; in 1983-1985: Post Doctoral Fellow, IRFMN,
Laboratory of Kidney Disease, Bergamo, Italy.
Areas of interest: experimental models of kidney diseases of immunological and non immunological
origin; vasoactive and inflammatory mediators of renal disease progression; role of proteinuria in
progressive kidney damage; protection of renal disease progression by a multidrug approach; novel
immunosuppressive and anti-inflammatory strategies for the treatment of lupus nephritis; role of
Shigatoxin in the pathogenesis of endothelial dysfunction in Hemolytic Uremic Syndrome.
Employement: since 1985 Scientist, IRFMN, Bergamo, Italy; in 1990-1994: Head, Unit of Experimental
Modelling for Human Renal Diseases, Laboratory of Kidney Diseases, IRFMN, Bergamo, Italy; since
1995: Head, Laboratory of Experimental Models of Kidney Diseases, IRFMN, Bergamo, Italy.
Selected publications
• C.Zoja, S. Angioletti, R. Donadelli, C. Zanchi, S. Tomasoni, E. Binda, B. Imberti, M. te Loo, L. Monnens, G.Remuzzi,
M. Morigi. Shiga toxin-2 triggers endothelial leukocyte adhesion and transmigration via NF-kB dependent up-regulation
of IL-8 and MCP-1. Kidney Int 2002;62:846-856.
• C. Zoja, D. Corna, D. Camozzi, D. Cattaneo, D. Rottoli, C. Batani, C. Zanchi, M. Abbate, G. Remuzzi. How to fully
protect the kidney in a severe model of progressive nephropathy: a multidrug approach. J Am Soc Nephrol
2002;13:2898-2908.
• R. Donadelli, C. Zanchi, M. Morigi, S. Buelli, C. Batani, S. Tomasoni, D. Corna, D. Rottoli, A. Benigni, M. Abbate, G.
Remuzzi, C. Zoja. Protein overload induces fractalkine upregulation in proximal tubular cells through NF-kB and p38
MAPK dependent pathways. J Am Soc Nephrol 2003; 14:2436-2446.
• C.Zoja, F.Casiraghi, S.Conti, D.Corna, D.Rottoli, R.A.Cavinato, G.Remuzzi, A.Benigni. Cyclin-Dependent kinase
inhibition limits glomerulonephritis and extends lifespan of mice with systemic lupus. Arthritis & Rheumatism 2007;
56; 1629-1637.
• C.Zanchi, C.Zoja, M. Morigi, F.Valsecchi, XY Liu, D. Rottoli, M. Locatelli, S. Buelli, A.Pezzotta, P.Mapelli, J.
Geelen, G.Remuzzi, J.Hawiger J. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to
Shiga toxin. J Immunol. 2008; 181:1460-9.
• M.Abbate, C.Zoja, D.Corna, D.Rottoli, C.Zanchi, N.Azzollini, S. Tomasoni, S.Berlingeri, M.Noris, M.Morigi,
G.Remuzzi. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J
Am Soc Nephrol. 2008; 19:1158-67.
Mauro Abbate obtained his M.D. degree in 1988 at the University of Brescia, Italy.
Educational training: in 1984-1988 Graduate Student, IRFMN, Bergamo, Italy; in 1988-1992 Post
Doctoral Fellow, IRFMN, Bergamo, Italy; in 1992-1994 Research Fellow, The Renal Unit,
Massachusetts General Hospital, HMS, Boston, USA.
Areas of interest: renal disease progression: the role of proteinuria, complement, and mediators of injury
in progressive kidney damage; mechanisms of glomerular injury; anti-GBM glomerulonephritis;
mechanisms of tubular injury; kidney fibrosis; the renal biopsy; membranous nephropathy.
Employement: in 1996 - 2000: Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Renal
Pathology and Immunopathology, IRFMN, Bergamo, Italy.
Principali pubblicazioni
• Abbate M, Zoja C, Morigi M, Rottoli D, Angioletti S, Tomasoni S, Zanchi C, Longaretti L, Donadelli R, Remuzzi G:
Transforming Growth Factor-beta 1 Is Up-Regulated by Podocytes in Response to Excess Intraglomerular Passage of
Proteins: A Central Pathway in Progressive Glomerulosclerosis. Am J Pathol.2002;161,2179-93.
• Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A,
Remuzzi G. Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target
for renoprotective intervention. Am J Pathol. 2006 Apr;168(4):1073-85.
• Abbate M, Zoja C, Remuzzi G. How does proteinuria cause progressive renal damage J Am Soc Nephrol. 2006
Nov;17(11):2974-84. Review
• Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G, Abbate M, Remuzzi G.
Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin J Am Soc Nephrol. 2008
Nov;3(6):1652-9.
• Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi A,
Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and
prolong survival in mice. Stem Cells. 2008 Aug;26(8):2075-82
• Abbate M, Zoja C, Corna D, Rottoli D, Zanchi C, Azzollini N, Tomasoni S, Berlingeri S, Noris M, Morigi M, Remuzzi
G. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J Am Soc
Nephrol. 2008 Jun;19(6):1158-67
270
ANNUAL REPORT 2008

IRFMN
Sistiana Aiello got the Biol.Sci. degree in 1993 at the University of Milano, Italy, and the Specialization
in Pharmacology Research in 1996, at IRFMN, Bergamo, Italy.
Educational training: in 1990-1993 research training, IRFMN, Bergamo; in 1993-2000 post doctoral
fellow, IRFMN, Bergamo.
Areas of interest: transplant immunology with a particular interest on dendritic cell biology and
mechanisms by which T regulatory cells arise and work; in vitro and in vivo studies on new compounds
with immunosuppressive capacity or capable to prevent ischemia/reperfusion tissue injury; vasoactive and
inflammatory mediators of progressive renal injury with a particular emphasis on platelet activating factor
(PAF) and nitric oxide (NO).
Employement: since 2000 Scientist within Laboratory of Immunology and Genetics of Rare disease and
Organ Transplantation; IRFMN, Bergamo; since 2006 Head, Unit of Cellular Biology of Autoimmunity
and Transplant Rejection, IRFMN, Transplant Research Center “Chiara Cucchi de Alessandri e Gilberto
Crespi”, Ranica.
Selected publications:
• Pezzotta A, Mister M, Monteferrante G, Cassis L, Azzollini N, Aiello S, Satta M, Benigni A, Remuzzi G, Noris M.
Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection in rat.
Transplantation. 2008; 85(10):1476-82.
• S. Aiello, P. Cassis, L. Cassis, S. Tomasoni, A. Benigni, A. Pezzotta, R.A. Cavinato, D. Cugini, N. Azzollini, M. Mister,
L. Longaretti, A.W. Thomson, G. Remuzzi, M. Noris. DnIKK2-transfected dendritic cells induce a novel population of
iNOS-expressing CD4 + CD25 - cells with tolerogenic properties. Transplantation 2007; 83:474-484.
• S. Tomasoni, S. Aiello, L. Cassis, M. Noris, L. Longaretti, R.A. Cavinato, N. Azzollini, A. Pezzotta, G. Remuzzi, A.
Benigni. Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent
CD4 + T regulatory cells. Transplantation 2005;79:1056-1061.
• L. Cassis, S. Aiello, M. Noris. Natural versus adaptive regulatory T cells. Contrib Nephrol 2005; 146: 121-131.
• M. Mister, M. Noris, J. Szymczuk, N. Azzollini, S. Aiello, A. Arduini, L. Trochimowicz, E. Gagliardini, M. Abbate, N.
Perico, G. Remuzzi. Propionyl-L-carnitine prevents renal function deterioration due to ischemia/reperfusion in isolated
perfused rat kidney and in kidney graft. Kidney Int 2002; 61:1064-1078.
• S. Aiello, M. Noris, G. Piccinini, S. Tomasoni, F. Casiraghi, S. Bonazzola, M. Mister, M.H. Sayegh, G. Remuzzi.
Thymic dendritic cells express inducible nitric oxide synthase and generate nitric oxide in response to self- and
alloantigens. J Immunol 2000;164:4649-4658.
Federica Casiraghi has obtained his degree in Industrial Chemistry in 1988, and the degree in Clinical
Monitoring and in Biochemical Research in 1993-1994 at IRFMN, Bergamo, Italy.
Educational Training: 1989-1994 research fellow, IRFMN, Bergamo.
Areas of interest: Transplant immunology with particular focus on pharmacological and cellular therapies
for induction and maintenance of transplantation tolerance. Characterization of regulatory T cells in renal
transplant patients and in experimental models of allograft tolerance. Impact of different
immunosuppressive drugs on T cell function in renal transplant patients. Vasoactive and inflammatory
mediators of progressive renal injury with a particular emphasis on arachidonic acid metabolites.
Employment: since 1994 Scientist within Laboratory of Immunology and Genetics of Rare Disease and
Organ Transplantation, IRFM, Bergamo; since 2006 Head, Unit of Cellular and Molecolar Biology of
Transplantation Tolerance, Transplant Research Center “Chiara Cucchi de Alessandri e Gilberto Crespi”,
Ranica.
Selected publications:
• Casiraghi F, Azzollini N, Cassis P, Imberti B, Morigi M, Cugini D, Cavinato RA, Todeschini M, Solini S, Sonzogni A,
Perico N, Remuzzi G, Noris M. Pretransplant infusion of mesenchymal stem cells prolongs the survival of a
semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol. 2008;181(6):3933-46.
• Noris M, Casiraghi F, Todeschini M, Cravedi P, Cugini D, Monteferrante G, Aiello S, Cassis L, Gotti E, Gaspari F,
Cattaneo D, Perico N, Remuzzi G. Regulatory T Cells and T Cell Depletion: Role of Immunosuppressive Drugs. J Am
Soc Nephrol. 2007; 18 (3):1007-1018.
• Cavinato RA, Casiraghi F, Azzollini N, Cassis P, Cugini D, Mister M, Pezzotta A, Aiello S, Remuzzi G, Noris M.
Pretransplant donor peripheral blood mononuclear cells infusion induces transplantation tolerance by generating
regulatory T cells. Transplantation, 2005;79(9):1034-9.
• Zoja C, Benigni A, Noris M, Corna D, Casiraghi F, Pagnoncelli M, Rottoli D, Abbate M, Remuzzi G. Mycophenolate
mofetil combined with a cyclooxygenase-2 inhibitor ameliorates murine lupus nephritis. Kidney Int. 2001; 60(2): 653-
63.
• Tomasoni S, Noris M, Zappella S, Gotti E, Casiraghi F, Bonazzola S, Benigni A, Remuzzi G. Upregulation of renal and
systemic cyclooxygenase-2 in patients with active lupus nephritis.
J Am Soc Nephrol.1998; 9(7): 1202-12.
271
ANNUAL REPORT 2008

IRFMN
• Casiraghi F, Ruggenenti P, Noris M, Locatelli G, Perico N, Perna A, Remuzzi G. Sequential monitoring of urine-soluble
interleukin 2 receptor and interleukin 6 predicts acute rejection of human renal allografts before clinical or laboratory
signs of renal dysfunction. Transplantation 1997; 63(10): 1508-14.
Miriam Galbusera got her Biol.Sci. degree in 1981 at the Università degli Studi di Milano.
Educational training: in 1981-1983 Post Doctoral Fellow, Istituto di Patologia Speciale Medica
dell'Università degli Studi di Milano, Italy; in 1985 - 1989 Post Doctoral Fellow, IRFMN, Bergamo,
Italy; in 1989-1991 Post Doctoral Fellow at Scripps Clinic and Research Foundation, Laboratory of
Thrombosis and Hemostasis, La Jolla, CA, USA; in 1991-1995 Post Doctoral Fellow, IRFMN, Bergamo,
Italy.
Areas of interest: ADAMTS-13 and VWF in thrombotic microangiopathies, VWF biochemistry,
xenotransplantation, platelet-endothelial cell interaction under flow condition, platelet pathophysiology in
uremia, receptor studies in kidney and platelets.
Employement: 1995 - 1999: Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Platelet-
Endothelial Cell Interaction, IRFMN, Bergamo, Italy.
Selected publications
• Tripodi, A., Chantarangkul, V., Bohm, M., Budde, U., Dong, J.-F., Friedman, K.D., Galbusera, M., Girma, J.-P., Moake,
J., Rick, M.E., Studt, J.-D., Turecek, P.L., Mannucci, P.M.: Measurement of von Willebrand factor cleaving protease
(ADAMTS-13): results of an international collaborative study involving 11 methods testing the same set of coded
plasmas. J Thromb Haemost 2004; 2:1601-1609.
• Galbusera, M., Morigi, M., Buelli, S., Gastoldi, S., Macconi, D., Testa, C., Angioletti, S., Remuzzi, G.: Xenogeneic
serum-induced thrombus formation on porcine endothelium is mediated by vitronectin receptor and P-selectin: role of
reactive oxygen species. Xenotransplantation 2005;12:110-120.
• Ruiz-Torres, M.P., Casiraghi, F., Galbusera, M., Macconi, D., Gastoldi, S., Todeschini, M., Porrati, F., Belotti, D.,
Pogliani, E.M., Noris, M., Remuzzi, G.: Complement activation: the missing link between ADAMTS13 deficiency and
microvascular thrombosis of thrombotic microangiopathies. Thromb Haemost 2005; 93:443-452.
• Noris, M., Bucchioni, S., Galbusera, M., Donadelli, R., Bresin, E., Castelletti, F., Caprioli, J., Brioschi, S., Scheiflinger,
F., Remuzzi, G.: Complement factor H mutation in familial thrombotic thrombocytopenic purpura with ADAMTS13
deficiency and renal involvement. J Am Soc Nephrol 2005; 16:1177-1183.
• Galbusera, M., Bresin, E., Noris, M., Gastoldi, S., Belotti, D., Capoferri, C., Daina, E., Perseghin, P., Scheiflinger, F.,
Fakhouri, F., Grunfeld, J-P., Pogliani, E., Remuzzi, G.: Rituximab prevents recurrence of thrombotic thrombocytopenic
purpura: a case report. Blood 2005;106:925-928.
• Rieger, M., Mannucci, P.M., Kremer Hovinga, J.A., Herzog, A., Gerstenbauer, G., Konetschny, C., Zimmermann, K.,
Scharrer, I., Peyvandi, F., Galbusera, M., Remuzzi, G., Böhm, M., Plaimauer, B., Lämmle, B., Scheiflinger, F.:
ADAMTS13 autoantibodies in patients with thrombotic microangiopathies and other immunomediated diseases. Blood
2005;106:1262-1267.
Susanna Tomasoni got her Biological Science degree in 1991 at the University of Milan.
Educational training: in 1989-1991 Graduate student, University of Milan; in 1991-1994 PhD student,
University of Milan; in 1994 Research Fellow, Renal Division, Brigham & Women’s Hospital, Harvard
Medical School, Boston, USA; in 1995 PhD degree in Physiological Science, University of Bologna;
1995-1998: Post Doctoral Fellow, IRFMN, Bergamo, Italy.
Areas of interest: construction of adenoviral vectors for gene therapy; gene transfer to the kidney in the
context of transplantation; transfection of dendritic cell for cell therapy; progression of renal disease.
Employement: in 1998-2000 Scientist, IRFMN, Bergamo, Italy; from 2000 Head, Unit of Gene Therapy,
IRFMN, Bergamo, Italy
Selected publications
• Allograft rejection: acute and chronic studies. Tomasoni S, Remuzzi G, Benigni A. Contrib Nephrol. 2008;159:122-34.
Review.
• Tomasoni S, Azzollini N, Casiraghi F, Capogrossi M C, Remuzzi G, Benigni A. CTLA4Ig gene transfer prolongs
survival and induces donor-specific tolerance in a rat renal allograft. J Am Soc Nephrol 2000; 11: 747-752.
• Tomasoni S, Benigni A. Gene therapy: How to target the kidney. Promises and pitfalls. Curr Gene Ther 2004; 4: 115-
122.
• Tomasoni S, Longaretti L, Azzollini N, Gagliardini E, Mister M, Buehler T, Remuzzi G, Benigni A. Favorable effect of
cotransfection with TGF-beta and CTLA4Ig of the donor kidney on allograft survival. Am J Nephrol 2004; 24: 275-283
• Tomasoni S, Aiello S, Cassis L, Noris M, Longaretti L, Cavinato R, Azzollini N, Pezzotta A, Remuzzi G, Benigni A.
Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent CD4+ T-
regulatory cells. Transplantation 2005; 79: 1056-1061.
• Benigni A, Tomasoni S, Turka LA, Longaretti L, Zentilin L, Mister M, Pezzotta A, Azzollini N, Noris M, Conti S,
Abbate M, Giacca M, Remuzzi G, Adeno-associated virus-mediated CTLA4Ig gene transfer protects MHC-mismatched
renal allografts from chronic rejection. J Am Soc Nephrol, 2006, 17: 1665-72.
272
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department of Molecular Medicine was established in 1999 at the Negri Bergamo
laboratories to coordinate the work of three laboratories and three units. The activities of the
Department of Molecular Medicine are strictly interrelated with those of the Department of
Renal Medicine of the Clinical Research Center for Rare Diseases Aldo e Cele Daccò.
The following major objectives have been pursued:
1) identification of mediators and mechanisms responsible for the relentless decline of renal
function in kidney diseases and development of therapeutic interventions to slow or even halt
the disease progression to end-stage renal failure;
2) understanding the mechanisms underlying endothelial cell dysfunction in thrombotic
microangiopathies and hyperacute rejection of xenograft
3) finding new strategies for modulating the immune response and preventing acute and chronic
rejection of kidney allograft as well as exploration of immunological pathways leading to donor
specific unresponsiveness and tolerance of the graft;
4) investigation of the molecular and genetic basis of rare diseases such as hemolytic uremic
syndrome/thrombotic thrombocytopenic purpura and pre-eclampsia and search for diseasesusceptibility
genes or gene polymorphisms predicting the patient's response to drug therapy in
more common and complex polygenic disorders.
Such goals have been pursued using various approaches: 1) experimental models of kidney
diseases of immunological and non-immunological origin mimicking human renal diseases to
study vasoactive and inflammatory mediators and to test novel antiproteinuric and
renoprotective drugs; 2) in vitro cultures of renal cells to address the toxicity of protein overload
reproducing the condition of exaggerated protein traffic of proteinuric progressive
nephropathies; 3) in vitro models to assess the interaction of vascular endothelial cells with
leukocytes and platelets under controlled flow conditions; 4) experimental models of kidney
allotransplant to study immunological processes responsible for acute and chronic rejection, the
nephrotoxicity of immunosuppressor drugs as well as to explore pathways responsible for
accomodation; 5) gene transfer of viral constructs carrying genes encoding immunomodulatory
molecules to overcome acute rejection of allotransplantation avoiding immunosuppression; 6)
identification of candidate genes with linkage analysis and search for mutations as well as
assessment of gene polymorphisms.
FINDINGS/MAIN RESULTS
Cord blood mesenchymal stem cells prolong survival of mice with acute kidney
injury.
Pre-transplant infusion of adult or embryonic mesenchymal stem cells prolongs
survival of heart transplant in mice through the generation of regulatory T cells.
C3 filtered in the urine induces renal damage.
Fractalkine induces micro vascular lesions typical of HUS because it induces leucocyte
adhesion to the endothelium.
273
ANNUAL REPORT 2008

IRFMN
Self proteins become antigenic in disease conditions and promote renal injury.
A cell cycle inhibitor is a potent immunosuppressor in transplant setting.
First description of fibronectin 1 mutations as a cause of a rare disease, the glomerulopathy with
fibronectin deposit.
Abnormal function of mutated MCP which predisposes to atypical HUS
Proof of concept that gene therapy with adenoviral vectors corrects ADAMTS13 deficiency in
Thrombotic Thrombocytopenic Purpura
NATIONAL COLLABORATIONS
Laboratorio di Terapia genica e cellulare, G. Lanzani, Divisione di Ematologia, Ospedali Riuniti
di Bergamo
Laboratorio di Tecnologie riproduttive, Istituto Sperimentale Lazzaro Spallanzani, Cremona
Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano
Dipartimento di Istologia Microbiologia e Biotecnologie Mediche, Università di Padova
International Centre for Genetic Engineering and Biotechnology, Molecular Medicine Group,
Trieste
U.O. di Ostetricia e Ginecologia, Ospedale San Gerardo di Monza
U.O. di Ostetricia e Ginecologia, Azienda Ospedaliera Ospedali Riuniti di Bergamo
U.O. di Ostetricia e Ginecologia, Azienda Ospedaliera Spedali Civili di Brescia
I.R.C.C.S. Policlinico San Matteo, Pavia
INTERNATIONAL COLLABORATIONS
Academisch Ziekenhuis Maastricht, Interne Geneeskunde, Maastricht, Olanda
Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, USA
Children's Hospital and Regional Medical Center, University of Washington, Seattle, USA
Departements of Pediatrics and Human Genetics, University of Michigan, Ann Arbor, USA
Deparment of Medicine, Division of Rheumatology, Washington University School of
Medicine, St. Louis, USA
Erasmus University of Rotterdam, Olanda
Hans-Knoll Institute for Natural Products Research, Jena, Germania
INSERM, Paris, Francia
Klinikum der Ludwig Maximillians Universitat Munchen, Germania
Max Delbruck Center for Molecular Medicine, Berlin, Germania
Max-Plank Gesellschaft zur Forderung der Wissenshaften, Hpi of experimental
endocrinology, Hannover, Germania
National Institute of Health, Bethesda, USA
Necker Hospital, Paris, Francia
Osaka University School of Medicine, Osaka, Giappone
Pediatric Nephrology and Hypertension, University of Utah, USA
Renal Transplant Unit, Hospital de Bellvitge, Barcelona, Spagna
Rosalind Franklin University of Medicine and Science, Chicago, USA
The Scripps Research Institute, La Jolla, USA
Universitaet Hamburg, Institut fur Molekulare Neuropathobiologie, Hamburg, Germania
274
ANNUAL REPORT 2008

IRFMN
University of Aarthus, Danimarca
University of Colorado Cardiovascular Institute, Denver, USA
University of Groningen, Olanda
University of Iowa, Department of Internal Medicine and Pediatrics, Iowa City, USA
University of Liverpool, School of Biological Sciences, UK
University of Oxford, UK
University of Oulu, Danimarca
University of Pittsburgh School of Medicine, Pittsburgh, USA
University of Zurich, Svizzera
Weizman Institute of Science, Rehovot, Israele
World Health Organization, Geneva, Svizzera
EDITORIAL BOARD MEMBERSHIP
Journal of American Society of Nephrology (Marina Noris)
PEER REVIEW ACTIVITIES
American Journal of Pathology
American Journal of Physiology
Cell Proliferation
Diabetologia
Hypertension
Immunology
Kidney International
Journal of Clinical Investigation
Journal of the Renin Angiotensin Aldosterone System
Journal of the American Society of Nephrology
Journal of Thrombosis and Haemostasis
Nature Medicine
Nephrology, Dialysis and Transplantation
Nephron Clinical Practice
New England Journal of Medicine
Pediatric Nephrology
Plos Medicine
The American Journal of Pathology
The Canadian Journal of Physiology and Pharmacology
The Journal of Experimental Medicine
The Lancet
The Open Urology & Nephrology Journal
Transplant Immunology
Trends in molecular medicine
275
ANNUAL REPORT 2008

IRFMN
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
Meeting per progetto GENECURE: Berlino, 16-17 gennaio 2008; Ranica (BG), 10-11 April
2008; Bruxelles, 15 Dicembre 2008
Nefropatie primitive e secondarie, Firenze, 21-22 November 2008
III Convegno Internazionale sulle cellule Staminali: le cellule staminali tra terapia e formazioni
dei tumori”, Agrigento-Favara, 20-22 November 2008
Workshop “Mesenchymal stem cells”, 18 November 2008, Milano
American Society of Nephrology, Philadelphia, 5-10 November 2008
“Annual GRIP Investigator Meeting, 9-10 November 2008, Philadelphia
"Rare diseases and Orphan Drugs" Istituto Superiore di Sanità, Roma dal 27-31 October 2008
Focus on Rare Diseases: the Genetic and Molecular basis of Rare kidney Disorders, Bergamo,
October 9-11 2008
XXII International complement workshop, Basilea, 28 settembre-2 October 2008
European Society for Pediatric Nephrology, Lione, 14 September 2008
Corso di aggiornamento "Renal Biopsy in Medical Diseases of the Kidneys", Columbia
University, New York, 10-13 September 2008
Meeting of the Dense Deposit Disease Focus group, Hinxton (London), 16-17 August 2008
Workshop “Kidstem Network”, 3-4 July 2008, Dresden
American Transplant Congress, Toronto, 31 May-4 June 2008
ERA-EDTA course on “Complement in renal disease and transplantation”, Leiden, 24-25 April
2008
Podonet Steering Committee meeting, Heidelberg, 20 April 2008
Advanced Workshop of Nephoprotection, 18-19 April 2008, Centro Daccò, Ranica (BG)
EuReGene Yearly Meeting- Symposium on Complex (Renal) Genetics, Nizza, 31 gennaio 2008
EuroKUP First Meeting, Bruxelles, 18-19 May 2008
Annual European Congress of Rheumatology “EULAR 2008”, Parigi, 13-14 June 2008
62 nd Annual Conference of the Council for High Blood Pressure Research, Atlanta, 17-20
September 2008
276
ANNUAL REPORT 2008

IRFMN
GRANTS AND CONTRACTS
Comitato Telethon Fondazione ONLUS
Commissione Europea FP6 e FP7
Fondazione Aiuti per la Ricerca sulle Malattie Rare (ARMR)
Fondazione ART per la Ricerca sui Trapianti ONLUS
Fondazione Cariplo
Fondazione La Nuova Speranza – Lotta alla Sclerosi focale ONLUS
Fondazione ROTRF/JDRF
Istituto Superiore di Sanità
ACRAF (Aziende Chimiche Riunite Angelini Francesco Spa)
Farmaceutici Damor Spa
Genzyme Corporation
Giuliani Spa
Pfizer Ltd
Sanofi-Aventis Spa
Speedel Pharma Ltd
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
M. Abbate, G. Remuzzi, C. Zoja. Role of proteinuria in progression. In: The Kidney: Physiology and
Pathophysiology, 4 th ed., chapter n. 89. Edited by R.J. Alpern, S.C. Hebert. Elsevier, London 2008, pp. 2563-2576.
M. Noris, G. Remuzzi, T.H.J. Goodship. Hemolytic uremic syndrome/Thrombotic thrombocytopenic purpura. In:
Handbook of Systemic Autoimmune Diseases, Vol. 7, chapter n. 14. Edited by J.C. Mason, C.D. Pusey. Elsevier,
London 2008, pp. 257-282.
F. Castelletti, R. Donadelli, F. Banterla, F. Hildebrandt. P.F. Zipfel, E. Bresin, E. Otto, C. Skerka, A. Renieri, M.
Todeschini, J. Caprioli, M.R. Caruso, R. Artuso, G. Remuzzi, M. Noris. Mutations in FN1 cause glomerulopathy with
fibronectin deposits. Proc Natl Acad Sci USA 2008;105:2538-2543.
M. Noris, G. Remuzzi. Translational Mini-Review Series on Complment Factor H: Therapies of renal diseases
associated with complement factor H abnormalities: atypical haemolytic uraemic syndrome and
membranoproliferative glomerulonephritis. Clin Exp Immunol 2008;151:199-209.
J. Caprioli, G. Remuzzi. A mouse model of non-Shiga toxin-associated haemolytic uraemic syndrome. (Editorial)
Nephrol Dial Transplant 2008;23:462-465.
R. Martinez-Barricarte, G. Pianetti, R. Gautard, J. Misselwitz, L. Strain, V. Fremeaux-Bacchi, C. Skerka, P.F. Zipfel,
T. Goodship, M. Noris, G. Remuzzi, S.R. de Cordoba on behalf of the European Working Party on the genetics of
HUS. The complement factor H R1210C mutation is associated with atypical hemolytic uremic syndrome. J Am Soc
Nephrol 2008;19:639-646.
A. Pezzotta, M. Mister, G. Monteferrante, L. Cassis, N. Azzollini, S. Aiello, M. Satta, A. Benigni, G. Remuzzi, M.
Noris. Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity and acute kidney allograft rejection
in rat. Transplantation 2008;85:1476-1482.
C. Zanchi, C. Zoja, M. Morigi, F. Valsecchi, X.Y. Liu, D. Rottoli, M. Locatelli, S. Buelli, A. Pezzotta, P. Mapelli, J.
Geelen, G. Remuzzi, J. Hawiger. Fractalkine and CX3CR1 mediate leukocyte capture by endothelium in response to
Shiga toxin. J Immunol 2008;181:1460-1469.
M.H. de Borst, A. Benigni, G. Remuzzi. Primer: strategies for identifying genes involved in renal disease. Nat Clin
Pract Nephrol 2008;4:265-276.
277
ANNUAL REPORT 2008

IRFMN
M. Abbate, C. Zoja, D. Corna, D. Rottoli, C. Zanchi, N. Azzollini, S. Tomasoni, S. Berlingeri, M. Noris, M. Morigi,
G. Remuzzi. Complement-mediated dysfunction of glomerular filtration barrier accelerates progressive renal injury. J
Am Soc Nephrol 2008;19:1158-1167.
S. Tomasoni, G. Remuzzi, A. Benigni. Allograft rejection: acute and chronic studies. In: Contribution to Nephrology:
Gene therapy for renal diseases and transplantation. Edited by C. Ronco, A. Benigni, G. Remuzzi. Karger, Basel 2008,
Vol. 159, pp. 122-134.
N. Perico, A. Benigni, G. Remuzzi. Present and future drug treatments for chronic kidney diseases: evolving targets in
renoprotection. Nature Reviews Drug Discovery 2008;7:936-953.
M. Morigi, M. Introna, B. Imberti, D. Corna, M. Abbate, C. Rota, D. Rottoli, A. Benigni, N. Perico, C. Zoja, A.
Rambaldi, A. Remuzzi, G. Remuzzi. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal
injury and prolong survival in mice. Stem Cells 2008;26:2075-2082.
P. Ruggenenti, P. Bettinaglio, F. Pinares, G. Remuzzi. Angiotensin converting enzyme insertion/deletion
polymorphism and renoprotection in diabetic and nondiabetic nephropathies. Clin J Am Soc Nephrol 2008;3:1511-
1525.
F. Casiraghi, N. Azzollini, P. Cassis, B. Imberti, M. Morigi, D. Cugini, R.A. Cavinato, M. Todeschini, S. Solini, A.
Sonzogni, N. Perico, G. Remuzzi, M. Noris. Pretransplant infusion of mesenchymal stem cells prolongs the survival of
a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol 2008;181:3933-3946.
P. Ruggenenti, M. Noris, G. Remuzzi. Thrombotic Microangiopathies. In: Therapy in Nephrology and Hypertension –
A companion to Brenner and Rector’s The Kidney (3 rd Edition), chapter n. 26. Edited by C.S. Wilcox. Saunders
Elsevier, Philadelphia 2008, pp. 294-312.
Azzollini N, Cugini D, Cassis P, Pezzotta A, Gagliardini E, Abbate M, Arduini A, Peschechera
A, Remuzzi G, Noris M. Propionyl-L-carnitine prevents early graft dysfunction in allogeneic rat kidney
transplantation. Kidney Int. 2008 Dec;74(11):1420-8. Epub 2008 Aug 13.
Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A. Assessment of in vitro differentiation of bovine
pancreatic tissue in insulin-expressing cells. JOP. 2008 Sep 2;9(5):601-11.
Caprioli J, Mele C, Mossali C, Gallizioli L, Giacchetti G, Noris M, Remuzzi G, Benigni A.
Polymorphisms of EDNRB, ATG, and ACE genes in salt-sensitive hypertension. Can J Physiol Pharmacol. 2008
Aug;86(8):505-10.
Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G,
Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin
J Am Soc Nephrol. 2008 Nov;3(6):1652-9. Epub 2008 Aug 6.
Remuzzi G, Cattaneo D, Perico N. The aggravating mechanisms of aldosterone on kidney fibrosis. J Am Soc Nephrol.
2008 Aug;19(8):1459-62. Epub 2008 Jun 11.
Tomasoni S, Remuzzi G, Benigni A. Allograft rejection: acute and chronic studies. Contrib Nephrol. 2008;159:122-
34. Review.
278
ANNUAL REPORT 2008

IRFMN
RESEARCH ACTIVITIES
Laboratory of Cell Biology and Xenotransplantation
Life-sparing effect of human cord-blood mesenchymal stem cells in
mice with acute kidney injury
In collaboration with the Laboratory of Experimental Models of Kidney Diseases
New approaches for the cure of acute kidney injury (AKI), a highly life-threatening
clinical setting, have identified in the transplantation of bone marrow-derived
mesenchymal stem cells (MSCs) a new tool for renal repair in experimental models. To
further explore innovative interventions for AKI, we investigated the potential of human
cord blood MSCs (CB-MSCs) in preventing cisplatin-induced AKI and prolonging
survival in an immunodeficient NOD/SCID mouse model.
NOD/SCID mice were subcutaneously injected with the nephrotoxic drug cisplatin (12.7
mg/kg) and one day later were intra vein injected with saline or human CB-MSCs (5x10 5
cells) stained with the fluorescent dye PKH26. Renal function was measured at different
time points as blood urea nitrogen. Mice were followed during time for survival studies
or sacrificed at 4 days after cisplatin. Kidneys were taken for ultrastructural analysis and
for studying proliferation and apoptosis. Intracellular pathways, involved in the
regenerative process induced by human CB-MSCs, were also investigated. Infusion of
human CB-MSCs in cisplatin mice with AKI ameliorated renal function and markedly
decreased proximal tubular epithelial cell injury and mortality (cisplatin mice: human
CB-MSCs 14% vs saline 100% mortality at 9 days p

IRFMN
the basis for graft tolerance. Therefore, we have studied whether the pre-treatment with ESC
could modulate tolerogenic properties in semiallogeneic ectopic heart transplantation (B6C3
into B6 recipients). Transplanted mice, pre-infused with ESC, 7 or 45 days before
transplantation, exhibited a prolongation of graft survival as compared to mice receiving
fibroblasts (P 100 days in 40% of
the animals). The study of the mechanism possibly involved in graft tolerance, by adoptive
transfer studies, showed that regulatory T cells are not involved.
Laboratory of Experimental Models of Kidney Diseases
Complement-mediated dysfunction of glomerular filtering barrier
accelerates progressive renal injury
Intrarenal complement activation leads to chronic tubulointerstitial injury in animal models of
proteinuric nephropathies and is candidate target of renoprotective therapy. The present study
investigates the role of abnormally filtered plasma proteins in this injury and mechanisms
responsible for C3 accumulation in kidney. We find that mice with C3 deficiency are protected
to a significant degree against the interstitial inflammatory response and tissue damage elicited
by protein overload. These effects are partly related to less severe podocyte injury and lower
proteinuria. Both C3 protein deposition and C3 mRNA upregulation in proximal tubule are
features in common with other progressive models. Here, antiproteinuric treatment with
angiotensin-converting enzyme inhibitor, lisinopril, limits filtered plasma protein and C3
accumulation by proximal tubular cells and lessens interstitial inflammation and damage in
wild-type mice. C3 deficient kidneys transplanted before induction of disease into wild type
mice develop glomerular injury, C3 accumulation in podocytes and proximal tubules, and
tubulointerstitial changes in response to protein overload; in contrast, wild type kidneys given to
C3-deficient mice reveal no abnormal C3 deposition and milder disease. These data show that 1.
the presence of C3 enhances susceptibility to injury of glomerular filtering barrier, 2.
ultrafiltered C3 has greater influence than locally synthesized C3 on tubulointerstitial damage
induced by protein overload, and 3. local C3 synthesis is irrelevant to the development of
proteinuria. Our results suggest that complement-targeted approaches should be combined with
interventions to minimize proteinuria as a way to more effectively prevent progression of renal
disease and associated cardiovascular events.
Fractalkine and CX3CR1 mediate leukocyte captare by endothelium in
response to Shiga toxin
In collaboration with the Laboratory of Cell Biology and Xenotransplantation
Shiga toxins (Stx) are the virulence factors of enterohemorrhagic E.coli O157:H7, a world-wide
emerging diarrheal pathogen, which precipitates post-diarrheal hemolytic uremic syndrome, the
leading cause of acute renal failure in children. Here we show that Stx2 triggered expression of
fractalkine, a CX3C transmembrane chemokine, acting as both adhesion counterreceptor on
endothelial cells and soluble chemoattractant. Stx2 caused in HUVEC expression of fractalkine
mRNA and protein, which promoted leukocyte capture, ablated by antibodies to either
endothelial fractalkine or leukocyte CX3CR1 receptor. Exposure of human glomerular
endothelial cells to Stx2 recapitulated its fractalkine-inducing activity and fractalkine-mediated
leukocyte adhesion. Both processes required phosphorylation of Src-family protein tyrosine
kinase and p38 MAPK in endothelial cells. Furthermore, they depended on nuclear import of
NF-kB and other stress-responsive transcription factors. Inhibition of their nuclear import with
280
ANNUAL REPORT 2008

IRFMN
the cell-penetrating SN50 peptide reduced fractalkine mRNA levels and fractalkine-mediated
leukocyte capture by endothelial cells. Adenoviral overexpression of IkB inhibited fractalkine
mRNA upregulation. The fractalkine-mediated responses to Stx2 were also dependent on AP-1.
In mice both virulence factors of Stx-producing E.coli –Stx and LPS- are required to elicit
hemolytic uremic syndrome. Here, fractalkine was detected within glomeruli of C57BL/6 mice
injected with Stx2, and further increased after Stx2 plus LPS co-administration. This was
associated with recruitment of CX3CR1-positive cells. Thus, in response to Stx2, fractalkine is
induced playing essential role in the promotion of leukocyte-endothelial cell interaction thereby
potentially contributing to the renal microvascular dysfunction and thrombotic microangiopathy
that underlie hemolytic uremic syndrome due to enterohemorrhagic E.coli O157:H7 infection.
Upon renal injury self-proteins generate antigenic peptides through a
proteasome-dependent pathway
In collaboration with the Laboratory of Renal Biophysics (Department of Bioengineering)
and the Laboratory of Analytic Biochemistry (Department of Environmental Health
Sciences)
In proteinuric nephropathies loss of the permselective properties of the glomerular membrane
leads to abnormal filtration of proteins, mainly albumin, that cause distress of the proximal
tubular cell inducing the release into the interstitium of chemokines responsible for the
recruitment and activation of inflammatory cells. The interstitial infiltrates are also
characterized by the presence of immune-competent cells including dendritic cells (DCs) and T
lymphocytes that accumulate within renal parenchyma even in absence of an immune insult. As
antigen presenting cells, DCs can initiate immune response or tolerance. The outcome depends
on context. The current view is that immunity is controlled by internal communication between
the tissue, where DCs are resident, and the immune-competent cells, so that tolerance is
maintained by the healthy organ, whereas immunity can be stimulated by a distressed organ.
Within the kidney, DCs are in close contact with the tubular epithelium and work as immune
sentinels to probe renal interstitium in search for foreign antigens. It is conceivable that upon
injury, the inflammatory stimuli released from the tubular cell represent danger signals that alert
DCs making them immunogenic instead of tolerogenic. This might favor the presentation of
normally ignored self-antigens triggering an immune response. The evidence that the antiproteinuric
effect of the angiotensin converting enzyme inhibitors is associated with a
significant reduction of interstitial infiltrates and renal damage further supports this hypothesis
and the idea that abnormally filtered plasmatic proteins represent self-antigens capable to trigger
an immune response. To verify our hypothesis, we investigated how albumin interacts with the
immune system. Rat proximal tubular cells exposed in vitro to autologous albumin
concentrations, mimicking those present in the ultrafiltrate in proteinuric conditions, release into
the supernatant peptides identified, by mass spectrometry, as N-terminal 24, 27, or 30 amino
acid albumin fragments. Given the abundance of the 24 amino acid peptide, we focused later
studies on this peptide we named Alb1-24. This fragment is not formed in the presence of
pepstatin A, suggesting that an acid protease with pepsin-like activity is responsible for the
selective N-terminal truncation of albumin at position 24. Alb1-24 is taken up by bone marrowderived
rat dendritic cells and processed, through a proteasome dependent pathway, into shorter
peptides that are loaded on the major histocompatibility complex class I (MHC-I) and presented
to syngeneic CD8 + T cells. The first encounter between Alb1-24-pulsed DCs primes naive CD8 +
T cells, so that when exposed a second time, CD8 + T cells become activated and release
interferon γ. In a cell-free system, purified proteasome cleaves albumin into peptides some of
them have a size compatible with MHC-I binding (8 to 10 amino acid long). Within the
products of in vitro proteasomal degradation of Alb1-24, the 15 to 22 amino acid peptide was
identified by bioinformatic tools, used to search for potential binders to MHC-I, as a top-ranked
8-mer ligand for H2Kb allele. The functional role of DC proteasome in generating potentially
antigenic peptides from albumin was next demonstrated in vivo in a model of non-immune
281
ANNUAL REPORT 2008

IRFMN
proteinuric nephropathy. Four weeks after the 5/6 nephrectomy of renal mass, a decrease in the
percentage of DCs in the kidney was paralleled by an enhancement of these cells in the renal
lymph nodes, suggesting migration of DCs from the renal interstitium. The CD8 + T cells
isolated from renal lymph nodes produced interferon γ, at the first encounter, when incubated
with Alb1-24- pulsed DCs, suggesting that they were educated in vivo during the course of the
disease. By contrast, the in vivo treatment with the proteasome inhibitor bortezomib, that
prevent the formation of peptides for recognition by CD8 + T cells, made these cells resistant to
activation by Alb1-24-pulsed DCs either in a primary or secondary culture. Our findings
indicate that albumin can become the source of potentially antigenic peptides upon renal injury
and outline the relationship between tubular cell biology and the role of the DC in processing
self-proteins through a proteasome-dependent pathway.
Laboratory of Immunology and Genetic of Rare Diseases and
Organ Transplantation
Effect of seliciclib (CYC202, R-roscovitine) on lymphocyte alloreactivity
and acute kidney allograft rejection in rat
T cell stimulation by alloantigens is followed by cell cycle progression, an event that is
critically dependent on cyclin-dependent kinases (CDKs). We conducted a study to evaluate
whether the CDK inhibitor seliciclib affected rat lymph node cell (LNc) activation and
proliferation induced by either concanavalin A or allogeneic splenocytes in vitro and studied
the mechanisms underlying the suppressive effect. We also investigated the immunosuppressive
properties of seliciclib in vivo.
Seliciclib completely inhibited in vitro proliferation of LNc and CD8 + Tcells, in response to
either concanavalin A or allogeneic splenocytes. The percentage of activated LNc was lower in
MLR added with seliciclib than in MLR added with vehicle. The percentages of viable and
apoptotic cells at the end of MLR with seliciclib were comparable to those of MLR with
vehicle. LNc pre-exposed in MLR to seliciclib did not respond to further stimulation with
alloantigens, and neither IL-2 nor IL-15 restored proliferation. These data indicate that the
inhibitory effect of seliciclib on T cell alloreactivity is not due to cytotoxic effect but is
associated with induction of profound T cell anergy. LNc harvested at the end of the primary
MLR with seliciclib did not suppress the proliferation of syngeneic LNc cells toward allogeneic
splenocytes, thus excluding that seliciclib induced the formation of regulatory cells. Finally,
seliciclib partially prolonged grafted animal survival in a rat model of fully MHC-mismatched
kidney transplantation.
Altogether these results document that seliciclib regulates lymphocyte reactivity and may exert
an immunosuppressive effect in vivo in the setting of transplantation.
Pretransplant infusion of mesenchymal stem cells prolongs the survival
of a semiallogeneic heart transplant through the generation of regulatory
T cells.
In this study, we investigated whether mesenchymal stem cells (MSC) had immunomodulatory
properties in solid organ allotransplantation, using a semiallogeneic heart transplant mouse
model, and studied the mechanism(s) underlying MSC tolerogenic effects. Either single (portal
vein, day -7) or double (portal vein, day -7 and tail vein, day -1) pretransplant infusions of
donor-derived B6C3 MSC in B6 recipients induced a profound T cell hyporesponsiveness and
prolonged B6C3 cardiac allograft survival. The protolerogenic effect was abrogated when
donor-derived MSC were injected together with B6C3 hematopoietic stem cells (HSC),
suggesting that HSC negatively impact MSC immunomodulatory properties. Both the induction
282
ANNUAL REPORT 2008

IRFMN
(pretransplant) and the maintenance phase (>100 days posttransplant) of donor-derived MSCinduced
tolerance were associated with CD4 + CD25 + Foxp3 + Treg expansion and impaired antidonor
Th1 activity. MSC-induced regulatory T cells (Treg) were donor-specific since adoptive
transfer of splenocytes from tolerant mice prevented the rejection of fully MHC-mismatched
donor-specific secondary allografts but not of third-party grafts. In addition, infusion of
recipient-derived B6 MSC tolerized a semiallogeneic B6C3 cardiac allograft, but not a fully
MHC-mismatched BALB/c graft, and expanded Treg. A double i.v. pretransplant infusion of
recipient-derived MSC had the same tolerogenic effect as the combined intraportal/i.v. MSC
infusions, which makes the tolerogenic protocol applicable in a clinical setting. In contrast,
single MSC infusions given either peritransplant or 1 day after transplant were less effective.
Altogether these findings indicate that MSC immunomodulatory properties require HSC
removal, partial sharing of MHC Ags between the donor and the recipient and pretransplant
infusion, and are associated with expansion of donor-specific Treg.
Mutations in FN1 cause glomerulopathy with fibronectin deposits.
Glomerulopathy with fibronectin (FN) deposits (GFND) is an autosomal dominant disease with
age-related penetrance, characterized by proteinuria, microscopic hematuria, hypertension, and
massive glomerular deposits of FN that lead to end-stage renal failure. The genetic abnormality
underlying GFND was still unknown. We hypothesized that mutations in FN1, which encodes
FN, were the cause of GFND. In a large Italian pedigree with eight affected subjects, we found
linkage with GFND at the FN1 locus at 2q32. We sequenced the FN1 in 15 unrelated pedigrees
and found three heterozygous missense mutations, the W1925R, L1974R, and Y973C, that
cosegregated with the disease in six pedigrees. The mutations affected two domains of FN
(Hep-II domain for the W1925R and the L1974R, and Hep-III domain for the Y973C) that play
key roles in FN-cell interaction and in FN fibrillogenesis. Mutant recombinant Hep-II
fragments were expressed, and functional studies revealed a lower binding to heparin and to
endothelial cells and podocytes compared with wild-type Hep-II and an impaired capability to
induce endothelial cell spreading and cytoskeletal reorganization. Overall dominant mutations
in FN1 accounted for 40% of cases of GFND in our study group. These findings may help
understanding the pathogenesis of proteinuria and glomerular FN deposits in GFND and
possibly in more common renal diseases such as diabetic nephropathy, IgA nephropathy, and
lupus nephritis. To our knowledge no FN1 mutation causing a human disease was previously
reported.
Membrane cofactor protein mutations in atypical hemolytic uremic
syndrome (aHUS), fatal Stx-HUS, C3 glomerulonephritis, and the HELLP
syndrome.
The hemolytic uremic syndrome (HUS) is a triad of microangiopathic hemolytic anemia,
thrombocytopenia, and renal impairment. Genetic studies demonstrate that heterozygous
mutations of membrane cofactor protein (MCP;CD46) predispose to atypical HUS (aHUS),
which is not associated with exposure to Shiga toxin (Stx). Among the initial 25 MCP
mutations in patients with aHUS were 2, R69W and A304V, that were expressed normally and
for which no dysfunction was found. The R69W mutation is in complement control protein
module 2, while A304V is in the hydrophobic transmembrane domain. In addition to 3 patients
with aHUS, the A304V mutation was identified in 1 patient each with fatal Stx-HUS, the
HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome, and
glomerulonephritis with C3 deposits. A major goal was to assess if these putative mutations
lead to defective complement regulation. Permanent cell lines expressing the mutated proteins
were complement "challenged," and membrane control of C3 fragment deposition was
monitored. Both the R69W and A304V MCP mutations were deficient in their ability to control
283
ANNUAL REPORT 2008

IRFMN
the alternative pathway of complement activation on a cell surface, illustrating the importance
of modelling transmembrane proteins in situ.
Unit of Gene Therapy
Gene therapy to correct Thrombotic Thrombocytopenic Purpura
Thrombotic thrombocytopoenic purpura (TTP) is a thrombotic microangiopathy, characterized
by anemia, thrombocytopenia and formation of microvascular platelet rich thrombi. TTP
patients show prevalent, but not exclusive, neurological symptoms and renal dysfunction due to
accumulation in the plasma of ultralarge von Willebrand multimers. In healthy subjects the
VWF-cleaving protease ADAMTS13 (a disintegrin-like and metalloprotease with
thrombospondin type I repeats) reduces the ultra large VWF multimers into smaller forms soon
after their secretion. Acquired and familial deficiency of ADAMTS13 causes TTP in humans.
Plasma infusion is the treatment of choice for TTP patients, however it exposes patients to a
high risk of infections, fluid volume overload and allergic responses. Only 5-6% of
ADAMTS13 activity is sufficient to avoid relapses of the disease. In light of these findings gene
therapy could be an alternative therapeutic strategy for patients with familial TTP. In order to
evaluate the efficacy of gene therapy we constructed two different viral vectors containing
human ADAMTS13 cDNA, an adeno-associated and an adenoviral vector. Both vectors were
able to infect human fibrosarcoma cells and to induce the synthesis of the recombinant protein.
The availability of Adamts13-/- knock out mice (Adamts13 -/- ) were instrumental to study if
systemic injection of the vectors was able to restore protein levels. Adamts13 -/- mice were given
1x10 11 vgu AAV-ADAMTS13 resulting in appreciable expression of rADAMTS13 mRNA into
the liver, the constitutive site of ADAMTS13 production. Time course experiments showed that
ADAMTS13 mRNA was expressed at 4, 8 and at much lower levels at 16 weeks from injection.
Antigen levels were measured in plasma of treated mice by ELISA but no protein was detected,
probably due to the low infectivity of the virus. It is well known that large cDNAs impair
correct packaging of AAV. The AAV serotype used in this experiment (AAV2) has an
insertional capacity similar to ADAMTS13 size. We are now going to test different seroptypes
with a larger insertional capacity.
We also tested the adenoviral vector in vivo. Systemic injection of 1x10 9 Ad-ADAMTS13 pfu
induced ADAMTS13 mRNA expression in the liver, kidneys and lung in Adamts13 -/- mice
sacrified 1 week after the treatment, yielding the secretion in the plasma of large amount of
protein. The released protein was functionally active even after 2 months from injection.
These results would indicate that gene therapy represents a promising therapeutic strategy to
correct ADAMTS13 deficiency in patients with familial TTP.
284
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF BIOMEDICAL
ENGINEERING
STAFF
Head Andrea REMUZZI, Eng. D.
Laboratory of Renal Biophysics
Head
Daniela MACCONI, Biol.Sci.D.
Laboratory of Biomedical Technologies
Head
Bogdan ENE-IORDACHE, Eng.D.
Unit of Tissue Engineering
Head
Marina FIGLIUZZI, Biol.Sci.D.
Unit of Medical Imaging
Head
Luca ANTIGA, Ph.D.
285
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Andrea Remuzzi got his degree in Mechanical (Biomedical) Engineering in 1979, Politecnico di Milano.
Research experience: 1980 Politecnico di Milano, Dipartimento di Ingegneria Biomedica; 1981 Istituto
Mario Negri (Milano), Laboratorio di Farmacologia Cardiovascolare; 1982-83 Massachusetts Institute of
Technology, Mechanical Engineering Department, Cambridge, USA.
Areas of interest: biological transport phenomena, mathematical models, renal pathophysiology, cellular
response to mechanical stimulation, tissue engineering, pancreatic islet transplantation, clinical databases,
computational fluid dynamics.
Chronology of appointment: From 1984 to 1986 Ricercatore Istituto Mario Negri (Bergamo), Laboratorio
di malattie renali, 1986-1989 Head, Unità di Bioingegneria, Istituto Mario Negri, 1989-1993 Head,
Laboratorio di Bioingegneria, Istituto Mario Negri, 1993-1999 Head, Dipartimento di Ricerca Renale,
Istituto Mario Negri, from 2000 Head, Dipartimento di Bioingegneria, Istituto Mario Negri. From 1998 to
2007 contract professor of Bioengineering, Politecnico di Milano. For 2007 Professor of Bioengineering,
University of Bergamo.
Selected publications
• Davies PF, Remuzzi A, Gordon EJ, Dewey CF Jr, Gimbrone MA Jr. Turbulent fluid shear stress induces vascular
endothelial cell turnover in vitro. Proc Natl Acad Sci U S A. 1986 Apr;83(7): 2114-7. PMID: 3457378
• Remuzzi A, Puntorieri S, Battaglia C, Bertani T, Remuzzi G. Angiotensin converting enzyme inhibition ameliorates
glomerular filtration of macromolecules and water and lessens glomerular injury in the rat. J Clin Invest. 1990
Feb;85(2):541-9. PMID: 1688888
• Noris M, Morigi M, Donadelli R, Aiello S, Foppolo M, Todeschini M, Orisio S, Remuzzi G, Remuzzi A. Nitric oxide
synthesis by cultured endothelial cells is modulated by flow conditions. Circ Res. 1995 Apr;76(4):536-43. PMID:
7534657
• Giavazzi R, Foppolo M, Dossi R, Remuzzi A. Rolling and adhesion of human tumor cells on vascular endothelium
under physiological flow conditions. J Clin Invest. 1993 Dec;92(6):3038-44. PMID: 7504697
• Antiga L, Ene-Iordache B, Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of
blood vessels from MR and CT angiography. IEEE Trans Med Imaging. 2003 May;22(5):674-84. PMID: 12846436
• Remuzzi A, Gagliardini E, Sangalli F, Bonomelli M, Piccinelli M, Benigni A, Remuzzi G. ACE inhibition reduces
glomerulosclerosis and regenerates glomerular tissue in a model of progressive renal disease. Kidney Int. 2006
Apr;69(7):1124-30.
• Ruggenenti P, Remuzzi A, Remuzzi G. Decision time for pancreatic islet-cell transplantation. Lancet. 2008 371:883-4.
• Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Bruno S, Remuzzi G, Remuzzi A. Computed tomography
evaluation of autosomal dominant polycystic kidney disease progression: a progress report. CJASN 2006 1:754-60.
Daniela Macconi got her Biol.Sci.D. degree in Milan in the 1983.
Research experience: 1977-81 CNR Institute of Neuroscience - Cell Mol Pharmacology - and Department
of Medical Pharmacology, University of Milan, Milan, Italy;1982-83 Laboratory of the Division of
Nephrology and Dialysis, Ospedali Riuniti di Bergamo, Bergamo, Italy; 1984-85 University of Michigan,
Medical School, Department of Pathology, Medical Science I, Ann Arbor Michigan, USA; 1985-89
Mario Negri Institute for Pharmacological Research, Laboratory of Kidney Disease, Bergamo, Italy.
Areas of interest: glomerular permeability, renal disease progression, podocytes, angiotensin II, reactive
oxygen species
Chronology of appointment: From 2000 Head Laboratory of Renal Biophisics, Department of
Biomedical Engineering; 1994-2000 Head, Unit of Inflammatory Mediator of Leucocyte Origin; 1989- 94
Scientist, 1985-89 post-doctoral fellow Mario Negri Institute for Pharmacological Research, Bergamo,
Italy; 1982-83 fellow Laboratory of the Division of Nefrology e Dialysis, Ospedali Riuniti di Bergamo,
Bergamo, Italy
Selected publications
• Macconi D, Remuzzi G. Candesartan and renal protection: more than blocking angiotensin type 1 receptor Kidney Int.
74:1112-4, 2008.
• Macconi D, Abbate M, Morigi M, Angioletti S, Mister M, Buelli S, Bonomelli M, Mundel P, Endlich K, Remuzzi A,
Remuzzi G: Permselective dysfunction of podocyte-podocyte contact upon angiotensin II unravels the molecular target for
renoprotective intervention. Am J Pathol 168:1073-85, 2006.
• Macconi D, Bonomelli M, Benigni A, Plati T, Sangalli F, Longaretti L, Conti S, Kawachi H, Hill P, Remuzzi G, Remuzzi
A. Pathophysiologic implications of reduced podocyte number in a rat model of progressive glomerular injury. Am J Pathol
68:42-54, 2006.
• Ruiz-Torres MP, Casiraghi F, Galbusera M, Macconi D, Gastoldi S, Todeschini M, Porrati F, Belotti D, Pogliani EM,
Noris M, Remuzzi G: Complement activation: the missing link between ADAMTS-13 deficiency and microvascular
thrombosis of thrombotic microangiopathies. Thromb Haemost 93:443-52, 2005.
286
ANNUAL REPORT 2008

IRFMN
• Morigi M, Macconi D, Zoja C, Donadelli R, Buelli S, Zanchi C, Ghilardi M, Remuzzi G: Protein overload-induced NFkappaB
activation in proximal tubular cells requires H(2)O(2) through a PKC-dependent pathway. J Am Soc Nephrol
13:1179-89, 2002
• Macconi D, Ghilardi M, Bonassi ME, Mohamed EI, Abbate M, Colombi F, Remuzzi G, Remuzzi A: Effect of angiotensinconverting
enzyme inhibition on glomerular basement membrane permeability and distribution of zonula occludens-1 in
MWF rats. J Am Soc Nephrol 11:477-89, 2000.
Bogdan Ene-Iordache got MSc in Mechanical Engineering in 1990 at the Oil & Gas Institute in Ploiesti
(Romania). In 1992 he joined the Bioengineering Laboratory at NegriBERGAMO Laboratories.
Main interests: renal research (hemodynamics and remodeling of arteriovenous fistula for vascular access,
morfometrical analysis of glomerular capillaries) and controlled clinical trials (data management and data
analysis for controlled clinical studies); biomedical informatics, web sites development, coordination of
IT activity in the Clinical Research Center for Rare Diseases ‘Aldo e Cele Daccò’; applied clinical
informatics (Nephrology, Diabetology and Hematology Units, Bergamo Hospital).
Roles: since January 2000 is head of the Biomedical Technologies Laboratory, Department of Biomedical
Engineering.
Selected publications
• Ene-Iordache B, Imberti O, Foglieni O, Remuzzi G, Bertani T and Remuzzi A. Effects of angiotensin-converting enzyme
inhibition on glomerular capillary wall ultrastructure in MWF/Ztm rats. J Am Soc Nephrol 5: 1378-1384, 1994.
• Ene-Iordache B and Remuzzi A. Numerical analysis of blood flow in reconstructed glomerular capillary segments.
Microvasc Res 49: 1-11, 1995.
• Remuzzi A and Ene-Iordache B. Capillary network structure does not affect theoretical analysis of glomerular size
selectivity. Am J Physiol 268: F972-F979, 1995.
• Ene-Iordache B, Mosconi L, Remuzzi G, Remuzzi A. Computational fluid dynamics of a vascular access case for
hemodialysis. J Biomech Eng 123(3): 284-292, 2001.
• Ene-Iordache B, Mosconi L, Antiga L, Bruno S, Anghileri A, Remuzzi G, Remuzzi A. Radial artery remodeling in response
to shear stress increase within arteriovenous fistula for hemodialysis access. Endothelium 10(2): 95-102, 2003.
• Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-Iordache
B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G for the Bergamo Nephrologic Diabetes
Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. NEJM 351(19): 1941-
1951, 2004.
• Ene-Iordache B, Carminati S, Antiga L, Rubis N, Ruggenenti P, Remuzzi G and Remuzzi A. Developing regulatorycompliant
electronic case report forms for clinical trials: experience with the DEMAND trial. JAMIA 15(2), 2009.
Marina Figliuzzi got her Biol.Sci.D. degree in Milan in the 1991.
Research experience :1991-94 Mario Negri Institute for Pharmacological Research, Bergamo, Italy.
Areas of interest: isolation of pancreatic islets from human, bovine , pig and rat pancreas, cell culture,
immunoisolation devices for pancreatic islets, differentiation of progenitor pancreatic cells in insulin
containing cells, immunhistochemistry.
Chronology of appointment: From 2000 Head Unit of Tissue Engineering, Department of Biomedical
Engineering; 1991-2000 fellow laboratory of Renal research, Mario Negri Institute for Pharmacological
Research, Bergamo, Italy.
Selected publications
• Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A.Assessment of in vitro differentiation of bovine
pancreatic tissue in insulin-expressing cells. JOP 9(5):601-11, 2008.
• Figliuzzi M, Plati T, Cornolti R, Adobati F, Fagiani A, Rossi L, Remuzzi G, Remuzzi A. Biocompatibility and function of
microencapsulated pancreatic islets. Acta Biomater. 2006 Mar;2(2):221-7.
• Figliuzzi M, Cornolti R, Plati T, Rajan N, Adobati F, Remuzzi G, Remuzzi A: Subcutaneous xenotransplantation of bovine
pancreatic islets. Biomaterials. 26:5640-47, 2005.
• Figliuzzi M, Zappella S, Morigi M, Rossi P, Marchetti P, Remuzzi A: Influence of donor age on bovine pancreatic islet
isolation. Transplantation. 70:1032-37, 2000
• Morigi M, Micheletti G, Figliuzzi M, Imberti B, Karmali MA, Remuzzi A, Remuzzi G, Zoja C. Verotoxin-1 promotes
leukocyte adhesion to cultured endothelial cells under physiologic flow conditions. Blood.: 86 4553-58, 1995.
• Zoja C, Morigi M, Figliuzzi M, Bruzzi I, Oldroyd S, Benigni A, Ronco P, Remuzzi G. Proximal tubular cell synthesis and
secretion of endothelin-1 on challenge with albumin and other proteins. Am J Kidney Dis.26: 934-41, 1995.
• Morigi M, Zoja C, Figliuzzi M, Foppolo M, Micheletti G, Bontempelli M, Saronni M, Remuzzi G, Remuzzi A. Fluid shear
stress modulates surface expression of adhesion molecules by endothelial cells. Blood. 85:1696-703, 1995.
287
ANNUAL REPORT 2008

IRFMN
Luke Antiga graduated in 1999 in Biomedical Engineering and got his PhD in Bioengineering in 2003,
Politecnico di Milano, having worked at the research laboratory of Biomedical Technology, Department
of Bioengineering Institute Mario Negri.
Training activities: 2003 Post-doctoral fellow at Imaging Research Laboratories, Robarts Research
Institute, London, Ontario.
Areas of interest: acquisition and image processing for medical and microscopy applications, numerical
modeling of transport phenomena.
Roles: from 2000 to 2002 Doctoral Student at the Laboratory of Biomedical Technology, Department of
Bioengineering, from 2002 to 2003 visiting scientist Robarts Institute for medical Imaging, London,
Ontario, Canada, from 2004 to 2006 resercher at the Laboratory of Biomedical Technology, Department
of Bioengineering, from 2007 Head Unit of Medical Imaging, Department of Bioengineering.
Selected publications
• Lee SW, Antiga L, Spence JD and Steinman DA. Geometry of the carotid bifurcation predicts its exposure to disturbed
flow. Stroke, in press, 2008.
• Antiga L, Piccinelli M, Fasolini G, Ene-Iordache B, Ondei P, Ruggenenti P, Remuzzi G and Remuzzi A. Computed
tomography evaluation of ADPKD progression: a progress report. Clinical Journal of the American Society of Nephrology
(CJASN), 1(4): 754-760, Jul 2006.
• Thomas JB, Antiga L, Che S, Milner JS, Hangan Steinman DA, Spence JD, Rutt BK and Steinman DA. Variation in the
carotid bifurcation geometry of young vs. older adults: Implications for "geometric risk" of atherosclerosis. Stroke, 36(11):
2450-2456, Nov 2005.
• Antiga L, Steinman DA. Robust and objective decomposition and mapping of bifurcating vessels. IEEE Transactions on
Medical Imaging, 23(6): 704-713, June 2004.
• Antiga L, Ene-Iordache B and Remuzzi A. Computational geometry for patient-specific reconstruction and meshing of
blood vessels from MR and CT angiography. IEEE Transactions on Medical Imaging, 22(5): 674-684, May 2003.
• Antiga L, Ene-Iordache B, Remuzzi G and Remuzzi A. Automatic generation of glomerular capillary topological
organization. Microvascular Research, 62: 346-354, June 2001.
288
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department of Bioengineering conducts research and development in biomedicine, both at
experimental and clinical level. Physiopathological processes are studied through the use of
engineering techniques and to develop innovative treatment strategies. Several lines of research
in basic, applied research and clinical are currently active within the Department . The main
instruments used for this research consist of: theoretical models; diagnostic imaging;
histological measures; physical and chemical parameters in both experimental and clinical
studies; cell culture techniques; biomaterials; technologies for archiving and processing of
clinical data. The ongoing studies relate to four main areas: 1) study of the mechanisms
involved in the progression of chronic nephropathy; 2) studies on the role hemodynamics in the
development of vascular diseases; 3) development of laboratory techniques for tissue
engineering, 4 ) developement of information systems to manage clinical data and images
generated in the context of controlled clinical trials and in routine clinical practice.
FINDINGS/MAIN RESULTS
We produced evidence demonstrating a possible relationship between the geometry of the
cerebral arterial vessels, hemodynamic conditions and location of cerebral aneurysms. These
findings are opening new perspectives in the understanding of the mechanisms responsible for
this disease.
Demonstration of a significant relationship between morphometric parameters of the renal
parenchyma, quantified through elaboration of CT images, and the loss of renal function in
patients with polycystic kidney disease.
Demonstration that the technique of islet immunoisolation does not preclude oxygenation and
consequently that this technique can be used for pancreatic islet transplantion.
Demonstration that the mechanism responsible for regeneration of the glomerular capillary,
induced by drugs that inhibit the angiotensin II, depends on the proliferation of parietal cells of
Bowman capsule and their migration on the glomerular capillary loops.
Implementation of a web-based system for the management, in compliance with GCP, of
clinical data generated in controlled clinical trials.
Set up a new network of specialists in Nephrology for data collection aimed at monitoring the
quality of treatment of chronic progressive nephropathy in current clinical practice.
NATIONAL COLLABORATIONS
Dipartimento di Bioingegneria, Politecnico di Milano, Milano.
Fidia Advanced Biopolymers, Abano Terme, Padova.
Istituto di Fisiologia Clinica CNR, Pisa.
Unità di Diabetologia, Ospedali Riuniti, Bergamo.
289
ANNUAL REPORT 2008

IRFMN
STMicroelectronics , Agrate Brianza, Milano
Università di Bergamo
Dipartimento di scienze Neurologiche e della visione, Università di Verona.
Dipartimento di Ingegneria Industriale e Dipartimento di Ingegneria dell’informazione e metodi
Matematici
Facoltà di Medicina e Chirurgia, Università degli studi di Milano
INTERNATIONAL COLLABORATIONS
Massachussetts Institute of Technology, Cambridge MA, USA.
National Alliance for Medical Imaging Computing, USA.
Harvard Medical School, Cambridge MA, USA.
Department of Mathematics and Computer Science, Emory University, Atlanta, Georgia, US.
Simula Laboratories, Oslo, Norway Academisch Medisch Centrum, Amsterdam, the
Netherlands
University of Toronto, Ontario, Canada.
Ghent University, Ghent, Belgium.
Technical University, Eindhoven, The Netherlands.
University Hospital, Maastricht, The Netherlands.
Universzitetni Klinikni Center Lubjana, Ljubljana, Slovenia.
The University of Sheffield, Sheffield, United Kingdom.
EDITORIAL BOARD MEMBERSHIP
International Journal of Artificial Organs, (Andrea Remuzzi)
PEER REVIEW ACTIVITIES
Annals of Biomedical Engineering ASME Journal of Biomechanical Engineering
Journal of Vascular Research Magnetic Resonance in Medicine Stroke
Journal of the American Society of Nephrology
Kidney International
American Journal of Kidney Diseases
American Journal of Pathology
American Journal of Physiology
Medical & Biological Engineering & Computing
New England Journal of Medecine
IEEE Transactions on Medical Imaging
IEEE Transactions on Biomedical Ingeneering
Medical Physics
Journal of Biomechanics
Medical Engineering and Physics
Artificial Organs
International Journal of Artificial Organs
Biomaterials
Contemporary Clinical Trials
290
ANNUAL REPORT 2008

IRFMN
Journal of Biomaterial Application
Journal of Endocrinological Investigation
EVENT ORGANIZATION
Seminar: “The Definition of human adult stem cells by substrate interactions” John A. Hunt,
University of Liverpool, December, Ranica, Bergamo.
Seminar: “Developments in neuroimaging with applications to Alzheimer's disease” Anna
Caroli, October, Ranica, Bergamo.
Seminar: “Hemodynamics in Atherogenesis and Thrombosis”, David Ku, Georgia Institute of
Technology, Luglio, Ranica, Bergamo.
Seminar: “MSC in pancreatic islets”, Thijs Molder, Maastricht University, July, Bergamo.
Seminar: “Model Based Drug Development and the FDA Critical Path Initiative”, Dr. Robert
Powell, Office of Translational Sciences, FDA, Silver Spring, MD, June, Ranica, Bergamo.
Seminario: “Tecniche avanzate di simulazioni numeriche in vasi sanguinei” Christian Vergara,
Università di Bergamo, April, Ranica, Bergamo.
Seminar: “Rotating versus perfusion bioreactor for the culture of engineered vascular constructs
based on hyaluronic acid”, Chiara Arrigoni, February, Ranica, Bergamo.
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
11th Asian Pacific Congress of Nephrology, Kuala Lumpur, Malaysia, May 2008. Results of the
program for early detection of chronic kidney disease and their risk factors in Mongolia.
29th Annual Meeting of the Society for Clinical Trials. St. Louis, May 2008. Randomized and
Non-Randomized patients in the Bergamo Nephrologic Diabetes Complications Trial
(BENEDICT).
14th International Symposium on Brain Edema and Brain Tissue Injury, June 2008 Warsaw,
Poland. Auto regulation of cerebral blood flow and autonomic drive.
ASN Renal Week 2008, November, Philadelphia, PA. GFR slopes underestimate renal function
decline in type 2 diabetic patients without overt nephropathy.
ASN Renal Week 2008, November, Philadelphia, PA. Community Screening and Intervention
for Hypertension, Diabetes and Chronic Kidney Disease in Poor Resource Setting in Eastern
Nepal.
291
ANNUAL REPORT 2008

IRFMN
ASN Renal week 2008, November, Philadelphia, PA. Structure and Function in ADPKD:
Correlation between GFR and Relative Volume of Hypo-Enhanced Parenchyma from Contrast-
Enhanced CT Imaging.
@neurIST Open Symposium agenda, September 2008, Barcelona, Spain. Translation of Virtual
Physiological Human concepts into clinical systems and services for disease understanding and
interventional planning: ARCH project.
Polycystic Kidney Disease Database Consortium Meeting, March 2008, Chicago, USA.
Discussion of PKD Clinical Database.
16th Congress of the European Society of Biomechanics, July 2008, Lucerne, Switzerland. Link
Between Vortex Structures And Voronoi Diagram In Cerebral Aneurysms.
16th Congress of the European Society of Biomechanics, July 2008, Lucerne, Switzerland. A
Open Source Parallel AMR FE Solver For Biological Flows Based On The Libmesh C++
Library.
National Alliance for Medical Image Computing, Fourth All Hands Meeting, Salt Lake City,
January 2008
National Alliance for Medical Image Computing, Summer Project Week, MIT, Boston, June
2008
ASME Summer Bioengineering Conference, Marco Island, FL, June 2008. Hands-on tutorial:
the Vascular Modeling Toolkit.
Fifth International Bio-Fluid Symposium and Workshop, Pasadena, CA, March 2008.
CBC Workshop on High-Performance Computing and Biomedical Flows, Oslo, Norway. May
2008.
GRANTS AND CONTRACTS
Research grant AIFA - controlled clinical trials (VARIETY, VALID, ATHENA, ARCADIA).
Research grant CHIESI Farmaceutici S.p.A - DEMAND trial “A multicenter, randomized,
prospective, double-blind study to evaluate the nephroprotective effect of delapril alone or
combined with manidipine in patients with type 2 diabetes”.
Research grant PKD foundation - ALADIN trial “Effect of long-acting somatostatin on disease
progression in ADPKD: a long-term three year follow-up study”.
Research grant Baxter – ASAP trial “Acute Start Access Programme”.
Research grant ISN per il Kidney Disease Data Center (KDDC ) del COMGAN.
Contributo Regione Lombardia per data management del “Centro di Coordinamento della Rete
Regionale per le Malattie Rare”.
292
ANNUAL REPORT 2008

IRFMN
Project - FP6 UE-STEPS "A system approach to tissue engineering products and processes"
FP6-500465
Project - FP7 UE - ARCH "Patient specific image-based computational modeling for
improvement of acute and long-term outcomes of vascular access for hemodialysis” FP7-
224390 Project Coordination.
Research grant PKD foundation - Effect of Long-acting Somatostatin on Disease Progression in
ADPKD: A Long-term Three Year Follow-up Study.
Project VASCOSILK, Fondazione Cariplo - N.536/5314 - Protesi vascolari in fibroina
elettrofilata per la rigenerazione in vivo di arterie di piccolo calibro.
Project LIGASILK, Regione Lombardia - N.534/5287 - Bioingegnerizzazione di tendini e
legamenti: impiego combinato di supporti tessili in seta e cellule staminali adulte.
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
• Cravedi P, Mannon RB, Ruggenenti P, Remuzzi A, Remuzzi G. Islet transplantation: need for a time-out
Nat Clin Pract Nephrol. 2008 Sep 23.
• Figliuzzi M, Adobati F, Cornolti R, Cassis P, Remuzzi G, Remuzzi A. Assessment of in vitro differentiation of
bovine pancreatic tissue in insulin-expressing cells. JOP. 2008 Sep; 9: 601-11.
• Morigi M, Introna M, Imberti B, Corna D, Abbate M, Rota C, Rottoli D, Benigni A, Perico N, Zoja C, Rambaldi
A, Remuzzi A, Remuzzi G. Human bone marrow mesenchymal stem cells accelerate recovery of acute renal
injury and prolong survival in mice. Stem Cells. 2008 Aug;26: 2075-82.
• Arrigoni C, Chittò A, Mantero S, Remuzzi A. Rotating versus perfusion bioreactor for the culture of engineered
vascular constructs based on hyaluronic acid. Biotechnol Bioeng. 2008 Aug;100:988-97.
• Ruggenenti P, Remuzzi A, Remuzzi G. Decision time for pancreatic islet-cell transplantation. Lancet. 2008 Mar
;371:883-4.
• Lee SW, Antiga L, Spence JD, Steinman DA. Geometry of the carotid bifurcation predicts its exposure to
disturbed flow. Stroke. 2008 Aug;39:2341-7.
• Antiga L, Wasserman BA, Steinman DA. On the overestimation of early wall thickening at the carotid bulb by
black blood MRI, with implications for coronary and vulnerable plaque imaging. Magn Reson Med. 2008
Nov;60:1020-8.
• Macconi D, Remuzzi G. Candesartan and renal protection: more than blocking angiotensin type 1 receptor
Kidney Int. 2008 Nov;74:1112-4.
• Ruggenenti P, Iliev I, Costa GM, Parvanova A, Perna A, Giuliano GA, Motterlini N, Ene-Iordache B, Remuzzi
G; Bergamo Nephrologic Diabetes Complications Trial Study Group. Preventing left ventricular hypertrophy by
ACE inhibition in hypertensive patients with type 2 diabetes: a prespecified analysis of the Bergamo
Nephrologic Diabetes Complications Trial (BENEDICT). Diabetes Care. 2008 Aug;31:1629-34.
• Ruggenenti P, Perticucci E, Cravedi P, Gambara V, Costantini M, Sharma SK, Perna A, and Remuzzi G. Role of
remission clinics in the longitudinal treatment of CKD. J Am Soc Nephrol. 2008 Jun;19:1213-24.
293
ANNUAL REPORT 2008

IRFMN
• Antiga L, Piccinelli M, Botti L, Ene-Iordache B, Remuzzi A, Steinman DA. An image-based modeling
framework for patient-specific computational hemodynamics.
Med Biol Eng Comput. 2008 Nov;46:1097-112.
RESEARCH ACTIVITIES
Laboratory of Renal Biophysics
Three dimensional reconstruction of the glomerular capillary network
We have recently documented that angiotensin II blockade not only retards the progression of
renal diseases, but also induces regression of the glomerular lesions. To quantify the extent of
the regression of sclerotic lesions and the potential regeneration of the glomerular capillary
induced by a therapy with angiotensin converting enzyme (ACE) inhibitors we are developing
three dimensional (3D) reconstruction of tissues by several approaches: one of them is the
vascular corrosion casting technique, using a polyurethane resin, that allows to obtain casts of
the vascular architecture of the glomerular capillary to be analyzed by scanning electron
microscopy. We are also setting up a new technique to study the morphology of the glomerular
capillary and its wall by electron microscopy, based on a combination of an electron beam with
an ionic one, in order to achieve sectioning of samples and acquisition of serial images for 3D
reconstruction of the ultrastructure by mathematical algorithms.
Study of the regression of sclerotic lesions and kidney regeneration in
response to a therapeutic treatment targeting angiotensin II
Clinical and experimental studies have documented that lowering proteinuria by
pharmacological interventions retards renal disease progression and even induces remission. We
have recently shown in MWF rats, a genetic model of spontaneous glomerular damage, that the
ACE inhibitor, lisinopril, induces regression of proteinuria and of existing glomerulosclerosis
and stabilizes renal function even when given in advanced stages of nephropathy. Reabsortion
of glomerular lesions is associated with remodeling of glomerular cell components which
results in a selective increase in podocyte number. These highly differentiated cells are key
determinants of perm-selective properties of the glomerular capillary barrier and their loss in the
disease correlates with the development of proteinuria and later on of glomerulosclerosis.
Beside podocyte restoration, enhanced endothelial cell volume density and reduced mesangial
hyperplasia were observed in response to treatment. The mechanism(s) underlying increased
podocyte number per glomerulus induced by ACE inhibition has not been established.
Podocytes have a limited capability to proliferate and this may be the cause of podocyte
depletion we observed in aged MWF rats and generally present in experimental models of
progressive nephropathies. However, in untreated MWF rats about 5% of WT1 positive cells
(WT1 is a podocyte-specific nuclear antigen) are also positive for Ki-67, a nuclear antigen
expressed during all active phases of the cell cycle (G1 to M), and this percentage increase in
response to ACE inhibition. At least two possibilities can be taken into account to explain this
finding. One is that some podocyte may re-engage and progress throughout the cell cycle. The
increase in WT1/Ki-67 positive cells in lisinopril treated MWF rats is associated with a reduced
expression of the cyclin-dependent kinase inhibitor p27, a negative modulator of the cell cycle
that induces cell cycle arrest. Thus, it is conceivable that reduced p27 may favor proliferation of
podocytes that have re-engaged the cell cycle. The second possibility is that WT1/Ki-67
positive cells may derive from podocyte precursor. Ultrastructural features characteristic of
podocytes (i.e. foot processes) have been documented in some parietal epithelial cells of the
294
ANNUAL REPORT 2008

IRFMN
Bowman’s capsule that express epitopes of mature visceral podocytes and retain the ability to
divide. In MWF rats the percentage of parietal podocytes over the total parietal epithelial cells
increase in response to ACE inhibition. Electron microscopy examination reveals a continuity
between the inner surface of the Bowman’s capsule and the outer glomerular capillary
membrane supporting the possibility for parietal cells to migrate from Bowman’s capsule to the
capillary tuft even in normal physiological condition. Moreover, occasional finding of parietal
epithelial cells protruding toward the tuft suggests that these cells can also migrate to the tuft
through cellular bridges in different areas than the vascular pole. Overall our results suggest that
remodeling of parietal epithelial cells of the Bowman’s capsule contributes to podocyte
restoration in response to ACE inhibition (in collaboration with the Department of Molecular
Medicine).
Application of confocal microscopy to the study of cells and tissues
Fluorescent probes to investigate the distribution and co-localization of proteins and other
specific antigens in cell monolayers and tissue specimens are commonly used in the majority of
experimental applications. Bidimensional images are not of high resolution to provide correct
information, thus the use of 3D reconstruction of fluorescent signal becomes mandatory. These
problems can not be solved by the classic fluorescent microscopy that has some limitations. For
these reasons we have optimized the laser confocal microscopy by using the LSM510 META
microscope from Zeiss (Germany). The microscope is equipped with four laser lines that excite
the sample within the whole visible spectrum plus the characteristic “META scan head” that
allow to analyze the emission spectrum of different fluorophores to optimally separate them.
The instrument is characterized by and used for its high specificity and for the excellence of the
revealed and elaborated signal.
Laboratory of Biomedical Technologies
Development of computerized systems for controlled clinical trials
Numerous clinical trials are conducted in the Clinical Research Center for Rare Diseases “Aldo
e Celè Daccò”. These studies must be carried out with respect to the GCP (Good Clinical
Practice) guidelines and require high quality of data. Every clinical study requires a paper case
report form (CRF) for collection of patients’ clinical observations. These data must be checked
for inconsistency by dedicated monitoring staff, and then recorded electronically. In our Lab we
have developed applications tailored for data management of clinical studies using relational
databases systems (RDBMS) and specific programs aimed to data elaboration, validation and
extraction for subsequent statistic analyses.
REIN, BENEDICT MYSS REIN2, DKG are just few of the trials concluded successfully and
published in prestigious medical journals. This accomplishment is, in part, due to the
contribution of our Lab. For the DEMAND study we have developed an innovative electronic
CRFs based on laptop computers. For the future studies, we are implementing also a web-based
framework for electronic data capture and clinical data management for clinical trials.
Data Management for the Registry for Rare Disease - Lombardy
There is in act a collaboration between our Lab and the Unit of Information Center for Rare
Diseases for the management of the Regional Network for Rare Diseases of Lombardy, Italy.
We are directly involved for the development and maintenance of the web site of the center and
management of a regional Registry for Rare Diseases. We have set-up the databases and
developed related web-pages for centers, disease archive, associations of patients and congenital
rare diseases. These are publicized on the homepage of the web-site
(http://malattierare.marionegri.it/).
295
ANNUAL REPORT 2008

IRFMN
The Registry for Rare Diseases was born in 2007 as collaboration between Mario Negri
Institute, Lombardia Informatica (LI) and Regione Lombardia. The aim was to create a regional
registry for rare diseases where all medical staff from Lombardy could register information
regarding rare diseases. The application (Sistema Malattie Rare - SMR) was developed by LI in
collaboration with our group. SMR application is using web-based technology and can be used
jointly with the health patient card. It is actually in use in almost all centers dedicated for rare
diseases in Lombardy. Our laboratory is involved in the maintenance of SMR, statistical
analyses of registry data and creation of a minimal set of data that are shared at national level by
the Istituto Superiore di Sanità (ISS). File download from LI is secured because data are
encrypted using an asymmetrical algorithm with public and private key at 2048 bit. Also data
transfer at ISS is performed securely by using the htpps Internet protocol.
KDDC – a coordinating center for data collection and surveillance of
prevention programs on non-communicable chronic diseases in emerging
countries
Chronic kidney diseases are emerging as a global threat to human health. Prevalence and
incidence of renal diseases in developing countries are not known, and this is an obstacle to the
adoption of preventive measures. Prevention is the only hope for these countries where
treatment options for end stage renal failure are simply not available to the vast majority of the
population because of their costs.
The International Society of Nephrology (ISN), through the Commission for Global
Advancement of Nephrology (COMGAN), has established a research committee in order to face
the problems about prevention of kidney diseases in developing countries. The coordination of
the team and intervention programs was committed to the Mario Negri Institute for
Pharmacological Research at the Clinical Research Center “Aldo e Cele Daccò”. The general
aim of the project is to define programs in developing countries to identify those subjects who
are at risk of developing a renal disease later in life, in order to design a prevention strategy on
national basis by means of interventions of the local ministries of health to governmental and
financial level.
The Kidney Disease Data Center (KDDC), headquartered in our Laboratory, is dedicated to data
management for the prevention programs underway in emerging countries. We have set up an
instrument to collect clinical data from different centres located world-wide. Data are stored in a
dedicated server in our Laboratory. Results of our epidemiological analyses, shared also with
medical staff of the center, allow us to have a general overview on the health of population
under study. The prevention program is underway and we have already collected data from
participating centers from Nepal, Mongolia, India, China, Moldova, Bolivia and Mozambique.
First results on screening on several thousands of subjects, confirm the need to proceed with
prevention programs in theses countries. Other countries are willing to start their programs, as
for example Mexico, Argentina, Philippines. Through the activity of KDDC it is possible to
monitor the course of the prevention programs and to tailor them to fit the needs of each
participating country.
Hemodynamics and vascular pathology
During the last ten years the presence of a close relationship between hemodynamics and
vascular pathology has been pointed out both in the biological and in the clinical field. Two
typical examples of such relationship are atherosclerosis and intracranial aneurysm disease. In
atherosclerosis, a pathology leading to lipid-rich and fibrotic plaque formation in artery walls,
lesions tend to form mainly in correspondence to bifurcations (e.g. carotid bifurcation) and great
curvature tracts (e.g. coronary arteries). In intracranial aneurysm disease, characterized by the
formation of one (or more) artery wall protrusions in the cerebral arterial circulation, cerebral
296
ANNUAL REPORT 2008

IRFMN
aneurysms tend to locate at the distal wall of main bifurcations and at the exteral wall of
segments characterized by large curvature.
Besides being involved in pathogenesis, hemodynamics also plays a role at a mechanical level
for the determination of atherosclerotic plaque rupture, responsible for heart attacks and stroke,
or cerebral aneurysm rupture with consequent intracranial hemorrhage.
It has been experimentally shown that endothelial cells (lining vessel walls) and smooth muscle
cells (the vasoactive component of the wall) exposed to flow exhibit changes in gene
expression, cytoskeleton reorganization and permeability according to the characteristics of the
imposed flow (e.g. laminar vs turbulent).
In spite of this data supporting the hypothesis of a close relationship between hemodynamics
and vascular pathology, the nature of such relationship is still under scrutiny, mainly due to the
difficulty of measuring in-vivo hemodynamics-related quantities.
Thanks to innovative technologies developed during the last few years in the medical imaging
and mathematical modeling fields, also within the Biomedical Engineering Department, it is
now possible to accurately reproduce patient-specific hemodynamic force distribution from
computed tomography (CT) or magnetic resonance (MR) acquisitions.
Within the Department of Biomedical Engineering such technologies have three main
applications: atherosclerosis (carotid bifurcation and renal artery level), intracranial aneurysm
disease and complications of vascular access in hemodialysis. In particular, the Department is
currently involved in two international collaborative projects: ARCH, 3-year project funded by
the European Union within the Seventh Framework Programme, starting from 2008, aimed at
improving the functionality of vascular access in hemodialysis patients, for which the
Department is the coordinator centre; ANEURISK, project funded by Siemens Medical and
Fondazione Politecnico, aimed at studying the role of hemodynamics in the pathogenesis of
intracranial aneurysms and in influencing their rupture risk.
Imaging and quantification in renal physiopathology
The use of imaging techniques such as CT, MR, and echography, and the application of
advanced image processing tools make it possible to perform non-invasive in-vivo quantitative
analysis of biological phenomena. Within the Department of Biomedical Engineering, this
approach is applied to the investigation of renal physiopathology. Through CT and MR imagebased
quantification, new therapies for autosomal dominant polycystic kidney disease
(ADPKD) are currently being evaluated. To this purpose, three ongoing clinical trials aimed at
reducing the overall kidney cyst volume, one of which funded by the Polycystic Kidney
Foundation, are currently relying on image-based quantification.
CT image quantification has recently led to the discovery of a high correlation between a
specific portion of polycystic kidney tissue and renal function, showing for the first time a likely
direct relationship between structure and function, thus opening the way to new therapeutic
targets.
Beyond ADPKD studies, new methodologies for noninvasive characterization of renal
functionality from diffusion-weighted MR images are currently under study.
Development of biomedical image analysis and computational modelling
tools
The employment of quantitative imaging and mathematical modeling techniques aimed at the
study of physiopathological processes is directly linked to medical image management and
processing methodologies. Within the Department, research activity related to the development
of new image processing algorithms and mathematical models for the numerical simulation of
biological phenomena, both theoretical and in terms of software development, is actively
performed.
The department contributes to the development of three main open-source projects in the
biomedical imaging field: Vascular Modeling Toolkit (www.vmtk.org), as main developer,
297
ANNUAL REPORT 2008

IRFMN
Insight Toolkit (www.itk.org), a state-of-the-art library for medical image analysis, and Slicer
3D (www.slicer.org), one of the main medical imaging applications. The Department is directly
involved in the development activities carried out within the National Alliance for Medical
Image Computing, an inter-university consortium gathering the main academic institutions of
the United States.
Development of devices for the transplantation of immunoisolated islets
The project’s main objective is to develop an immunisolation device for pancreatic islets that
can be implanted in diabetic patients permitting allo-islet transplantation without the use of
pharmacological immunosuppression and avoiding allosensitization of the patient. The study
started from the design and characterization of the semi-permeable membrane used for the
device construction. The device that we are developing can be implanted with minimally
invasive surgical procedures and easy to retry. This system is a device made using polysulfone
hollow fibers or a tubular membrane in polivinil-alcool. The aims of our studies in the next
months will be to improve functionality using nanotechnologies for materials characterization.
Moreover we will develop new kinds of device and we will test new implantation sites.
Effect of immunoisolation on oxygen consumption by pancreatic islets
Immunoisolated pancreatic islet transplantation open new perspectives in the treatment of type 1
diabetes without the use of immunosoppressive drugs. In the last years, our group has been
interested in the development of two immunoisolation devices for the transplantation of bovine
pancreatic islets in streptozotocin induced diabetic rats. The first device is formed by alginate
capsules produced with techniques pointed in our laboratory. The second system is constituted
from a device of polysulfone hollow fibers. Both devices must allow adequate diffusion of
oxygen to maintain islet viability and function. In the attempt to understand whether
immunoisolated islets have adequate oxygen supply in these devices, we have measured their
consumption of oxygen. To this purpose, we have set up an experimental technique to measure
oxygen consumption rate using a Clark’s electrode inserted in a glass thermostated chamber
connected to a data recorder and acquisation system. We have then estimated oxygen
consumption rate of free and encapsulated islets in alginate capsules and in polysulfone hollow
fibers. Both immunoisolation devices allowed oxygen consumption values comparable to that
measured in free islets. The data suggests that islet encapsulation in alginate gel and in
polysulfone hollow fibers allows an adequate oxygenation to maintain islet function and
viability.
Mesenchymal stem cells improve vascolarization in pancreatic islet
transplantation
Type I diabetes is a chronic metabolic disorder that results from the progressive destruction of
the ß cells leading to insulin insufficiency and hyperglycemia which is the main determinant of
chronic vascular complications. Pancreatic islet transplantation represents a cure for type I
diabetes. Recently developed protocols enhanced the success rate of islet transplantation but
there are still big limitations including the lack of metabolic capacity of transplanted islets in the
long-run. This phenomenon must be mainly attributed to delayed and insufficient islet
revascularization that can deprive newly transplanted islets of oxygen, resulting in permanent
cell death. Promotion of islet revascularization through locally increased expression of growth
factors, such as vascular endothelial growth factor (VEGF), could improve the efficiency of islet
transplantation. On this basis, we elected to investigate whether rat MSCs co-transplanted with
pancreatic islets may serve as cell therapy to promote therapeutic angiogenesis ultimately
leading to an effective metabolic activity of islet grafts. 2,000 syngenic islets alone or in
combination with MSCs were transplanted under the kidney capsules of diabetic Lewis rats.
Animals transplanted with islets alone never reached normoglycemia. In contrast, in rats
298
ANNUAL REPORT 2008

IRFMN
transplanted with islets and MSCs, glycemia gradually fell after transplantation and
normoglycemia was maintained for a long time. In transplanted animals, islet vascularization
was quantified by morphometrical analysis. Mean capillare density was significantly increased
in islet co-transplanted with MSCs indicating that co-transplantation of MSCs with pancreatic
islets improves islet graft function by promoting graft vascularization. (In collaboration with
Laboratory of Cellular Biology and Xenotransplantation).
Vascular tissue engineering
Alternative strategies have been conceived in recent years to develop artificial vascular
prothesis to restore the correct transport of blood in the circulation. Currently available synthetic
vascular grafts are limited to large internal diameter grafts because of frequent thrombosis and
occlusion. The alternative is represented from the use of autologous vascular graft, but they are
not always available in patients affected from vascular pathology. Vascular tissue engineering
has the objective to generate cellularized vascular prosthesis made of biodegradable materials
that can be colonized by endothelial cells. For this purpose (in collaboration with Stazione
Sperimentale della Seta in Milano and Politecnico di Milano), we have studied an innovative
strategy for the vascular prosthesis realization using biodegradable material, the fibroina of the
silk. Tubular structures have been produced, through elettrospinning of the fibrin of the silk, and
they will be implanted in small animals (rats) to replace abdominal aorta and, therefore, to
evaluate the functionality of the prosthesis.
The effect of flow on renal tubular cells
ADPKD (Autosomal Dominant Policystic Kidney Disease) is one of the major congenital renal
pathologies and results in the development of cysts in the tubular segment, that leads to renal
failure. This pathology is due to the modification of the gene encoding for policystin 1, which is
contained in the cilia of tubular cells. In the literature it has been shown that cilia work as
mechanosensory organs. The bending of the cilium, caused by tubular flow, leads to the
activation of the policystin 1 and to the generation of a peak of intracellular calcium
concentration. This increase in intracellular calcium activates signalling pathways that modify
cellular proliferation and other cellular functions. In pathologic conditions, policystin
modification impairs the mechanosensitive function of cilia, altering tubular cellular functions.
The aim of this project is the in vitro study of renal tubular cells (MDCK2) in laminar flow
conditions, to investigate the role of mechanical stimulation in the pathology development.
Preliminary results showed that the application of a constant laminar flow to MDCK2 causes a
different tridimensional organization of the cellular layer, as compared to static control.
Furthermore, the inhibition of intracellular calcium increase impairs this reorganization when
flow is applied.
299
ANNUAL REPORT 2008

IRFMN
300
ANNUAL REPORT 2008

IRFMN
LABORATORY OF BIOLOGY AND
THERAPY OF METASTASIS*
STAFF
Head
Raffaella GIAVAZZI, Biol.Sci.D., Ph.D.
Head
Giulia TARABOLETTI, Biol.Sci.D.
* Research activities of this Laboratory are listed in the Department of Oncology section (pag. 7)
301
ANNUAL REPORT 2008

IRFMN
302
ANNUAL REPORT 2008

IRFMN
Aldo and Cele Daccò Center
Ranica (Bg)
ANNUAL
REPORT 2008
departments and laboratories
303
ANNUAL REPORT 2008

IRFMN
304
ANNUAL REPORT 2008

IRFMN
DEPARTMENT OF RENAL MEDICINE
STAFF
Head
Piero RUGGENENTI, M.D.
Laboratory of Biostatistics
Head
Annalisa PERNA, Stat.Sci.D.
Unit of Drug Monitoring
Head
Giulia GHERARDI
Laboratory of Clinical Chemistry
Head
Flavio GASPARI, Chem.D.
Laboratory of Advanced Development of Drugs
Head
Norberto PERICO, M.D.
Unit of Pharmacology and Pharmacogenetics
Head
Dario CATTANEO, Chem.Pharm. D., Ph.D
Unit of Early Clinical Evaluation of Drugs
Head
Aneliya PARVANOVA, M.D.
305
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Piero Ruggenenti got his Medicine degree in 1983 at the University of Milan, Italy; he got his
specialization in Cardiology in 1985 and in Clinical Nephrology in 1989 at the same University; he
specialized in Pharmacological Research in 1988 at IRFMN.
Educational training: in 1980-1983 researcher at "Centro di Fisiologia Clinica e Ipertensione, Clinica
Medica IV", Università degli Studi di Milano; in 1984 Researcher at IRFMN, Bergamo, Italy in 1987-
1988 Honorary Registrar of the Unit for Metabolic Medicine, Division of Medicine (University of
London) of Guy's and St. Thomas's Hospitals, London; in 1988-1989 Assistant Professor of the Division
of Nephrology and Dialysis of the Ospedali Riuniti di Bergamo.
Areas of interest: mechanisms of chronic renal disease progression, diabetes and diabetic complications,
clinical transplantation, thrombotic microangiopathies, cardiovascular complications of chronic renal
disease, clinical trials, clinical pharmacology.
Employment: from 1990 Assistant Professor of the Division of Nephrology and Dialysis of the
Ospedali Riuniti di Bergamo; in 1994-1999 Head, Unit of Advanced Development of Drugs, Daccò
Center, Ranica, Bergamo, Italy; since 2000 Head, Department of Renal Medicine, Daccò Center,
Bergamo, Italy.
Selected publications
• P. Ruggenenti, A. Perna, L. Mosconi, M. Matalone, G. Garini, M. Salvadori, C. Zoccali, F. Scolari, Q. Maggiore, G.
Tognoni, G. Remuzzi (for The GISEN Group). Randomised placebo-controlled trial of effect of ramipril on decline in
glomerular filtration rate and risk of terminal renal failure in proteinuric, non-diabetic nephropathy. Lancet
1997;349:1857-1863
• P. Ruggenenti, A. Perna, G. Gherardi, F. Gaspari, R. Benini, G. Remuzzi, on behalf of GISEN. Renal function and
requirement for dialysis in chronic nephropathy patients on long-term ramipril: REIN follow-up trial. Lancet
1998;352:1252-1256.
• P. Ruggenenti, A. Perna, G. Gherardi, G. Garini, C. Zoccali, M. Salvadori, F. Scolari, F.P. Schena, G. Remuzzi.
Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet
1999;354:359-364.
• G. Remuzzi, C. Chiurchiu, M. Abbate, V. Brusegan, M. Bontempelli, P. Ruggenenti. Rituximab for idiopathic
membranous nephropathy. Research Letter. Lancet 2002;360:923-924.
• P. Ruggenenti, A. Fassi, A. Parvanova, S. Bruno, I. Iliev, V. Brusegan, N. Rubis, G. Gherardi, F. Arnoldi, M. Ganeva, B.
Ene-Iordache, F. Gaspari, A. Perna, A. Bossi, R. Trevisan, A.R. Dodesini, G. Remuzzi for the Bergamo Nephrologic
Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J
Med 2004;351:1941-1951
• G. Remuzzi, P. Cravedi, A. Perna, B.D. Dimitrov, M. Turturro, G. Locatelli, P. Rigotti, N. Baldan, M. Beatini, U.
Valente, M. Scalamogna, P. Ruggenenti Dual Kidney Transplant Group. Long-term outcome of renal transplantation
from older donors. N Engl J Med 2006;354:343-352.
Flavio Gaspari got his Chemistry degree in 1977 at the University of Milano, Italy, and the
specialization in the same University in 1979.
Educational training: in 1981-1985 Fellow and Researcher at IRFMN, Milan; in 1985-1991 at IRFMN,
Bergamo, Italy.
Areas of interest: pharmacokinetics and the metabolism of xanthines in different animal species; drug
pharmacokinetics in uremic patients and in subjects with different degrees of renal function; analytical
methods to measure the most important immunosuppressive drugs to determine their pharmacokinetics in
kidney, heart, and liver transplant recipients; evaluation of the renal function by using different
approaches, in the study of renal disease progression, and in the comparison of different methods for
albuminuria determination.
Employement: He is Chief of Laboratory of Pharmacokinetics and Clinical Chemistry since January
2000 and he was Chief of this Unit since 1991.
Selected publications
• Gotti E, Perico N, Gaspari F, Cattaneo D, Lesti MD, Ruggenenti P, Segoloni G, Salvadori M, Rigotti P, Valente U,
Donati D, Sandrini S, Federico S, Sparacino V, Mourad G, Bosmans JL, Dimitrov BD, Iordache BE, Remuzzi G. Blood
cyclosporine level soon after kidney transplantation is a major determinant of rejection: insights from the Mycophenolate
Steroid-Sparing Trial. Transplant Proc. 2005 Jun;37(5):2037-40.
• Perico N, Gaspari F, Remuzzi G. Assessing renal function by GFR prediction equations in kidney transplantation. Am J
Transplant. 2005 Jun;5(6):1175-6.
306
ANNUAL REPORT 2008

IRFMN
• D. Cattaneo, F. Gaspari, S. Zanoni, S. Baldelli, E.Gotti, A. Perna, N. Perico, G. Remuzzi. Two-hour post-dose
cyclosporine monitoring does not fit all in kidney transplantation. Therapy 2005;2:95-105.
• Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-
Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G; Bergamo Nephrologic Diabetes
Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J Med. 2004
Nov 4;351(19):1941-51. Epub 2004 Oct 31.
• Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M,
Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G, Perico N; MY.S.S. Study Investigators.
Performance of different prediction equations for estimating renal function in kidney transplantation. Am J Transplant.
2004 Nov;4(11):1826-35.
• Cattaneo D, Merlini S, Pellegrino M, Carrara F, Zenoni S, Murgia S, Baldelli S, Gaspari F, Remuzzi G, Perico N.
Related Articles, Links Therapeutic drug monitoring of sirolimus: effect of concomitant immunosuppressive therapy and
optimization of drug dosing. Am J Transplant. 2004 Aug;4(8):1345-51.
Norberto Perico got his Medicine degree in 1983 at the University of Milano, Italy. He got his
specialization in Pharmacological Research in 1986 at IRFMN, Bergamo and in Clinical Nephrology in
1989 at the University of Verona, Italy.
Educational training: in 1982 Fellow, Department of Pharmacology, New York Medical College,
Valhalla, New York, USA; in 1984-1988 Post Doctoral Fellow, Laboratory of Kidney Diseases, IRFMN,
Bergamo, Italy; in 1988-1989 Researcher in the same laboratory.
Areas of interest: pathophysiology and pharmacology of cyclosporine nephrotoxicity; new
immunosuppressive strategies to prevent renal graft rejection; innovative approach to induce tolerance to
organ transplantation; mechanism(s) and management of progression of chronic renal diseases.
Employment: in 1990-1994 Head, Renal Physiology Unit, Laboratory of Kidney Diseases, IRFMN,
Bergamo, Italy; in 1990-2000 Assistant Professor, Division of Nephrology and Dialysis, Ospedali Riuniti
di Bergamo, Italy; in 1994 –1999 Head, Laboratory of Transplant Immunology, IRFMN, Bergamo, Italy;
from January 2000 Head, Laboratory of Drug Development, Department of Renal Medicine, IRFMN,
Bergamo, Italy; from September 2000 Health Director, Daccò Center, IRFMN, Bergamo, Italy. From
October 2002 he’s Member, ISN-COMGAN Research Committee of the International Society of
Nephrology.
Selected publications:
• E. Gotti, N. Perico, A. Perna, F. Gaspari, D. Cattaneo, R. Caruso, S. Ferrari, N. Stucchi, G. Marchetti, M. Abbate, G.
Remuzzi. Renal transplantation: can we reduce calcineurin inhibitor/stop steroids Evidence based on protocol biopsy
findings. J Am Soc Nephrol 2003;14:755-766.
• N. Perico, P. Ruggenenti, G. Remuzzi. Losartan in diabetic nephropathy. Expert Rev. Cardiovasc. Ther. 2004; 2(4): 473-
483.
• Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S,
Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P. mofetil
versus azathioprine for prevention of acute rejection in renal transplantation (MYSS): a randomised trial. Lancet. 2004
Aug 7;364(9433):503-12.
• Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M,
Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G, Perico N; MY.S.S. Study Investigators. of different
prediction equations for estimating renal function in kidney transplantation. Am J Transplant. 2004 Nov;4(11):1826-35.
• Perico N, Cattaneo D, Sayegh MH, Remuzzi G. Delayed graft function in kidney transplantation. Lancet. 2004 Nov
13;364(9447):1814-27.
• Cattaneo D, Merlini S, Zenoni S, Baldelli S, Gotti E, Remuzzi G, Perico N. Influence of co-medication with sirolimus or
cyclosporine on mycophenolic acid pharmacokinetics in kidney transplantation. Am J Transplant. 2005 Dec;5(12):2937-
44.
Annalisa Perna got her Statistical Sciences degree in 1984 at the University of Bologna, Italy.
Educational training: She completed her research training at IRFMN, Bergamo Labs. and at the Daccò
Center.
Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology,
statistical methods for calculating sample size and for meta-analytic techniques. She is also involved in
performing systematic reviews for the Cochrane Collaboration – Renal Review Group.
Employment: she is Head of the Laboratory of Biostatistics - Department of Renal Medicine at Daccò
Center, Ranica (Bergamo).
307
ANNUAL REPORT 2008

IRFMN
Principal publications:
• G. Remuzzi, P. Cravedi, A. Perna, B.D. Dimitrov, M. Turturro, G. Locatelli, P. Rigotti, N. Baldan, M.
Beatini, U. Valente, M. Scalamogna, P. Ruggenenti. Dual Kidney Transplant Group. Long-term outcome of
renal transplantation from older donors. N Engl J Med 2006;354:343-352.
• Ruggenenti P, Perna A, Loriga G, Ganeva M, Ene-Iordache B, Turturro M, Lesti M, Perticucci E, Chakarski IN,
Leonardis D, Garini G, Sessa A, Basile C, Alpa M, Scanziani R, Sorba G, Zoccali C, Remuzzi G for the REIN-2 Study
group. Blood-pressure control for renoprotection in patients with non-diabetic chronic renal disease (REIN-2):
multicentre, randomised controlled trial. Lancet 365:939-946, 2005.
• Schieppati A, Perna A, Zamora J, Giuliano AG, Braun N, Remuzzi G. Immunosuppressive treatment for idiopathic
membranous nephropathy in adults with nephrotic syndrome. Cochrane Database Syst Rev. Oct 18(4):CD004293, 2004
• Ruggenenti P, Fassi A, Ilieva AP, Bruno S, Iliev IP, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-
Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G. Preventing microalbuminuria in type 2
diabetes. N Engl J Med 351(19)1941-51, 2004.
• The BENEDICT Group. The Bergamo NEphrologic Diabetes Complications Trial (BENEDICT): design and baseline
characteristics. Control Clin Trials 24(4): 442-461, 2003.
• Plata R, Cornejo A, Arratia C, Anabaja A, Perna A, Dimitrov BD, Remuzzi G, Ruggenenti P for the Commission on
Global Advancement of Nephrology (COMGAN), Research Subcommittee of the International Society of Nephrology.
Angiotensin-converting-enzyme inhibition therapy in altitude polycythaemia: a prospective randomised trial. Lancet 359:
663-66, 2002.
Dario Cattaneo got the Pharmacy degree in 1996 at the University of Milan, and the specialisation in
Pharmacology (2001) awarded by the same University. In 2000 he got the specialization in
“Pharmacological Research” at the Mario Negri Institute for Pharmacological Research (IRFMN) and in
2005 he has been awarded the PhD degree by the Open University of London, UK.
Educational Training: in 1997 Post Doctoral Fellow, IRFMN, Laboratory of Pharmacokinetics and
Clinical Chemistry; in 2000 beneficiary of the Fellowship “Girola” and from 2001 to 2005 recipient of
the “Monzino” Fellowship for his research activity done at the IRFMN.
Areas of interest: pharmacology (pharmacokinetics, pharmacodynamics and pharmacogenetics) of
immunosuppressants, antiviral agents and hypolipidemic drugs; study of secondary forms of
dyslipidemia; polypharmacological approaches for the treatment of chronic kidney diseases; assessment
of humoral response as predictor of acute or chronic rejection after organ transplantation.
Employement: in 1997 researcher, IRFMN, Laboratory of Pharmacokinetics and Clinical Chemistry,.
Since 2006, Head of the Unit of Pharmacology and Pharmacogenetics, IRFMN, Ranica Bergamo.
Component of the Ethical Committee (2003) of the Hospital “Bolognini” (Seriate, Italy) and the
Hospital “E.Medea” (Bosisio Parini, Italy) since 2006. Member of the editorial board of Current
Clinical Pharmacology and affiliate of the International Association of Therapeutic Drug Monitoring
and Clinical Toxicology (IATDMCT) since 2005.
Selected publications
• Cattaneo D, Merlini S, Zenoni S, Baldelli S, Gotti E, Remuzzi G, Perico N. Influence of co-medication with sirolimus or
cyclosporine on mycophenolic acid pharmacokinetics in kidney transplantation. Am J Transplant. 2005 Dec;5(12):2937-
44.
• Cattaneo D, Gotti E, Perico N, Bertolini G, Kainer G, Remuzzi G. Cyclosporine formulation and Kaposi's sarcoma after
renal transplantation. Transplantation. 2005 Sep 27;80(6):743-8.
• Perico N, Cattaneo D, Sayegh MH, Remuzzi G. Delayed graft function in kidney transplantation. Lancet. 2004 Nov 13-
19;364(9447):1814-27.
• Cattaneo D, Merlini S, Pellegrino M, Carrara F, Zenoni S, Murgia S, Baldelli S, Gaspari F, Remuzzi G, Perico N.
Therapeutic drug monitoring of sirolimus: effect of concomitant immunosuppressive therapy and optimization of drug
dosing. Am J Transplant. 2004 Aug;4(8):1345-51.
• Cattaneo D, Perico N, Gaspari F, Gotti E, Remuzzi G. Glucocorticoids interfere with mycophenolate mofetil
bioavailability in kidney transplantation. Kidney Int. 2002 Sep;62(3):1060-7.
Giulia Gherardi got her Scientific High School Diploma on 1989 at the Liceo Scientifico Marie Curie in
Zogno (Bergamo), the Nurse Diploma on 1995 at the Scuola per Infermieri Professionali, Ospedali
Riuniti, Bergamo.
Educational training: Clinical Research Nurse Diploma on 1997 at IRFMN –Daccò Center.
Areas of interest: statistical methodology of long-term randomised clinical trials in nephrology,
diabetology; the organisation and the monitoring of clinical trials.
Emplyment: In1997-2003 involved as co-organazing, speaker, co-speaker and tutor for the Clinical
Research Course for Nurse at IRFMN – Daccò Center (Ranica – Bergamo). Several training activities for
Nurses in Clinical Research area. In 1997-1999, Clinical Research Monitor at IRFMN – Daccò Center;
since 2000 Monitoring Unit Chief.
308
ANNUAL REPORT 2008

IRFMN
Selected pubblications
• Gaspari F, Ferrari S, Stucchi N, Centemeri E, Carrara F, Pellegrino M, Gherardi G, Gotti E, Segoloni G, Salvadori M,
Rigotti P, Valente U, Donati D, Sandrini S, Sparacino V, Remuzzi G and Perico N on the behalf of the MY.S.S. study
investigators. Performance of different prediction equations for estimating renal function in kidney transplantation.
American Journal of Transplantation. 2004; 4: 1826-1835.
• Ruggenenti P, Fassi A, Parvanova Ilieva A, Bruno S, Petro Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi A, Ganeva
M, Ene-Iordache B, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini AR, Remuzzi G, for the Bergamo Nephrologic
Diabetes Complications Trial (BENEDICT) Investigators. Preventing microalbuminuria in type 2 diabetes. N Engl J
Med. 2004; 351: 1941-51.
• Remuzzi G, Lesti M, Gotti E, Ganeva M, Dimitrov BD, Ene-Iordache B, Gherardi G, Donati D, Salvadori M, Sandrini S,
Valente U, Segoloni G, Mourad G, Federico S, Rigotti P, Sparacino V, Bosmans JL, Perico N, Ruggenenti P, for the
MY.S.S. Group. Mycophenolate mofetil versus azathioprine for prevention of acute rejection in renal transplantation
(MYSS): a randomized trial. Lancet. 2004 Aug 7; 364: 503 – 12.
• Ruggenenti P, Perna A, Gherardi G, Benini R, Remuzzi G. Chronic proteinuric nephropathies: outcomes and response
to treatment in a prospective cohort of 352 patients with different patterns of renal injury. Am J Kidney Dis. 2000 Jan;
35 (6): 1155 – 65.
• Ruggenenti P, Perna A, Zoccali C, Gherardi G, Benini R, Testa A, Remuzzi G. Chronic proteinuric nephropathies. II.
Outcomes and response to treatment in a prospective cohort of 352 patients: differences between women and men in
relation to the ACE polymorphism. Gruppo Italiano di Studi Epidemiologici in Nefrologia. J Am Soc Nephrol. 2000
Jan; 11 (1): 88 – 96.
• Ruggenenti P, Perna A, Gherardi G, Garini G, Zoccali C, Salvadori M, Scolari F, Schena FP, Remuzzi G.
Renoprotective properties of ACE-inhibition in non-diabetic nephropathies with non-nephrotic proteinuria. Lancet.
1999 Jul 31; 354 (9176): 359 – 64.
Aneliya Parvanova Ilieva got her Medical Doctor degree at the Faculty of Medicine, Thracian
University (former Higher Medical Institute), Stara Zagora, Bulgaria in 1988, and the specialisation in
Pharmacology in Department of Pharmacology, Faculty of Medicine, of the same university in 1992.
Educational training: 1989-1998: Teaching of 3 rd , 4 th and 5 th -year medical students and 2 nd and 3 rd -year
clinical nurses in a general pharmacology and clinical pharmacology, Thracian University, Stara Zagora,
Bulgaria. Examiner of these students in theoretical and practical, oral and written exams and tests. 1993:
Course on investigation of isolated organs – Bulgarian Academy of Sciences, Sofia. 1998: Visiting
scientist, IRFMN, Ranica, Bergamo, Italy. 1998: Proficiency in the methods for insulin sensitivity
evaluation (hyperinsulinemic euglicaemic clamp technique) and glomerular filtration rate evaluation
(plasma clearance of iohexol).
Areas of interest: primary and secondary prevention of the chronic microvascular diabetic complications
(diabetic nephropathy, diabetic retinopathy and diabetic neuropathy); role of insulin resistance and
hyperhomocysteinemia in these pathologies.
Employment: She participated as investigator in several clinical studies. She is Chief Unit of Early
Clinical Evaluation of Drugs at IRFMN since 2000. She is a member of the Union of Bulgarian Doctors
(since 1989), of the Union of Pharmacologists in Bulgaria (since 1990), and member of the Union of
Scientists in Bulgaria (since 1991).
Selected publications
• Parvanova A, Chiurchiu C, Ruggenenti P, Remuzzi G. Inhibition of the renin-angiotensin system and cardio-renal
protection: focus on losartan and angiotensin receptor blockade. Expert Opinion on Pharmacotherapy 2005 Sep; 6
(11):1931-1942.
• Ruggenenti P, Fassi A, Parvanova A, Bruno S, Iliev I, Brusegan V, Rubis N, Gherardi G, Arnoldi F, Ganeva M, Ene-
Iordache, Gaspari F, Perna A, Bossi A, Trevisan R, Dodesini A, Remuzzi G. Preventing Microalbuminuria in Type 2
Diabetes. NEJM 2004;351(19):1941-51.
• Parvanova A, Iliev I, Filipponi M, Dimitrov BD, Vedovato M, Tiengo A, Trevisan R, Remuzzi G, Ruggenenti P. Insulin
resistance and proliferative retinopathy: a cross-sectional, case-control study in 115 patients with type 2 diabetes. J Clin
Endocrinol Metab 2004 Sep; 89(9):4371-6.
• The BENEDICT Group. The Bergamo Nephrologic DIabetes Complications Trial (BENEDICT): design and baseline
characteristics. Controlled Clinical Trials 2003; 24:442-461.
• Parvanova A, Iliev I, Dimitrov BD, Arnoldi F, Zaletel J, Remuzzi G, Ruggenenti P. Hyperhomocysteinemia and
increased risk of retinopathy: a cross-sectional, case-control study in patients with type 2 diabetes. Diabetes Care 2002;
25 (12): 2361.
309
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE DEPARTMENT'S ACTIVITIES
The Department of Renal Medicine was established on 1999 at the Clinical Research Center for
Rare Diseases “Aldo e Cele Daccò” – Villa Camozzi, Ranica to coordinate the activities of three
Laboratories and two Units.
The activities of the Department are mainly focused on the study of the mechanisms of
progression of chronic nephropathies, of new prevention and intervention strategies for diabetic
nephropathy, non diabetic chronic nephropathies, chronic allograft dysfunction, of
cardiovascular complications of diabetes, chronic renal disease, dialysis and transplantation and
of thrombotic microangiopathies.
The main aims of these activities are:
1. To identify screening and intervention strategies aimed to prevent the onset of nephropathy
and of other chronic complications of diabetes and/or hypertension.
2. To define intervention strategies to prevent or slow the progression of chronic nephropathies
and eventually obtain remission/regression of renal dysfunction.
3. To optimize immunosuppressive protocols in kidney transplantation and to define new donor
selection criteria in order to expand the pool of available organs.
These aims will be pursued through the following modalities:
1. Pilot pathophysiology and clinical pharmacology studies fully finalized at the Clinical
Research Center to test new pathogenetic hypotheses and new treatment modalities.
2. National and international networks and multicenter trials aimed to verify the efficacy of
treatments of potential interest identified as described at point 1.
3. Meta-analyses and probabilistic models to test new risk factors and treatments in large
samples of patients and to transfer this information at individual level.
Many of these activities rest on the possibility of a tight cooperation with the Department of
Molecolar Medicine, the Department of Bioengineering and the Public-Private Department of
Specialist and Transplant Medicine. This cooperation allows to plan the research activities of
the Department on the basis of new information derived from basic research and of problems of
major clinical relevance emerging from routine clinical activities.
FINDINGS/MAIN RESULTS
Definition and validation of specific treatments aimed to prevent the development and
progression of nephropathy and related micro and macrovascular complications in subjects with
type 2 diabetes
Definition and validation of new integrated treatment protocols aimed to slow the progression
and/or to achieve remission/regression of diabetic and non-diabetic chronic nephropathies
Institution of a standardized protocol “on line” (The “Remission Clinics”) finalized to achieve
regression/remission of chronic nephropathies and limit overall renal and radiovascular risk in
hospital practice in the setting of a multicenter Network
Characterization of the antiproteinuric, nephroprotective and cardioprotective effect of
maximized and polypharmacologic renin-angiotensin system inhibition, intensified blood
pressure and lipid control and identification of novel treatments to reduce the blood pressure
and ameliorate insulin sensitivity in subjects at increased cardiovascular risk
310
ANNUAL REPORT 2008

IRFMN
Identification of acquired or congenital risk factors for chronic complications of diabetes and
cardiovascular morbidity and mortality
Identification and validation of early markers of acute kidney failure and of methods for direct
and indirect measurements of kidney function and GFR decline
Definition and validation of new, specific treatments for idiopathic membranous nephropathy
and for HUS forms associated with genetic defect of complement factors including the
standardization of combined liver and kidney transplantation to prevent post transplant
recurrence of genetic associated HUS.
Definition and validation of new laboratory procedures and predictive models to help
monitoring and optimizing immunosuppressive therapy in clinical transplantation with
particular focus on pharmacokynetic markers of drug exposure and genetic predictors of drug
tolerability and efficacy
Definition and validation of selection and allocation criteria of kidneys from marginal and oldvery
old donors to increase the donor pool and the transplant activity
Finalization and activation of multicenter clinical trials aimed to prevent onset and progression
of diabetic and non-diabetic chronic nephropathies, to achieve remission of the nephrotic
syndrome in primary glomerular diseases, minimize maintenance immunosuppression in kidney
transplantation and prevent cardiovascular morbidity and mortality in chronic hemodialysis.
Computerization of data acquisition and monitoring procedures for the conduction of controlled
clinical trials
NATIONAL COLLABORATIONS 2008
Lombardia
- Ospedale C. Cantù, Abbiategrasso (MI)
- Ospedale Civile di Asola, Asola (MN)
- Ospedale Fenaroli, Alzano Lombardo (BG)
- Azienda Ospedaliera OO.RR., Bergamo
- Ospedale Caduti Bollatesi, Bollate (MI)
- Azienda Ospedaliera Spedali Civili, Brescia
- Ospedale San Biagio, Clusone (BG)
- Ospedale S. Anna, Como
- Azienda Ospedaliera Istituti Ospedalieri, Cremona
- Ospedale di Desio (MI)
- Ospedale Briolini, Gazzaniga (BG)
- Azienda Ospedaliera di Melegnano, Melegnano – Vizzolo Predabissi (MI)
- Ospedale San Leopoldo Mandic, Merate (LC)
- Ospedale Maggiore Policlinico, Milano
- Ospedale Provinciale San Carlo Borromeo, Milano
- Azienda Ospedaliera - Polo Universitario L. Sacco, Milano
- Ospedale Fatebenefratelli, Milano
- Ospedale Niguarda Ca' Granda, Milano
- Clinica Pediatrica “G. e D. De Marchi’, Milano
- Ospedale San Raffaele, Milano
- Ospedale Pediatrico di Montichiari, Montichiari (BS)
311
ANNUAL REPORT 2008

IRFMN
- Ospedale San Gerardo, Monza (MI)
- Istituti Clinici Zucchi, Monza (MI)
- Università degli Studi di Pavia, Dipartimento di Medicina Interna e Terapia Medica, Pavia
- Centro Antidiabetico, Ponte San Pietro (BG)
- Azienda Ospedaliera Ospedale Treviglio Caravaggio, Romano di Lombardia (BG)
- Istituto Clinico Humanitas, Rozzano (MI)
- Ospedale Bolognini, Seriate (BG)
- Ospedale Civile, Ivrea
- Azienda Ospedaliera Ospedale Treviglio Caravaggio, Treviglio (BG)
- A.O. della Valtellina e della Valchiavenna, Sondrio
- Azienda Ospedaliera Santa Croce e Carli, Cuneo
- Ospedale Regionale di Circolo Fondazione Macchi, Varese
- USL 60, Unità Operativa di Nefrologia e Dialisi, Vimercate (LC)
- Ospedale Bassini, Cinisello Balsamo, Milano
- IRCCS Multimedia, Sesto San Giovanni, Milano
Piemonte
- A.S.O. Maggiore della Carità, Novara
- Azienda Ospedaliera San Giovanni Battista, Torino
- Ospedale Mauriziano Umberto I, Torino
- Ospedale Regina Margherita, Torino
- Ospedale Martini, Torino
- Ospedale Luigi Einaudi, Torino
Veneto, Trentino Alto-Adige e Friuli Venezia Giulia
- Casa di Cura Abano Terme, Abano Terme (PD)
- Ospedale Civile, Belluno
- Ospedale S. Giacomo Apostolo, Castelfranco Veneto, Treviso
- Ospedale Provinciale Umberto I, Mestre (VE)
- Ospedale Giustinianeo, Padova
- Università degli Studi di Padova, Istituto di Anatomia Patologica, Padova
- Ospedale Civile, Padova
- Ospedale S. Camillo dé Lellis, Schio (VI)
- Ospedale Regionale Santa Maria dei Battuti, Treviso
- Ospedale Ca’ Fondello, Divisione Nefrologia e Dialisi, Treviso
- Ospedale Civile Maggiore Borgo Trento, Verona
- Ospedale Policlinico Borgo Roma, Verona
- Ospedale Civile San Bortolo, Vicenza
- Ospedale Santa Chiara, Trento
- Istituto Scientifico per l'infanzia Burlo Garofalo, Trieste
- Ospedale S. Antonio, S. Daniele del Friuli, Udine
- Università degli Studi di Udine, Centro Trapianti Fegato-Rene-Pancreas, Udine
Liguria, Emilia Romagna e Toscana
- Azienda Ospedaliera San Martino, Genova
- Istituto “G. Gaslini”, Genova
- Ospedale S. Orsola Malpighi, Bologna
- Ospedale Policlinico, Modena
- Istituto di Clinica Medica e Nefrologia, Parma
- Ospedale Santa Maria delle Croci, Ravenna
- Arcispedale Santa Maria Nuova, Reggio Emilia
312
ANNUAL REPORT 2008

IRFMN
- Ospedale Santa Maria Annunziata, Bagno a Ripoli, Firenze
- Azienda Ospedaliera Careggi-Monna Tessa, Firenze
- Ospedale Nuovo “S. Giovanni di Dio”, Firenze
- Azienda Ospedaliera Meyer, Firenze
- Ospedale di S. Miniato, S. Miniato (FI)
- Azienda Ospedaliera Cisanello, Pisa
- Ospedale di Pistoia, Pistoia
- AUSL Rimini, Rimini
- ASL Ravenna, Ravenna
- Ospedale G.B. Morgagni, Forlì
- Ospedale Civile, Imola Bologna
- Università di Pisa, Ospedale S. Chiara, Pisa
Marche
- Ospedale Regionale Torrette, Torrette di Ancona, Ancona
- Ospedale I.N.R.C.A., Ancona
- Azienda Ospedaliera S. Salvatore, Pesaro
Lazio, Basilicata e Campania
- Ospedale Polispecializzato, Anzio, Roma
- Ospedale Fatebenefratelli, Roma
- Ospedale Pediatrico Bambino Gesù, Roma
- Policlinico Gemelli, Roma
- Ospedale Policlinico Umberto I, Roma
- Ospedale San Camillo Forlanini, Roma
- Università Cattolica del Sacro Cuore, Roma
- Dipartimento di Biopatologia Umana, Università La Sapienza, Roma
- Ospedale Grande degli Infermi, Viterbo
- Ospedale Riuniti, Matera
- Azienda Ospedaliera Ospedale Civile, Caserta (NA)
- Università Federico II di Napoli, Cattedra di Nefrologia, Napoli
- Università di Napoli, Policlinico Nuovo, Napoli
- Azienda Ospedaliera “S. G. di Dio e Ruggi d’Aragona”, Salerno
Abruzzo
- Ospedale G. Bernabeo, Ortona, Chieti
- Presidio Ospedaliero “San Massimo”, Penne (PE)
- Presidio Ospedaliero "G.Mazzini", Teramo
Puglia, Calabria, Sicilia e Sardegna
- Ospedale Regionale “Miulli”, Acquaviva delle Fonti, Bari
- Ospedale Pediatrico “Giovanni XXIII”, Bari
- Ospedale Policlinico, Bari
- Azienda Ospedaliera V.Fazzi, Lecce
- Ospedale Casa Sollievo dalla Sofferenza, S.Giovanni Rotondo (FG)
- Presidio Ospedaliero di Martina Franca, Martina Franca, Taranto
- A.U.S.L. TA/1 - Presidio Ospedaliero, Taranto
- Azienda Ospedaliera Ospedale Pugliese Ciaccio, Catanzaro
- Ospedale dell'Annunziata, Cosenza
- Centro di Fisiologia Clinica del CNR, Divisione di Nefrologia, Reggio Calabria
- Azienda Ospedaliera “Bianchi-Melacrino-Morelli”, Reggio Calabria
- Ospedale “N. Giannettasio”, Rossano Calabro, Cosenza
313
ANNUAL REPORT 2008

IRFMN
- Nuovo Presidio Ospedaliero, Acireale, Catania
- Azienda Ospedaliera "Ferrarotto", Catania
- Ospedale Zonale Maggiore, Modica (RG)
- Ospedale Civico, Palermo
- Ospedale “V. Cervello”, Palermo
- Azienda Ospedaliera "Umberto I", Siracusa
- Azienda Sanitaria G. Brotzu, Ospedale San Michele, Cagliari
- Istituto di Clinica e Biologia dell’Età Evolutiva, Cagliari
- Ospedale A. Segni, Ozieri, Sassari
- Ospedale SS. Annunziata, Sassari
- Istituto di Patologia Speciale Medica dell'Università degli Studi di Sassari
- Ospedale Policlinico, Sassari
- Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione (IsMeTT), Palermo
- A.O. Universitaria “Ospedali Riuniti”, Foggia
Umbria
- Azienda Ospedaliera di Perugia, Perugia
INTERNATIONAL COLLABORATIONS 2008
- University Medical Center, Ljubljana Slovenia
- University Hospital Ziekenhuius, Edegem Antwerpen, Belgium
- Clinique de Nephrologie-Dialyse Chu Brugmann, Bruxelles, Belgium
- University Ziekenhuius Gent, Gent, Belgium
- U.Z. Gasthuisberg, Leuven, Belgium
- General Hospital Maria Middelares, Sint Niklaas, Belgium
- University of Groningen, AV Groningen, The Netherlands
- Academisch Ziekenhuis, Maastricht, The Netherlands
- Thracian University, Stara Zagora, Bulgaria
- The Birmingham Children's Hospital, Birmingham, UK
- Guy’s Hospital, London, UK
- Manchester Children's Hospital, Manchester, UK
- Nottingham City Hospital, Nottingham, UK
- Aalborg Hospital, Aalborg, Denmark
- Nephrological Department, University of Copenaghen, Copenaghen, Denmark
- Steno Diabetes Center, Gentofte, Denmark
- Department of Nephrology, Odense University Hospital, Odense, Denmark
- Department of Nephrology, Sahlgrenska University Hospital, Goteborg, Sweden
- Ospedale San Giovanni, Bellinzona, Switzerland
- Department of Nephrology, University of Wien, Wien, Austria
- Carl Thiem Klinikum, Cottbus, Germany
- Klinikum der Johann Wolfgang, Frankfurt am Main, Germany
- Arbeitsgruppe fyr Biomolekulare Medizin, Hamburg, Germany
- Univeristatklinik Heidelberg, Heidelberg, Germany
- Medizinische Klinik, Mannheim, Germany
- Luitpold Krankenhaus Med. Universitatklinik/Dialyse, Wurzburg, Germany
314
ANNUAL REPORT 2008

IRFMN
- Hospital Ntra Sra. de Sonsoles, Avila, Spain
- Hospitalet de Llobregat, Institut Català de la Salut, Barcellona, Spain
- Fundacion Jimenez Diaz, Madrid, Spain
- Hospital Clinico Martin Logas, Madrid, Spain
- Hospital 12 de Octubre, Madrid, Spain
- Hospital Gregorio Maranon, Madrid, Spain
- Hospital La Paz, Madrid, Spain
- Hospital Puerta de Hierro, Madrid, Spain
- Hospital Ramon y Cajal, Madrid, Spain
- Hospital Severo Ochoa, Leganes, Madrid, Spain
- Hospital Universitario de Tarragona Joan XXIII, Tarragona, Spain
- Hospital Garcia de Orta, Almada, Portugal
- Brigham & Women's Hospital, Boston, USA
- Hennepin County Medical Center, Minneapolis, USA
- SIU School of Medicine, Springfield, USA
- The Toronto Hospital, Toronto, Canada
- INCUCAI, Buenos Aires, Argentina
- Hospital Italiano de Buenos Aires, Buenos Aires, Argentina
- Hospital Regional de Valdivia, Valdivia, Cile
- Soroka Medical Center, Beer Sheva, Israel
EDITORIAL BOARD MEMBERSHIP 2008
Journal of Nephrology (Piero Ruggenenti)
Current Diabetes Reviews (Piero Ruggenenti)
Clinical Journal of the American Society of Nephrology (Piero Ruggenenti)
Current Pharmacology Reviews (Dario Cattaneo)
PEER REVIEW ACTIVITIES
American Journal of Kidney Diseases
American Journal of Physiology
American Journal of Transplantation
Biomed Central Edition
Brazilian Journal of Medical and Biological Research
Circulation
Clinical Journal of the American Society of Nephrology (CJASN)
Clinical Nephrology
Clinical Transplantation
315
ANNUAL REPORT 2008

IRFMN
Current Diabetes Reviews
Diabetic Nephropaty
Diabetologia
Dove Press
Expert Opinion on Emerging Drugs
Informa Healthcare
Journal of Acta Diabetologica
Journal of Clinical Investigation
Journal of Nephrology
Journal of the American Society of Nephrology (JASN)
Kidney International
Nature Clinical Practice Nephrology
Nephrology Dialysis and Transplantation
Nephron Clinical Practice
New England Journal of Medicine
Pediatric Nephrology
Plos Medicine
Talanta
The Brazilian Journal
The Lancet
The Open urology &Nephrology Journal
Translational Research
Transplantation
Vascular Pharmacology
PARTICIPATION IN EVENTS IN WHICH THE DEPARTMENT WAS
INVOLVED
“Pharmacokinetics of mycophenolate sodium and comparison with the mofeil formulation in
stable kidney transplant recipients”. In “oportunidades para maximizar o sucesso do doente”,
Lisboa, Portugal, February 23, 2008.
“Pharmacogenetics of immunosuppressants: progress and promises”. In GENECURE Agenda,
Ranica, Italy, 10-11 April 2008.
“Statins, dyslipidemia and novel hypolipidemic agents”. Advanced Workshop for Turkish
Nephrologists, Ranica, April 18-19, 2008.
“La farmacogenetica”. Forum annuale del Comitato di Bioetica, organizzato dall’Azienda
Ospedaliera Ospedali Riuniti di Bergamo, 12 May 2008.
“Polymorphisms in ABCB1, the gene encoding for p-glycoprotein, predicts recovery of graft
function early after kidney transplantation”. Oral presentation for the American Transplant
Congress, Toronto, May 31 – June 4, 2008.
“La ricerca in campo genetico”. Azienda Ospedaliero-Universitaria di Parma, Parma 11 July
2008.
316
ANNUAL REPORT 2008

IRFMN
“Strategie farmacologiche di renoprotezione”. All’interno del Simposio “Terapia conservativa,
dietetica e farmacologica della malattia renale cronica”, Pisa 26 September 2008.
“Valutazione del filtrato glomerulare e utilizzo dell’esame delle urine nella pratica clinica”, 2°
Convegno sulla funzione renale, Centro Congressi Villa Gualino, Torino 27 June 2008
Society for Clinical Trials, St. Louis, Missouri, USA, May 18-21 2008
Institute for Health Metrics and Evaluation, Global Health Metrics and Evaluation: current state
and future directions. Research Conference, Seattle, Washington, USA, April 9-11 2008
‘Geni, proteine e malattie’. 20 November 2008, Borsa Merci Bergamo
‘Prima Giornata Europea delle Malattie rare’, 29 February 2008, Centro Congressi Giovanni
XXIII, Bergamo
XLV ERA-EDTA Congress, Stoccolma, 10-13 May 2008
Convegno “La ricerca indipendente sui farmaci promossa dall’AIFA”. Roma, 30 September
2008
Course in Renal Endocrinology. Stoccolma, 06-07 October 2008
49° Congresso Nazionale della Società Italiana di Nefrologia. Rimini, 08-11 October2008
Convegno “Nefropatie Primitive e Secondarie”, Firenze, 21-22 November 2008
40° Corso di Aggiornamento in Nefrologia e Metodiche Dialitiche, Milano, 05-08 December
2008
Corso di Aggiornamento “La Remission Clinic nella Pratica Clinica”, Ranica (BG), 16
December 2008
GRANTS AND CONTRACTS
AIFA (Agenzia Italiana del Farmaco)
PKD Foundation
Abbott GmbH & Co.
Astrazeneca SPA
ACRAF Spa (Aziende Chimiche Riunite Angelini Francesco)
BOHERINGER INGELHEIM Italia Spa
Chiesi Farmaceutici
Dompè Spa
Genzyme Europe BV
Roche SpA
Sanofi-Aventis Spa
Sigma Tau Spa
Solvay Pharmaceuticals
317
ANNUAL REPORT 2008

IRFMN
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
G. Remuzzi, N. Perico, M.E. De Broe. Tubulointerstitial Diseases. In: Brenner and Rector's The Kidney, 8 th ed., Vol.
II, chapter n. 33. Edited by B.M. Brenner. Saunders Elsevier, Philadelphia 2008, pp. 1174-1202.
S.K. Sharma, N. Perico, P. Ruggenenti, G. Remuzzi. Prevention of chronic renal diseases in the elderly. In: The Aging
Kidney in Health and Disease, chapter n. 13. Edited by J.F. Macias Nunez, J.S. Cameron, D.G. Oreopoulos. Springer,
New York 2008, pp. 231-255.
P. Ruggenenti, I. Iliev, G.M. Costa, A. Parvanova, A. Perna, G.A. Giuliano, N. Motterlini, B. Ene-Iordache, G.
Remuzzi, and the BENEDICT Study Group. Preventing left ventricular hypertrophy by ACE inhibition in
hypertensive patients with type 2 diabetes. Diabetes Care 2008;31:1629-1634.
P. Ruggenenti, E. Perticucci, P. Cravedi, V. Gambara, M. Costantini, S.K. Sharma, A. Perna, G. Remuzzi. Role of
remission clinics in the longitudinal treatment of CKD. J Am Soc Nephrol 2008;19:1213-1224.
P. Ruggenenti, A. Remuzzi, G. Remuzzi. Decision time for pancreatic islet-cell transplantation. (Comment) Lancet
2008;371:883-884.
M. Cortinovis, D. Cattaneo, N. Perico, G. Remuzzi. Investigational drugs for diabetic nephropathy. Expert Opin
Investig Drugs 2008;17:1487-1500.
N. Perico, A. Benigni, G. Remuzzi. Present and future drug treatments for chronic kidney diseases: evolving targets in
renoprotection. Nature Reviews Drug Discovery 2008;7:936-953.
P. Ruggenenti, P. Bettinaglio, F. Pinares, G. Remuzzi. Angiotensin converting enzyme insertion/deletion
polymorphism and renoprotection in diabetic and nondiabetic nephropathies. Clin J Am Soc Nephrol 2008;3:1511-
1525.
F. Casiraghi, N. Azzollini, P. Cassis, B. Imberti, M. Morigi, D. Cugini, R.A. Cavinato, M. Todeschini, S. Solini, A.
Sonzogni, N. Perico, G. Remuzzi, M. Noris. Pretransplant infusion of mesenchymal stem cells prolongs the survival of
a semiallogeneic heart transplant through the generation of regulatory T cells. J Immunol 2008;181:3933-3946.
P. Ruggenenti, M. Noris, G. Remuzzi. Thrombotic Microangiopathies. In: Therapy in Nephrology and Hypertension –
A companion to Brenner and Rector’s The Kidney (3 rd Edition), chapter n. 26. Edited by C.S. Wilcox. Saunders
Elsevier, Philadelphia 2008, pp. 294-312.
Saland JM, Ruggenenti P, Remuzzi G; Consensus Study Group. Liver-Kidney Transplantation to Cure Atypical
Hemolytic Uremic Syndrome. J Am Soc Nephrol. 2008 Dec 17.
Ruggenenti P, Cravedi P, Remuzzi G. Rituximab for membranous nephropathy and immune disease: less might be
enough. Nat Clin Pract Nephrol. 2008 Dec 2
Cravedi P, Mannon RB, Ruggenenti P, Remuzzi A, Remuzzi G. Islet transplantation: need for a time-out Nat Clin
Pract Nephrol. 2008 Dec;4(12):660-1. Epub 2008 Sep 23.
Cravedi P, Mannon RB, Remuzzi G. Lymphocyte depletion for kidney transplantation: back to the past Nat Clin
Pract Nephrol. 2008 Oct;4(10):534-5. Epub 2008 Aug 26.
Braun N, Schmutzler F, Lange C, Perna A, Remuzzi G, Risler T, Willis NS. Immunosuppressive treatment for focal
segmental glomerulosclerosis in adults.
Cochrane Database Syst Rev. 2008 Jul 16;(3):
Ruggenenti P, Cravedi P, Sghirlanzoni MC, Gagliardini E, Conti S, Gaspari F, Marchetti G,
Abbate M, Remuzzi G. Effects of rituximab on morphofunctional abnormalities of membranous glomerulopathy. Clin
J Am Soc Nephrol. 2008 Nov;3(6):1652-9. Epub 2008 Aug 6.
Remuzzi G, Cattaneo D, Perico N. The aggravating mechanisms of aldosterone on kidney fibrosis. J Am Soc Nephrol.
2008 Aug;19(8):1459-62. Epub 2008 Jun 11.
318
ANNUAL REPORT 2008

IRFMN
Cattaneo D, Bitto A, Baldelli S, Cortinovis M, Gotti E, Perico N, Remuzzi G. Pharmacokinetic/pharmacodynamic
drug interaction between rosiglitazone and mycophenolate mofetil in kidney transplantation: a case report.
Transplantation. 2008 Mar 27;85(6):921-2.
Fassi A, Rubis N, Parvanova A, Iliev I, Zamora J, Giuliano GA, Ene-Iordache B, Perna A, Anabaya A, Motterlini N,
Ruggenenti P, Remuzzi G for the BENEDICT Study Investigators. Randomized and non-randomized patients in the
Bergamo Nephrologic Diabetes Complications Trial (BENEDICT). Abstract. 29 th Annual Meeting of the Society for
Clinical Trials St Louis, Missouri, May 18-21, 2008
RESEARCH ACTIVITIES
Laboratory of Biostatistics
European Study for Preventing by Lipid-lowering agents ANd Aceinhibition
Dialysis Endpoints (ESPLANADE) clinical trial: final analyses.
ESPLANADE is a randomised, multicenter trial aimed to test the antiproteinuric effects of
statins added to combined ACE-inhibitor (ACEi) and angiotensin II receptor antagonist (ATA)
therapy in proteinuric, chronic nephropathies. The primary aim of the study is to assess whether
combined therapy of ACEi, ATA and statins is more effective than ACEi and ATA alone in
reducing proteinuria. During 2008 we completed data collection and statistical analysis of final
data.
‘Acetyl-carnitine in insulin resistance’ clinical trial: final analyses.
‘Acetyl-carnitine in insulin resistance’ clinical trial is a pilot, longitudinal, prospective cohort
study. It is aimed to evaluate the short-term effects of acetyl-carnitine on insulin resistance and
on metabolic sindrome in subjects with increased risk of type II diabetes. During 2008 we
completed data collection and statistical analysis of final data.
BErgamo NEphrologic DIabetes Complications Trial (BENEDICT) – Phase
A and B: comparison of baseline characteristics of randomised and non
randomised subjects.
BENEDICT is a randomised, double-blind, multicentre clinical trial, aimed to assess the effects
of an ACE inhibitor, alone or in combination with a non diidropiridinic calcium channel
blocker, on progression to microalbuminuria in hypertensive, normoalbuminuric, type II
diabetic subjects (Phase A) and on progression to macroalbuminuria in hypertensive,
microalbuminuric, type II diabetic subjects (Phase B). During 2008 we completed statistical
analysis of baseline characteristics in randomised and non randomised subjects.
BErgamo NEphrologic DIabetes Complications Trial (BENEDICT) – Phase
A and PPARγ2 P12A polymorphism: final analyses.
BENEDICT is a randomised, double-blind, multicentre clinical trial performed in 1204 subjects
with normoalbuminuria, hypertension and type II diabetes. Primary efficacy analyses have
shown that Trandolapril and Verapamil in combination and Trandolapril alone significantly
slowed the progression to microalbuminuria as compared to placebo. On the contrary
Verapamil alone did not significantly differ from placebo (Phase A). Data on PPARγ2 P12A
polymorphism were collected and it was explored whether the above effects on progression to
microalbuminuria were regulated by the PPARγ2 P12A. During 2008 we performed statistical
analysis of final data.
319
ANNUAL REPORT 2008

IRFMN
MYcophenolate Steroid Sparing Study (MYSS) clinical trial and ABCB1 -
exons 21 and 26 genotype: final analyses.
MYSS is a randomised, multicentre, clinical trial which showed that in cadaveric renal
transplanted patients with immunosuppressive therapy with ciclosporine Neoral®,
Mychophenolate Mofetil was not more effective than Azathioprine in preventing acute
rejections at 21 months after transplant. It was also evaluated whether adverse events associated
to the use of cyclosporine during the MYSS trial were regulated by ABCB1 – exons 21 and 26
genotype. During 2008 we performed statistical analysis of final data.
Long-term renal survival in recipients of grafts from old (aged 60-69 years)
and very old donors (aged >70 years): final analyses.
Aim of this prospective, longitudinal, controlled study is to compare long-term renal survival in
a cohort of recipients of one or two kidneys from donors aged 60 to 69 years (N=67) with that
of kidney recipients from donors aged 70 years or older (N=71). During 2008 we completed
data collection and statistical analysis of final data.
Laboratory of Clinical Chemistry
Increase of urinary excretion of NGAL (neutrophil gelatinase-associated
lipocalin) as an early marker in predicting acute renal dysfunction in
patients with solid organ cancer given cisplatin as chemotherapeutic
agent
Cisplatin has a major impact in cancer medicine and is widely used for therapeutic management
of several tumors, such as those of ovary, testes, and the head and neck. However, while several
anti-neoplastic agents frequently exhibit nephrotoxicity, the platinum derivatives are among the
most frequent compounds leading to renal injury. Indeed, approximately 25-35% of patients
develop evidence of nephrotoxicity following an initial dose of cisplatin and these findings
indicate that there is a pressing need for way to protect the kidney while administering effective
chemotherapeutic agents such as cisplatin. Unfortunately, creatinine is an unreliable indicator
during acute changes in kidney function and among compounds that might serve as a novel
biomarkers for the initiation phase of acute renal failure, neutrophil gelatinase-associated
lipocalin (NGAL) has been identified as one of the most strikingly upregulated genes and
overexpressed proteins in the kidney after ischemia. Markedly increased NGAL concentrations
were easily detected in urine early after renal ischemia in mouse and rat models. Recent studies
have demonstrated that NGAL is a useful early predictor of acute renal failure also in humans.
Taken altogether these findings indicate that urinary and/or serum NGAL concentration
monitoring may represent a sensitive, specific, and highly predictive early biomarker for acute
renal failure. So far, however, no data are available on the reliability of NGAL to predict the
development of acute renal dysfunction in cancer patients receiving cisplatin treatment.
To this purpose, we set up a study in 46 patients with solid tumors aimed to evaluate whether
serum and/or urinary NGAL concentrations soon after cisplatin infusion (determined by ELISA
assay) were suitable to predict development of subsequent acute renal dysfunction.
Our findings indicated that urinary NGAL is an early and reliable marker of development of
acute renal dysfunction in cancer patients receiving cisplatin treatment.
Among the 46 patients receiving cisplatin, acute renal failure developed in 12, as documented
by the increase in serum creatinine concentration that occurred between day 3 to 7 after cisplatin
infusion.
In all patients developing acute renal failure a mild increase in serum NGAL was observed at
day 1 post-cisplatin treatment. Thereafter the concentration of the marker progressively declined
320
ANNUAL REPORT 2008

IRFMN
below baseline values. However, the serum profile of NGAL was comparable in both patients
with and without acute renal injury.
At variance, the concentration of NGAL in the urine progressively increased from day 1 to day
7 post-cisplatin administration only in patients who had the increase in serum creatinine and
thus developed acute renal failure.
Thus, urinary but not serum NGAL concentrations were proven to be a useful surrogate marker
to predict patients who may develop acute kidney injury after single cisplatin infusion. Increase
of urinary NGAL concentration more than 3500% over baseline value at day 2 post-drug
infusion allows to identify a subgroup of patients with persistent renal dysfunction.
Evaluation of renal function decline determined by 13 different GFR
estimating formulas in type 2 diabetic patients
Evaluation of glomerular filtration rate (GFR) is of critical importance in the clinical
management of both clinic and outclinic diabetic patients since diabetic nephropathy affects 25-
40% of the patients, and diabetes is the leading cause of end-stage renal disease. However, the
measurement of the true GFR either by the renal clearance of the gold standard inulin or by
plasma clearance of contrast agents is time consuming, difficult to perform and cannot be easily
implemented in clinical daily practice. Furthermore, creatinine clearance is inconvenient and
sometimes inaccurate due to variable creatinine metabolism and tubular secretion.
To circumvent these drawbacks, a number of equation has been developed to provide an
estimate of renal function from serum creatinine and demographic characteristics.
Recently we demonstrated in both transplant and diabetic patients that prediction formulas were
inaccurate in predicting true GFR especially in subjects with renal function values higher than
60 ml/min/1.73m 2 .
However, their performance and suitability to monitor renal function decline in patients with
type 2 diabetes have not been assessed so far.
Thus, we evaluated the decline of renal function by calculating the regression line vs. measured
GFR (by iohexol plasma clearance) or GFR estimated by 13 different prediction equations in
337 normo- and microalbuminuric subjects, enrolled in the BENEDICT and in the DEMAND
study, who had at least 4 serial renal function determinations. Each patient had 4-18 GFR
determinations performed in a 0.8 to 8 years period. The measured renal function declined by -
3.95 ml/min/1.73m 2 /year. All the prediction formulas markedly underestimated the GFR decline
and neither baseline albuminuria (normo vs. micro) nor follow-up duration (more or less than
3.5 years) affected the results. Estimated GFR decline ranged from -0.53 (Ibrahim equation) to -
1.55 ml/min/1.73m 2 /year (Hull formula). In diabetics patients with baseline GFR >120
(hyperfiltering) or between 80 and 120 ml/min/1.73m 2 GFR declined by 4.37+0.55 and
3.51+0.38 ml/min/1.73m 2 /y, respectively. In the two cohorts estimated slopes under-estimated
GFR decline by 6.14 and by 3.33 folds, respectively.
These findings showed that none of the 13 GFR models do not reliably predict GFR decline in
diabetics with normo or microalbuminuria, in particular in those at increased risk because of
hyperfiltration. Failure to early detect progressive kidney dysfunction may delay the institution
of renoprotective therapies in this population. This may have major clinical implications since
glomerular hyperfiltration has a pathogenic role in onset and progression of diabetic renal
disease and early intervention to ameliorate hyperfiltration may prove renoprotective in the long
term.
Development of a new equation to predict creatinine clearance in Cuban
population
Determination of glomerular filtration rate (GFR) provides the measure of the filtering capacity
of the kidneys and is considered the best overall index of renal function currently used to
determine the effectiveness of therapies designed to slow the progression of chronic renal
321
ANNUAL REPORT 2008

IRFMN
diseases. However, the measurement of the true GFR either by the renal clearance of the gold
standard inulin or by plasma clearance of contrast agents is time consuming, difficult to perform
and cannot be easily implemented in clinical daily practice and particularly in poor or third
world countries. Urinary creatinine clearance is widely used to measure GFR, but it is
inconvenient and sometimes inaccurate due to variable creatinine metabolism and tubular
secretion. To overcome these limitations, the National Kidney Foundation-Kidney Disease
Outcomes Quality Initiative recommends the estimation of GFR using prediction equations
based on serum creatinine, demographic and anthropometric data. A relevant number of
prediction formulas developed mainly in caucasic subjects have been published so far, but none
of them correctly estimated renal function in Cuban population.
Thus, together with the National Institute of Nephrology of Havana City, we aimed to develop a
new equation to predict creatinine clearance in Cuban population. The database consisted of
9014 subjects who attended the Cuban Institute of Nephrology during the period 1985-2005.
64.3% of the subjects were male and 6408 were older than 18 years. For all the subjects body
weight, height, body surface area age, and gender were collected together with the measurement
of serum creatinine and creatinine clearance.
The new developed equation, suitable to estimate creatinine clearance in adult population,
included age, weight, height, gender and serum creatinine as independent variables.
The correlation between measured creatinine clearance and predicted values by the new
equation was good, (Spearman correlation coefficient r = 0.794) and better than with both
Cockcroft-Gault and MDRD formula (r = 0.555 and 0.667, respectively). The accuracy in
creatinine clearance estimation was markedly higher using the new equation: the number of
values predicted with an error within 10% were 50% more than with Cockcroft-Gault and
MDRD formulas. In particular, at variance with the results obtained with Cockcroft-Gault and
MDRD equations, the new developed formula was proven to be more accurate in predicting
creatinine clearance in both male and female subjects with renal function higher than 60
ml/min/1.73m 2 GFR.
Laboratory of Drug Development
Blood cell count and lipid lowering therapy affect sirolimus exposure in
kidney transplant recipients on calcineurin inhibitor-free regimen
Sirolimus (SRL) is an immunosuppressive agent characterized by a narrow therapeutic index.
Studies in organ transplant recipients given this drug in combination with cyclosporine (CsA)
have shown that SRL exposure is extremely variable. However, no data are available on the
pharmacokinetics of SRL in calcineurin inhibitor-free protocols.
Fifty-five pharmacokinetics profiles were collected from 20 kidney transplant patients given
SRL over a 36 months follow-up. None of them were on CsA or tacrolimus. Large interindividual
variability in the daily distribution of SRL concentrations was documented. Patients
with low blood cell count showed a significant decrease in daily SRL exposure, requiring 35%
increments in the daily dose to reach drug concentration targets. At variance, concomitant
administration of omega-3 polyunsaturated fatty acids increased SRL AUC 0-24 by 25%
compared to values found in normolipidemic patients. Altogether, these information can be used
to optimize SRL dose-adjustments in the routine clinical practice. These results have been
presented at the meeting of the International Association of Therapeutic Drug Monitoring and
Clinical Toxicology (Nice, September 2007) and are now submitted for publication in the
Journal of Clinical Pharmacology.
322
ANNUAL REPORT 2008

IRFMN
Abcb1 genotypes predict cyclosporine-related adverse events and graft
outcome in kidney transplantation: a pre-specified analysis of the
mycophenolate steroids sparing (Myss) study
Cyclosporine A (CsA) is a substrate of P-glycoprotein, an efflux transporter encoded by the
ABCB1 gene. As compared to carriers of the wild-type gene, carriers of T allelic variants in
exons 21 or 26 have reduced P-glycoprotein activity and secondarily increased CsA intracellular
concentration. Thus, carriers of T variants might be at increased risk of CsA-related events. We
evaluated the associations between ABCB1 genotypes in exon 12, 21 and 26 and CsA-related
outcomes in 147 renal transplant recipients on CsA-based immunosuppression included in the
Mycophenolate Steroids Sparing study and followed prospectively for a median of 65.5 months.
As compared to carriers of the wild genotype, carriers of T allelic variants in exons 21 or 26 had
a three-fold excess risk of delayed graft function (p=0.011 and p=0.019, respectively), a trend to
slower renal function recovery and lower glomerular filtration rate at study end, significantly
higher incidence of new-onset diabetes and cytomegalovirus reactivation. T variants in both
exon 21 (p=0.022) and 26 (p=0.034) were independently associated with 3.8 and 3.5 folds
excess of DGF. Incidence of acute rejections and mean CsA dose and blood levels were
comparable in genotype groups. Renal transplant recipients with T allelic variants in exon 21 or
26 of the ABCB1 gene are at increased risk of CsA-related adverse events. Genetic evaluation
may help identify patients at risk and modulate CsA therapy to optimize graft and patient
outcomes.
These results have been presented at the meeting of the American Society of Transplantation
(Toronto, June 2008) and are now submitted for publication in the Journal of the American
Society of Nephrology.
323
ANNUAL REPORT 2008

IRFMN
324
ANNUAL REPORT 2008

IRFMN
LABORATORY OF COORDINATION OF
DIAGNOSIS AND INFORMATION ON
RARE DISEASES
STAFF
Head
Arrigo SCHIEPPATI, M.D.
Information Centre for Rare Diseases
Head
Erica DAINA, M.D.
325
ANNUAL REPORT 2008

IRFMN
CURRICULA VITAE
Arrigo Schieppati got his degree in Medicine at the University of Milan in 1978 and the specialisation in
Medical Nephrology in 1984 at the same University.
He performed his training at the Mario Negri Bergamo Laboratories with Dr. Remuzzi, and completed it
with stages at the laboratories of prof. Patrono (Catholic University in Rome), prof. John Gordon
(Cambridge, GB), and at the Division of Renal Diseases - University of Colorado Medical School,
directed by Dr. Schrier (Denver, USA). Since 1982 he works at the Division of Nephrology and Dialysis
– Riuniti Hospital – Bergamo, where he is in charge of Outpatients Clinic and Day Hospital. From 1992
to 1995 he was Head of the Information Center for Rare Diseases and since 1996 he is Head of the
Laboratory for Coordination of Information and Diagnosis of Rare Disease at the Clinical Research
Center for Rare Diseases Aldo e Cele Daccò of the Mario Negri Institute.
Areas of interest: diagnosis and therapy of chronic renal diseases, hypertension and rare kidney diseases.
Affiliations: ethical committee Riuniti Hospital - Bergamo; member of the working group of the regional
network for rare diseases in Lombardy; scientific committee Bolognini Hospital – Seriate (BG); member
of the Task Force on Rare Diseases (DG Health and Consumer Protection); International Society of
Nephrology; American Society of Nephrology; Editorial Board Journal of Nephrology.
Selected publications
• Schieppati A, Henter J I, Daina E, Aperia A. Why rare diseases are an important medical and social issue. Lancet. 2008
June 14; 371:2039-55.
• Schieppati A, Remuzzi G. Chronic renal diseases as a public health problem: epidemiology, social, and economic
implications. Kidney Int Suppl. 2005 Sep;(98):S7-S10.
• Remuzzi G, Schieppati A, Boissel JP, Garattini S, Horton R. Independent clinical research in Europe. Lancet. 2004 Nov
6-12;364(9446):1723-6.
• Schieppati A, Perna A, Zamora J, Giuliano GA, Braun N, Remuzzi G. Immunosuppressive treatment for idiopathic
membranous nephropathy in adults with nephrotic syndrome. Cochrane Database Syst Rev. 2004 Oct 18;(4):CD004293.
• Remuzzi G, Schieppati A, Ruggenenti P. Clinical practice. Nephropathy in patients with type 2 diabetes. N Engl J Med.
2002 Apr 11;346(15):1145-51.
• Schieppati A, Remuzzi G, Garattini S. Modulating the profit motive to meet needs of the less-developed world. Lancet.
2001 Nov 10;358(9293):1638-41.
Erica Daina got his degree in Medicine at the University of Milan in 1987 and the specialisation in
Medical Nephrology in 1990 at the same University.
She performed her training at the II° Medical Division - San Raffaele Hospital - Milan, and at the
Division of Nephrology and Dialysis - Riuniti Hospital - Bergamo.
In March 1988 she started her collaboration with the Mario Negri Institute and since June 1993 she works
as full-time clinical researcher at the Clinical Research Center for Rare Diseases Aldo e Cele Daccò.
Since 1996 she is Unit Head – Information Center for Rare Diseases. Areas of interest: rare diseases,
Takayasu arteritis, Hemolytic Uremic Syndrome/Thrombotic Thrombocytopenic Purpura, Fabry’s
disease, Alport’s syndrome. Since January 2002 she is representative of Coordinating Centre - Regional
Network for Rare Diseases - and collaborates to didactics activity at the University of Turin (School of
Clinical Pathology Specialization - 2 nd Level Master in Rare Diseases).
Selected publications
• Bresin E, Daina E, Noris M, Castelletti F, Stefanov R, Hill P, Goodship T H J, Remuzzi G, International Registry
Recurrent Familial HUS/TTP. Outcome of renal transplantation in patients with non-Shiga toxin-associated Hemolytic
Uremic Syndrome: Prognostic significance of genetic background. Clinical Journal American Society Nephrology 2006;
1: 88-99
• Galbusera M, Bresin E, Noris M, Gastoldi S, Belotti D, Capoferri C, Daina E, Perseghin P, Scheiflinger F, Fakhouri F,
Grünfeld JP, Pogliani E, Remuzzi G. Rituximab prevents recurrence of thrombotic thrombocytopenic purpura: a case
report. Blood. 2005 Aug 1;106(3):925-8.
• Vanoli M, Daina E, Salvarani C, Sabbadini MG, Rossi C, Bacchiani G, Schieppati A, Baldissera E, Bertolini G; Itaka
Study Group. Takayasu's arteritis: A study of 104 Italian patients. Arthritis Rheum. 2005 Feb 15;53(1):100-7.
• Remuzzi G, Galbusera M, Noris M, Canciani MT, Daina E, Bresin E, Contaretti S, Caprioli J, Gamba S, Ruggenenti P,
Perico N, Mannucci PM; Italian Registry of Recurrent and Familial HUS/TTP. von Willebrand factor cleaving protease
(ADAMTS13) is deficient in recurrent and familial thrombotic thrombocytopenic purpura and hemolytic uremic
syndrome. Blood. 2002 Aug 1;100(3):778-85.
• Daina E, Schieppati A, Remuzzi G. Mycophenolate mofetil for the treatment of Takayasu arteritis: report of three cases.
Ann Intern Med. 1999 Mar 2;130(5):422-6.
326
ANNUAL REPORT 2008

IRFMN
INTRODUCTION TO THE LABORATORY’S ACTIVITIES
Rare Diseases (RD) represent about ten percent of all human medical illnesses and infirmities. It
is difficult to define what exactly is intended as a RD. The US Congress in the Orphan Drug Act
has given the first definition in 1983. Under this law it is considered rare a disease that affects
less than 200 000 Americans (prevalence 0.75 per 1 000).
Recently, the European Parliament adopted a more strict definition; they consider rare a
condition that affects not more than five individuals per 10 000 in the European Community
(prevalence 0.5 per 1 000). Besides the epidemiological parameter, RD have certain
characteristics in common: 1) most of them are of genetic origin; 2) rarity often brings a
difficult and/or late diagnosis; 3) generally, RD are heavy social burdens both to the family and
community and cause early mortality.
The greatest barrier to prevention, diagnosis and treatment of RD is inadequate knowledge.
Once a diagnosis of RD is put, a major complaint of patients and those involved in their care is
the difficulty to obtain pertinent information about causes, symptoms and either established or
experimental treatments. Often, patients with RD are willing to participate in clinical studies,
but they do not know where and how, and physicians or health authorities are seldom able to
help them.
RD is not a very attracting field for basic and clinical investigators for several reasons: it is
difficult to find adequate animal models for many rare disorders; clinical trials may require
more patients than available; financial support is insufficient.
Few countries have a central body or system to disseminate information on RD. Accurate
information on the incidence and prevalence of RD is extremely important for both basic and
clinical investigators. Invaluable help to research advances in RD would come from the
availability of registries and databases containing diagnostic, clinical and biological data of
patients with rare disorders.
A pilot experience has been established at the Clinical Research Centre for Rare Diseases “Aldo
and Cele Daccò” since 1992. This Centre is unique with its integration of an Information Centre
for Rare Diseases, clinical facilities for the implementation and development of clinical studies,
educational activities for physicians, nurses and patients.
In 2001 it has been nominated Coordinating Centre of the Regional Network for Rare Diseases
in the Lombardy Region, an area of 9 million people in Northern Italy. As Coordinating Centre
is also working with the National Centre of Rare Diseases at Istituto Superiore di Sanità. All the
up-to-date information regarding the activities of the Coordinating Centre are available at the
web site: http://malattierare.marionegri.it
FINDINGS/MAIN RESULTS
The database of the Information Centre for Rare Diseases contains data about 10991 patients
affected by 882 different rare disorders.
In the Bank of biological materials, samples from 1394 patients with rare conditions and their
families have been collected.
The Centre has established contacts with 327 Italian Associations for rare diseases. It was even
possible that patients with 88 different rare diseases - for which no Associations have been
established in Italy yet - to meet among themselves.
327
ANNUAL REPORT 2008

IRFMN
In July 2000, the European Commission recognized the Laboratory as a site for "Postgraduate
training on rare diseases" (contract No. QLK4-1999-50547).
In December 2001 (Delibera della Giunta Lombarda N. 7328), the Centre was identified as
"Coordinating Centre of the Regional Network for Rare Diseases".
The Laboratory coordinates the International Registry of Recurrent and Familial Hemolytic
Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP), since 1996. The
research projects developed in collaboration with the Laboratory of Immunology and Genetic of
Rare Diseases and Organ Transplantation have allowed to better comprehend the pathogenesis
of these diseases and to identify some genetically determined forms of HUS, associated to
defects of complement regulatory factors, and of TTP, associated to congenital deficiency of
ADAMTS13 enzyme.
NATIONAL COLLABORATIONS
Italian National Institute of Health
G. Bosco Hospital, Turin, Regional Coordinating Center for Rare Diseases
Riuniti Hospital, Bergamo
Italian Takayasu's Arteritis Study Group - ITAKA Group
Italian Registry of the Membranoproliferative Glomerulonephritis
Italian Registry of the Familial Focal Segmental Glomerulosclerosis
Biotechnology Laboratory, IRCCS Policlinico San Matteo, Pavia
Assessorato alla Sanità, Lombardia Region
University of Torino, School of Clinical Pathology, Faculty of Medicine and Surgery
Italian Network for Promotion of Folic Acid to Prevent Birth Defects
University of Turin, Department of Experimental Medicine and Oncology, 2 nd Level Master in
Rare Diseases
Italian Society of Neonatology (Lombardy section), Rare Congenital Respiratory Diseases
Study Group
University of Milan, 1 st Level Master in Clinical Research
Department of Molecular Biology, Unit of Medical Genetics, Policlinico “Le Scotte”, Siena
“BergamoScienza” Association
INTERNATIONAL COLLABORATIONS
International Registry of Recurrent and Familial Hemolytic Uremic Syndrome and Thrombotic
Thrombocytopenic Purpura
Information Centre for Rare Diseases and Orphan Drugs – ICRDOD, Bulgaria
Podonet: Consortium for Clinical, Genetic and Experimental Research into Hereditary Diseases
of the Podocyte – Coordinator: Heidelberg University Hospital, Germany
International Network and Registry for Thrombotic Microangiopathy (TMA) – Coordinator:
Schneider Children’s Hospital, Albert Einstein College of Medicine, USA
EDITORIAL COMMITTEE MEMBERSHIP
Journal of Nephrology (Arrigo Schieppati)
Quaderni di Farmacoeconomia (Erica Daina)
328
ANNUAL REPORT 2008

IRFMN
NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP
Network for Rare Diseases – Lombardy Region (Delibera Regione Lombardia N°7328,
11/12/2001)
Task Force on Rare Diseases (established by DG Health and Consumer Protection on 21
January 2004)
Scientific Committee A.O. Bolognini di Seriate
Ethical Committe, A.O. Ospedali Riuniti di Bergamo
Working group “Classification and coding of rare diseases” coordinated by Italian National
Institute of Health
EVENT ORGANIZATION
“La Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena incontra il
Centro di Ricerche Cliniche per le Malattie Rare Aldo e Cele Dacco’”
Clinical Research Centre for Rare Diseases “Aldo and Cele Daccò” – Mario Negri Institute
Ranica (Bergamo), January 24, 2008
“Una Speranza per le malattie rare, La Pre-Eclampsia”
Clinical Research Centre for Rare Diseases “Aldo and Cele Daccò” – Mario Negri Institute
Ranica (Bergamo), May 16, 2008
Focus on Rare Diseases “The Genetic and Molecular Basis of Rare Kidney Disorders”
Bergamo, October 9-11, 2008
PARTICIPATION IN EVENTS IN WHICH THE LABORATORY WAS
INVOLVED
11° Convegno “Patologia Immune e Malattie Orfane”
Torino, January 24-26, 2008
Prima Giornata Europea delle Malattie Rare “Un giorno raro per persone molto speciali”
Bergamo, February 29, 2008
Prima Giornata Europea delle Malattie Rare “Sviluppi della Rete Regionale per le Malattie
Rare”
Milan, February 29, 2008
XXI Convegno Nazionale Associazione Italiana per le Mucopolisaccaridosi e Malattie Affini -
AIMPS
San Donato Milanese (Milan), march 28-30, 2008
II Convegno Nazionale Associazione Italiana Sindrome di Shwachman-Diamond “Dal difetto
genetico alla terapia: venti anni nella conoscenza di una malattia rara”
Rimini, April 18-19, 2008
329
ANNUAL REPORT 2008

IRFMN
Convegno : Le malattie Croniche in età pediatrica
Brescia, April 19, 2008
Malattie Rare Impegno Comune
Rome, June 24, 2008
XXII International Complement Workshop
Basilea, September 28 – October 2, 2008
Corso di aggiornamento “Malattie Polmonari: percorsi diagnostici e terapeutici”
Milan, October 16, 2008
III Congresso Nazionale Associazione Pediatri in Gruppo - APeG
Florence, October 17-18, 2008
Congresso Internazionale Rare Diseases and Orphan Drugs
Rome, October 27-31, 2008
Workshop Consorzio MIA “Lupus Eritematoso Sistemico”
Milan, November 14, 2008
Progetto PARTECIPASALUTE: Le Associazioni definiscono le priorità della ricerca: uno
scenario possibile in Italia
Milan, December 2, 2008
Progetto di formazione sul campo: registro Regionale Malattie Rare
Presidi Rete Regionale, 2008
European Commission
"Fondazione Ricerca Malattie Rare", Bergamo
Lombardy Region
Fondazione Telethon
GRANTS AND CONTRACTS
SELECTION OF SCIENTIFIC PUBLICATIONS FROM 2008
A. Schieppati, J.I. Henter, E. Daina, A. Aperia
Why rare diseases are an important medical and social issue
Lancet; june 14 2008, vol. 371, pp. 2039-55
F. Castelletti, R. Donadelli, F. Banterla, F. Hildebrandt, P. Zipfel, E. Bresin, E. Otto, C. Skerka, A. Renieri, M.
Todeschini, J. Caprioli, M.R. Caruso, R. Artuso, G. Remuzzi, M. Noris
Mutation in fn1 cause glomerulopathy with fibronectin deposits
Proc Natl Acad Sci Usa. 2008, vol. 105, pp 2538-2543
A. Schieppati G. Remuzzi
Novel Therapies of Lupus Nephritis
330
ANNUAL REPORT 2008

IRFMN
Curr. Opin. Nephrol. Hypertens. 2008, Vol. 17, pp 156-61
OTHER PRODUCTS PUBLISHED IN 2008
Schieppati A, Daina E, Gamba S
Parla anche con me. L’esperienza di 18 anni del Centro di Informazione per le Malattie Rare
In: “Dire, fare, curare – Parole tra medici e malati”
Collana Salute e Società, FrancoAngeli Editore, novembre 2008
Cattaneo D, Daina E
Malattie Autoimmuni
In: “Antagonisti recettoriali dell’Endotelina: quali composti e quali patologie”
Il Pensiero Scientifico Editore, dicembre 2008
RESEARCH ACTIVITIES
Information Centre for Rare Diseases
In 1992, in the frame of the Clinical Research Centre for Rare Diseases “Aldo e Cele Daccò”
was established an Information Centre for Rare Diseases.
The Information Service is available to patients with rare diseases, their families and doctors. It
offers information, update and useful addresses free of charge with particular emphasis on
etiology, pathogenesis, genetics, treatments and availability of referral research centres.
One of the most difficult issues to address when studying rare diseases are those, related to the
recruitment of a sufficient number of patients with a given disease. The database of the
Information Centre for Rare Diseases has become a useful tool for identification of potentially
eligible patients for clinical studies. Till now, the Information Centre for Rare Diseases has
collected patients’ clinical data for about 200 different rare conditions with 10 or more cases.
Furthermore, intensive collaboration with groups with the same interest from Italy and abroad
provides many possibilities for starting of clinical projects with a multicentral design.
Bank of biological samples and description of hereditary nephropathies
The aim of this project is to collect clinical data and biological samples from patients and their
families with rare genetic conditions. A database with clinical data and a Bank for biological
samples collection and preservation has been created.
The availability of clinical data and biological samples is useful to perform new biochemical
and genetic tests within specific research projects aimed to better reveal the mechanisms of the
diseases, their manifestation and therapeutic opportunities.
In particular, the attention is focused on rare genetic disorders of the kidney. A thorough clinical
evaluation, including clinical data collection, medical physical examination, renal
ultrasonography, laboratory tests of blood and urine is offered to patients, affected by hereditary
nephropathies (Alport syndrome, Fabry disease, Familial Focal Glomerulosclerosis,
Glomerulopathy with Fibronectin deposits, Membranoproliferative glomerulonephritis,
Medullary Cystic Kidney disease, Cystinuria), who are addressing our Centre. After obtaining a
written informed consent, biological samples from patients and their relatives are collected,
labelled with specific codes to assure the anonymity and conserved in the Bank for biological
samples. In case the responsible gene for a hereditary nephropathy is known (Alport syndrome,
Fabry disease, Medullary Cystic Kidney disease, Cystinuria), the blood samples are redirected
to the relevant Laboratory of reference. For other nephropathies, where the identification of the
331
ANNUAL REPORT 2008

IRFMN
gene mutation is unknown or still in course, the blood samples are conserved with the aim to be
used in specific future research projects.
Evaluation of the long term efficacy of enzyme replacement therapy in
Fabry disease
Fabry disease is an X-linked disorder of the glycosphingolipid catabolism caused by the
deficient activity of the lysosomal hydrolase alfa-galactosidase A (A-gal A) in tissues and fluids
of affected hemizygous males. Most heterozygous females have an intermediate level of
enzymatic activity.
The enzymatic defect leads to a systemic deposition of glycosphingolipid in the heart, kidneys,
eyes and other organs and tissues. Preliminary studies of enzyme replacement therapy
demonstrate that periodic infusions of recombinant human Alfa-galactosidase A are safe (in
terms of infusion reactions) and effective in patients with Fabry disease.
The purpose of this study is to evaluate the long term efficacy of enzyme replacement therapy in
patients with Fabry disease and renal involvement.
International Registry of Recurrent and Familial Hemolytic Uremic
Syndrome and Thrombotic Thrombocytopenic Purpura
Hemolytic Uremic Syndrome (HUS) and Thrombotic Thrombocytopenic Purpura (TTP) are
rare diseases of microangiopathic haemolytic anemia and thrombocytopenia with signs of renal
(most prevalent in HUS) and cerebral (most prevalent in TTP) damage. There are extremely rare
forms of HUS and TTP, often occurring in families, in which the patients relapse even after
complete recovery of the first episode with permanent renal and neurological sequelae. In the
familial and recurrent forms of HUS and TTP, the attention is concentrated mainly on the
genetic predisposition to the disease.
The Laboratory coordinates the International Registry of Recurrent and Familial HUS/TTP,
since 1996. The Registry has collected more than 540 cases of HUS/TTP referred from 100
Italian and 80 European and extra-European Centres. Clinical and laboratory data of all patients
referred to the Registry are collected by a uniform data extraction form. The family history and
also the personal data of the unaffected relatives are collected, when possible. Biological
samples are collected from all patients and available relatives, for the biochemical and genetic
analyses. Many research projects in collaboration with the Laboratory of Immunology and
Genetic of Rare Diseases and Organ Transplantation and the Laboratory of Cellular Biology –
Negri Bergamo are developed and have still allowed to identify some genetically determined
forms of HUS and TTP. The maintenance of a centralised bank of biological samples ensure the
availability of clinical material for new investigative approaches as they will be developed.
Rituximab in relapsing and chronic thrombotic thrombocytopenic purpura
In most cases, TTP is related to an acquired deficiency of ADAMTS13 activity, due to the
presence of autoantibodies anti-ADAMTS13. The absence of ADAMTS13 activity leads to the
accumulation of von Willebrand Factor ultralarge multimers (normally cleaved to smaller
molecular forms by ADAMTS13 enzyme), which probably induces platelets aggregation and
the ensuing process of thrombotic microangiopathy.
The purpose of the project is to evaluate the use of a monoclonal anti-CD20 antibody
(Rituximab) in patients with relapsing and chronic thrombotic thrombocytopenic purpura,
related to the presence of anti-ADAMTS13 antibodies. Rituximab is a humanized monoclonal
antibody that targets the CD20 molecule on B cells (white cells producing antibodies).
Administration of Rituximab leads to a selective B cell depletion that usually lasts 6 to 9
months. Inhibition of B cell activation or growth by rituximab treatment limit the uncontrolled
synthesis of autoantibodies, that may predispose to chronic relapsing TTP.
332
ANNUAL REPORT 2008

IRFMN
Rituximab is infused intravenously once weekly for a total of four infusions. After Rituximab
treatment, the patients are periodically controlled with medical examination, blood and urine
exams, and with ADAMTS13 plasma activity assay and research of anti-ADAMTS13
autoantibodies, in collaboration with the Laboratory of Cellular Biology – Negri Bergamo.
Observational period lasts 12 months.
Evaluation of urinary podocyte excretion
Recent evidence recognize an important role for podocytes in the progression of renal diseases.
Podocytes contribute to glomerular permeability and are important target cell for renal disease
progression. Glomerular injury is usually associated with the leakage of protein across the filter
into the urine and with the disappearance of podocyte foot process. Injuried podocytes either
undergo apoptosis or detach from the glomerular basement membrane, with subsequent
appearance in the urine. Urinary excretion of podocytes has been reported as possible predictor
of disease progression in a number of progressive glomerular diseases, such as IgA
nephropathy, focal segmental glomerulosclerosis, diabetic nephropathy. During the past years
the Clinical Research Center for Rare Diseases, in collaboration with the Department of
Molecular Medicine at Mario Negri Institute, has established an indirect immunohistochemistry
method for the detection of podocytes in urinary sediments. This method has allowed to identify
and quantitate the extent of urinary podocyte loss in patients with glomerular disease. The
present study has the aim to evaluate excretion of urinary podocytes in rare genetic renal
diseases. This methodology, applied on a larger number of patients, could provide a non
invasive way of indexing the extent of glomerular damage and a useful instrument to evaluate
the response to treatment.
Effects of an intensified treatment with ACE-inhibitors, Angiotensin II
receptor antagonists and Statins in Alport syndrome
Alport syndrome (AS) represents a form of progressive hereditary nephritis in which the genetic
defect resides in the synthesis of one of several subunits of type IV collagen, the predominant
constituent of basement membranes in renal glomeruli. Renal impairment occurs with time and
severe renal failure with hypertension and uremia represent the end stage of the disease, even if
a high variability in the rate of progression is described. The prognosis is variable. Males are
usually affected by a progressive form of the disease. Affected females with X-linked syndrome
usually have a good prognosis with a mild renal impairment. Other clinical manifestations are
anterior lenticonus which occur in about one third of patients. Other findings are myopia, lens
opacities and retinal pigment abnormalities and corneal vesicles or erosions. The disease is also
associated to a sensor neural deafness which can occur in approximately half of the patient
affected and usually correlates with renal impairment.
No definite treatment exists in order to delay the time of dialysis or a kidney transplant. Many
studies showed that Angiotensin converting enzyme (ACE) inhibitors slow glomerular filtration
rate (GFR) decline and limit progression to end stage renal disease (ERDS) and dialysis in
several chronic nephropathies associated with proteinuria. The combination of ACE-I with
Angiotensin II receptor antagonists may reduce proteinuria more effectively than the two drugs
alone. Moreover the addition of Statins may synergize the antiproteinuric effects of ACE-I and
ATAII antagonists in experimental models of chronic renal diseases.
The purpose of this study is to evaluate the effect of a standardized multimodal
nephroprotection intervention (Remission Clinic) in Alport patients with renal involvement.
Nine patients with Alport syndrome and macroalbuminuria will be enrolled in this study.
Recruitment of normo or microalbuminuric patients will be stopped and the analysis will be
performed on the patients that will be available when the recruitment phase of
macroalbuminuric patients will be completed.
333
ANNUAL REPORT 2008

IRFMN
Update of the Congenital Respiratory Malformations database
This Project originated from the collaboration between the Study Group on Respiratory Disease
of the newborn and the Coordinating Centre for Rare Diseases.
The Congenital Respiratory malformations database was conceived to facilitate the diagnostic
and assistance procedures for paediatricians facing with more frequent respiratory
malformations. The update consisted of a review of the lists of malformation categories and of
all syndromes previously included; more syndromes have been added.
Furthermore, in the brief description, more detailed than the previous version, the Code for Rare
Disease was added, allowing to address the patient to the Referral Centres for Lombardy.
A link with the OMIM database is provided for each syndrome for a better description of the
disease.
Identification of new genes associated to the Autosomal Dominant
Familial form of Focal Segmental Glomerulosclerosis
Focal segmental glomerulosclerosis (FSGS) is a pathological entity, and is a significant cause of
end-stage renal disease (ESRD). Glomerular disease is the third leading cause of ESRD, and
FSGS comprises a significant proportion of this subgroup: up to 5% of adults and 20% of
children with ESRD.
The diagnosis of FSGS is based on renal pathology. The clinical hallmarks include proteinuria,
nephrotic syndrome, occasional hematuria and frequent progression to ESRD. Hypertension is
also a common finding. At present, there are no consistently reliable treatments for FSGS and
response rates to available treatments have been estimated at

IRFMN
Complement abnormalities in primary Membranoproliferative
glomerulonephritis
Primary membranoproliferative glomerulonephritis (MPGN) is an uncommon cause of chronic
renal disease that occurs principally in children and young adults, and that often has an
unfavourable prognosis.
Data on incidence and prevalence of MPGN are scanty, thus a first aim of this project is to
collect through a Registry clinical data and biological samples from well characterized patients
with primary MPGN. Thus, a case report form has been designed in order to obtain
homogeneous clinical data for all recruited patients. The family history will be also recorded
and biological samples will be collected from the patients to perform biochemical and genetic
analyses. Genetic counselling will be provided to all patients and relatives.
The pathogenesis of primary MPGN is ill defined. The available data indicate that excessive
activation of complement due to autoimmune and genetic abnormalities plays a role in inducing
damage to the kidney and other organs. The complement is a complex system that mediates the
immune response against bacteria and viruses. In normal conditions human cells are protected
from complement attack by several inhibitors that restrict complement activation to the surface
of pathogens. Such regulatory system is defective in MPGN. Thus a major goal of this project,
in collaboration with the Laboratory of Immunology and Genetic of Rare Diseases and Organ
Transplantation, will be the identification of the genetic and acquired defects underlying
complement activation in MPGN and to establish whether specific complement abnormalities
are associated with disease progression, response to therapy and recurrence on a transplanted
kidney.
335
ANNUAL REPORT 2008

IRFMN
336
ANNUAL REPORT 2008

IRFMN
The Transplant Research Center
Chiara Cucchi De Alessandri e
Gilberto Crespi
The Transplant Research Center (CRT) was set up in 2002 to support and promote the work of
outstanding research scientists throughout the world and to carry out major organ transplant
research programs.
The Center is housed in the Villa Camozzi, at Ranica, under the same roof as the Mario Negri
Institute in Bergamo and is managed in collaboration with the Institute.
The Center’s staff is mainly made up of senior and junior researchers that were trained in the
laboratories of the Mario Negri Institute in Bergamo, focusing on transplant immunology,
research for less toxic immunosuppressant drugs, and new gene therapy techniques to prevent
acute rejection of transplanted organs.
Information on the Center’s activities can be found in the sections addressed to the Department
of Molecular Medicine (Laboratory of Immunology and Genetics of Organ Transplantation and
Rare Diseases) and the Department of Renal Medicine (Laboratory of Pharmacokinetics and
Clinical Chemistry).
337
ANNUAL REPORT 2008

IRFMN
338
ANNUAL REPORT 2008

IRFMN
EDUCATIONAL ACTIVITIES
Dean, Mario Salmona, PH.D.
The Institute's training programs fall under the heading of biomedical science, and are part of
the Lombardy Region's professional training schemes. There are courses for specialized
laboratory technicians, and for graduates intending to do research. In 1999 the Institute has set
up a Ph.D. course, in collaboration with the Open University of London. This degree is
recognised throughout Europe and in the USA. In 2006 the Institute also set up a First Level
master Course in Clinical Research, in collaboration with the Milan University and a Second
Level master Course in rare Diseases, in collaboration with the University of Torino.
In 2008 the Institute started a two-years Advanced School in Applied Pharmacology (SAFA).
The course provides advanced cultural education and practical experience with experimental
work in the laboratories.
Students enrolled in formal courses receive one month preparatory training, after which they are
assigned study grants. Between 1963 and 2007 the Mario Negri Institute awarded 6,578
grants, 701 of them to foreign researchers who came to the Institute for special training.
Everything possible is done to help students find work once they finish the course.
The main feature of these courses is that technicians and researchers receive their training "on
site". They work full-time in research programs of a high scientific standard, using advanced
equipment and learning the latest methods, in regular contact with colleagues in different
countries. Besides its scientific value, this approach provides an excellent preparation on the
human and personal scale.
Students are usually assigned to one of the Institute’s Laboratories, where they gradually gain
specialized skills by working on specific research projects. They are expected to attend lessons,
seminars, courses and congresses and learn to make full use of the Institute’s well-stocked
library. Students all have access to the internet and to biomedical database and can print out
copies of articles they need to consult, from major international journals. Should the opportunity
arise, students are expected to be available for trips abroad, to participate in conferences or
courses.
The Pharmacological Research Specialists and Biochemical Research Technicians will receive
diplomas issued officially by the Lombardy Region and the Mario Negri Institute for
Pharmacological Research. These have legal value throughout Italy, and are recognised in
competitions for public posts, where they are worth a certain number of points. Mario Negri
Institute diplomas are widely considered a guarantee of an excellent theoretical and practical
training. The Open University of London Ph.D. degree earned at the Institute has legal value
throughout Europe and in the USA.
Once they have their diploma or Phd., graduates who want to continue doing research at the
Institute may be offered a chance to spend a year or two abroad.
339
ANNUAL REPORT 2008

IRFMN
The SAFA course is aimed at preparing young researchers and enabling them to specialize and
work for the pharmaceutical industry, for other research institutes and for public institutions,
such as the Regions, for instance. Some of our students, after the SAFA course, decide tu enroll
the PhD programs offered by our Institute, while others decide to study abroad.
At the moment these courses are available:
• Three-year course for graduates, in Milan or Bergamo, leading to a diploma as
Pharmacological Research Specialist.
• Three-year course for diploma-holders, in Milan or Bergamo, leading to a diploma as
Biochemical Research Technician.
• Research doctorates (Ph.D.), run under an agreement with the Open University of
London. And the Universities of Maastricht and Groningen (NL).
• First Level Master in Clinical Research , in collaboration with the University of Milano.
• Second Level Master Course in Rare Diseases, in collaboration with the University of
Torino.
• Two-years Advanced School in Applied Pharmacology for graduate students, in Milano
and Bergamo.
Other training opportunities
PREPARING A THESIS FOR A DEGREE:
Students can prepare their thesis in scientific subjects at the Institute, with the approval of their
university faculty. These students must work at the Institute for at least two years.
SUMMER STUDENTS
In June and July each year the Institute accepts a certain number of students in their last two
years at high school, to give them experience as part of school/work programs.
340
ANNUAL REPORT 2008

IRFMN
STAFF
Executive Offices
Services and Offices
341
ANNUAL REPORT 2008

IRFMN
342
ANNUAL REPORT 2008

IRFMN
Prof. Silvio Garattini
SIlvio Garattini was born in Bergamo (Italy) on 12 November 1928. He earned a diploma in Chemistry,
then a degree in Medicine and was appointed lecturer in Chemotherapy and Pharmacology. He held the
post of Assistant then Deputy Professor at the Milan University Institute of Pharmacology, until 1962.
A founder of the Mario Negri Institute for Pharmacological Research, when it opened in 1963 he was its
director. The Institute now has four locations (Milan, Bergamo, Ranica (Bg), S. Maria Imbaro (Ch)) and
more than 950 people work there. Professor Garattini is a member of the Gruppo 2003 (a group of the
most cited Italian scientists in international scientific literature) and has hundreds of publications in
Italian and English in international scientific journals, and texts on pharmacology. He was a founder of
the European Organisation for Research and Treatment of Cancer (EORTC).
In the last ten years Professor Garattini has acted in various organizations, including the Italian National
Research Council (CNR) - Committee on Biology and Medicine; the National Health Council, the
Committee for Italian Research Policy, set up by the Presidency of the Council of Ministers; the Ministry
of Health Commissione Unica del Farmaco (CUF). He has held the following posts: President of the
UICC Committee on Antitumoral Chemotherapy, President of the European Organisation for Research
and Treatment of Cancer (EORTC), Consultant to the World Health Organisation; President of the
European Society of Biochemical Pharmacology; member of the Committee for Proprietary Medicinal
Products (CPMP) of the European Agency for the Evaluation of Medicinal Products (EMEA); member of
the Committee of Experts of Research Policy – CEPR - at the Ministry for University and Scientific and
Technological Research; member of the Scientific Committee of the Lega Italiana per la Lotta Contro i
Tumori; member of the Board of the Istituto Superiore di Sanità; Chairman of the Committee for
Research of the Italian Agency for Drugs (AIFA). Other appointments include: Vice-president of the
Consiglio Superiore di Sanità; President of the Research and Development Commission of the Agenzia
Italiana del Farmaco (AIFA); President of the Angelo and Angela Valenti Foundation and the "Via di
Natale" Foundation; President of the Technical Commission for Pharmaceutical Assistance of the
Regione Autonoma of Sardinia. He is a member of the Strategic Committee for Welfare, Regione
Lombardia, the Consiglio Superiore di Sanità and the Comitato Nazionale di Bioetica, the International
Scientific Committee, Centro di Riferimento Oncologico, Aviano, and the Scientific Committee of
AISLA, Member Comitato Scientifico, Dipartimento Politiche Antidroga Presidenza del Consiglio dei
Ministri, Member, Advisory Board ADAMO Onlus, Milan, Member of Committee of the
Recommendation for the Call, Wemos Foundation, Amsterdam, , Member, Development Advisory
Board of the International Center for Biomedical Sciences (ICBMS), Luxu Town, Wujiang City, China.
Silvio Garattini is a Fellow of the New York Academy of Sciences, the American Association for the
Advancement of Science, Honorary Fellow of the Royal College of Physicians (Pharmaceutical
Medicine), London, Honorary Fellow of the Italian Society of Pharmacology and a member of numerous
other Italian and international scientific societies. He has received many awards for his work, including
the French Legion d'Honneur for scientific merit, and the Grand Ufficiale della Repubblica Italiana, and
holds honorary degrees from the Universities of Bialystok in Poland, and Barcelona in Spain. Recent
awards include the Ippocrate Prize, 2003; Mens Sana in Corpore Sano; Nuova Spoleto, 2003; Angelo
dell’anno; Alkmeon International Prize; International Prize Sant’Agostino Città di Bergamo; Il Campione
della Scienza; Natta Medal; Coppola Prize, Scienza e Società in the framework of the Premio Città di
Firenze and Premio Rana d’Oro, Casalbeltrame (Novara). In its 40-plus years, the Mario Negri Institute
for Pharmacological Research, under Professor Garattini's leadership, has published more than 11,000
scientific papers and more than 250 books, on topics ranging from cancer and its treatment to tumour
immunology, neuropsychopharmacology, and cardiovascular and renal pharmacology. More than 4000
young Italian and 600 foreign graduates and technicians have obtained specialist qualifications at the
Institute.
343
ANNUAL REPORT 2008

IRFMN
344
ANNUAL REPORT 2008

IRFMN
Prof. Giuseppe Remuzzi
Giuseppe Remuzzi was born in Bergamo, Italy in 1949. Upon completion of his medical training at the
University of Pavia in 1974, he received specialty training in Hematology and Nephrology at the
University of Milan.
Since 1975, he has pursued his academic career at the Ospedali Riuniti of Bergamo, where he was
appointed Professor of Nephrology and Director of the Department of Medicine and Transplantation in
1996. Since 1999, he is Director of the Department of Nephrology and Dialysis of the same hospital. The
Negri Bergamo Laboratories, which he has directed since 1984, is a group of basic scientists,
physiologists, pharmacologists, molecular and cellular biologists, pathologists and clinicians devoted to
the study of renal disease. As Research Coordinator of the Negri Bergamo Laboratories and the affiliated
Clinical Research Center for Rare Diseases "Aldo e Cele Dacco", both divisions of the Mario Negri
Institute for Pharmacological Research, he conduct a diverse team of researchers studying human renal
diseases and their corresponding experimental models from the perspective of pathophysiology and
therapeutic intervention. He touched major advances in many areas of nephrology, with particular focus
on platelet endothelial interactions, vascular prostaglandin biology, coagulation and renal disease,
progression of renal disease, experimental models of glomerular damage, and transplant immunology and
tolerance. Particularly his contributions to the understanding of the pathophysiology of hemolytic uremic
syndrome, prostaglandin metabolism in pregnancy, renal vascular biology in uremia, the role of protein
trafficking in renal disease progression, the induction of graft tolerance by intrathymic injection of donor
antigens, the role of the co-stimulatory CD28-B67 pathway in transplant rejection and the prevention of
renal and cardiovascular damage in diabetes.
Prof. Remuzzi serves on editorial boards of numerous journals including the prestigious New England
Journal of Medicine and is member of the International Advisory Board of The Lancet.
In recognition of his achievements, he has been awarded in 1998 honorary memberships of the
Association of American Physicians and the British Royal College of Physicians. In 2001 he was
nominated Chairman of the Research Committee of the COMGAN (Commission on Global Advancement
of Nephrology) of the International Society of Nephrology (ISN). He received an Honorary Professorship
at the University of Maastricht in 2003.
In 2005 during the World Congress of Nephrology in Singapore he received the ISN Jean Hamburger
Award and in November 2007 he received during the annual congress of the American Society of
Nephrology the precious John P. Peters Award.
On August 2008, he was appointed "Honoris Causa Professor" by the Catholic University of Cordoba
(Argentina).
Prof. Remuzzi has authored and co-authored more than 979 scientific articles, reviews and monographs.
345
ANNUAL REPORT 2008

IRFMN
346
ANNUAL REPORT 2008

IRFMN
Mario Negri Institute Milan
Executive Office
Director
Prof. Silvio GARATTINI, M.D.
Administrative Office
Maria Grazia PEZZONI, Chief
Technical Office
Fabio BRIGHENTI, D. Arch.
General Maintenance
Emanuele SINELLI
Studies Office
Armanda JORI, Pharm.D.
Press Office
Isabella BORDOGNA, Phil. D.
Public Relations Office
Claudio PANTAROTTO, Comm.
Prevention and Safety Office
Emilio BENFENATI, Chem.D.
Annamaria SEGALINI, Phys.D.
English Style Editor
Judi BAGGOTT
Photography and Audio-Visual Service
Felice DE CEGLIE
Purchasing Office
Eufrasia COVIELLO
Director’s Office
Rosanna MAPELLI, Chief
General Secretariat
Elena POZZOLI
347
ANNUAL REPORT 2008

IRFMN
348
ANNUAL REPORT 2008

IRFMN
Negri Bergamo Laboratories
Executive Offices
Director
Research Coordinator
Scientific Secretariat
Prof. Silvio GARATTINI, M.D.
Giuseppe REMUZZI, M.D.
Ariela BENIGNI, Biol.Sci.D., Ph.D.
Administration of Research Projects/Admn. Assistance
Daniela MELACINI, Biol.Sci.D.
Press and Communication Office
Francesca Di Fronzo, Mod. Lit. D.
Audio-Visual Service
Antonella PICCINELLI, Biol.Sci.D.
Prevention and Safety Office
Chief
Annamaria SEGALINI, Phys.D.
Library
Chief
Anna BOZZALE
Valeria MIGLIOLI
General Maintenance
Giancarlo GASPARI
Director’s Office
Antoinette van ENGELEN, Chief
349
ANNUAL REPORT 2008

IRFMN
350
ANNUAL REPORT 2008

IRFMN
Centro Aldo e Cele Daccò Ranica (Bg)
Executive Offices
Director
Research Coordinator
Scientific Secretariat
Health Director
Prof. Silvio GARATTINI, M.D.
Giuseppe REMUZZI, M.D.
Ariela BENIGNI, Biol.Sci.D., Ph.D.
Norberto PERICO,M.D.
Press and Communication Office
Francesca Di Fronzo, Mod. Lit. D.
Prevention and Protection Office
Chief
Annamaria SEGALINI, Phys.D.
Paola BOCCARDO, Biol.Sci.D.
Library ‘Mansueto Astori’
Chief
Anna BOZZALE
Monica MINALI
Director’s Office
Barbara REMONTI, Oriental Languages D.
Clinical Trials Office
Paola BOCCARDO, Biol.Sci.D.
Custodian/general maintenace
Giampiero CUGUSI
351
ANNUAL REPORT 2008