FNPS Japan 2007 - Fetal and Neonatal Physiological Society

35th Annual Meeting ofThe Fetal and Neonatal Physiological Society (FNPS)Hotel Van der Valk, Maastricht, the Netherlands22 – 25 June 2008Maastricht, the Netherlands, awaits you in 2008!During this FNPS meeting we will celebrate the careers ofJelte de Haan and Carlos Blanco,both of whom will have retired by 2008MaastrichtIn June the weather will be mild and sunny in Maastricht. The oldest and most Burgundian city in theNetherlands offers many ways to relax (terraces, pubs and restaurants). Located in the hilliest part ofthe Netherlands, its hills and marlstone caves offer great views and numerous possibilities for nicewalks, biking, horse-riding, etc. Situated in the center of Europe, Maastricht is a one-hour drive fromBrussels or Düsseldorf, a two-hour drive from Amsterdam. The city also has its own airport withconnections to London, Amsterdam, Pisa, etc.Hotel Van der ValkKnown for its spacious rooms and reasonable prices, the Hotel van der Valk in Maastricht is a luxury5-star hotel and only a 10-minute walk from the centre of town. The 2007 price for a standard room is €85 and a business suite is € 132,50. When sharing a room, this conference will certainly offer “valuefor money.”Society Sport EventThe classic FNPS Sport Event will not be forgotten. With the Dutch organizing things, you can be surethat we will have a real competition!Organizing CommitteeThe familiar faces from the Netherlands (i.e. Bilardo, Blanco, De Haan, Mulder, De Vries, Erwich,Hasaart, Nijhuis, Oei, Van den Berg, Visser, etc.) will participate in the Organizing Committee to createa meeting that is successful - both scientifically and socially.Contact for the Dutch 2008 FNPS MeetingJan G. NijhuisConference Agency Maastrichte-mail: jnij@sgyn.azm.nlinfo@conferenceagency.comT +31 43 387 47 64 T +31 43 361 91 92P.O. Box 5800 P.O. Box 1402NL-6202 AZ MaastrichtNL-6201 BK MaastrichtPage 7

Previous meetings of the FNPS1974 Oxford, United Kingdom1975 Oxford, United Kingdom1976 Malmö, Sweden1977 Oxford, United Kingdom1978 Nijmegen, The Netherlands1979 Paris, France1980 Oxford, United Kingdom1981 Maastricht, The Netherlands1982 London, Canada1983 Malmö, Sweden1984 Oxford, United Kingdom1985 Haifa, Israel1986 Banff, Canada1987 Groningen, The Netherlands1988 Cairns, Australia1989 Reading, United Kingdom1990 Pacific Grove, USA1991 De Eemhof, The Netherlands1992 Niagara-the-Lake, Canada1993 Plymouth, United Kingdom1994 Palm Cove, Australia1995 Malmö, Sweden1996 Arica, Chile1997 S.Margherita Ligure, Italy1998 Lake Arrowhead, USA1999 Vlieland, The Netherlands2000 Southampton, United Kingdom2001 Auckland, New Zealand2002 Prague, Czech Republic2003 Banff, Canada2004 Tuscany, Italy2005 Glenelg, South Australia2006 Cambridge, UKPage 8

Social EventsMonday, August 27Sporting Event16:00-Activity Room 3F 「Matsushima Minami」, Hotel Sendai PlazaDress code: CasualNintendo Wii WILL OFFER UNPRECEDENTED VIDEO GAME EXPERIENCES FOR EVERYONEAn afternoon of fun, the highlight of which will be the fun sporting game.2006 - For more than 20 years, video game players have used body language to "help" them play.With Nintendo's upcoming Wii? console, those movements become a real part of the play. Aftergrabbing the Wii Remote for the first time, hesitation gives way to concentration. Confidence builds.Excitement morphs into pure delight. And everyone watching says the same thing: "Hey - let metry!"What drives this phenomenon? A remarkable controller and games that enhance the experience …realizing that the swing of your arm - not the movement of your thumb - causes a baseball to leavethe park or a sword to find its mark. The Wii console introduces the next leap in gaming, one whereplayers not only control their characters on the screen, but they also become them.Why not test your ability and take part? Participants will be divided into groups. The winning teamwill receive a prize.Monday, August 27Dinner Sendai Night19:30-Typical Japanese restaurant in Sendai cityDress code: CasualEnjoy a typical Japanese restaurant, which is called “Izakaya” in Japanese.Izakaya are drinking places that offer a variety of small dishes, such as robata (grilled food), saladsand finger food. It is probably the most popular restaurant type among the Japanese people.Izakaya tend to be informal, and the people at one table usually share all dishes, rather thanordering and eating individually.Page 9

Social Events continuedTuesday 28 th AugustExcursion14:00-20.30Matsushima sightseeingZuiganji was founded in 828 as a temple of the Tendai sect. It is now one of the Tohoku's mostfamous Zen temples, well known for its beautifully painted sliding doors (fusuma). Zuiganji Templewas built by Lord Date Masamune. The main hall was built in the traditional study room style ofarchitecture known as shoinzukuri. The roof of its magnificent, single-story main structure is tiled inthe hongawarabuki style, in which round and square tiles are laid down alternately. Many parts ofthe temple have been designated as national treasures or important cultural properties. The mainhall, priests' quarters and corridors are designated national treasures.Matsushima Bay CruiseDress code: very casualThe fun excursion cruise will be held in the afternoon. You can enjoy talking with other friends andcolleagues with some foods and drinks on a ship for 2 hours.Matsushima, a small bay dotted with more than 260 islands beautifully covered with evergreens,is one of Japan's celebrated Three Views. Poems have been composed on Matsushima since oldendays. "Matsushima is the most scenic place in Japan," wrote Matsuo Basho, the master haiku poet."… Some islands soar, pointing to the heavens, while others crouch, creeping on the waves….Thepines are deep green, and their branches and needles have been bent by exposure to the seabreeze….The landscape is enchanting, like an embellished beauty's face."Wednesday 29 th AugustGala dinner19.30-21.30Main Hall 3FDress code: Lounge suitsThis promises to be the social highlight of the meeting and is definitely an evening not to be missed.Entertainment will be supplied by Show Asano who is a brilliant shamisen player.shamisen is a stringed instrument shaped like a banjo, but its outstanding feature is that it has nofrets. It is played by striking its three strings with a plectrum and is used as accompaniment forKabuki and bunraku plays and for folk songs. It came from China through Okinawa and spreadthroughout Japan, in which it developed independently from the original style. It became therepresentative instrument of Japan since the Edo Period(1603-1867).Page 10

Notes for speakersOral presentationOral presentations will take place in the Main Hall at the Hotel SendaiPlaza.All oral presentations will be given in Microsoft Powerpoint Office 2000/2002/2003 on a PC.Speakers are asked to submit their presentations on CD or a memory stick to the technician on thePC preview desk at least one hour before their presentation.If you wish to use Macintosh computer or Windows VISTA or moving images, please bring your owncomputer.Oral presentations will be 10 minutes in length followed by a 5 minute discussion. Due to the timeschedule, speakers must ensure their presentation does not exceed this time limit.There will be a prize for the best oral presentation, which will be presented at the Gala dinner onAugust 29th (Wed).Poster presentationPosters can be displayed throughout the meeting. However, there will be a 1.5 hour formal postersession(Wednesday 29th August 11:00 - 12:30).These will be held in Poster Presentation Room(Matsushima Higashi)..There will be a prize for the best poster presentation, which will be presented at the Gala dinner onAugust 29th (Wed).Set-up and Removal Hours:Set-up: Sunday, August 26, 10:00-13:00Removal: Wednesday, August 29, 16:45-17:00Poster Guidelines:Poster presenters are expected to be present at their display during thefollowing hours:Wednesday, August 29, 11:00-12:30 (Poster Session)Please Note:*Posterboard size: 180*180cm*A presentation number to be placed at the top left of the poster will beprovided by the secretariat. Each author is requested to indicate "the title,""the authors' names"and "the authors' affiliations" at the top right of the panel witin anarea measuring 160cm wide by 20cm high.*Posters are attached to the boards with thumbtacks, which will beprovided by the Secretariat. No paste, glue, staples or nails are permitted.*Presenters are asked to remain with their posters during the aboveassigned hours to answer questions and expand on their material for visitors.Page 11

General InformationRegistrationRegistration Desk will be located on the 3 rd floor at Hotel Sendai Plaza.Opening hours:Satuday 25 th August 17:00-19:00Sunday 26 th August 9:00-17:00Monday 27 th August 7:30-19:30Tuesday 28 th August 8:00-14:00Wednesday 29 th August 8:00-19:30It will be manned throughout the meeting and should be the first port of call for delegates andaccompanying persons requiring any assistance.Internet accessInternet access will be available, free of charge on the 3 rd floor at Hotel Sendai Plaza. Out ofconsideration to others, delegates are kindly requested to limit their access time to 10 minuteseach.Trade ExhibitorsTrade Exhibitors will be located on the 3 rd floor at Hotel Sendai Plaza.Please take the time to visit them as they have been generous sponsors of this meeting.RefreshmentsAfternoon tea will be served in the poster presentation room(Matsushima Higashi) on theLunch will be served in the activity room(Matsushima Minami).Page 12

ACCESS TO SENDAIPage 13

SENDAI HOTEL MAPHOTEL SENDAI PLAZA2-20-1 HON-CHOU AOBA-KU, SENDAI-CITY, MIYAGI, 980-0014 JAPANTelephone: 022-262-7111 Facsimile: 022-262-8169Page 14

HOTEL SENDAI PLAZA 3FPage 15

AT A GLANCEPage 16

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ProgramPresenting authors are underlinedSATURDAY, AUGUST 25, 200717:00-19:00 Registration19:00 You can freely participate in the banquet of JSMES* meetingSUNDAY, AUGUST 26, 20079:00- Registration12.00 – 12.45 LunchPlenary lecture 1 :ChairpersonTsuyomu Ikenoue13:30-14:00 Fetal Heart Rate Monitoring: Is it Time to Establish International Consensus onIntrapartum Management?Julian T. Parer14:00-14:05 DiscussionPlenary lecture 2 :ChairpersonsShiro Kozuma14:05-14:35 Developmental Origins of Disease - a Japanese problem and a Japaneseanswer?Mark Hanson14:35-14:40 DiscussionPlenary lecture 3 :ChairpersonYoshio Matsuda14:40-15:10 An excess of circulating antiangiogenic factors: the cause of the maternalhypertensive syndrome of preeclampsia Richard Levine15:10-15:15 Discussion15.15 – 15.45 Afternoon teaPlenary lecture 4 :ChairpersonHiroshi Chisaka15:45-16:15 The affect of maternal hyperglycemia on fetal outcomeHuixia Yang16:15-16:20 DiscussionPage 21

SUNDAY, AUGUST 26, 2007 (continued)Plenary lecture 5 :ChairpersonTakashi Okai16:20-16:50 The pathogenesis of preterm brain injuryLaura Bennet16:50-16:55 DiscussionDINNER(We offer a meal coupon. You can choose one of the restaurant in Hotel Sendai Plaza.)MONDAY, AUGUST 27, 20077.30 – RegistrationSESSION 1 : Fetal GrowthChairpersonsGraham JenkinAlison Forhead8:00-8:30 Keynote speakerAbstract1: Impact of low birthweight on developing organs: separating the effectsof IUGR and preterm birth.(including Discussion)Richard Harding8:30-8:40 Abstract2: Postnatal high-fat feeding impacts on metabolic profiles andgluconeogenic capacity during fasting and this is exaggerated by prenatalundernutrition in adolescent lambsAnna Hauntoft Kongsted and Mette Olaf Nielsen8:40-8:45 Discussion8:45-8:55 Abstract3: Maternal Isocaloric High-protein Diet Ameliorates the Elevation in bothBlood Pressure and Cardiac Remodeling in the Adult Mice Offspring withMaternal Calorie Restriction Possible Involvement of Branched-Chain AminoAcids-Hiroaki Itoh, Makoto Kawamura, Shigeo Yura, Haruta Mogami,Fujii Tsuyoshi and Norimasa Sagawa8:55-9:00 Discussion9:00-9:10 Abstract4: Maternal ovine cortisol concentrations are important for key endocrinefactors involved in fetal growthEllen Jensen, Bennet L, Wood CE, Gunn AJ and Keller-Wood MPage 22

MONDAY, AUGUST 27, 2007 (continued)9:10-9:15 Discussion9:15-9:25 Abstract5: Diet-Induced Obesity and Prenatal Undernutrition Lead to CentralLeptin Resistance by Different MechanismsMhoyra Fraser, Charisma K Dhaliwal, Stefan Krechowec,Mark Vickers and Bernhard H Breier9:25-9:30 Discussion9:30-9:40 Abstract6: Impact of maternal exposure to constant light during late gestation onpostnatal adrenal function in the capuchin monkey: long terms effects on DHASand transient effects on cortisol.Claudia Torres-Farfan, N Mendez, H Richter, Gj Valenzuela and M Seron-Ferre9:40-9:45 Discussion9:45-9:55 Abstract7: Maternal signals control the fetal biological clockHidenobu Ohta, Xu Shanhai, Takahiro Moriya, Masayuki Iigo, Tatsuya Watanabe,Norimichi Nakahata, Hiroshi Chisaka, Tadashi Matsuda, Toshihiro Ohura,Yoshitaka Kimura, Nobuo Yaegashi, Shigeru Tsuchiya, Hajime Tei and Kunihiro Okamura9:55-10:00 Discussion10:00-10:10 Abstract8: Dietry protein restriction of pregnant mouse induces alteredmethylation of Oct-4 and Sphk-1 gene promoters in the liver of offspring.Jun Murotsuki, Kunihiro, Okamura, Hiroi Kaku, Yoshihiko Araki,Hiroshi Yoshitake, Mark A. Hanson and Felino R. Cagampang10:10-10:15 Discussion10:15-10:25 Abstract9: Transcriptional suppression of IGF-2 mediated by altered histonemodifications in a maternal protein restricted IUGR mouse modelMichiyo Nakamura, Sharif J, Ito T, Chisaka H, Kimura Y, Mitsuya K and Okamura K10:25-10:30 Discussion10:30-11:00 Morning tea (poster viewing)SESSION 2 : NeonateChairpersonsR Usha RajIan Wright11:00-11:30 Keynote speakerAbstract10: The state of the art in head cooling for neonatal encephalopathyAlistair J Gunn(including Discussion)Page 23

MONDAY, AUGUST 27, 2007 (continued)11:30-11:40 Abstract11: Cerebral oxygenation measured by spatially resolved spectroscopy innewborn lambsFlora Wong, Theodora Alexiou, Brodecky V, Samarasinghe T,Wilkinson M and Walker AM11:40-11:45 Discussion11:45-11:55 Abstract12: Treatment of fetal lung hypoplasia with antenatal corticosteroids:effects on pulmonary circulation and respiration in the newborn sheepKeiji Suzuki, Hooper SB and Harding R11:55-12:00 Discussion12:00-12:10 Abstract13: Clinical presentation, causes and outcome of Persistent PulmonaryHypertension of Newborns (PPHN) in a tertiary care hospital of Karachi.Samana Ali, Parkash Jai, Feroze Asher, Atiq Mehnaz and Khan Iqtidar12:10-12:15 Discussion12:15-12:25 Abstract14: Disease spectrum term neonates admitting in Neonatal ICUSamana Ali and Salat Sohail12:25-12:30 Discussion12:30-12:40 Abstract15: Prenatal Diagnosis of Schizencephaly with Septo-optic Dysplasia byUltrasound and Magnetic Resonance ImageJeng-Hsiu Hung, Shu-Huei Shen, Wan-You Guo, Chih-Yao Chen,Kuan-Chong Chao, Ming-Jie Yang and Chia-Yi Selena Hung12:40-12:45 Discussion12:45-13:45 LunchSESSION 3 : Perinatal medicineChairpersonsJulian T. ParerTomoaki Ikeda13:45-13:55 Abstract16: A possibility of applying system-engineering methods to amniotic fluidvolume changes.Yoshiyasu Hombo and Michio Ooshita13:55-14:00 DiscussionPage 24

MONDAY, AUGUST 27, 2007 (continued)14:00-14:10 Abstract17: Pregnancy after Stillbirth and Home Fetal Heart Rate Monitoringvia the InternetJason H Collins14:10-14:15 Discussion14:15-14:25 Abstract18: Prediction of fetal coarctation of the aorta from the three vessel andtracheal viewHelena Gardiner, Hikoro Matsui , Lucia Pasquini, Anna Seale,Mats Mellander, Michael Roughton and Siew Yen Ho14:25-14:30 Discussion14:30-14:40 Abstract19: Development of new digital fetal monitor "Behaviogram" for earlytomid pregnancyNorio Shinozuka14:40-14:45 Discussion14:45-14:55 Abstract20: Comparison of long axis cardiac function assessed by Vector VelocityImaging and conventional tissue Doppler and M-mode methods in the humanfetusHelena Gardiner, Hikoro Matsui and Ioannis Germanakis14:55-15:00 Discussion15:00-15:10 Abstract21: The genetic analysis of deep vein thrombosis during pregnancy andperinatal associated disorders in japaneseReiko Neki, N. Yamada, Y. Tokito, N. Iwanaga, K. Ueda, K. Yamanaka,M. Nozawa, T. Ikeda, T. Fujita, J. Ishikawa, Y. Sato, T. Miyata, O. Fukui and N,Suehara15:10-15:15 Discussion15:15-15:25 Abstract22: A new monitoring method for FGR fetusesKazunao Suzuki, Motoi Sugimura and Naohiro Kanayama15:25-15:30 Discussion16:00- SPORTING EVENT19:30- DINNER SENDAI NIGHTPage 25

TUESDAY, AUGUST 28, 20078.00 – RegistrationSESSION 4 : BrainChairpersonsLaura BennetMhoyra Fraser8:30-9:00 Keynote speakerAbstract23: Growth hormone: A neurotrophic factor during chick embryogenesis.Steve Harvey, Marie-Laure Baudet and Esmond J Sanders(including Discussion)9:00-9:10 Abstract24: Maternal administration of dexamethasone causes significantalterations in brain activity in the preterm fetal sheep.Joanne Davidson, Lindsea Booth, Josine Quaedackers,Alistair J Gunn and Laura Bennet9:10-9:15 Discussion9:15-9:25 Abstract25: Therapeutic Hypothermia Changes the Prognostic Value of ClinicalEvaluation of Neonatal Encephalopathy.Alistair Jan Gunn, Andrew Whitelaw, John Barks, John S. Wyatt,Denis Azzopardi, Charlene M. Robertson and Marianne Thoresen9:25-9:30 Discussion930-9:40 Abstract26: Can erythropoietin be used to protect the fetal sheep brain fromrepeated exposure to endotoxin?Robert De Matteo, L Cardamone, V Stacy, N Blasch, M Probyn,N Hale, S Rees and R Harding9:40-9:45 Discussion9:45-9:55 Abstract27: Anti-inflammatory effects of Sulfasalazine in an ovine model of inutero infection: a prospect for neuroprotection?Burcu Saglam, Graham Jenkin, Suzanne L. Miller and Euan M. Wallace9:55-10:00 Discussion10:00-10:10 Abstract28: Fetal brain injury in single umiblical artery ligation (SUAL) inducedIUGRSuzanne L. Miller, Low HM, Supramaniam VG,Castillo-Meléndez M, Jenkin G and Wallace EM10:10-10:15 DiscussionPage 26

TUESDAY, AUGUST 28, 2007 (continued)10:15-10:25 Abstract29: Perinatal DiI tracing of developing hypothalamic connectionsIrina G. Makarenko and Alpeeva E.V10:25-10:30 Discussion10:30-11:00 Morning tea (poster viewing)SESSION 5 : HypoxiaChairpersonsBrian KoosAlistair Gunn11:00-11:30 Keynote speakerAbstract30: Chronic hyoxia : Consquence and Treatment.Suzanne L. Miller , Veena G. Supramaniam, Euan M. Wallaceand Graham Jenkin(including Discussion)11:30-11:40 Abstract31: Proliferation in the subventricular zone (SVZ) after severe hypoxiaand induced hypothermia in preterm fetal sheepSherly George, Robert Barrett, Laura Bennet,Justin Dean and Alistair Jan Gunn11:40-11:45 Discussion11:45-11:55 Abstract32: The ontogeny of chemoreflex and hemodynamic responses toprolonged umbilical cord occlusion in fetal sheepGuido Wassink, Laura Bennet, Lindsea C. Booth, Ellen C. Jensen,Bert Wibbens, Justin M Dean, and Alistair J Gunn11:55-12:00 Discussion12:00-12:10 Abstract33: Effect of acute and chronic hypoxia on amniotic fluid gases in chickembryogenesisM. V. Nechaeva, H. Toenhardt and D. Marquardt12:10-12:15 Discussion12:15-12:25 Abstract34: Maternal treatment with allopurinol diminishes fetal cardiac oxidativestress following repeated episodes of ischaemia-reperfusion in sheepJan B Derks, H Torrance , MA Oudijk, AS Thakor,T Cindrova-Davies, F van Bel, GHA Visser, GJ Burton and DA GiussaniPage 27

TUESDAY, AUGUST 28, 2007 (continued)12:25-12:30 Discussion12:30-12:40 Abstract35: Pre-existing hypoxia is associated with a delayed but more sustainedrise in T/QRS ratio during prolonged umbilical cord occlusion in near-term fetalsheepLaura Bennet, Bert Wibbens, Jenny A. Westgate,Harmen H. De Haan, Guido Wassink, and Alistair J. Gunn12:40-12:45 Discussion12:45- Lunch14:00- Free or excursion (including cruising dinner)WEDNESDAY, AUGUST 29, 20078.00 – RegistrationSESSION 6 : PlacentaChairpersonsEllen JensenAlan Bocking8:30-9:15 Invited lectureRole of PEG10 in mammalian placenta development and evolution.Fumitoshi Ishino(including Discussion)9:15-9:25 Abstract36: High incidence of ketosis and hyper-homocysteinemia in the lasttrimester of Japanese mothers.without any complicationsHideoki Fukuoka, Hiroko Watanabe, Yasushi Nagai,Chiyoko Ogasawara and Shigetaka Asano9:25-9:30 Discussion9:30-9:40 Abstract37: Genomic imprinting in the placenta involves histone tail modificationsindependent of DNA methylationKohzoh Mitsuya and Kunihiro Okamura9:40-9:45 DiscussionPage 28

WEDNESDAY, AUGUST 29, 2007 (continued)9:45-9:55 Abstract38: Lactobacilli supernatant inhibits TNF-α production and COX2expression in LPS-activated placental trophoblastsAlan Bocking, M Yeganegi, C Watson, 2 S Kim, G Reid and J Challis9:55-10:00 Discussion10:00-10:10 Abstract39: Interaction between Lipoxygenase Pathways and Progesterone onthe Regulation of 11 -Hydroxysteroid Dehydrogenase type 2 in Cultured HumanTerm Placental TrophoblastsKazuyo Sato, Hiroshi Chisaka, John R.G.Challis and Kunihiro Okamura10:10-10:15 Discussion10:15-10:25 Abstract40: Distribution of the thyroid hormone transporter, MCT8, in fetal andplacental tissues of murine, human and ovine speciesAlison J Forhead, Jenny L Macrae, Theo J Visser,Graham J Burton, FB Peter Wooding and Abigail L Fowden10:25-10:30 Discussion10:30-11:00 Morning tea (with poster session)11:00-12:30 Poster sessionFor a list of poster presentation, please refer to posters12:30-13:45 LunchSESSION 7 : Circulation, CardiovascularChairpersonsJun MurotsukiJan Derks13:45-14:30 Invited lectureCell sheet engineering for tissue and organ regenerationTeruo Okano(including Discussion)14:30-14:40 Abstract41:Determinants of the number of cardiomyocytes in sheep: theinfluence of fetal and postnatal growthRichard Harding, Victoria Stacy, Megan Probyn,Robert DeMatteo, Nigel Wreford and M. Jane Black14:40-14:45 DiscussionPage 29

WEDNESDAY, AUGUST 29, 2007 (continued)14:45-14:55 Abstract42: Modulation of Pulmonary Vascular Smooth Muscle Cell (PVSMC)Phenotype in Hypoxia: Role of cGMP-Dependent Protein Kinase (PKG)J Usha Raj, Weilin Zhou, Chiranjib Dasgupta and Sewite Negash14:55-15:00 Discussion15:00-15:10 Abstract43: Human fetal cardiovascular baro-reflex responses in maternal steroidadministration analyzed by pulsed Doppler minute tissue displacementmeasurementNorio Shinozuka15:10-15:15 Discussion15:15-15:25 Abstract44: Effects of maternal betamethasone administration on regional bloodflow in the intrauterine growth restricted (IUGR) ovine fetusVeena Supramaniam, Jenkin G, Wallace EM and Miller SL15:25-15:30 Discussion15:30-15:40 Abstract45: A Novel extraction method of fetal electrocardiogram from thecomposite abdominal signal.Yoshitaka Kimura,Takuya Ito, Michiyo Nakamura, Kaori Uchida, Hiroshi Chisaka,Norihiro Katayama, Mitsuyuki Nakao and Kunihiro Okamura15:40-15:45 Discussion15:45-15:55 Abstract46: The effects of the tocolytics atosiban and nifedipine on fetalmovements, heart rate, and blood flow.Jan B Derks, Roel de Heus, Eduard J.H. Mulder and Gerard H.A. Visser15:55-16:00 Discussion16:00-16:10 Abstract47:The effect of Sulfasalazine on LPS-induced changes in fetaloxygenation and carotid blood flowBurcu Saglam, Graham Jenkin, Suzanne Miller and Euan Wallace16:10-16:15 Discussion16:15-16:45 Afternoon teaGeoffrey Dawes Lecture16:45-17:45 Avian influenza and pandemic threat(including Discussion)ChairpersonKunihiro OkamuraHitoshi OshitaniPage 30

WEDNESDAY, AUGUST 29, 2007 (continued)17:45-18:45 Business Meeting/AGM19:30- GALA DINNERPage 31

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LIST OF POSTER PRESENTATIONP1: The role of the neural sympathetic and parasympathetic systems in diurnal and sleep statecardiovascular rhythms in the late gestation ovine fetus.Ellen Jensen, Laura Bennet, Sarah-Jane Guild,Lindsea C. Booth, Jenny A. Westgate and Alistair J. GunnP2: Np95, a master regulator of the epigenome, is essentially required for embryonic developmentSharif Jafar, Muto M., Nakamura M., Takebayashi S.,Okano M., Koseki H., Nagamune T., Mitsuya K. and Okamura KP3: Evaluation of the placental vascularity using fractal parameters: could they help in earlydiagnosis of fetal growth restriction?Caterina Guiot, E. Piccoli, F. Saccomandi and T. TodrosP4: Fetal and perinatal growth curves: what can we learn from them?Caterina Guiot, Tullia Todros, Antonio Gliozzi and Pier Paolo DelsantoP5: Nitric Oxide and CD4+25+Regulatory T cell production from iNOS knockout mice and their F1mice treated with LPS.Hidenori Takahashi, Toshiaki Okawa, Keiya Fujimori and Akira SatoP6: Nitric Oxide production in placental tissue from iNOS knockout mice with LPS and theiroffsprings.Hidenori Takahashi, Toshiaki Okawa, Keiya Fujimori and Akira SatoP7: Maternal severe undernutrition during both late gestation and lactation period inducehypertension in male rat offspringHidenori Takahashi, Toshiaki Okawa, Keiya Fujimori and Akira SatoP8: Regulation of maternal feeding during lactation period may control adulthood hypertension.Hidenori Takahashi, Toshiaki Okawa, Keiya Fujimori and Akira SatoP9: Maternal regulation of high fat nourishment during lactation period reduce a hypertension ofmale offspringHidenori Takahashi, Toshiaki Okawa, Keiya Fujimori and Akira SatoP10: Epigenetic stability and developmental plasticity in cloned placentae during somatic cellnuclear tranfer.Naomi Matsuda, Jafar Sharif, Kimiko Inoue, Narumi Ogonuki,Hiromi Miki, Hiroshi Chisaka, Kunihiro Okamura, Atsuo Ogura and Kohzoh MitsuyaP11: Male disadvantage? Gender and fetal cardiovascular responses to asphyxia in preterm fetalsheepLaura Bennet, Bert Wibbens, Jenny A. Westgate,Harmen H. De Haan, Guido Wassink and Alistair J. GunnPage 35

P12: Fetal brain, liver and renal blood flow assessment using 3D power Doppler ultrasoundFong –Ming Chang, Chiung-Hsin Chang, Chen-Hsiang Yu,Lin Kang, Pei-Ying Tsai and Huei-Chen KoP13: Molecular Genetic Study on Angiotensin-Converting Enzyme Gene and Preeclampsia inTaiwanese: Two polymorphisms Tested and as HaplotypesFong –Ming Chang, Ming-Hui Chen, Chiung-Hsin Chang,Chia-Fu Li , Chen-Hsiang Yu and Huei-Chen KoP14: Clinical application of electromechanical delay time for indirect evaluation of fetal bloodpressure during labor and delivery. A preliminary study of 18 full-term infants.Yasuyuki Kawagoe, Hiroshi Sameshima and Tsuyomu IkenoueP15: Temporal Changes in Mediators of Microvascular Tone are Influenced by Sex andGlucocorticoid Exposure in Preterm InfantsIan Wright, Stark MJ and Clifton VLP16: The effect of carperitide, human atrial natriuretic peptide, on acute heart failure caused byendocardial cushion defect with atrioventricular block in a premature infant: a case reportYuichiro Miura, Tadashi Matsuda, Masaki Sato, Shizuko Akiyama,Ryuta Kitanishi, Takushi Hanita, Tatsuya Watanabe and Hidenobu OhtaP17: Effect of Maternal Stress on Fetal Heart Rate Assessed by Vibroacoustic StimulationIkuko Makino, Yoshio Matsuda, Marie Yoneyama , Kyoko Hirasawa ,Koichiro Takagi, Hiroaki Ohta and Yukuo KonishiP18: Non-Invasive long-term fetal ECG monitoringJan B. Derks, Graatsma E.M., Mulder E.J.H.and Visser G.H.AP19: The Relationship of Microchimerism to AbortionTomoko Sato, Keiya Fujimori, Akira Sato, Hitoshi Ohto, Kenich Hata,Hiroko Sasaki, Mihoko Yazawa, Shizuka Honda and Mayumi NoguchiP20: The effect of cerebral hypothermia on cortisol and ACTH responses after umbilical cordocclusion in preterm fetal sheep.Joanne Davidson, Mhoyra Fraser, Andrew S. Naylor,Vincent Roelfsema, Alistair J. Gunn and Laura BennetP21: The prevention of fetal brain and lung injuries based on interleukin-6 levels in amniotic fluidbefore birthTakushi Hanita, Tadashi Matsuda, Masaki Sato, Shizuko Akiyama,Ryuta Kitanishi, Yuichiro Miura, Tatsuya Watanabe, and Hidenobu OhtaP22: Effect of intrauterine inflammation on induction of antenatal periventricular leukomalacia inchronically instrumented fetal sheepMasatoshi Saito , Tadashi MatsudaPage 36

P23: Maternal Low Protein Diet Aggravates Fetal White Matter Damage Caused by InfectionTakuya Ito, Kaori Uchida, Michiyo Nakamura, HiroshiChisaka, Yoshitaka Kimura and Kunihiro OkamuraP24: Prenatal diagnosis of bradycardia using fetal electrocardiogram via maternal abdomenYoshitaka Kimura, Michiyo Nakamura, Kaori Uchida, Takuya Ito,Hiroshi Chisaka and Kunihiro OkamuraP25: Endothelin-1 inhibits both L-type Ca2+ current and ATP-sensitive K+ current in neonatal ratventricular myocytes.Yasuhiro Katsube, Nobuko Suzuki, Makoto Watanabe, Masanori Abe,Miharu Hajikano, Mitsuhiro Kamisago, Ryuji Fukazawa and Shunichi OgawaP26: Placenta accreta/increta in unscarred uterusKozo Akagi, Shima T, Ishigaki N, Oota S, Hayasaka A, Fujita N, Asano K and Wada YP27: QT interval during pregnancy in Long QT syndromeYuri Tokitou, Akiko Omoto, Naoshi Yamada, Naoko Iwanaga, Kaoru Yamanaka,Keiko Ueda, Masayo Nozawa, Reiko Neki, Wataru Shimizu and Tomoaki IkedaP28: High altitude chronic hypoxia modifies cardiovascular responses in newborn sheep dependingon the time and duration of the hypoxic insult during gestation.EA Herrera, G Ebensperger,RA Riquelme, C Torres-Farfan,VR Reyes, JT Parer, DA Giussani and AJ Llanos.Page 37

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Abstract Plenary lecture 1Fetal heart rate monitoring: Is it time to establish internationalconsensus on Intrapartum management?J T ParerDepartment of Obstetrics, Gynecology & Reproductive SciencesUniversity of California San FranciscoDespite numerous attempts in the past several yearsthere is no broad agreement on FHR pattern management.In order to develop a framework we have classified fetalheart rate monitor patterns according to risk of fetalacidemia, and risk of evolution to a more serious pattern,and we have used this information to construct astandardized process for FHR pattern management, withthe ultimate aim of minimizing newborn acidemia withoutexcessive obstetric intervention.Each FHR pattern has been color-coded, from no threat offetal acidemia (green, no intervention required) to severethreat of acidemia (red, rapid delivery recommended).Three intermediate categories (blue, yellow and orange)require escalated informing of appropriate individuals forintervention and resuscitation (obstetrician, anesthesiologist,and neonatal resuscitator) and preparation for urgentdelivery, e.g. staff and surgical suite availability, andconservative techniques to ameliorate the FHR patterns.We have identified 134 FHR patterns classified bybaseline rate, baseline variability, and type ofdeceleration. Based on the best available evidence wehave assigned a risk of newborn acidemia or low 5 minApgar score to these patterns. We have also evaluatedeach pattern for the risk that the pattern would furtherevolve into a pattern with a higher risk of acidemia.This framework is potentially applicable to the institutionswhere it was developed, and will need to be modified forother situations depending on the logistics, facilities andpersonnel available. This may provide a framework fordeveloping algorithms for standardized management ofFHR patterns during labor, which can be tested for validity.NotesPage 40

Plenary lecture 2Developmental Origins of Disease – a Japanese problem and aJapanese answer?AbstractMark HansonInstitute Of Developmental Sciences, University of Southampton, UKIt is now well-established that low birthweight is linked toan increased risk of many chronic, non-communicableadult diseases. But size at birth is a poor proxy measureof the prenatal environment, so that smallness does notnecessarily mean that pathological processes havestarted even before birth. Moreover, animal studies showthat stimuli such as unbalanced nutrition in pregnancy canproduce effects on the offspring which resemble manyaspects of the human metabolic syndrome, but withoutinvariably reducing size at birth. Changes in the trajectoryof development of a range of tissues, organs and controlsystems is part of the ‘strategy’ which the developingembryo, and then the fetus, adopt to optimise thephenotype developed to match the environment predictedfor postnatal life. The processes involved in thisdevelopmental matching (1) have presumably beenselected through evolutionary pressure in many species,including our own, to maximise survival to reproduce.Negative consequences of this strategy may emerge inlater life, and increasing longevity in many populations willincrease their incidence. When mismatch occurs, the riskof disease is increased. This may be due to inaccuratecues about the environment, transmitted from the mother,or because the environment has changed within ageneration.Historically, Japanese people were very long-lived. This ischanging as diets and physical activity patterns change,increasing potential mismatch. Other factors in Japan alsoaffect development. Falling family size increasesprimiparous pregnancy and childbearing age is increasing.Both of these increase maternal constraint of fetaldevelopment. Whilst Japan was highly effective inreducing the incidence of low birthweight and pretermbirth in the 1970s and 80s, mean birthweight has nowfallen over about the last 20yr, especially in cities. Thisappears to be due to dieting in young women to loseweight, perhaps to smoking, and advice from clinicians torestrict weight gain in pregnancy to low levels.Unbalanced diet and low BMI send inappropriate cues tothe developing fetus, increasing risk of mismatch.Coupled with a tendency to deposit abdominal fat whichmay be partly genetic, there are a range of factors whichmust raise great concern about the future incidence ofchronic disease in Japan. But perhaps Japan can teachother countries lessons about large scale educationalinterventions too – the Basic Law on Shokuiku enacted inJune 2005 promotes knowledge about food and the abilityto make food choices, at multiple levels in the population.Will this Japanese problem have a Japanese solution?MAH is supported by the British Heart Foundation1. Gluckman PD and Hanson MA. Mismatch – why ourworld no longer fits our bodies. OUP 2006.NotesPage 41

Abstract Plenary lecture 3An Excess of Circulating Anti-Angiogenic Factors: the Cause ofthe Hypertensive Syndrome of PreeclampsiaRichard J. Levine, MDDivision of Epidemiology, Statistics, and Prevention Research, National Institute of Child Healthand Human Development, Department of Health and Human Services, Bethesda, MD, USAThe maternal syndrome of preeclampsia may be causedby release of excessive quantities of two anti-angiogenicproteins from the placenta into maternal blood: solublefms-like tyrosine kinase 1 (sFlt1) and soluble endoglin(sEng). High circulating concentrations of both moleculesare usually required to cause preeclampsia. Exogenousgene transfer of sFlt1 and sEng into rats using anadenoviral vector produced hypertension, proteinuria,glomerular endotheliosis, and a HELLP-like syndromewith hemolysis, elevated circulating levels of liverenzymes, low platelet counts, and severe restriction offetal growth. sFlt1 acts through binding and neutralizingvascular endothelial growth factor (VEGF) and placentalgrowth factor (PlGF), preventing them from attaching tocell-surface receptors. sEng is thought to interfere withbinding of TGF-β to endoglin cell-surface co-receptors.Circulating concentrations of sFlt1 and sEng are elevatedweeks, even months, before onset of preeclampsia; andthey increase considerably during the last two months ofnormal pregnancy. High levels are associated with earlypreeclampsia onset and the delivery of a small-forgestational-ageinfant. Risk factors for preeclampsia –such as multiple gestation (increases risk) and cigarettesmoking (decreases risk) – appear to operate by alteringangiogenic factor levels. I hypothesize that 1) an excessof circulating anti-angiogenic factors is both necessaryand sufficient to cause clinical preeclampsia and that 2)rising levels of circulating anti-angiogenic factors play animportant role in concluding pregnancy and triggering theonset of labor. In the future I believe that successfultreatment will become available for the prevention ortherapy of preeclampsia by antagonizing or removingsFlt1 and/or sEng or by administering VEGF or PlGF.NotesPage 42

Plenary lecture 4Effects of maternal hyperglycemia on both mothers and theirinfantsAbstractHuixia YangDepartment of Obstetrics and Gynecology, Peking University First Hospital, Beijing, ChinaWith the improved nutrition and continued industrializationover the past decade, both of obesity and diabetes inChina are increasing quickly. Gestational DiabetesMellitus (GDM) is a growing problem in China. Aprospective population-based study of 16,286 pregnantwomen has been conducted in 18 cities of China. Theoverall prevalence of abnormal carbohydrate metabolismwas 10.4% when using ADA criteria, the prevalence inSouth of China was higher than that of in the North.Many studies have showed that GDM is linked to a varietyof separate conditions, including: preterm labor,preeclampsia, increased cesarean section rates,macrosomia, birth injury, stillbirth, and neonatalhypoglycemia and hyperbilirubinemia. With tight maternalblood glucose controlled, pregnancy outcomes in womenwith GDM have been improved.Glucose intolerance diagnosed during pregnancy,identifies women at high risk for Type 2 diabetes later inlife. Recent studies have also shown GDM to beassociated with increased risk of obesity and metabolicsyndrome later in life for both mothers and their babies.So, it will be very important to early diagnose and managethese women with GDM in order to improve both mothersand their infants’ outcomes.NotesPage 43

Abstract Plenary lecture 5The pathogenesis of preterm brain injury: understanding timingLaura BennetFetal Physiology and Neuroscience Group, Dept PhysiologyThe University of Auckland, Auckland, New ZealandA viable neuroprotection treatment will soon be availablefor the term newborn infant suffering fromhypoxic-ischaemic encephalopathy: that of cerebralhypothermia. Currently, however, we do not know whetherthis treatment will be effective for the more vulnerablepreterm infant. Application of hypothermia, or any othertreatment, requires that we understand the mechanismsmediating neural injury and when injury occurs.While it is likely that many of the mechanisms mediatinginjury at term are the same for the preterm, aspects of thepatterns and evolution of injury are quite different. Preterminfants have a high rate of neurodevelopmental handicap.Recent imaging studies have revealed that adverseoutcomes are strongly associated with reduced braingrowth and neural complexity later in life. Increasing datasuggest that these chronic deficits primarily reflect acuteneuronal and glial injury sustained during adverse eventsin utero, such as exposure to severe hypoxia-ischaemiaand inflammation. This chronic impairment appears to bepartly due to upregulation of physiological apoptosis,related to input deprivation and output isolation secondaryto acute white and grey matter damage and axonal injury.However, progenitor cells in the subventricular zone arealso vulnerable to injury, and loss of this critical populationlikely further compromises brain development. Acutehypoxic brain damage in premature infants has beenprincipally related to a supposed unique cellularvulnerability to glutamate and free radicals. However, it isunlikely that in vivo these damaging processes occur inisolation, and that the brain is simply a passive ‘immature’target for toxic factors.In this talk I will examine recent evidence that survival ofvulnerable, injured neurons and immatureoligodendrocytes after acute injury is determined by adynamic balance between damaging events such asincreased glutamate receptor activity, and active neuralprotection by endogenous neural inhibitors, which occursduring the first few hours after the insult (the ‘latent’phase). I will briefly discuss whether treatments likehypothermia can reduce acute neuronal and glial injury inthe preterm, and thus potentially improve the chronicdevelopment of the brain.NotesPage 44

Geoffrey Dawes LectureAvian influenza and pandemic threatHitoshi OshitaniDepartment of Virology, Tohoku University, Graduate School of MedicineSince December 2003, avian influenza A(H5N1) has swept through poultry populationsacross Asia. It has now spread to Europe, Middle-East and Africa. The outbreaks are historicallyunprecedented in scale and geographical spread. Their economic impact on the agricultural sectorof the affected countries has been enormous. Evidence shows that the H5N1 virus is most likelyendemic in many parts of Asia. It has established an ecological niche in poultry, making it extremelydifficult to prevent outbreaks. Outbreaks have recurred despite aggressive control measures,including the culling of more than 160 million poultry. During this period, more than 300 humancases with an overall fatality rate around 60%, have been confirmed in 12 countries. Most of humaninfections can be linked to contact with infected poultry, but isolated instances of inefficienthuman-to-human transmission may have occurred.When the virus acquires capability of efficient human-to-human transmission, globalinfluenza pandemic in human population can occur. It is expected that the next pandemic wouldresult in huge impact on human health, economy and whole society of the world. Influenza virusesare genetically unstable and their behavior cannot be predicted. For these reasons, the currentoutbreaks of influenza A(H5N1) in poultry and humans has moved the world closer to a pandemicthan any time since 1968, when the last such event occurred.Immediate actions should be taken by all relevant partners to minimize the risk of nextpandemic by implementing various control measures in poultry and human. It is necessary to beprepared for emergence of a virus with pandemic potential. If early signs of potential pandemic aredetected rapidly, early containment of the virus may be possible by implementing aggressivemeasures including mass drug administration in affected areas. All countries should also beprepared for the worst case scenario, i.e. influenza pandemic by improving their pandemicpreparedness. By implementing various measures including the use of vaccines and antivirals andother public health interventions, the impact of a next pandemic can be minimized.Page 46

Abstractinvited lectureRole of PEG10 in mammalian placenta development andevolutionFumitoshi Ishino 1 , Ryuichi Ono 1 , Sunsuke Suzuki 1 and TomokoKaneko-Ishino 21 Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University.2 Tokai University, School of Health Sciences.Eutherian chorioallantoic placenta is an evolutionary neworgan that enables fetal growth and development inmother for a relatively long pregnancy period. Manygenes are known to be involved in placenta formation,however, it is still unknown how this new organ evolved inthe eutherians. PEG10 (paternally expressed 10) is animprinted gene that is derived from a sushi-ichi relatedretrotransposon that is highly conserved in eutherianmammals but not in birds and fish. We have recentlydemonstrated that lack of Peg10 causes early embryoniclethality in mice due to sever placental defects lackingspongiotrophoblast and labyrinth layers. Thus, Peg10plays an essential role in mouse development in spite ofits exogenous origin.Comparative genome study among three mammaliangroups, monotremes, marsupials and eutherians, hasdemonstrated that PEG10 also exists in wallaby (amarsupial species), but not in platypus (a monotremespecies). Interestingly, marsupial PEG10 locates exactlythe same position to that of eutherian PEG10, betweenSGCE and PPP1R9A genes, and its amino acid sequenceis conserved. These results indicate that integration ofthe original PEG10 retrotransposon occurred in one of thecommon ancestors of marsupials and eutherians, andtransformed to the present PEG10 form presumably bymultiple mutations before divergence of these two groups.Conservation of PEG10 means that it has high selectiveadvantage in mammalian development under naturalselection, therefore, the event of PEG10 acquisition maybe one of the driving forces in mammalian placentalevolution.NotesPage 48

invited lectureAbstractCell Sheet Engineering for Tissue and Organ RegenerationTeruo OkanoInstitute of Advanced Biomedical Engineering and Science Tokyo Women’s Medical UniversityIdeal tissue construction in vitro and in vivo can beachieved by cell sheet engineering that has beenestablished with three core technologies: First,noninvasive harvest of cultured cells as cell sheets can beachieved using previously developed temperatureresponsiveculture dishes. The surfaces are hydrophobicat 37°C, but become hydrophilic below 32°C. Therefore,various cell types adhere, spread, and proliferate on thesurfaces similarly to on commercial tissue culture dishes.By temperature reduction, cells spontaneously detachfrom the surfaces and confluent cells are recovered as asingle contiguous monolayer sheet with intact cell-celljunctions and deposited extracellular matrix. Second,the harvested viable cell sheets can be transferred toother surfaces, such as culture dishes or devices (2D cellsheet manipulation) because of the adhesive extracellularmatrix proteins associated with the basal side of cellsheets. Thus, tissue regeneration with cell sheetengineering can be accomplished by transplantation ofsingle cell sheets, as with skin, cornea and periodontalligaments. Finally, recovered cell sheets can be layered toreconstruct complex stratified tissue architectures such asliver lobules, kidney glomeruli, and cardiac patches (3Dcell sheet manipulation). For example, layeredcardiomyocyte sheets harvested from temperatureresponsive dishes show synchronous pulsations anddiffuse gap junction formation. When transplanted into thesubcutaneous tissues of nude rats, spontaneous beatingscould be macroscopically observed and maintained forover 1 year. We believe that these 2D and 3D cellmanipulations using cell sheet tissue engineering, willbecome new and revolutionary tools for tissueengineering.NotesPage 49

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Abstract1Keynote speakerImpact of low birthweight on developing organs: separating theeffects of IUGR and preterm birth.Richard Harding PhD, DSc.,Department of Anatomy and Cell Biology, Monash University, Melbourne 3800, AustraliaNumerous epidemiological studies have shownassociations between low birthweight (LBW) and adversehealth outcomes. However most such studies do notdistinguish between the effects of IUGR and preterm birth,both of which can cause LBW and alter organdevelopment. The brain, lungs, heart and kidneys, forexample, develop both before and after birth, andtherefore could be affected by IUGR and preterm birth.The sheep is a good model as birth occurs at a similarstage of organ development as in humans.the effects of preterm birth on the development of theseorgans are relatively unexplored.As 8-10% of births are affected by IUGR, and similarnumbers occur prematurely, it is important that theseparate effects of IUGR and preterm birth on criticalorgan development are defined; appropriate animalmodels are essential. It is also important thatepidemiological studies distinguish between LBW due toIUGR and preterm birth.Using sheep, we have investigated the separate effects ofIUGR and preterm birth on lung development and lungstructure through to maturity. LBW induced by IUGRduring late gestation impairs alveolar formation, resultingin reduced alveolar number, thicker alveolar walls andreduced airway support in adult animals 1,2 ; airway wallstructure was not affected 3 . In contrast, LBW due tomoderately preterm birth has no persistent effects onalveoli 4 , but adult airways were more reactive 5 . Our datasuggest that IUGR predominantly affects lungparenchyma whereas preterm birth primarily affects theairways. In recent studies we have also shown how IUGRaffects kidney 6 , brain 7,8 and heart 9 development; however1. Maritz et al. Pediatric Pulmonology 32: 201-210; 2001.2. Maritz et al. Pediatric Research 55: 287-295; 2004.3. Wignarajah et al. Pediatric Research 51: 681-688;2002.4. Cock et al. Pediatric Pulmonology 40: 336-348; 2005.5. Snibson et al. Experimental Lung Research, 32:215-228; 20066. Mitchell et al. Pediatric Research 55: 769-773; 2004.7. Rees & Harding. Neuroscience Letters 361: 111-114;2004.8. Duncan et al. J Neuropath & Exp Neurol 63, 1131-43;2004.9. Bubb et al. J Physiology 578: 871-881; 2007.NotesPage 52

Postnatal high-fat feeding impacts on metabolic profiles andgluconeogenic capacity during fasting and this is exaggeratedby prenatal undernutrition in adolescent lambs.Mette Olaf Nielsen & Anna Hauntoft KongstedDepartment of Basic Animal and Veterinary Sciences, The Faculty of Life Sciences,Copenhagen University, Grønnegårdsvej 7, DK-1870 Frederiksberg C, Denmark.Abstract2Liver function and metabolic profiles during fasting and agluconeogenic (propionate) challenge was studied inlambs exposed to different prenatal and postnatalnutritional levels and diets. For the last 6 weeks prepartum20 twin-pregnant Shropshire ewes were assigned to eithera High (ME and protein according to requirements) orLow (50% of High) diet. Three days post-partum the twinlambs were assigned to each their feeding: CONV (milkreplacer and dried hay to achieve 250g weight gain/d) orHCHF (High-Fat-High-Carbohydrate: 38% cream + milkreplacer + maize ad libitum). When approx. 6 months oldthe lambs were fasted for 47 hours, subjected to apropionate challenge between hours 45-46 and refed athour 47.From hours 0-44 plasma glucose levels were similar in allgroups. During the propionate challenge HCHF lambs hadslightly slower glucose production and cleared glucoseslower than CONV. HCHF lambs also had fatty livers andhigher plasma gamma glutamyl transferase (GGT) levels.For the prenatal Low diet the same trend was seen as forHCHF, but less pronounced. Low/HCHF lambs thustended to have the slowest, while High/CONV had thefastest glucose clearance after the propionate challenge.Plasma triglyceride levels declined markedly in HCHFlambs over the first 16 hours of fasting upon cessation ofcream feeding, but triglyceride as well as NEFA levelsremained consistently higher in HCHF compared toCONV lambs throughout fasting and propionate challenge.NEFA gradually increased in all groups during fasting,rapidly declined after the propionate-injection, andincreased again 30-60 minutes after, ie. the reversepattern of that observed for glucose.Plasma BOHB levels were initially similar in all groups.After 16 hours of fasting, a marked and steady increasewas observed in HCHF lambs, in comparison with adelayed and very modest increase in CONV lambs. TheBOHB increases during fasting were further exaggeratedin Low lambs.In conclusion, HCHF lambs showed indications ofreduced liver (gluconeogenic) function, impairedglucose-clearance and a more ketogenic metabolismduring fasting. This appeared to be exaggerated in lambsexposed to prenatal undernutrition (Low), ie. as aconsequence of a pre- and postnatal nutrition mismatch.NotesPage 53

Abstract3Maternal Isocaloric High-protein Diet Ameliorates the Elevation inboth Blood Pressure and Cardiac Remodeling in the Adult MiceOffspring with Maternal Calorie Restriction Possible Involvement ofBranched-Chain Amino Acids–Hiroaki Itoh 1 , Makoto Kawamura 2 , Shigeo Yura 2 , Haruta Mogami 2 , FujiiTsuyoshi 2 , Norimasa Sagawa 31 Osaka National Hospital, Osaka; 2 Departmemt of Gynecology and Obstetrics Kyoto UniversityGraduate School of Medicine, Kyoto, 3 Department of Obstetrics and Gynecology Mie UniversitySchool of Medicine, Tsu, JapanBackground;Epidemiological evidences have elucidated thatundernutrition in utero is a risk factor for cardiovasculardisorders (CVD) in adulthood. However, the mechanismis yet be fully understood. Recently, we demonstrated amouse animal model of intra uterine growth restriction(18% decrease in pup weight) with maternal calorierestriction (30%) using regular chow diet (RCD) (CellMetab 2005, 1;371). After birth, the undernourishedoffspring (UN offspring) showed a significant increase insystolic blood pressure and cardiac remodeling at 16 wks,compared to normally nourished offspring (NN offspring)(Endocrinology 2007,148:1218).Aim;The aim of the present study was to investigate the effectof maternal high protein intake, isocaloric base, on thedevelopment of risk factors of CVD in adult offspring.Methods;The maternal food restriction was applied to pregnant micewith either RCD or isocaloric high-protein diet (HPD).Amino analysis was carried out in the fetal plasma, whichwas obtained by uterine section at 18.5 dpc. Systolic bloodpressure and cardiac remodeling-associated morphologicalparameters, i.e. perivascular fibrosis of the coronary artery,cardiomegaly and cardiomyocyte enlargement, weremeasured in the adult offspring (P-UN offspring) incomparison with UN and NN offspring.Results;Maternal isocaloric HPD augmented fetal plasmaconcentrations of branched-chain amino acids (BCAA:valine, leucine and isoleucine). At 8 and 16 wks,significant systolic blood pressure increase was observedin UN offspring, but not in P-UN offspring, as comparedwith NN offspring. In the similar manner, all of cardiacremodeling-associated parameters measured weresignificantly elevated in UN offspring, but not in P-UNoffspring, as compared with NN offspring at 16 wks.Conclusion;It is suggested that maternal isocaloric HPD may alleviatethe deterioration in systolic blood pressure as well as incardiac remodeling in the adult offspring with maternalcalorie restriction, probably at least partly via exposure toBCAA in utero.NotesPage 54

AbstractMaternal ovine cortisol concentrations are important for keyendocrine factors involved in fetal growth4Jensen EC 1 , Bennet L 1 , Wood CE 2 , Gunn AJ 1 and Keller-Wood M 3 .1 Department of Physiology, Faculty of Medical and Health Sciences, The University ofAuckland, Auckland, New Zealand, 2 The Department of Physiology and Functional Genomicsand 3 The Department of Pharmacodynamics, College of Medicine, The University of Florida,Gainesville, Florida, USAThe normal elevation of maternal plasma corticosteroidconcentrations during pregnancy is important for thesupport of fetal growth and development. Both elevatedmaternal cortisol levels and reduced maternal cortisolbelow normal levels reduce fetal growth, and elevatedmaternal cortisol levels cause fetal hypertension. Themechanisms mediating these changes in fetal growthremain unknown. We examined the hypothesis thatchronic alterations in maternal plasma cortisolconcentrations reduce insulin-like growth factor (IGF)mRNAs and alter binding protein (BP) mRNA, and soreduce fetal growth.Ewes of mixed breeds carrying singleton pregnancies wereoperated on between 112 and 116 days of gestation (115 ±0.4, term=147 days). Animals were randomly assigned intothree groups before surgery. The first group consisted of sixcontrol animals, the second group consisted of five ewesthat were administered cortisol by continuous intravenousinfusion (1 mg/kg/day) (high cortisol), and the thirdconsisted of five ewes that were adrenalectomized andreplaced with 0.5-0.6 mg cortisol/kg/day and 3 µgaldosterone/kg/day in order to produce cortisolconcentrations similar to endogenous concentrations inintact nonpregnant animals (low cortisol). Fetuses werestudied from the day after surgery until 130 (± 0.2) days ofgestation. Fetal liver was snap frozen in liquid nitrogenand stored at -80 o C for further mRNA analysis.Fetal liver IGF-II and IGFBP-3 mRNA were decreased inthe low cortisol group compared to controls (p

Abstract5Diet-Induced Obesity and Prenatal Undernutrition Lead to CentralLeptin Resistance by Different MechanismsCharisma K Dhaliwal, Stefan Krechowec, Mark Vickers, Bernhard H Breier,Mhoyra FraserNational Research Centre for Growth and Development and Liggins Institute, Faculty of Medical andHealth Sciences, The University of Auckland, Auckland, New ZealandBackground:We have previously reported that prenatal undernutritionleads to a dysregulation of appetite suppressive effectsthrough alterations in neuropeptide gene expression ofwhole hypothalami, which may be associated with centraland peripheral leptin resistance [1]. In the current study,we expand our initial observations and investigate theneuroendocrine transcriptional response and leptinsensitivity in the arcuate nucleus (ARC) of rats exposed toprenatal undernutrition (UN) or a postnatal high-fat diet(DIO).Methods:Pregnant Wistar rats were fed a standard chow diet eitherad libitum (AD) or at 30% of AD intake throughoutgestation (UN). At weaning, female offspring were fedeither a chow [C], high fat [HF] (30% fat wt/wt) or calorierestricted [CR] (70% of standard chow intake) diet adlibitum for the remainder of the study. At 142 ± 5 days, ADand UN offspring received either recombinant rat leptin(2.5µg/g/day) or saline for 14 days, subcutaneously [2].Results:Using quantitative reverse transcription-polymerase chainreaction (qRT-PCR), we found that both NPY and ObRbmRNA expression in the ARC was significantly increasedin UN animals (P

Impact of maternal exposure to constant light during lategestation on postnatal adrenal function in the capuchinmonkey: long terms effects on DHAS and transient effects oncortisol.Abstract61 Torres-Farfan C, 1 Mendéz N, 2 Richter H, 3 Valenzuela G and 1,4 Seron-Ferre M.ICBM, Fac Medicina, Universidad de Chile; 2 Fac Medicina, Universidad Austral de Chile, 3 Dpt ofWomen’s Health, Arrowhead Regional Medical Center, Colton, CA. 4 Universidad de Tarapacaand CIHDE, Arica, Chile.Several adult diseases are linked with alterations of thehypothalamus-pituitary-adrenal axis in utero. Maternalmelatonin suppression (by exposure to constant lightduring last third of gestation) induced precociousmaturation of the fetal adrenal (JPineal Res 41:58-66,2006), increased plasma cortisol concentrations in thenewborn soon after birth (JPhysiol 554:841-856, 2004);and resulted in an increased cortisol response to ACTH atfirst month of postnatal age (2006 FNPS P:4). Incontrast, at one month of age, basal DHAS and the DHASresponse to ACTH were decreased. To assess whetherthe effects of maternal exposure to constant light on thenewborn adrenal were permanent, we measuredresponsiveness to ACTH at 8 months of ageProtocols:Four pregnant capuchin monkeys were maintained inconstant light (LL) from 63% gestation and other fourremaining in light:dark cycle (14:10; Control). Afterdelivery the LL newborn and its mother returned to 14:10photoperiod. At 8 month postpartum the newbornsreceived dexamethasone (2.5mg/kg, im) and twelve hourslater, blood samples were drawn at 0-30-60 and 120 minpost ACTH 1-24 (125 μg/kg, iv). Cortisol and DHAS wasmeasured by RIA.Results and conclusion:Maternal exposure to constant light during gestation hadlong lasting effects on DHAS and transient effects oncortisol. At 8 months of age, the decrease in DHAS and inthe DHAS response to ACTH was maintained, whereascortisol response to ACTH returned to normal. Thepresent data suggest that exposure to constant lightduring pregnancy selectively impairs postnatal DHASproduction, probably by decreasing the number of DHASproducing cells. These results are consistent with thepreviously suggested role of maternal melatonin duringgestation as trophic factor for the DHAS producing cells ofthe fetal adrenal gland.Supported-Fondecyt1050833-SBMC.NotesPage 57

Abstract7 Maternal feeding controls fetal biological clockXu Shanhai 2 , Hidenobu Ohta 1,2,3 , Takahiro Moriya 4 , Masayuki Iigo 5 , TatsuyaWatanabe 1,3 , Norimichi Nakahata 4 , Hiroshi Chisaka 1,2 , Tadashi Matsuda 1,3 ,Toshihiro Ohura 3 , Yoshitaka Kimura 6 , Nobuo Yaegashi 2 , ShigeruTsuchiya 3 , Hajime Tei 7 , and Kunihiro Okamura 1,21 Center for Perinatal Medicine, Tohoku University Hospital, Sendai, Japan, 2 Department ofObstetrics and Gynecology, Tohoku University Hospital, Sendai, Japan, 3 Department ofPediatrics, Tohoku University Hospital, Sendai, Japan, 4 Department of Cellular Signaling,Tohoku University, Sendai, Japan, 5 Department of Applied Biochemistry, Faculty of Agriculture,Utsunomiya University, Utsunomiya, Japan, 6 Tohoku University Biomedical EngineeringResearch Organization, Sendai, 980-8574, Japan , 7 Research Group of Chronogenomics,Mitsubishi Kagaku Institute of Life Sciences, Tokyo, JapanCircadian physiological rhythms of the fetus are affectedby oscillators in the maternal brain that are coupled to theenvironmental light cycle. Circadian gene expressionsoccur in the fetal brain and organs during pregnancyalthough it is unclear how the fetal biological clock iscoupled to the maternal clock. To study the link betweenfetal and maternal biological clocks, we investigated theeffects of cycles of food availability (which exert powerfulentraining effects on maternal behavior) on the rhythms ofgene expression in the fetal suprachiasmatic nucleus(SCN). We exposed pregnant rats to a restricted-feeding(RF) regimen, in which food was available only for 4 hoursduring the light portion of a 12-hour:12-hour LD cycle.With a transgenic rat model in which the mouse Period1(a circadian clock gene) promoter has been linked to aluciferase reporter, we continuously monitored therhythmic expression of this "clock gene" by recording lightemission from tissues in vitro. While rhythmicity in thematernal SCN remained phase-locked to the light-darkcycle, restricted feeding entrained the fetal SCN, shiftingits rhythm by 5 hours within the 22-day pregnancy (ad libfed group, n=5; RF fed group, n=5). Our resultsdemonstrate that feeding cycles can entrain the fetal SCNindependently of both maternal SCN and light cycle, andthe results also suggest the need to reexamine thematernal-fetal communication mechanism..NotesPage 58

AbstractDietary protein restriction of pregnant mouse induces alteredmethylation of Oct-4 and Sphk-1 gene promoters in the liver ofoffspring8Jun Murotsuki 1 , Kunihiro Okamura 1 , Hiroi Kaku 2 , Yoshihiko Araki 3 , HiroshiYoshitake 3 , Mark A. Hanson 4 , Felino R. Cagampang 4Department of Obstetrics and Gynecology, Tohoku University School of Medicine 1 , andDepartment of Obstetrics and Gynecology, Iwate Medical University School of Medicine 2 ,Institute for Environmental and Gender-Specific Medicine, Juntendo University GraduateSchool of Medicine 3 , Centre for Developmental Origins of Health and Disease, University ofSouthampton 4The developmental origins hypothesis postulates thatduring critical ontogenetic periods, transientenvironmental stimuli perturb developmental pathwaysand induce permanent changes in gene expression,metabolism, and chronic disease susceptibility. Onelikely mechanism is via early nutritional influences onepigenetic gene modification consisting of the presence ofa methyl group on the carbon 5 of a cytosine residue.This modification is responsible for an important form ofgene regulation in eukaryotes. In the present study, wehave tested the hypothesis that maternal low-protein dietaltered epigenetic regulation of specific gene of theoffspring. C57BL/6 female mice were mated and on theday the plug was detected, these females were thenrandomly allocated to be fed isocaloric diets consisting18% protein or 9% protein. At delivery, offspring werekilled and the livers were removed immediately, frozen inliquid nitrogen and stored at -80C. Genomic sequencingafter bisulfite modification is used to study site-specificDNA methylation. DNA methylation status of Oct-4 andSphk-1 gene upstream regions in the mouse liver wasanalyzed. Hepatic Oct-4 or Sphk-1 promoter methylationwas not significantly different between both groups.However, DNA methylation pattern of the genomic DNA isspecific in low-protein diet group. Aberrant Oct-4 andSphk-1 gene expression may cause perturbations in celldifferentiation. We suggest that the epigeneticmechanism consisting of DNA methylation underlies thefetal programming theory.NotesPage 59

Abstract9Transcriptional suppression of Igf2 mediated by altered histonemodifications in a maternal protein restricted IUGR mouse modelNakamura M 1 , Sharif J 1,2 , Ito T 3 , Chisaka H 1 , Kimura Y 3 , Mitsuya K 3 , andOkamura K 11 Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine;2 Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo;3 Tohoku University Biomedical Engineering Research OrganizationAIMSDietary restriction during pregnancy may provide cues formetabolic disorders such as hypertension and glucoseintolerance. Although prenatal programming can beexplained by certain epigenetic modifications, the precisemechanisms remain obscure. We have investigated theeffect of maternal nutrition on the fetus.MethodsMice were fed a control (C) diet or a protein restricted(PR) diet from 7 days prior to conception to throughoutpregnancy. Another two groups of mice were prepared byreversing the diet of the C and PR at mid-gestation (d10)to measure the effects of nutrition at early and lategestational stages. Dams were sacrificed at d17 and fetaland placental weight was measured. Expression ofpromoter specific Igf2 mRNA in fetal liver was analysed byquantitative real-time RT-PCR. Histone modifications inrespective promoter regions of Igf2 (P1-P3) was analysedto elucidate the epigenetic mechanism behind theobserved phenotypes.RESULTSProtein restricted (PR) diet throughout pregnancy did notaffect placental and fetal weight. Interestingly,administration of PR diet during late gestation (d10-d17)caused significant reduction of fetal weight, a hallmark ofintrauterine growth restriction (IUGR). In this group, theexpression of P1 promoter specific Igf2 expression wasalso reduced by approximately 50%. However, the P2 andP3 transcripts did not exhibit any notable change in Igf2expression. ChIP analyses revealed that the P1 promoterregion was depleted of H3K4me3 and H4K20me3permissive modifications and enriched in H3K27me3repressive modification. These histone modifications wereunaltered in the P2 and P3 promoter regions.CONCLUSIONSWe report a novel epigenetic regulatory mechanism thatrepresses promoter specific Igf2 expression in fetal liverduring maternal protein restriction. Our results suggestthat the fetus is capable of adapting to an environmentalcue if it is provided from the onset of gestation. But if theexternal insult is applied during a critical period ofdevelopment, a mismatch between the expected andexperienced environment occurs which affects the fetalability to maintain its natural developmental program. Ourworks will provide new insights into the epigenetic andmolecular basis of fetal programming and the response ofthe fetus to the early nutritional environment.NotesPage 60

AbstractKeynote speaker10 The state of the art in head cooling for neonatal encephalopathyAlistair Jan GunnFetal Physiology and Neuroscience Group, Dept of Physiology, The University of Auckland,Auckland, New Zealand.The possibility that hypothermia started during or afterresuscitation at birth might reduce brain damage andcerebral palsy has tantalized clinicians for a long time.The key insight was that transient severehypoxia-ischemia can precipitate a complex biochemicalcascade leading to delayed neuronal loss. There is nowstrong experimental and clinical evidence that mild tomoderate cooling can interrupt this cascade, and improvenumbers of infants surviving without disability in themedium-term. The key remaining issues are to find betterways of identifying babies who are most likely to benefit,to define the optimal mode and conditions of hypothermiaand to further improve the effectiveness of treatment.Existing trial data suggests a number of possible avenues.These trials have confirmed that outcome is stronglyinfluenced by severity of neonatal encephalopathy. In theCoolCap study, infants with the most severeelectrographic changes did not seem to respond tohypothermia. In contrast, there was a similar relativebenefit for hypothermia in infants with clinically definedmoderate and severe encephalopathy, raising thepossibility that aEEG monitoring may be more effective inidentifying infants at a treatable time after birth.Intriguingly four days after rewarming infants withcontinuing moderate encephalopathy who were treatedwith hypothermia had a significantly higher rate offavorable outcome than controls, suggesting thathypothermia may change the prognostic value of clinicalassessment. Further, the CoolCap trial has suggested forthe first time that greater birth weight in control infants washighly associated with adverse outcome, even afteradjustment for pyrexia and severity of encephalopathy,however, larger infants also showed greater benefit fromcooling. Thus, potentially the biology of neonatal injurymay differ between larger and smaller infants.NotesPage 62

AbstractCerebral oxygenation measured by spatially resolved spectroscopyin newborn lambs11Wong F 1,2 , Theodora Alexiou 1 , Brodecky V 1 , Samarasinghe T 1 , WilkinsonM 1 , Walker AM 11Ritchie Centre for Baby Health Research, Monash University, Clayton, Victoria, Australia;2Newborn Services, Monash Medical Centre, Clayton, Victoria, AustraliaBackground:Loss of cerebral autoregulation in preterm infants hasbeen associated with adverse outcome 1 . Howeverassessment of cerebral autoregulation is problematic dueto difficulty with continuous measurement of cerebralblood flow (CBF). Spatially Resolved Spectroscopy (SRS)enables continuous cotside measurement of meancerebral oxygen saturation, expressed as a tissueoxygenation index (TOI). Theoretically, if cerebral oxygenconsumption is constant, changes in TOI reflect changesin CBF. As yet, the CBF-TOI relationship has not beenestablished experimentally. We aimed to evaluate therelationship between simultaneous measurements of CBFand TOI in the newborn lamb brain, as a suitable animalmodel for the preterm human brain.Method:In four newborn lambs (2-6 d of age), TOI was measuredcontinuously with SRS (Hamamatsu NIRO-200). CBF wasmeasured with an ultrasonic flow probe (TransonicTM)implanted around the superior sagittal sinus. Mean arterialpressure (MAP) was recorded via an axillary intra-arterialcatheter and pressure transducer. An inflatable siliconrubber cuff was positioned around the commonbrachiocephalic artery; inflation of this cuff reduced arterialpressure distal to the cuff and hence cerebral perfusionpressure. Relationships between changes in CBF and TOIwere evaluated in the time and frequency domain.Coherence analysis was performed to quantify correlation ofCBF and TOI in a frequency-specific manner; a coherencescore ≥ 0.5 indicates a significant relationship between thewaveforms of CBF and TOI in a given frequency rangeResults:CBF and TOI showed concordant changes during MAPfluctuations (Fig.1). In time domain analysis, there issignificant correlation between ∆TOI(%) and ∆CBF(%)TOI(%)CBF(ml/min)MAP(mmHg)6050400 2 4 6 8 1010500 2 4 6 8 10806040200 2 4 6 8 10Time (min)Figure 1. TOI and CBF during MAP fluctuations(R 2 =0.53, p

Abstract12Treatment of fetal lung hypoplasia with antenatal corticosteroids:effects on pulmonary circulation and respiration in the newbornsheep*Suzuki K, Hooper SB, Harding R*Center for Maternal, Fetal, and Neonatal Medicine, Saitama Medical Center, Saitama MecicalUniversity, Kawagoe, Saitama, JapanDepartment of Physiology, Monash University, Melbourne, Victoria, AustraliaAims:We have previously shown that fetal lung hypoplasia (LH)has more influence on the pulmonary circulation than onventilation in newborn lambs (Suzuki K et al; Pediatr Res57:530-6, 2005). The objective of this study was todetermine the effects of antenatal maternal corticosteroidadministration on the pulmonary circulation andrespiration in a sheep model of LH.Methods:LH was induced in 12 ovine fetuses by tracheo-amnioticshunt and amniotic fluid drainage beginning at ~105 daysof gestation (term ~147days). In 6 of these fetuses(LH+S group), one dose of betamethasone (11.4 mg im)was given to the mother 24 hours before elective delivery.At ~140 days, after vascular catheterizations andultrasonic flow probe implantation, lambs were deliveredand ventilated for 2 hours under sedation, during whichwe recorded systemic and pulmonary artery pressures,left pulmonary artery blood flow, airway pressure andventilatory air-flow.Results:Control LH LH+S(n=6) (n=6) (n=6)Lung weight (g/kg BW) 29.8+/-2.7 *21.7+/-1.3 **19.6+/-2.1Lung DNA (mg/kg BW) 186+/-13 *145+/-7 **132+/-11FiO 2 at 2h 0.38+/-0.03 0.35+/-0.02 0.33+/-0.01Crs at 2h0.57+/-0.04 *0.40+/-0.03 **0.41+/-0.03(mL/cmH 2 O/kg BW)PVR at 2h(mmHg/mL/min*kg BW) 0.32+/-0.04 **0.76+/-0.06 ## 0.44+/-0.04Crs: total respiratory system compliancePVR: pulmonary vascular resistancedifference between control and LH (*p

Clinical presentation, causes and outcome of PersistentPulmonary Hypertension of Newborns (PPHN) in a tertiary carehospital of Karachi.Abstract13Ali Samana, Parkash Jai, Feroze Asher, Atiq Mehnaz, Khan IqtidarDepartment of Pediatrics. Aga Khan University Karachi, PakistanIntroduction:PPHN is a cardiopulmonary disorder characterized bysystemic arterial hypoxemia secondary to elevatedpulmonary vascular resistance with resultant shunting ofpulmonary blood flow to the systemic circulation.Objectives:All studies related to PPHN available in literature are ofdeveloped world where advanced treatment is easilyavailable. There is no data found to see the clinical courseand outcome of PPHN in developing world. Our objectivewas to see clinical presentation, common causes andoutcome of PPHN in a tertiary care hospital of Pakistan.Study Design:A retrospective descriptive study is conducted in Neonatalintensive care unit of Aga Khan Hospital from July 2000 toJuly 2005.All babies < 30 days of life with respiratory distress wereincluded who were diagnosed as PPHN by either one ofthe following laboratory results:• Echocardiography showing Pulmonary ArterialHypertension.• Failed Hyperoxia test.• Significant difference of pre and Post ductal bloodgases (> 20 mmHg).All neonates with cyanotic heart disease or in the absenceof above mentioned inclusion criteria were excluded.Results:Total 8o charts were pulled through ICD system of medicalrecord. Out of which 16 cases were excluded due topresence of cyanotic structural heart diseases andmissing charts. 64 cases were included in our study. 67%patients were presented before 24 hours of life. 72%patients were male. 57% patients were AGA. 90% patientsrequired mechanical ventilation. 70% patients requiredventilation for 6 days. Pneumonia was the most commoncause (42%) found. Sepsis and Meconium aspirationsyndrome were the second and third common causes ofPPHN. Hypothermia found in 25% cases. Hypoglycemiafound in 14% cases. Blood Culture was +ve in 26%cases.25% developed Pneumothorax. 50% patientsexpired while 45% survived and remaining were shifted toother hospital. Patients presenting

Abstract14 Disease spectrum of term neonates admitting in Neonatal ICUAli Samana, Salat SohailDepartment of Pediatrics, Aga Khan University, Karachi, PakistanIntroduction:Newborn babies born after 36 weeks of gestation areconsidered as term neonates. In comparison to pretermbabies, term babies have a smooth course and do notrequire intensive care admission. Some of them still needintensive care admission.In developed countries the common problems of newbornbabies admitted in NICU are infections, prematurity, birthasphyxia, and congenital anomalies. We don’t know thedisease spectrum of neonates admitting in NICU ofdeveloping country. Multiple studies have beenpublished to know the disease spectrum and outcome ofpreterm babies but no significant data found for termbabies both with in and out side our country.ObjectivesOur objectives were:• To know the type of diseases acquired by term babiesrequiring intensive care hospitalization.• To assess the outcome of full term babies in terms ofrecovery/discharges, expiries or shifts to otherhospitals.MethodologySetting:• This study is done in neonatal intensive care unit(NICU) of the Aga Khan University Hospital Karachi(AKUH) from Jan 2005 to Dec 2005.Sampling Technique:• It was purposive sampling. All term neonates admittedin NICU during study period will be included.Inclusion Criteria:• All term (>36 weeks) neonates (0 to 28th day of life)born inside or outside AKUH born admitted in NICUwere included in study.Exclusion Criteria:• All preterm neonates

AbstractPrenatal Diagnosis of Schizencephaly with Septo-opticDysplasia by Ultrasound and Magnetic Resonance Image15Jeng Hsiu HungDepartment of Obstetrics and Gynecology, Taipei Veterans General Hospital, 201, Section 2,Shih-Pai Rd, Taipei 112, TaiwanAbstractsA 28-year-old gravida, G1 A0, menarche at 12 years old,was referred to our fetal diagnosis unit at 28 +2 gestationalweeks because of suspected fetal cerebral anomalies. Ontwo-dimensional ultrasonography, the cephalic axial viewshowed multiple hypoechoic spaces in the fetal brain,both cerebral cortex and occipital lobe showed bilateraldefects, and the septum pellucidum was absent. Multipleirregularly shaped cystic lesions connected withsubarachnoid spaces were observed by three-dimensionalultrasonography in surface rendering mode. Septo-opticdysplasia with dysgenesis of corpus callosum was furtherconfirmed by prenatal MRI. Since schizencephaly withbilateral clefts involve bilateral frontal, parietal andoccipital lobes, learning disability, epilepsy and cerebralpalsy are often present after birth. The flaccid mobility ofall four extremities of the fetus, demonstrated prenatallyby real-time ultrasound and functional MRI, can forecastthe risk of postnatal spastic quadriplegia.NotesPage 67

NotesPage 68

Abstract16A possibility of applying system-engineering methods toamniotic fluid volume changes.Yoshiyasu Hombo, Michio OoshitaKanazawa, JapanAmniotic fluid volume (AFV) during pregnancy is usuallysurveyed using amniotic fluid index (AFI). However, thisone-dimensional index is unable to predict the actual AFV.We then assumed each pocket’s shape as a half-ellipsoidor as a crescentric-cylinder and calculated its volume (ml)using distances measured along three axes. We reportedelsewhere that the sum of all volumes of the pockets wasa good predictor of AFV (pAFV) in near-tem pregnancy.We have extended the use of the pAFV to the secondtrimester and obtained 15005 measurements of pAFVfrom 12 to 42 weeks in 1352 pregnancies. The mean ofpAFV increased at the rate of 40-55ml/week from 12 to 22weeks, reached a peak of 590ml in 27 weeks anddecreased at the rate of 20-30ml/week after 31 weeks.The regression equation of pAFV by weeks was:pAFV=0.0033W 4 -0.3633W 3 +12.702W 2 -137.71W +437.68(W; gestational weeks, R 2 =0.994).Other investigators have also presented a regressionequation for estimation of fetal body weight (EFBW) byweek. We added the pAFV to EFBW and obtained volumeof amniotic cavity (VOAC) for each week. The regressionequation was; VOAC=-0.0032W 4 +0.2177W 3 -1.8459W 2+22.675W-263.2(R 2 =0.9997), which showed that theamniotic cavity continued to expand in a smooth manner.Therefore, the inner pressure of the amniotic fluid shouldpush outwards and stretch amniotic membrane.Investigators have shown that major sources of amnioticfluid are fetal urine and lung exudates and major removalroutes are fetal swallowing and the intra-membranouspathway.Taking these into consideration, we have derived twodifferential equations describing AFV and its osmolarity,which correspond to the state and output ones in field ofsystem engineering, respectively. This suggests thatAFV-changes during pregnancy can be analyzed usingsystem-engineering means.NotesPage 70

AbstractPregnancy after Stillbirth and Home Fetal Heart Rate Monitoringvia the Internet.17Jason H Collins MDObstetrics & Gynecology, Collins ClinicSlidell, LA, U.S.AStillbirth affects 30,000 pregnancies each year in theUnited States. Umbilical Cord Accident (UCA) accountsfor 20%-25% of all stillbirths or 2-4 deaths/1000 live births.Recent studies have shown pregnancy after stillbirth hasa recurrent stillbirth risk of 5-10 fold. The specific risk ofUCA recurrence is unknown. It is hypothesized that theuse of Home Fetal Heart Rate Monitoring can identify atrisk fetuses for UCA. Those fetuses with umbilical cordcompression patterns can be evaluated and impendingrecurrent stillbirth prevented.Aims:To prospectively follow 40 patients with a history of UCAstillbirth and their subsequent pregnancy by using homefetal heart rate monitoring daily via the internet. Todetermine the presence of umbilical cord compressionpatterns and evaluate the delivery record for recurrentUCA.Methods:Patients with a UCA stillbirth history contacted PregnancyInstitute (PI) through the internet web site(www.preginst.com). Patients were interviewed about theirpregnancy loss and apparent cause. Patients who decideto repeat pregnancy were offered an evaluation at 28-30weeks at PI. This visit consisted of a 30 min Fetal HeartRate Recording and an Ultrasound for UCA recurrence.Patients were instructed on home use of a hospital gradeFDA approved fetal monitor (Analogic:Boston, MA).Patients monitored for 30 minutes every night. Recordingswere sent via the internet to PI .The software wasdeveloped by Remote Care and is FDA approved andHIPPA compliant. Delivery findings were noted throughpatient interview.Results:29 out of 40 patients (study cases) had UCA recurrence at28-30weeks. 20 patients delivered with a recurrent UCAwith 1of these a recurrent UCA Stillbirth. All patientsdelivered by 37 weeks. 4 patients had expeditedC/Sections for FHR decelerations during labor.Conclusion: UCA can repeat in a subsequent pregnancyafter UCA related stillbirth. Home Fetal Heart RateMonitoring can detect umbilical cord compression andpossibly offer an effective method of identifying the fetusat risk of repeat UCA stillbirth.NotesPage 71

Abstract18Prediction of fetal coarctation of the aorta from the three vesseland tracheal viewHikoro Matsui 1 , Lucia Pasquini 1 , Anna Seale 2 , Mats Mellander 1,2 , MichaelRoughton 2 , Siew Yen Ho 2 , Helena Gardiner 1,21 Faculty of Medicine, Imperial College at Queen Charlotte's & Chelsea Hospital,2 Royal Brompton Hospital, London, UKBackground: Prenatal diagnosis of isolated coarctationof the aorta (CoA) suffers from high false positive andfalse negative rates. It is a duct-dependent lesion that isfatal if not treated early. Suspicion of isolated CoA atscreening relies on sonographic disproportion at fourchamber and/or great arterial views. We aimed to developZ-scores for the aortic isthmus and measure theisthmal:duct ratio (I:D) in normal fetuses and test thesescores in fetuses referred with suspected CoA. IsolatedCoA was defined as hearts with normal situs andconnections, with or without a VSD and a narrowtransverse aortic arch or isthmus. Cases with bicuspidaortic valve were included.Methods: The aortic isthmal diameter, immediatelyproximal to the insertion of the arterial duct, wasmeasured prospectively in the transverse plane (threevessel and tracheal view) in 221 normal fetuses at 18 to37 weeks’ gestation. The ductal diameter (D) wasmeasured immediately before it entered the descendingaorta (DAo) in the same view. Four examiners contributedto the data collection. All measurements were repeatedthree times by a single investigator and averaged.Z-scores were created relating isthmal and ductaldiameters to femoral length and gestational age and theI:D ratio recorded.These scores were tested retrospectively on archiveddigital data in 200 fetuses we knew to have normaloutcomes and in 31 with suspected CoA. All hadpostnatal follow-up for at least one year. Thesemeasurements were examined by a single observer,blinded to outcome and ROC curves created.Results: Inter-observer mean difference of isthmalZ-scores was -0.04mm (95% confidence interval -0.8,0.71). The I:D ratio remained constant mean (SD) 0.99(0.13) 95% CI normal values (0.74, 1.23). ROC curvesshowed AUC Isthmus of 0.98 and I:D ratio 0.985. Serialisthmal Z-scores improved to > -2 in most normalneonates by term. (Figure)ConclusionsThe isthmal Z scores and I:D ratio distinguishes thosewith coarctation from fetuses with a normal aortic arch atfirst scan. In those with arch hypoplasia I:D ratio was abetter indicator of need for surgery.NotesPage 72

AbstractDevelopment of new digital fetal monitor “Behaviogram” forearly to mid pregnancy19Norio ShinozukaDepartment of Obstetrics and Gynecology, Japan Red Cross Medical CenterObjectivesChange in fetal behavior and heat rate reflects fetalcondition in utero. However , at early to mid trimesterpregnancy there has been a methodological limitation toobtain these information in quantitative terms. Tomonitor heart rate at beat to beat quality with fetalbehavioral parameter such as FBM, we devised newsystem named “Behaviogram”Method / HardwareBased the previous studies (Ref), the multi-transducer /multi-channel (7 Tr x 18ch ) Doppler system wasdeveloped. The whole transducer size is same as that ofan ordinal cardiotocogram. With this system, multi-pointtissue Doppler signals from fetus are measured. FHR iscalculated from Doppler digital phase signal using twodimensional correlation analysis. FBM is recorded fromminute tissue displacement signals. Fetal gross bodymovements (FGM) are expressed as the power ofDoppler shift signals.To examine the clinical feasibility and to optimize dataprocessing protpcol of devised system, the clinical data offetuses from pregnancy 15 to 37 weeks (15-19w:n=18,20-24w:n=16, 25-29w; n=23,35w-:n=13) were analyzed.Results Example of the display indicates FHR. FBM, FGMwas shown in Fig 1. FHR was measured with nearly beatto beat quality from fetus at 15w of gestation. From 20wonward, simultaneous FHR with behavior (FBM. FGM)were successfully recorded (Fg1).ConclusionProposed system showed a potential to be a nextgeneration fetal monitoring device for early to midtrimester.Ref:Shinozuka N et al. J Ultrasound Med,1994:131:19-25Yamakoshi Y et al. Ultrasonics,1996 34:769-775.Fig1 Example display of “Behaviogram”NotesPage 73

Abstract20Comparison of long axis cardiac function assessed by VectorVelocity Imaging and conventional Tissue Doppler and M-modemethods in the human fetusHikoro Matsui 1,2 Ioannis Germankis 1,2 , Helena Gardiner 1,21 Faculty of Medicine, Imperial College at Queen Charlotte's & Chelsea Hospital;2 Royal Brompton Hospital, London, UKBackground:We present preliminary data on the feasibility ofmeasuring long axis fetal cardiac function using a newspeckle-tracking algorithm. This allows evaluation ofmyocardial motion from two dimensional echocardiographicdigital images. We aimed to compare long axis cardiacfunction measurements using vector velocity imaging(VVI) with measurements of tissue velocities (TDI) andmaximal displacement of the atrioventricular ring recordedusing the established methods of pulsed Doppler andM-mode in human fetuses.Methods:Conventional and VVI measurements were recorded inthe four chamber view in 12 fetuses aged 17 to 34 weeks’gestation. Seven had normal hearts, 3 had structuralmalformations: hypoplastic left heart syndrome,atrioventricular septal defect and pulmonary stenosis.One pair had twin-twin transfusion syndrome. All valuesrepresent the mean of 3 measurements at the left freewall, septal and right free wall of each atrioventricular ring.Off-line analysis was performed prospectively usingSyngo-VVI software (Siemens) creating a tracking movieautomatically. The average frame rate of the images was89fps (52-146fps). We allowed only 10 minutes fortracking and considered recordings that lay outside thistime limit unsuitable for analysis. We created atime-velocity curve of the atrioventricular ring andmeasured myocardial long axis shortening (Sm-VVI) andlengthening (Em-VVI and Am-VVI). We calculated themaximal amplitude of atrioventricular ring from thetracking coordinates (Amp-VVI). Conventional recordingsof right and left ventricular myocardial long axis shortening(Sm-TDI) and lengthening (Em-TDI, Am-TDI) were madeusing pulsed Doppler and maximum amplitude of theatrioventricular ring displacement from M moderecordings as described previously (Amp-M).Results:Automatic tracking was not feasible in 4 ventricles (16%)but was possible using conventional methods in all but theleft ventricle in the case of hypoplastic left heart syndrome.In line with conventional methods, VVI myocardialvelocities increased with gestational age and were higherin RV free wall than in LV free wall. However, while therewas good correlation using both methods, particularlyAmp-M and Amp-VVI (R=0.91, p

THE GENETIC ANALYSIS OF DEEP VEIN THROMBOSISDURING PREGNANCY AND PERINATAL ASSOCIATEDDISORDERS IN JAPANESER. Neki 1 , N. Yamada 1 , Y. Tokito 1 , N. Iwanaga 1 , K. Ueda 1 , K. Yamanaka 1 , M.Nozawa 1 , T. Ikeda 1 , T. Fujita 2 , J. Ishikawa 3 , Y. Sato 3 , T. Miyata 3 , O. Fukui 4 ,N,Suehara 41 Department of Perinatology, National Cardiovascular Center, Suita, Japan 2 Department ofMaternal Medicine, Osaka Medical Center and Research Institute for Maternal and ChildHealth, Izumi, Japan 3 Research Institute, National Cardiovascular Center, Suita, Japan4 Department of Obstetrics, Osaka Medical Center and Research Institute for Maternal and ChildAbstract21Health, Izumi, JapanIntroduction:Deep vein thrombosis (DVT) and pulmonarythromboembolism (PTE) are easily occurred duringpregnancy because of its hypercoagulation status. It isreported that DVT or PTE is closely related with geneticfactors. Furthermore in Europe and the United States,thrombophilia-associated pregnancy wastage is reported.Our purpose is to examine the relation of genetic factorsto DVT/PTE during pregnancy and pregnancy wastage inJapanese.Methods:We sequenced the entire coding regions of 3anticoagulant genes, PROS1 (protein S), PROC (proteinC), and SERPINC1 (antithrombin) in 178 complicatedpregnant cases: 12 cases of DVT/PTE, 124 cases ofinfertility including habitual abortion, 42 cases of othercomplications including intrauterine growth restriction,intrauterine fetal death, pregnancy induced hypertension,abruptio placenta.Results:Three cases with DVT/PTE had genetic mutations inPROS1,one case in PROC that could affect the activity.The genetic mutations in PROS1, PROCand SERPINC1that could affect the activity have been identified in each2cases with infertility. The genetic mutations in PROS1have been identified in 2 cases with other complications.Thus, genetic mutations of 3 genes have been observedin 33.3% in DVT/PTE cases, 4.8% in infertility cases, and4.8% in other complicated cases.Conclusions:Our intensive sequence analysis of 3 anticoagulant genesshowed that 12out of 178 cases (6.7%) carried geneticmutations that may influence the functional activity. Weconcluded that we Japanese also have the genetic factorsto DVT/PTE during pregnancy and pregnancy wastage.NotesPage 75

Abstract22 A NEW MONITARING METHOD OF FGR FETUSKazunao Suzuki, Motoi Sugimura, Naohiro KanayamaDepartment of Obstetrics & Gynecology, Hamamatsu University School of Medicine,Hamamatsu, Shizuoka, JapanThe management of FGR is becomoing one of the majorproblems in medicine from the aspect of fetal origin ofadult disease. Near-infrared spectroscopy (NIRS) hasbeen used as a noninvasive method for monitoring thereal -time oxygenation status in area such as the brainand striated muscle. Because the oxygenation status ofthe placenta is closely related to the fetal condition, theevaluation of placental oxygen condition would contributefetal management. We developed transabdominalmonitoring of the oxygenation of the placenta by NIRS in15 normal fetus and 15 FGR fetus. We succeeded inobtaining oxyhemoglobin and deoxyhemoglobin datathrough the maternal abdomen. We calculated tissueoxygenation index (TOI) from these data . The TOI levelsof normal pregnancy were not changed during 2nd and3rd trimester. The TOI value of normal fetus was70.1+/-0.6. Interestingly the TOI value of FGR was79.1+/-2.8. The TOI value of FGR was significantly higherthan that of normal pregnancy. This results could beexplained that impairment of trophoblastic functiondecreases the absorption of oxygen of maternaloxyhemoglobin from intervillous space to fetal circulationresulting in high level of TOI in FGR placenta. Thisphenomenon seems to be similar to TOI changes in lowtemperature therapy for the brain. Measurement ofPlacental TOI by transabdominal NIRS could be a newand useful indicator for management of FGR.NotesPage 76

Abstract23Keynote speakerGROWTH HORMONE: A NEUROTROPHIC FACTOR DURINGCHICK EMBRYOGENESISSteve Harvey, Marie-Laure Baudet, Esmond J. SandersDepartment of Physiology, University of Alberta, Edmonton, Alberta, T6G 2H7, CanadaEmbryogenesis and fetal growth largely occur in theabsence of the pituitary gland and are traditionallyconsidered to be growth-without-growth hormonesyndromes. However, while pituitary somatotrophs donot appear ontogenetically until at least ED (embryonicday) 14 of the 21 day incubation period of chickembryogenesis, extrapituitary tissues express the GHgene much earlier in development. Neural tissues, forinstance, (particularly projecting neurons of thefacial/acoustic, glossopharangeal, trigeminal and vagalnerves and from retinofugal and tectotubular tracts)contain abundant GH immunoreactivity and GH mRNA byED3 and the overlapping distribution of GH receptor(GHR) immunoreactivity and GHR mRNA suggests GHinvolvement in early neurogenesis. This possibility issupported by antiapoptotic actions of retinal GH thatpromote the survival of retinal ganglion cells (RGCs),since the immunoneutralization of endogenous GH inRGCs or the siRNA knockdown of endogenous GH mRNAresults in an increase in RGC apoptosis. Retinal GHmay also act as a growth factor during neurogenesis,since GH is found in the retinofugal tract (optic fibre layer,optic nerve head, optic chiasm, optic tract) prior to thesynapsing of these neurons with their target sites in thevisual centre of the brain. The growth of these neuronsmay therefore be due to the anterograde translocation ofthe GH along RGC axons. This possibility is supportedby a loss of GH immunoreactivity in the optic fibre layerafter ED 12 of development, following the synapsing of theretinofugal nerves within the optic tectum of the brain.This possibility is also supported by a trophic effect ofexogenous chicken GH on RGC neurite outgrowth.Neurotrophic actions of GH within the visual systemduring embryogenesis are also indicated by its inductionof insulin-like growth factor (IGF)-1 in the neural retinaand by the expression of a GH-response gene (GHRG)-1(a specific marker of GH action in birds) in the optictectum and other visual centres of the brain. Chickembryogenesis is therefore not a growth-without-GHsyndrome and neurogenesis within the visual system, inparticular, appears to be dependent upon local autocrineor paracrine actions of extrapituitary GH that promoteneural development.Supported by NSERC of CanadaNotesPage 78

Maternal administration of dexamethasone causes significantalterations in the electrocortical activity of the preterm fetalsheepJoanne Davidson, Lindsea Booth, Josine Quaedackers, Alistair J Gunnand Laura Bennet.Fetal Physiology and Neuroscience Group, Dept of Physiology, The University of Auckland,Auckland, New Zealand.Abstract24Antenatal corticosteroid therapy enhances fetal lungmaturation in pregnancies at risk for preterm delivery, butmay lead to injury or impaired development of otherorgans such as the brain. This study examined the effectsof a single course of maternally administereddexamethasone (DEX) on fetal brain activity. Pregnantewes at 103 days gestation, received two intramuscularinjections of either DEX (n = 8) or vehicle (n = 7) 24 hoursapart. Fetal electrocorticogram (ECoG) activity wasmonitored continuously from 24h before until 120h afterthe first injection. DEX injection one, led to a significantrise in ECoG amplitude, peaking around 12h. A second,smaller rise occurred after injection two with a return tonormal seen after 30 hours. Further analysis of the ECoGsignal showed that the rise in amplitude was mediated byvery high amplitude electrocortical discharges (up to400uV compared to normal background of 50uV) whichwere epileptiform in nature (Figure). ECoG frequency(spectral edge) was not significantly different during thistime, but was significantly elevated compared to controlfrom 17 hours after injection 2. In conclusion,dexamethasone appears to induce short-term epileptiformactivity, and long-term alterations in ECoG frequency inthe preterm fetal sheep, at an age which is equivalent tothe 28-30 week human in terms of neural maturation.Studies show that glucocorticoids can facilitate asustained calcium influx through glutamate receptorswhich may mediate the epileptiform activity we observed.The increased spectral edge may reflect dexamethasoneinduced advanced cortical maturation. Further study isrequired to determine whether dexamethasone causedneural injury.ABCDFigure 1. Fetal ECoG activity after maternaldexamethasone treatment (A, B, C) vs vehicle treatment (D)NotesPage 79

Abstract25Therapeutic Hypothermia Changes the Prognostic Value ofClinical Evaluation of Neonatal Encephalopathy.Alistair J. Gunn, Andrew Whitelaw, John Barks, John S. Wyatt, DenisAzzopardi, Charlene M. Robertson, Marianne Thoresen on behalf of theCoolCap Study GroupObjectives:to evaluate whether therapeutic hypothermia alters theprognostic value of clinical grading of neonatalencephalopathy.Study Design:Secondary analysis of a multicenter study of 234 terminfants with neonatal encephalopathy randomized to headcooling for 72 h starting within 6 h of birth, with rectaltemperature maintained at 34.5±0.5°C, followed byrewarming over 4 h, or standard care at 37.0±0.5°C.Severity of encephalopathy was measured prerandomizationand on day 4, after rewarming, in 177infants; 31 infants died before day 4 and data weremissing for 10 infants. The primary outcome was death orsevere disability at 18 months of age.Results:Milder pre-randomization encephalopathy, greaterimprovement in encephalopathy from randomization today 4, and cooling were associated with favorableoutcome in multivariate binary logistic regression.Hypothermia did not affect severity of encephalopathy atday 4, however, among infants with moderateencephalopathy at day 4, those treated with hypothermiahad a significantly higher rate of favorable outcome (31/45infants, 69%, p=0.006) compared with standard care(12/33, 36%).Conclusions:Infants with moderate encephalopathy on day 4 may havea more favorable prognosis after hypothermia treatmentthan expected after standard care.NotesPage 80

AbstractCan erythropoietin be used to protect the fetal sheep brain fromrepeated exposure to endotoxin?261 R De Matteo, 1 L Cardamone, 1 V Stacy, 1 N Blasch, 2 M Probyn, 3 N Hale,3 S Rees, 1 R Harding1 Department of Anatomy and Cell Biology, Monash University, Melbourne, Australia2 School of Biomedical Sciences, University of Queensland, Brisbane, Australia3 Department of Anatomy and Cell Biology, University of Melbourne, Melbourne, AustraliaBackground and aim:Intrauterine inflammation and infection have been linkedto preterm birth and fetal brain injury. In human infants,the brain injury is particularly seen in white matter.Exposure of immature fetal sheep to the bacterialendotoxin lipopolysaccharide (LPS) causes fetalhypoxemia, hypotension and white matter injury (Duncanet al 2002). Erythropoietin (EPO) has neuroprotectiveproperties and could alleviate the brain injury induced byintra-uterine infection. In the present study our aim wasto determine the physiological effects of EPO in the ovinefetus exposed to LPS.Methods:At 0.7 of gestation (term ~147 days) ewes and fetuseswere anesthetized and fetuses chronically catheterized.Following 5±1 days of recovery fetuses received either: i)an i.v. bolus dose of LPS (~0.9µg/kg) on 3 consecutivedays (n=9); ii) i.v. bolus of LPS followed 1 hour later by5000IU of recombinant human Epo (rhEpo; n=15); iii)rhEpo alone (n=3); iv) saline alone (n=3). Fetal arterialblood gases and mean arterial pressure were monitored.Six days after the study period ewes and fetuses wereeuthanased and the fetal brains examined histologically.Results:Fetal exposure to saline alone or rhEpo alone did not alterfetal arterial blood gases or blood pressure. Three fetuses(33%) did not survive treatment with LPS alone, and sevenfetuses (47%) died after receiving LPS plus rhEpo. Insurviving fetuses, repeated administration of LPS resultedin hypoxemia and an increase in hematocrit (Hct).Fetuses treated with LPS plus rhEpo become significantlymore hypoxemic, displayed higher plasma lactate levels,became more hypotensive and tended to be tachycardicthan fetuses treated with LPS alone (p

Abstract27ANTI-INFLAMMATORY EFFECTS OF SULFASALAZINE IN ANOVINE MODEL OF IN UTERO INFECTION: A PROSPECT FORNEUROPROTECTION?Burcu Saglam 1 , Graham Jenkin 2 , Suzanne L. Miller 3 , and Euan M. Wallace 11 Department of Obstetrics and Gynaecology, 2 Monash Immunology and Stem Cell Laboratoriesand 3 Deparment of Physiology, Monash University, Clayton, Victoria.Background:There is a causal relationship between inflammation,arising from intrauterine infection, and injury to the fetalbrain. Preterm ovine fetal exposure to lipopolysaccharide(LPS) is a commonly used model for exploring themechanisms underlying inflammation- induced fetal braininjury. Inflammation caused by LPS is mediated by thetranscription factor nuclear kappa-B (NFκB). The aim ofthis study was to investigate the effects of oraladministration of Sulfasalazine (SSZ), an NFκB inhibitor,on circulating tumour necrosis factor-α (TNF-α)concentrations and fetal brain injury.Methods:6 twin-pregnant ewes were randomly assigned to receiveSSZ tablets (n=2) or placebo sugar tablets (n=4). Surgerywas performed at 104-106 days gestation for catheterplacement in the femoral artery and vein of both of twinfetuses. Each ewe received SSZ or placebo, orally. Onefetus received three daily i.v. infusions of LPS (100ng/kg,placebo-LPS or SSZ-LPS); the control co-twin(placebo-Saline or SSZ-Saline) received saline injectionsof identical volume. Fetal plasma samples were taken tomeasure TNF-α and fetal brains were collected at postmortem for immunohistochemical evaluation of apoptoticcells by terminal deoxynucleotidyl transferase-mediateddUTP-nick end labelling (TUNEL).Results:There was an increase in plasma TNF-α concentrations(from undetectable levels to 85.77±15.84ng/ml) in theplacebo-LPS fetuses following the first LPS administration,which returned to baseline within 6 hours post-LPSexposure. There was an attenuation of the TNF-αresponse with subsequent LPS exposure. TNF-α wassignificantly increased in the SSZ-LPS fetuses (fromundetectable levels to 42.16±3.86ng/ml) but to a lesserextent than in the Placebo-LPS group. TNF-α was notdetectable in either of the Saline treated groups.TUNEL-positive staining cells were present in the brainsections of Placebo-LPS fetuses and there appeared tobe a decrease in TUNEL-positive staining in the SSZ-LPSbrains.Conclusion: Sulfasalazine reduces concentrations ofTNF-α in the fetal circulation and there was a decrease inthe presence of TUNEL-positive cells within these fetalbrains. We propose that SSZ may be neuroprotectivefollowing inflammation.NotesPage 82

AbstractFETAL BRAIN INJURY IN SINGLE UMBILICAL ARTERYLIGATION (SUAL) INDUCED IUGR28Low HM 1 , Supramaniam VG 1 , Castillo-Meléndez M 2 , Jenkin G 1 , Miller SL 1 ,Wallace EM 3 .Monash Immunology and Stem Cell Laboratories 1 , Departments of Physiology 2 and Obstetricsand Gynaecology 3 , Monash University, Victoria, 3800, AustraliaIntrauterine growth restriction (IUGR) is associated withincreased risk of premature delivery, risks of death andmorbidity and increased risks of neurological andcognitive impairments in IUGR infants. We haveestablished a chronic fetal sheep hypoxia model of singleumbilical artery ligation (SUAL) with brain sparing andsignificantly elevated brain to body weight ratio. Weexamined the morphological changes in the subcorticalwhite matter, striatum, thalamus and hippocampusregions of the brain following SUAL.Methods:Pregnant singleton ewes underwent surgery at 105-110days gestation (dGA). Fetuses were instrumented withfemoral artery, jugular vein and amniotic fluid catheters.Single umbilical artery ligation (SUAL) was performed toinduce IUGR (n=5) while the cord remained intact incontrols (n=4). Electromyography (EMG) leads weresutured to the external wall of the uterus. SUAL animalswere sacrificed when an increased EMG activity,indicative of labour onset, was observed. Controls weresacrificed at 135 dGA. At post mortem, fetal brains wereperfused with 0.9% saline followed by 4%PFA. The fixedbrains were sectioned into blocks and paraffin embedded.White matter injury was assessed using cresyl violet andacid fuchsin (CV-AF; neuronal injury), lectin (activatedmicroglia/macrophages), Tau (neurofibrillary tangles) andalbumin (blood brain barrier; BBB) immunohistochemistry.Results:Fetal hypoxaemia and IUGR was established via SUAL.IUGR fetuses showed significant brain sparing comparedto controls with brain to body weight ratios of 18.09±0.49and 13.45±0.13 respectively. Albumin and Tau positivecells were present in the subcallosal bundle, cingulategyrus, thalamic nuclei, periventricular thalamic nuclei andinternal capsule of SUAL fetuses but not in controls.Lectin positive cell numbers were elevated in the mostseverely affected SUAL fetus (11.34±2.489 cells per fieldof view) and minimal in other SUAL fetuses (0.96±0.32)and control fetuses (0.56±0.23). Histochemical stainingusing CV-AF identified a slight increase in pyknotic cells inSUAL fetuses (20.87±1.45) compared to controls(18.65±1.77) in the cingulate gyrus, ventral striatum andseptal nucleus.Conclusion:The brains of SUAL induced IUGR fetuses displayedmarked neuronal damage, activation of microglia/macrophages, presence of neurofibrillary tangles and BBBcompromise, which may lead to significant morbidity afterbirth.NotesPage 83

Abstract29 Perinatal DiI tracing of developing hypothalamic connections.Makarenko I.G., Alpeeva E.V.N.,K. Koltzov Institute of Developmental Biology RAS, Moscow, Russian FederationOBJECTIVE:Hypothalamus plays an important role in regulation ofbasic visceral, somatic and emotional functions that canbe realized by its connections with numerous brainstructures. There is a notion that disruption ofhypothalamic development may be unsuspected cause ofdisorders such as obesity and high blood pressure,emotional or neurological disturbances. Unfortunatelyformation of axonal connections is one of the mostunknown events of brain development. The aim of ourstudy was to describe the time schedule andconsequence of the development of the hypothalamicprojection systems.METHODS:Carbocyanine dye tracing method was used on the datedrats from E14 to P10. Crystals of DiI were inserted into thedifferent parts of hypothalamus, septum, midbrain andpituitary lobes on the prefixed with 4% paraformaldehydebrains. After the storage in the same fixative for 4-16month labeled nerve cells and fibers were revealed onthick vibratome sections using fluorescent and confocalmicroscopy.RESULTS:Retrogradely labeled neurons were visualized in the brainand carefully described after all kinds of the DiIapplications and thus revealed the sources of distinctprojection systems. Early prenatal development beginningfrom E14-E15 was specific for hypothalamo-posteriorpituitary, mammillo-tegmental and septohypothalamicconnections. It was shown that septohypothalamicconnections are reciprocal and more numerous with thepreoptic region and anterior hypothalamus than withmedial and caudal hypothalamic regions. All these fibersystems are well developed before the birth. Hypothalamicprojections to the intermediate pituitary lobe were revealedfirst close to the end of intrauterine life and differentiatedprogressively from P5 to P20. Mammillothalamic tracteven organized by the collateral branches of themammillotegmental tract grows from E18, reach unilateralanteromedial thalamic nucleus on E20-E21 and innervatesall anterior thalamic nuclei on postnatal stages (from P2 toP10). Functional activity of the thalamic synapses wasconfirmed by the immunocytochemical visualization ofsynapsin on P4-P5.CONCLUSIONS:Thus we have described specific time schedule of thenormal development of several hypothalamic projectionsystems. This work was supported by RFBR grant07-04-00798.NotesPage 84

AbstractKeynote speaker30 Chronic Fetal Hypoxia : Consequences and TreatmentSuzanne L. Miller 1,2 , Veena G. Supramaniam 2 , Euan M. Wallace 3 , GrahamJenkin 2Department of 1 Physiology, 2 Monash Immunology and Stem Cell Laboratories, and 3 Obstetricsand Gynaecology, Monash University, Victoria, AustraliaChronic fetal hypoxia, together with intrauterine growthrestriction (IUGR), continue to present serious challengesin modern obstetrics, both in terms of diagnosis andtreatment options. IUGR is strongly associated withincreased risks of perinatal mortality and significant shortand long term morbidity. To explore possible treatmentstrategies, we use the technique of single umbilical arteryligation (SUAL) in ovine fetuses. This model inducesplacental insufficiency thereby producing chronic hypoxia,cardiovascular redistribution and asymmetric fetal growthrestriction, as occurs in human IUGR fetuses. A notablecomplication of IUGR pregnancies is preterm birth andtherefore glucocorticoids (including betamethasone; BM)are often administered to mature the fetal lungs. However,in our SUAL model, BM is potently vasoactive causingsignificant cerebral hypoperfusion, followed by a largereperfusion in SUAL fetuses which is not observed inhealthy fetuses. The degree of rebound perfusion iscorrelated with fetal brain cellular lipid peroxidation andapoptosis, suggesting that BM administration could havesignificant adverse effects in the IUGR fetus. IUGR is alsoassociated with significant fetal oxidative stress andinflammation. Mitigation of these factors may lead toimproved perinatal and long term outcomes. Using ourSUAL model, we are currently exploring a number ofselect treatment regimens, including phosphodiesteraseinhibitors to improve uterine blood flow, anti-inflammatoryagents and antioxidants to reduce adverse effects ofIUGR, thereby improving neonatal wellbeing.Unfortunately, administration of the type 5phosphodiesterase inhibitor sildenafil citrate (Viagra),does not improve uteroplacental blood flow in IUGRfetuses. Outcomes from the maternal administration ofanti-inflammatory or antioxidant agents in IUGR look morepromising as possible future therapies.DR SUZANNE L MILLERQualifications:BSc (Hons), PhD, Department of Physiology, Faculty of Medicine, Nursing and Health Sciences Monash University.Current Appointment:Senior Research Officer, Department of Physiology and Monash Immunology and Stem Cell Laboratories, MonashUniversity. Research Fellow, Royal Australian and New Zealand College of Obstetricians and Gynaecologists(RANZCOG).PI on NHMRC Program Grant: “Control of Reproductive Processes”CI on NHMRC Project Grant “The effects of maternal glucocorticoid administration in growth restricted fetuses”Research Summary:After completing my PhD at Monash University in 1999, in which I studied the regulation of uterine blood flow duringpregnancy, I gained a Postdoctoral Fellowship at University College London, where I worked with Professors MarkHanson and Donald Peebles in the highly regarded Fetal and Neonatal Research Group. My postdoctoral researchaddressed brain growth, development and the use of potential neuroprotective strategies to ameliorate the effects ofreduced oxygen availability and infection in a fetal sheep model. I returned from the UK in 2001 and, since then, havebeen working with a team that is exploring fetal growth, development and adaptation in response to chronic fetal hypoxiaor infection in pregnancy. Presently, I am funded by the NHMRC and the RANZCOG on projects that investigate the fetaladaptations observed in response to acute hypoxia, chronic hypoxia, reduced blood flow, infection and glucocorticoidexposure from the cellular level through to gross developmental and cognitive effects, with a view to implementingclinically useful neuroprotective strategies for the at risk fetus and neonate.NotesPage 86

AbstractProliferation in the subventricular zone (svz) after severehypoxia and induced hypothermia in preterm fetal sheep31Sherly George, Robert Barrett, Laura Bennet, Justin Dean, Alistair JanGunn.Fetal Physiology and Neuroscience Group, Dept Physiology, The University of Auckland,Auckland, New ZealandProlonged, moderate cerebral hypothermia is the firstclinically proven therapy to improve recovery fromhypoxia-ischaemia at term. Recent experimental datasuggest that cooling is also neuroprotective after severehypoxia in preterm animals, but that it may reduceproliferation of brain cells in the periventricular whitematter, at a critical period in the development of the brain.Although there is significant ongoing proliferation in theperiventricular regions, new cells are primarily producedin the subventricular zone (SVZ). In a chronicallyinstrumented sheep model of preterm asphyxia, weinvestigated the effects of cerebral hypothermia onproliferation in the SVZ. Preterm (0.7 gestation) fetalsheep received complete umbilical cord occlusion for 25min followed by cerebral hypothermia starting 90 min afterreperfusion and continuing until 72 h after occlusion.There was a significant increase in numbers of cellslabeled with proliferating cell nuclear antigen (PCNA), butnot of cells labeled with KI-67, in the hypothermiaocclusiongroup compared with normothermia-occlusionanimals and sham controls. In conclusion, despiteprofound suppression of proliferation in the vulnerableperiventricular region after severe hypoxia and treatmentwith hypothermia in the developing brain, proliferation inthe SVZ was not suppressed, either by hypoxia or byprolonged hypothermia. The apparent differences in effectof hypothermia suggested by the two markers forproliferation likely reflect the more restricted binding ofPCNA in the cell cycle. Further studies are needed toevaluate the long-term impact of hypoxia and hypothermiaon cell division and differentiation in the developing brain.NotesPage 87

Abstract32The ontogeny of chemoreflex and haemodynamic responses toprolonged umbilical cord occlusion in fetal sheepGuido Wassink, Laura Bennet, Lindsea C. Booth, Ellen C. Jensen, BertWibbens, Justin M Dean, and Alistair J Gunn.Fetal Physiology and Neuroscience Group, Dept Physiology, The University of Auckland,Auckland, New ZealandIt has been hypothesized that blunted chemoreflexmediated responses to hypoxia may compromise the abilityof the preterm fetus to adapt to hypoxic or asphyxial stress.However, there are only limited quantitative data on theontogeny of chemoreflex and hemodynamic responses tosevere asphyxia. Chronically instrumented fetal sheep at0.6 (n=12), 0.7 (n=12) and 0.85 (n=8) of gestational age(ga; term = 147 days) were exposed to 30, 25 or 15minutes of complete umbilical cord occlusion, respectively.At all ages occlusion was associated with early onset ofbradycardia, profoundly reduced femoral blood flow andconductance, and hypertension. The 0.6ga fetuses showeda significantly slower and lesser fall in femoral blood flowand conductance compared with the 0.85ga group, with acorrespondingly reduced relative rise in mean arterial bloodpressure. As occlusion continued, the initial adaptation wasfollowed by loss of peripheral vasoconstriction andprogressive development of hypotension in all groups. The0.85ga fetuses showed significantly more sustainedreduction in femoral conductance but also more rapid onsetof hypotension than either of the younger groups.Electroencephalographic (EEG) activity was suppressedduring occlusion in all groups but the rate and degree ofsuppression were least at 0.6ga. In conclusion, thenear-midgestation fetus shows attenuated initial(chemoreflex) responses to severe asphyxia comparedwith more mature fetuses, but more sustainedhemodynamic adaptation and reduced suppression ofEEG activity during continued occlusion of the umbilicalcord. These results are consistent with greater cardiacand cerebral tolerance to profound hypoxianear-midgestation.NotesPage 88

AbstractEffect of acute and chronic hypoxia on amniotic fluid gases inchick embryogenesis.33M.V. Nechaeva 1 , H. Toenhardt 2 , D. Marquardt 2 .1 Institute of Developmental Biology RAS, Moscow, Russia, Mnechaeva2003@yahoo.com.2 Free University, Institute of Veterinary Physiology, Berlin, Germany.Objective:The prenatal hypoxia induces a complex response, andnumerous functional systems are involved to prevent thedamage of embryo. The mechanism of the developmentalresponse to hypoxia is not entirely clear. We studied thechanges of oxygen tension in amniotic fluid (AF) duringdevelopment of chick embryo and under the influence ofacute and chronic hypoxia.Methods:Chick eggs were incubated at 37.5°C. Po 2 , Pco 2 , and pHwere measured in samples of AF using blood gasanalyzer Radiometer Copenhagen ABL 605 on incubationdays 10 (D10), 12 and 14 in control and after the eggexposure to acute hypoxia (10%O 2 for 10min) or chronichypoxia (15%O 2 since D6).Results:In control, the Po 2 in AF was about 55 mmHg andremained relatively constant during this period. The Pco 2significantly increased from 19.85 ± 0.52 to 30.22 ± 1.87mmHg, while pH significantly decreased from 6.91 ± 0.03to 6.58 ± 0.05 since D10 to D14.Acute hypoxia reduced the Po 2 in AF by 25; 23 and 16%from the control values on D10, 12 and 14, respectively.The Pco 2 increased, but the effect was significant only onD12. The pH decreased and this effect was morepronounced on D14.Chronic hypoxia increased the Po 2 in AF by 20% from thecontrol value on D10, and did not change it significantly onD12 and D14. The Pco 2 decreased on D10, D12, butdidn’t change on D14. The pH decreased on D10 and D12,but rose significantly on D14.Conclusion:The effects of acute and chronic hypoxia on gases valuesin AF were different. These results are discussed inregards to the role of oxygen in AF for the embryometabolism during hypoxia and for the function ofamniotic membrane, which is not vascularised and mayuse oxygen from amniotic fluid. This work was supportedby RFBR grant 05-04-49434.NotesPage 89

Abstract34Maternal treatment with allopurinol diminishes fetal cardiacoxidative stress following repeated episodes of ischaemiareperfusionin sheepH Torrance 1 , JB Derks 1 , MA Oudijk 1 , AS Thakor 2 , T Cindrova-Davies 2 ,F van Bel 1 , GHA Visser 1 , GJ Burton 2 & DA Giussani 21 Department of Perinatology, University Medical Centre Utrecht, The Netherlands and2 Department of Physiology, Development & Neuroscience, University of Cambridge, UK.Introduction The prevention and managementof perinatal asphyxia remain major concerns inobstetric practice today. Umbilical cordcompressions (UCC) not only induce fetalasphyxia but also episodes of ischaemiareperfusion(I/R). I/R promotes the productionof reactive oxygen species (ROS), for instancevia activation of the xanthine oxidase (XO)pathway, which may lead to oxidative stress inthe fetal cardiovascular system. While treatmentwith allopurinol of severely asphyxic humanneonates reduced free radical formation andimproved cardiovascular status (1), allopurinoltreatment started postnatally was deemed toolate to prevent oxidative damage (2). As a result,in pregnancy complicated with severe fetalasphyxia, recommendations to treat the fetusvia the mother, rather than the neonate, withallopurinol are currently being entertained. Thisstudy investigated the effects of maternaltreatment with allopurinol on indices of oxidativestress in the fetal heart following repeated UCCin late gestation sheep.Methods Under halothane anaesthesia, 10sheep fetuses and their mothers wereinstrumented at 0.8 of gestation with vascularand amniotic catheters, an inflatable occluderaround the umbilical cord, and a Transonic flowprobe around an umbilical artery. At least 5 dayslater, all fetuses were submitted to an I/Rchallenge produced by 5 x 10 min UCC at 10min intervals, each designed to reduce umbilicalblood flow (UBF) by 80-90% from baseline andto lead to a progressive fall in fetal pHa to 6.9. In5 fetuses, the I/R challenge was induced duringmaternal i.v. treatment with allopurinol (SigmaLtd., 30 mg.kg -1 over 20 minutes). In theremaining 5 fetuses, the I/R challenge wasinduced during maternal infusion with salinevehicle at the same rate. Infusion started 10 minbefore the 4 th UCC and finished immediatelyafter the end of the 4 th UCC. The fetal heartswere collected 48h after I/R and snap frozen formeasurement of oxidant and antioxidantproteins by Western blot. Hearts collected from5 non-instrumented fetal sheep at 0.8 gestationserved as controls. Statistical comparisons weremade using one-way ANOVA with the Tukeypost-hoc test. Significance was accepted whenp

Pre-existing hypoxia is associated with a delayed but moresustained rise in T/QRS ratio during prolonged umbilical cordocclusion in near-term fetal sheepLaura Bennet, Bert Wibbens, Jenny A. Westgate, Harmen H. De Haan,Guido Wassink, and Alistair J. GunnFetal Physiology and Neuroscience Group, Dept of Physiology, The University of Auckland,Auckland, New Zealand.Abstract35There is limited information on whether pre-existing fetalhypoxia alters hemodynamic responses and changes inT/QRS ratio and ST waveform shape during subsequentsevere asphyxia. Chronically instrumented near-termsheep fetuses (124±1 days) were identified as eithernormoxic PaO 2 > 17 mmHg (n=9) or hypoxic PaO 2 ≤ 17mmHg (n=5), and then received complete occlusion of theumbilical cord for 15-min. Umbilical cord occlusion lead tosustained bradycardia, severe acidosis and transienthypertension followed by profound hypotension in bothgroups. Pre-existing hypoxia did not affect changes inmean arterial blood pressure, but was associated with amore rapid initial fall in femoral blood flow and vascularconductance and with transiently higher fetal heart rate at2 minutes and from 9 to 11 minutes of occlusioncompared with previously normoxic fetuses. Occlusionwas associated with a significant but transient rise inT/QRS ratio; pre-existing hypoxia was associated with asignificant delay in this rise (maxima 3.7 ± 0.4 vs. 6.2 ± 0.5minutes), but a slower rate of fall. There was a similarelevation in troponin-T levels 6 hours after occlusion in thetwo groups (median (range) 0.43 (0.08, 1.32) vs 0.55(0.16, 2.32) µg/L, N.S.). In conclusion, mild pre-existinghypoxia in normally grown singleton fetal sheep isassociated with enhanced centralisation of circulationafter umbilical cord occlusion and delayed elevation of theST waveform and slower fall, suggesting that chronichypoxia improves myocardial dynamics during asphyxia.NotesPage 91

NotesPage 92

Abstract36High incidence of ketosis and hyper-homocysteinemia in thelast trimester of Japanese mothers. without any complicationsFukuoka Hideoki*, Watanabe Hiroko**, Nagai Yasushi***, OgasawaraChiyoko***, Asano Shigetaka** Institute for Epigenetic Regulation of Fetal Development Waseda University, Tokyo, Japan** Dapt of Nursing Science, the Graduate School of Medicine Kyoto University***Nagai Clinic, Saitama, Japan.In these 2 or 3 decades, the Japanese rate of low birthweight infant has been growing to 9.44 % in 2004, and theaverage birth weight of boys and girls has been loweringunder 3000g in 2005. The next generation has beensuspected to face the high risk of adult diseases in future.To study the maternal nutritional state, in one urbandistrict hospital on the outskirts of Tokyo, we studied 198mothers at the gestational age 12th, 20th, 32nd and 36thweek, with DHQ (Diet History Questionnaire) and serumhomocystein and ketone bodies, under the approval of theEthical Committees of University of Tokyo. In Japan, theestimated energy requirement for 20’s pregnant mothersis 2550 Cal for the last trimester. DHQ study, however,disclosed no change of energy intake from 1st, 2nd until3rd trimester (average energy intake: 1723, 1754, 1792Cal, respectively), and ,by comparison, that of nonpregnant ladies was 1722 Cal. Many mothers weresuspected to be in the severe energy deficit and abnormalmetabolic state. Under this doubt, we measured theketone bodies. ( acetoacetate, 3βhydroxbutylate andacetone). High ketone bodies were detected in 11.7 and29.3 % at 12th and 32nd gestational week. In the lasttrimester, 89% mothers had the folate intake under 400 μgand 13% showed serum homocystein over 5.5±0.4 μg/L.High homocystein means the possibility of high3βadenosylhomocystein(3βADH) inside cells. TheJapanese maternal nutritional state is not so ideal withlong lower energy and folate intake and high incidence ofketosis, inducing lowering the birth weight. The averagebirth weight in no ketosis mothers was higher than theketosis group in each BMI without any clear relation offolate and energy intake. But low intake of folate withhigher homocystein suggests possibility of disruptedturnover of one carbon metabolism and DNA methylationin the fetus.NotesPage 94

AbstractGenomic imprinting in the placenta involves histone tailmodifications independent of DNA methylation37Kohzoh Mitsuya 1 and Kunihiro Okamura 21Biofunctional Science, Tohoku University Biomedical Engineering Research Organization(TUBERO)2Department of Obstetrics and Gynecology, Tohoku University School of MedicineImprinted genes are expressed from only one of theparental chromosomes and are marked epigenetically byDNA methylation and histone modifications. Theimprinting center 2 (IC2) on mouse distal chromosome 7is flanked by several paternally repressed genes, with themore distant ones imprinted exclusively in the placenta.We found that most of these genes lack parent-specificDNA methylation, and genetic ablation of methylationdoes not lead to loss of their imprinting in the trophoblast(placenta). The silent paternal alleles of the genes aremarked in the trophoblast by repressive histonemodifications (dimethylation at Lys9 of histone H3 andtrimethylation at Lys27 of histone H3), which are disruptedwhen IC2 is deleted, leading to reactivation of the paternalalleles. Thus, repressive histone methylation is recruitedby IC2 (potentially through a noncoding antisense RNA) tothe paternal chromosome in a region of at least 700 kband maintains imprinting in this cluster in the placenta,independently of DNA methylation. We propose that anevolutionarily older imprinting mechanism limited toextraembryonic tissues was based on histonemodifications, and that this mechanism was subsequentlymade more stable for use in embryonic lineages by therecruitment of DNA methylation. A possible role forhistone tail modifications in epigenetic inheritance will befurther discussed.NotesPage 95

Abstract38LACTOBACILLI SUPERNATANT INHIBITS TNF-α PRODUCTIONAND COX 2 EXPRESSION IN LPS-ACTIVATED PLACENTALTROPHOBLASTS1,3 M Yeganegi, 3 C Watson, 2 S Kim, 2 G Reid, 1 J Challis and 1,3 A Bocking1 Dept. Physiology & Ob/Gyn, U Toronto, Canada; 2 Dept. Microb. & Immun., U Western Ontario,London, Canada and 3 SLRI, Mt. Sinai Hosp., Toronto, CanadaObjective:Bacterial Vaginosis (BV) is characterized by absence ofendogenous Lactobacillus and an increased risk ofpreterm birth (PTB). BV-associated pathogenic bacteriaupregulate pro-inflammatory cytokines, leading toincreases in prostaglandins. Lactobacilli are known toinhibit pro-inflammatory cytokines in mouse macrophages.We hypothesized that Lactobacillus rhamnosus GR-1interferes with the cascade leading to prostaglandinsynthesis by downregulating pro-inflammatory cytokineproduction and COX 2 protein expression in humanplacental trophoblasts.Methods:Placental trophoblasts were isolated from term electivec-section placentae and divided into six groups: 1) Notreatment; 2) Treatment with LPS; 3,4) Treatment withlactobacilli supernatant or MRS media; 5,6) Pretreatmentwith lactobacilli supernatant or MRS followed by LPStreatment. COX 2 expression was measured by WesternBlot and cytokine concentrations by ELISA. Cell viabilitywas tested by LDH analysis.Results:LPS stimulation caused a marked increase in TNF-αproduction and COX 2 expression in placental trophoblasts.Pretreatment with lactobacilli supernatant downregulatedthese increases. Treatment with supernatant alone had noeffect on cytokine production or COX 2 expression. Therewere no changes in IL-1β concentrations with anytreatment. LDH analysis showed minimal apoptosis inlactobacilli treated cells.Conclusion:Probiotic lactobacilli inhibit both TNF-α production andCOX 2 expression in placental trophoblasts. This studyprovides evidence for potential mechanisms by whichlactobacilli reduces the risk of BV-associated PTB.NotesPage 96

INTERACTION BETWEEN LIPOXYGENASE PATHWAY ANDPROGESTERONE ON 11β-HYDROXYSTEROIDDEHYDROGENASE TYPE 2 IN CULTURED HUMAN TERMPLACENTAL TROPHOBLASTSK Sato 1,2 , H Chisaka 2 , K Okamura 2 , JRG Challis 11 Department of Physiology, Obstetrics, Gynecology and Medicine, University of Toronto,Ontario, Canada 2 Department of Obstetrics and Gynecology, Tohoku University GraduateSchool of Medicine, Sendai, JapanAbstract39Objective:Placental 11β-hydroxysteroid dehydrogenase type 2(11β-HSD2) is localized to placental syncytiotrophoblastsand plays an important role in pregnancy maintenanceand fetal maturation by inactivating excess glucocorticoids.In the event of intrauterine infection, lipoxygenase (LOX)metabolites are produced in the placenta and contribute topreterm labor and adverse fetal outcomes. On the otherhand, LOX metabolites are involved in production ofprogesterone, which is required for pregnancymaintenance. In this study, we evaluated the interactionsbetween LOX pathway and progesterone on 11β-HSD2.Specifically, we hypothesized that LOX metabolites wouldalter 11β-HSD2 and this effect would be mediated byprogesterone.Methods:We cultured human term placental trophoblasts in thepresence/absence of LOX inhibitors, Nordihydroguaiareticacid (NDGA), AA861 and Baicalein; LOX metabolites,Leukotriene B 4 (LTB4) and 12(S)-hydroxyeicosatetraenoate(12-HETE); progesterone (P4) and progesterone receptorantagonist RU486. We used radiometric conversion assay,real-time RT-PCR, Western blot analysis and ELISA.Results:LOX metabolites down-regulated 11β-HSD2 activity andmRNA expression. LOX inhibitors up-regulated 11β-HSD2activity and mRNA/protein expression, and these effectswere attenuated by addition of LOX metabolites. Netprogesterone output was increased by LOX metabolitesand decreased by LOX inhibitors in a dose dependentmanner. 11β-HSD2 activity and mRNA/protein expressionwere down-regulated by progesterone, and these effectswere blocked by RU486. Both mRNA and proteinexpression of 11β-HSD2 were up-regulated by RU486.The suppressive effect of 12-HETE on 11β-HSD2 activitywas reversed by RU486.Conclusion:We conclude that 11β-HSD2 in human placentaltrophoblasts is decreased by progesterone and increasedby inhibition of endogenous LOX metabolites, and that acomponent of the effect of LOX metabolites on 11β-HSD2is mediated by their stimulation of endogenousprogesterone output.NotesPage 97

Abstract40Distribution of the thyroid hormone transporter, MCT8, in fetal andplacental tissues of murine, human and ovine speciesJenny L Macrae, Theo J Visser*, Graham J Burton, F B Peter Wooding,Abigail L Fowden and Alison J ForheadDepartment of Physiology, Development and Neuroscience, University of Cambridge,Cambridge CB2 3EG, UK; *Department of Internal Medicine, Erasmus Medical Center,Rotterdam, The Netherlands.Membrane transporters, such as monocarboxylatetransporter MCT8, allow thyroid hormones access tonuclear receptors in target tissues. Thyroid hormonesare important regulators of fetal growth and development;however, MCT8 expression in fetal tissues is unknown.This study investigated the distribution of MCT8 in fetaland placental tissues from murine, human and ovinespecies.Mouse embryos and placentae were collected at 16 and19d of gestation (term 20d). Tissues were also obtainedfrom newborn and adult mice. Human placentae in thefirst trimester (5-6w) and at term (40w), and embryos at9w, were collected during clinical procedures. A varietyof tissues were obtained from sheep fetuses at 130 and144d of gestation (term 145d). All animal and clinicalsamples were obtained according to UK legislation.Tissue expression of MCT8 was determined byimmunohistochemistry using a polyclonal rabbit antibodyagainst human MCT8 and visualised with 3-3’-diaminobenzamine. In all species, immunostaining wasabolished by co-incubation with blocking peptide.In mouse embryos, MCT8 was localised to cardiac andskeletal myocytes, hepatocytes, brown adipocytes, bonecells, pulmonary epithelium, intestinal epithelium, renaltubules, dorsal root ganglia, skin epidermis and choriodplexus. In the murine placenta, MCT8 was localisedspecifically to the yolk sac. There were no differences inlocalisation between embryonic, neonatal and adulttissues.In partial sections of human embryos, similar expressionof MCT8 protein was identified in cardiac and skeletalmyocytes, bone cells, skin epidermis and areas of thebrain. In the human placenta, MCT8 was expressed inendometrial gland epithelium in the first trimester, and infetal villous trophoblast in the first trimester and at term.In ovine fetuses, MCT8 was detected in all tissuesexamined. Expression was observed in cardiac andskeletal myocytes, hepatocytes, brown adipocytes,pulmonary epithelium, renal tubules, adrenal gland,pancreatic islets and skin epidermis. In the ovineplacenta, MCT8 appeared to be localised to the fetaltrophoblast layer. There was no difference in tissuelocalisation of MCT8 between the gestational agesstudied.Therefore, in murine, human and ovine species, MCT8 iswidely expressed in the fetus and placenta, especially intissues sensitive to thyroid hormones before birth.Supported by The Royal Society and Isaac Newton Trust.NotesPage 98

Abstract41DETERMINANTS OF THE NUMBER OF CARDIOMYOCYTES INSHEEP: THE INFLUENCE OF FETAL AND POSTNATALGROWTHVictoria Stacy, Megan Probyn, Robert DeMatteo, Nigel Wreford, RichardHarding, M. Jane BlackDepartment of Anatomy and Cell Biology, Monash University, VIC 3800, AustraliaAims:The muscle cells of the heart (cardiomyocytes) becometerminally differentiated and cease proliferating soon afterbirth; therefore, the ultimate number of cardiomyocytes isdetermined during fetal and early postnatal life. Our aimwas to determine the effects of fetal and postnatal growthin singleton and twin lambs on the number of ventricularcardiomyocytes.Methods:Hearts were collected at necropsy from both singleton(n=5) and twin (n=5) lambs at 9 weeks of age, whencardiomyocytes are terminally differentiated. The walls ofthe left and right ventricles were systematically sampled.The number of cardiomyocytes was stereologicallyestimated in the left and right ventricles and septum usingan unbiased optical dissector / fractionator technique.Results:At birth, twins were 17% lighter than singletons and 28%lighter at 9 weeks (both p

Modulation of Pulmonary Vascular Smooth Muscle Cell(PVSMC) Phenotype in Hypoxia: Role of cGMP-DependentProtein Kinase (PKG) .Weilin Zhou, Chiranjib Dasgupta, Sewite Negash, J Usha Raj.Division of Neonatology, Harbor-UCLA Medical Center, Los Angeles Biomedical ResearchInstitute at Harbor-UCLA, Torrance, CA 90502.Abstract42Chronic hypoxia triggers pulmonary vascular remodeling,which is associated with changes in PVSMC phenotypefrom a contractile, differentiated to a synthetic,de-differentiated type. We have reported previously thatacute hypoxia results in decrease in PKG activity inPVSMC (AJP, 2004), hence we investigated whetheraltered expression of PKG in hypoxia explain SMCphenotype modulation. Hypoxia-induced reduction in PKGexpression correlated strongly with the repressedexpression of SMC phenotypic markers myosin heavychain, calponin, vimentin, α-smooth muscle actin andthrombospondin, indicating that exposure to hypoxiaresulted in phenotype modulation to de-differentiatedstate and PKG may be involved in SMC phenotypemodulation. The results from PKG-specific smallinterfering RNA (siRNA) transfection and treatment withDT-3 (a peptide inhibitor of PKG, 30μM) in FPVSMCindicate that the expression of SMC phenotype proteinexpression decreased by hypoxia was also similarlyimpacted by PKG inhibition. Over-expression of PKG inFPVSMC by transfection of FPVSMC with a full lengthPKG construct tagged with GFP (PKG-GFP) reversed theeffect of hypoxia on the expression of SMC phenotypemarker proteins. These results suggest that PKG could beone of the determinants for the expression of SMCphenotype maker proteins and may be involved in themaintenance of the differentiated phenotype in pulmonaryvascular SMCs in hypoxia.NotesPage 101

Abstract43Human fetal cardiovascular baro-reflex responses in maternalsteroid administration analyzed by pulsed Doppler minutetissue displacement measurementNorio SHINOZUKADepartment of Obstetrics and Gynecology, Japan Red Cross Medical CenterObjectivesInvestigation of the cardiovascular baro-refelx response inhuman fetus has been methodologically limited. Dopplerminute tissue displacement measurement, we devised,enables to measure fetal aortic pulses at microns order.To clarify fetal baro-reflex changes in human fetus, thechanges in aortic pulse pressure amplitude and heart rateare analyzed in the fetus treated with trans-maternalsteroid administrationMethodsHardware: We developed the multi-channel Dopplerdevice with high accuracy tissue displacementmeasurement system that enabled to calculate minutetissue displacement at the order of micrometer. A ColorDoppler Ultrasound machine(Mochida SonoColor) wasmodified to handle multi-depth Doppler raw signals withdigital quadrature detector. This system can measure 32channels of tissue Doppler signals at 1.5mm intervalstoward the beam direction simultaneously (Freq.5MHz:Rep.Freq:1.6kHz SampleFreq.400Hz).Measurements: Twelve fetuses treated with antenataltrans-maternal steroid administration(range26w0d-32w0d)were analyzedProtocol of trans-maternal steroid administration :Dexamethasone 12mg x 2Fetal descending aortic wall pulse wave was measured atpre-steroid and 24h after steroid administrationThe amplitude of aortic wall pulse wave (AWPW) andheart rate were calculated (60 second data obtained atquiet state )Fig 1ResultsThe baseline FHR was decreased from 154±10 to144±10bpm (p < 0.05) and AWPW amplitude wasincreased from 579±123 to 890±174μm (p

EFFECTS OF MATERNAL BETAMETHASONE ADMINISTRATIONON REGIONAL BLOOD FLOW IN THE INTRAUTERINE GROWTHRESTRICTED (IUGR) OVINE FETUSSupramaniam VG 1 , Jenkin G 1 , Wallace EM 2 , Miller SL 3Monash Immunology & Stem Cell Laboratories, Departments of Obstetrics and Gynaecology 2and Physiology 3 Monash University, Victoria, 3800, AustraliaAbstract44Glucocorticoid administration to pregnant women at riskof delivering preterm has beneficial effects on fetal lungmaturation. However, glucocorticoids may also causenon-pulmonary effects, particularly on the fetalcardiovascular system. These effects may beexacerbated in IUGR fetuses. The aim of this study wasto investigate the effect of maternal betamethasoneadministration on regional blood flow in the IUGR fetus.Methods:Pregnant sheep carrying twins underwent surgery at105-110 days gestation. Both twins were instrumentedwith catheters in the carotid artery, and femoral arteryand vein. Single umbilical artery ligation (SUAL) wasperformed in one twin to induce IUGR. On day 5 aftersurgery, betamethasone (11.4mg) was administered imto the ewe (BM1) and repeated 24 hours later (BM2). Toassess blood flow, coloured microspheres were injectedinto the fetal venous circulation at time points relative toBM1: -24h, -1h, +5h, +11h and +33h. Animals weresacrificed after BM2 injection.Results:SUAL fetuses weighed less than the controls(1.4±0.01kg versus 1.69 ± 0.03kg) and had a greaterbrain-to-body weight ratio (22.64±1.19 versus17.83±1.75), indicative of brain sparing. Mean basalblood flow to the brain was 900ml/min/100g in controlsand 740ml/min/100g in SUAL fetuses. At +5h therewas a 20% fall in total brain blood flow in SUAL fetusesand a 7% fall in controls. However, at +33h blood flowincreased by 97% (SUAL) and 21% (control). The trendfor a decrease in blood flow at +5h was observed in allbrain regions examined; i.e. white matter, grey matter,hippocampus, thalamus, brainstem and cerebellum. Inthe SUAL fetuses only, regional blood flow wasincreased at +33 h (11h after BM2) in all areas, rangingfrom 60% to 110% above basal levels. There weresignificant differences between control and SUALfetuses in blood flow in the heart, lungs andplacentomes.Conclusions:Blood flow to the brain of SUAL fetuses, in response tobetamethasone, differs from that of control fetuses, Thesignificant reperfusion after a second BM administrationin all regions may be detrimental to the alreadycompromised fetal brain.NotesPage 103

Abstract45A Novel extraction method of fetal electrocardiogram from thecomposite abdominal signal.Yoshitaka Kimura *1 ,Takuya Ito *1 , Michiyo Nakamura *2 , Kaori Uchida *2 ,Hiroshi Chisaka *2 ,Norihiro Katayama, Mitsuyuki Nakao and KunihiroOkamura *2*1 Tohoku University Biomedical Engineering Research Organization*2 Department of Gynecology and Obstetrics, Tohoku UniversityGraduate School of MedicineIn contrast to the ultrasound measurement of fetal heartmotion , the electrocardiogram (ECG) provides clinicallysignificant information concerning the electrophysiologicalstate of a fetus. The problem of the fetal ECG extractionfrom maternal skin electrode signals was not only thesevere noise contained in the measured signal but alsothe non-linear property of fetal or maternal ECG itself. Inthis paper, a novel extraction method of fetal ECG fromabdominal composite signals is proposed. This methodconsists of the cancellation of the mother’s ECGcomponents and blind source separation with referencesignal (BSSR). The BSSR is a fixed-point algorithm theLagrange function of which includes the higher ordercross-co-relation between the extracted signal and thereference signal as the cost term rather than a constraint.By practical application, the proposed method has beenshown to be able to extract the P and T waves in additionto the R wave. The reliability and accuracy of this methodwas confirmed by comparing the extracted signals withthe directly recorded ECG at the second stage of labor.The gestational age dependency of the physiologicalparameters of the extracted fetal ECG also coincided wellwith that of the magneto-cardiogram, which proves theclinical applicability of this proposed method.NotesPage 104

AbstractThe effects of the tocolytics atosiban and nifedipine on fetalmovements, heart rate, and blood flow.46Roel de Heus, MD, Eduard J.H. Mulder, MSc, PhD, Jan B. Derks, MD, PhD,Gerard H.A. Visser, MD, PhDDepartment of Woman and Baby, University Medical Centre Utrecht, The NetherlandsObjective: to study the direct fetal effects of tocolysiswith atosiban or nifedipine combined with a course ofbetamethasone.Methods Women with preterm labour (n=40), gestationalage between 25 and 33 weeks and no previous tocolytictreatment, were prospectively randomised for treatmentwith atosiban or nifedipine combined with a course ofbetamethasone. One-hour recordings of fetal heart rate(FHR) and its variation and fetal movements were madeon each of five successive days (day0- day4) starting inthe early morning. Fetal blood flow velocity patterns werestudied daily by Doppler ultrasound.Main outcome measures: Primary outcome measureswere effects on FHR and its variation, short term variation(STV). Secondary endpoints were effects on fetalmovement and fetal blood flow parameters, umbilicalartery (UA) and medial cerebral artery (MCA).Results: Baseline characteristics of 31 women notdelivered at day 0 and no escape tocolysis, did not differbetween the study groups. Multilevel analysis showed nosignificantly contribution of either tocolytic to the models ofFHR parameters and fetal movement parameters. Therewas a strong significant time course in basal FHR andSTV during day 0 to 4 of the morning recordings.Compared to baseline (day 0) a significant rise in STVoccurred on day 1 (p=0.01). Basal FHR decreasedsignificantly on day 1 (p< 0.01) and day 2 (p< 0.01)compared to baseline (day 0). The use of tocolytics didnot contributed significantly to the models of the UApulsatility index (PI) (p=0.91) and MCA PI (p=0.62). Nosignificant changes in time course of PI parameters werefound.Conclusion This study demonstrates for the first time thedirect effects of atosiban on fetal movement, fetal heartrate and fetal blood flow. Tocolysis with either atosiban ornifedipine combined with betamethasone administrationappears to have no adverse direct adverse fetal effects.Figure. Mean morning values ± SEM of the study parameters. Open circles represent women treated withatosiban and closed circles women treated with nifedipine. The first arrow indicates start of tocolytic treatmentand betamethasone administration. The second arrow indicates the second dose betamethasone.NotesPage 105

Abstract47THE EFFECT OF SULPHASALAZINE ON LPS-INDUCEDCHANGES IN FETAL OXYGENATION AND CAROTID BLOODFLOWBurcu Saglam 1 , Graham Jenkin 2 , Suzanne L. Miller 3 , and Euan M. Wallace 11 Department of Obstetrics and Gynaecology, 2 Monash Immunology and Stem Cell Laboratoriesand 3 Deparment of Physiology, Monash University, Victoria, Australia.Background:Intrauterine infection is associated with fetal brain injurythought to be caused, at least in part, by the resultinginflammatory response. However, infection is alsoassociated with fetal hypoxia and it is possible that thiscontributes to the resulting brain injury, either directly onvia changes in cerebral blood flow and subsequentoxidative stress. We have used an ovine model of pretermfetal infection, using exposure to lipopolysaccharide (LPS),to investigate the effect of the administration ofsulfasalazine (SSZ), an NFκB inhibitor, on LPS-inducedchanges in fetal cerebral blood flow.Methods:Seven twin-pregnant ewes were randomly assigned toreceive either SSZ (n=3) or placebo (n=4) orally. Surgerywas performed at ~105 days gestation for placement of afemoral artery and vein catheter and a carotid artery flowprobe into each fetus. In each twin pair, whether SSZ orplacebo, one fetus received three daily i.v. infusions ofLPS (100ng/kg) and the co-twin received the samevolume of saline. Carotid blood flow, fetal blood gases andpH were monitored daily.Results:(1) The administration of SSZ was associated with analmost two-fold increase in baseline fetal carotid bloodflow in both saline and LPS-adminsitered groups. (2) Theadministration of LPS was associated with fetal acidaemia,hypoxaemia and hypercapnia within 4-6 hours in fetusesof both placebo and SSZ ewes. In the fetuses of ewespre-treated with SSZ the degree of these changes wassignificantly worse than in fetuses of placebo treated ewes.(3) LPS administration was associated with significantincreases in fetal carotid blood flow, by 70% in placebotreated ewes and by 85% in SSZ treated ewes. Thisresponse in fetal carotid blood flow to LPS was similar inthe two groups.Conclusion:These preliminary observations suggest that maternalSSZ administration alters fetal cerebrovascular function,which may be detrimental. Further, LPS administrationsignificantly increases fetal carotid blood flow – an effectnot mitigated by SSZ. Indeed, the maternal administrationof SSZ worsens the fetal acideaemic, hypoxaemic andhypercapnic response to LPS. Together, theseobservations suggest that SSZ would not be suitable as aanti-inflammatory in the management of intrauterineinfection.Fetal Carotid Blood FlowFetal carotid blood flow (ml/min)10080604020LPS #1LPS #2 LPS #3SSZ+LPSPlacebo+LPSSSZ-SalineSSZ-Placebo0-12 0 12 24 36 48 60 72Time (hours)NotesPage 106

Poster1The role of the neural sympathetic and parasympatheticsystems in diurnal and sleep state cardiovascular rhythms inthe late gestation ovine fetusEllen C. Jensen, Laura Bennet, Sarah-Jane Guild, Lindsea C. Booth, JennyA. Westgate, and Alistair J. GunnDepartment of Physiology, Faculty of Medical and Health Sciences, The University of Auckland,Auckland, New ZealandDiurnal and sleep state rhythms for fetal heart rate (FHR)and FHR variability (FHRV) are well established by lategestation in mammalian species. However, despite thecommon use of FHR and FHRV for fetal surveillance, theunderlying efferent mechanisms controlling these rhythmsremain incompletely understood. In the present study weevaluated the contributions of the neural parasympatheticand sympathetic nervous systems in chronicallyinstrumented fetal sheep at a mean (SE) of 121.6 (0.9)days gestation.Eight fetuses underwent chemical sympathectomy using6-hydroxydopamine the day after surgery, 8 fetusesunderwent vagotomy at surgery and 8 fetuses were shamcontrols. Three days after surgery FHR, FHRV, meanarterial blood pressure (MAP), carotid blood flow, andelectroencephalogram (EEG) activity were measuredcontinuously for 24 hours. Changes between sleep states,i.e. high voltage slow activity and low voltage fast activity,were determined in a 6 h interval.Control fetal sheep showed consistent diurnal rhythms inFHR and FHRV, with maximal activity close to midnight.Both the sympathectomy and vagotomy groups alsoshowed diurnal rhythms in FHR and FHRV, butsympathectomy shifted the peak in FHR to much later(0500 h, p

Np95, a master regulator of the epigenome, is essentiallyrequired for embryonic development2Sharif J. 1,2 , Muto M. 3 , Nakamura M. 2 , Takebayashi S. 4 , Okano M. 4 , KosekiH. 3 , Nagamune T. 1 , Mitsuya K. 5 and Okamura K. 21 Department of Chemistry & Biotechnology, School of Engineering, The University of Tokyo,Tokyo, Japan, 2 Department of Obstetrics & Gynecology, Tohoku University School of Medicine,Sendai, Miyagi, Japan, 3 RIKEN Research Center for Allergy and Immunology, Yokohama,Japan, 4 RIKEN Center for Developmental Biology, Kobe, Hyogo, Japan and 5 TohokuUniversity Biomedical Engineering Research Organization, Sendai, Miyagi, JapanPosterAIMSAcquirement and maintenance of a correct epigeneticstate of the genome is indispensable for biologicalprocesses such as germ cell development, fertilizationand embryogenesis. The insights gained from assistedreproductive technology (ART) and somatic cell nucleartransfer clearly indicate the importance of epigenetics inembryonic and postnatal development. We havepreviously identified a novel epigenetic regulatory protein,Np95. In this report, we discuss the role of Np95 inembryogenesis and mammalian development.METHODSWe have studied genomewide DNA methylation level inthe Np95 KO ES cells by bisulphite DNA conversiontechniques. We generated Np95-null homozygoticembryos (B6 x JF1 cross) and examined any possibleimprinting defects. We investigated the expression ofrepeat associated transcripts in Np95 mutant embryos byquantitative real-time PCR. We also examined cell cycledependent cellular localization of Np95 by immunostaining.RESULTSDigestion with methylation specific enzymes revealed thatNp95 KO ES cells are severely hypomethylated. TheNp95-null homozygotic embryos were found to beembryonic lethal at d9.5. IAP, LINE and otherheterochromatin associated repeat elements constitutemore than 40% of the mouse genome. In Np95 mutantembryos, IAP, LINE L1 and SINE B1 transcripts weredramatically upregulated. This suggested that Np95 isessential for heterochromatin regulation. To examine therole of Np95 in modulation of single copy genes in theeuchromatin, we studied the imprinted H19/Igf2 andLit1/Kcnq1ot1 loci. We found that in both these lociparent-of-origin specific expression of H19 and Lit1 waslost. The adjacent Igf2 and Cdkn1c genes werecompletely repressed. Loss of imprinting was associatedwith severe loss of CpG methylation in the H19-DMD andDlk1/Gtl2 IG-DMR and the histone modifications in theseloci were also affected. These results indicated that Np95is essential to regulate the euchromatin along with theheterochromatin. Coimmunoprecipitation andimmunostaining analyses revealed that Np95 physicallyinteracts with Dnmt1 during the middle-S1 phase andsuggested a possible mechanism that involved Dnmt1.CONCLUSIONWe show that Np95 acts as a master regulator of themammalian epigenome by regulating both theheterochromatin and euchromatin compartments of thegenome. We also describe an essential role for Np95 inmammalian development.NotesPage 109

Poster3EVALUATION OF THE PLACENTAL VASCULARITY USINGFRACTAL PARAMETERS: COULD THEY HELP IN EARLYDIAGNOSIS OF FETAL GROWTH RESTRICTION?C. Guiot, E. Piccoli^, F. Saccomandi & T. Todros^Dept Neuroscience, University of Torino, Torino, Italy and ^Dept of Gynecologic and ObstetricDisc., University of Torino, Torino, ItalyPlacental vascularization has been extensivelyinvestigated in the past (Giles et al, 1985, Salafia et al,1997) . Most of the studies referred to villous arteries andarterioles, and compared placentas from ‘normal’,uneventful pregnancies at term with cases of Fetal GrowthRestrictions (FGR) placentas. Studies of the placentalcapillaries were also performed. (Mayhew et al, 2004,Macara et al, 1996; Krebs et al, 1996; Todros et al, 1999).Results were conflicting.All the previous studies were performed by countingmethods, i.e. based only on the comparison of thevascular density evaluated on placental samples, andrequired a certain amount of placental tissue. Otherparameters, such as the ‘fractal dimension D and thelacunarity L , were proposed to evaluate thecharacteristics of the vascular networks, i.e. their fractalproperties, which are expected to portrait some specific,intrinsic characteristics of the system (.Mandelbrot 1967,1982). The main advantage of such parameters is thatthey are invariant in self-similar structures, i.e. D has thesame value independently of the number of generations ofbranches developed by the tree.Up to now only one paper related to the placenta andusing fractal parameters was published.( Bergman & Ullberg, 1998), confirming the self-similarityof the placental vascular tree.In the present study we investigate placentas deliveredfrom normal and FGR pregnancies. Biopsy on placentaltissue fixed and paraffin included after performing aimmunoistochemical reaction with anto-CD31, which is amarker specific for vascular endothelium, are investigatedat magnification x40 (Neurolucida, MBF Bioscience), andvessels up to 2-5 microns can be observed, includingplacental capillaries. D and L are estimated using Fraclac,a downloadable computing code (http://rsb.info.nih.gov/ij/)on the 2D specimen (a 3D extension is already understudy) .Results show that, although in a very narrow range,the parameter values are related to fetal growth. Theyencourage further investigation aimed at proposing a newtool for early detection of FGR on histologic probes fromchorial villi samples withdrown for genetic tests.NotesPage 110

Fetal and perinatal growth curves: what can we learn fromthem?4Caterina Guiot, Tullia Todros ^, Antonio Gliozzi * , Pier Paolo Delsanto*Dept Neuroscience and ^ Dept Gynecol Obstet Disciplines, University of Torino, Torino,Italy*Dept Physics, Politecnico of Torino, Torino, ItalyPosterMany different mathematical models, based onobservational datasets, have been proposed for decades.Various parameters X k ( mass, height, long bones length,biparietal diameter, etc) have been investigated bothlongitudinally, i.e. on single individuals, and transversally,i.e., averaged over a given homogeneous population.According to a recent formalism [1] developed on thebasis of the ontogenic growth model by West andcollaborators [2], fetal and perinatal datasets can beclassified and fitted in the framework of a completelygeneral scheme. As Fig 1 shows, plotting the logarithmicgrowth rate ( a = d ln X k / dt ) vs. its time derivative ( b=da/dt ) , curves are obtained which result from acombination of a polynomial expansion and complete orpartial loops. The former account for an overall growthterm as predicted by the classification scheme of ref. [1],while the latter are due to intrinsic cyclicities in the growthprocess. They are usually related to various feedbackmechanisms, as already observed qualitatively [3] , andpossibly due to adaptation to the surrounding environment,hormonal control cycles, etc.A thorough interpretation of plots such as in Fig. 1, canlead to a deeper understanding of the variousmechanisms involved. In addition, the model could beutilized to discriminate abnormal growth processes dueto intrinsic (e.g. genetic) factors from the ones resultingfrom a mismatch with environmental conditions.[1] P. Castorina, P.P. Delsanto, C. Guiot, Phys Rev Letter 96,188701 (2006)[2] G. B. West, J. H. Brown, B. J. Enquist, Nature 413, 628 (2001)[3] C.B. Davenport. J Gen Physiol 10,205 (1926)NotesPage 111

Poster5Nitric Oxide and CD4+25+Regulatory T cell production fromiNOS knockout mice and their F1 mice treated with LPS.Hidenori Takahashi, Toshiaki Okawa, Kiya Fujimori, Akira SatoFukushima Medical University, Ob/Gyn, Fukushima, Japan,OBJECTIVE: To evaluate the effect of inherited iNOS onNitric Oxide (NO) production by placental tissue andCD4+25+ regulatory T cell (Treg) from spleen by using ofiNOS knockout (iNOSKO: iNOS-/-) mice and their F1 femalemice half inherited iNOS(paternal(+p) or maternal(+m))were treated with lipopolysaccharide (LPS).STUDY DESIGN: Pregnant mice with genetic compositionof placenta for mating as (A) male iNOSKO× femaleiNOSKO; iNOS-/-, (B) male iNOSKO × femaleC3H(iNOS+/+) ; iNOS-/+m, (C) male C3H×femaleiNOSKO; iNOS+p/- day 14 of gestation were sacrificed 6hours after intraperitoneal injection(i.p.) of LPS (400 μg/kg) or vehicle (n=8 per group). Uterine rings from eachmice were equilibrated in Krebs-Henseleit solution. Insome rings, a piece of placenta was left attached to theuterine wall . After 60 min incubation one ml sample of thesolution was taken and analyzed for NOx production.Their F1 female mice were sacrificed 6 hours after LPSi.p.(400 μg/ kg). Their blood serum were analyzed for NOxand mononuclear cells isolated from spleen were for Tergby FACS. Statistical analysis was performed using oneway ANOVA.RESULTS: (1)NOx Production of uterine rings attachedwith 3 types placenta: Increasing order of magnitude ofNOx production without LPS; (A) iNOS(-/-) =(C)iNOS(-/+m)< (B) iNOS(+p/-) (p

Nitric Oxide production in placental tissue from iNOS knockoutmice with LPS and their offsprings.6Hidenori Takahashi, Toshiaki Okawa, Kiya Fujimori, Akira SatoFukushima Medical University, Ob/Gyn, Fukushima, Japan,PosterOBJECTIVE: To evaluate the effect of inherited iNOS onNitric Oxide (NO) production by placental tissue by usingof iNOS knockout (iNOSKO: iNOS-/-) mice and their F1female mice half inherited iNOS(paternal(+p) ormaternal(+m)) were treated with lipopolysaccharide(LPS).STUDY DESIGN: Pregnant mice with genetic compositionof placenta for mating as (A) male iNOSKO× femaleiNOSKO; iNOS-/-, (B) male iNOSKO × femaleC3H(iNOS+/+) ; iNOS-/+m, (C) male C3H×femaleiNOSKO; iNOS+p/- day 14 of gestation were sacrificed 6hours after intraperitoneal injection(i.p.) of LPS (400 μg/kg) or vehicle (n=8 per group). Uterine rings from eachmice were equilibrated in Krebs-Henseleit solution. Insome rings, a piece of placenta was left attached to theuterine wall . After 60 min incubation one ml sample of thesolution was taken and analyzed for NOx production.Their F1 female mice were sacrificed 6 hours after LPSi.p.(400 μg/ kg). Their blood serum were analyzed for NOx.Statistical analysis was performed using one way ANOVA.RESULTS: (1)NOx Production of uterine rings attachedwith 3 types placenta: Increasing order of magnitude ofNOx production without LPS; (A) iNOS(-/-) =(C)iNOS(-/+m)< (B) iNOS(+p/-) (p

Poster7Maternal severe undernutrition during both late gestation andlactation period induce hypertension in male rat offspring.Hidenori Takahashi, Toshiaki Okawa, Keiya Fujimori, Akira SatoDepartment of Obstetrics and Gynecology Fukushima Medical University School of Medicine,FukushimaOBJECTIVE:Our objective was to investigate the effects of eithersevere undernutrition during late gestation or lactationperiod on blood pressure and the development of vascularfunction in male rat offspring.STUDY DESIGN:We use normal pregnant Wistar rats (Group A),nutritionally restricted by feeding with 30% of the normalgestation-matched dietary intake from day 17 of gestationto delivery (Group B) and 30% restricted after delivery tothe end of lactation period (Group C).The offspring was measured blood pressure at 12 and 24weeks by using indirect tail-cuff method.Rings of thoracic aorta with intact endothelium from themale offspring of A and B at 8 weeks, were equilibrated at2 g passive tension in organ chambers filled withKrebs-Henseleit solution. Concentration-responserelationships to Norepinephrine (NE) and angiotensinⅡ(ATⅡ) were obtained in the absence or presence ofN(omega)-nitro-L-arginine methyl ester (L-NAME) or aselective AT Ⅱ type-1 receptor blocker (Valsartan).Responses to cumulative concentrations of sodiumnitroprusside (SNP) and to 10-5M oxyhemoglobin (Hb,nitric oxide scavenger) were also determined.RESULTS: Body weight was significantly reduced in Boffspring compared to A and C in male offspring at day 1(p C >A.NE concentration-dependently stimulated tension ofaortic rings from in A and B offspring, which was notsignificantly (n=6). Maximal contractions to NE weresignificantly stimulated by L-NAME in A (p

Regulation of maternal feeding during lactation period maycontrol adulthood hypertension.8Hidenori Takahashi, Toshiaki Okawa, Keiya Fujimori, Akira SatoDepartment of Obstetrics and Gynecology Fukushima Medical University School of Medicine,FukushimaPosterOBJECTIVE:Exposure to undernutrition during fetal life has beenproposed as an underlying cause of adult hypertension,but the effect of either high fat nourishment orundernutrition during lactation period on blood pressure isunclear.Our objective was to investigate the most effectivematernal nourishment and feeding period for offspringinduced adulthood hypertension in using high-fat diet(HFD).STUDY DESIGN:We use 5 types pregnant Wistar rats as fed with normalnutrition (Group A), nutritionally restricted by feeding with30% of the normal gestation-matched dietary intake fromday 17 of gestation to delivery (Group B), 30% restrictedafter delivery to the end of lactation period (Group C), witha high fat diet (HFD) during gestation to lactation period(Group D) and with HFD nutritionally 30% restricted fromthe day of delivery to the end of lactation period (GroupE).The offspring was measured Body Weight (BW) andmeasured blood pressure at 12, 24 and 60 weeks byusing indirect tail-cuff method. Statically analysis wasperformed using one-way ANNOVA.RESULTS: BW was significantly reduced in B offspringcompared to another (A, C, D, E) male offspring at day 1(p B > C>A. (pD>E>C>A. (p

Poster9Maternal regulation of high fat nourishment during lactationperiod reduce a hypertension of male offspringHidenori Takahashi, Toshiaki Okawa, Keiya Fujimori, Akira SatoDepartment of Obstetrics and Gynecology Fukushima Medical University School of Medicine,FukushimaOBJECTIVE:Exposure to undernutrition or high fat nourishment duringfetal life has been proposed as an underlying cause ofadult hypertension, but the effect of maternal feedingregulation during lactation period on blood pressure ofoffspring is unclear.Our objective was to investigate the effects of eitherhigh-fat diet (HFD) during gestation to lactation period orrestrictive fed a HFD during lactation period on bloodpressure in male rat offspring.STUDY DESIGN:We use 3 types pregnant Wistar rats as fed with normalnutrition (Group A), with a high fat diet (HFD) duringgestation to lactation period (Group B) and with HFDnutritionally restricted by feeding with 30% of the normallactation-matched dietary intake from the day of deliveryto the end of lactation period (Group C).The male offspring was measured blood pressure at 12,24 and 60 weeks by using indirect tail-cuff method.Statically analysis was performed using one-wayANNOVA.RESULTS: Body weight was significantly reduced in Coffspring compared to A and B in male offspring at day 28after delivery (p C>A. (p

PosterEpigenetic stability and developmental plasticity in clonedplacentae during somatic cell nuclear transfer10Naomi Matsuda 1 , Jafar Sharif 1 , Kimiko Inoue 2 , Narumi Ogonuki 2 , HiromiMiki 2 , Hiroshi Chisaka 1 , Kunihiro Okamura 1 , Atsuo Ogura 2 , and KohzohMitsuya 31Department of Obstetrics and Gynecology, Tohoku University School of Medicine, 1-1Seiryomachi, Aobaku, Sendai 980-8574, Japan2Bioresource Engineering Division, RIKEN Bioresource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki305-0074, Japan3Biofunctional Science, Tohoku University Biomedical Engineering Research Organization(TUBERO), 2-1 Seiryomachi, Aobaku, Sendai 980-8575, JapanEarly developmental process largely involves epigeneticgene regulation. One of the most remarkablereprogramming of the epigenome can be seen duringsomatic cell nuclear transfer (SCNT) whereby the nucleusfrom a differentiated cell could be reset by oocytecytoplasm, resulting in the production of live offspring. Inthis study, we have focused on the epigenetic aspects ofthe placentae derived from cloned mice, in which morethan 3-fold enlargement was observed in placental weightat term. All the twelve cloned placentae derived fromcloned conceptuses exhibit loss of genomic imprintingwhich is closely linked to histone tail modificationsindependent of DNA methylation, implying a possible rolefor histone modification remodeling in nuclearreprogramming. Moreover, we have identified multipletranscripts that are repressed specifically in the clonedplacentae irrespective of the degree of enlargement andof the presence of embryo proper. It has also been evidentthat the imprinted Dlk1 transcripts were less abundant inthe cloned placentae that were delivered with fetus butwere extensively silenced in all the four placentaerecovered without fetus. Our results contrast with theprevious observations indicating the variegated epigeneticanomalies in cloned animals and highlight considerablestability of the epigenome during SCNT anddevelopmental plasticity throughout early embryogenesisand placentation.NotesPage 117

Poster11Pre-existing hypoxia is associated with a delayed but moresustained rise in T/QRS ratio during prolonged umbilical cordocclusion in near-term fetal sheepLaura Bennet, Bert Wibbens, Jenny A. Westgate, Harmen H. De Haan,Guido Wassink, and Alistair J.Gunn Fetal Physiology and Neuroscience Group, Dept of Physiology, The University ofAuckland, Auckland, New Zealand.There is limited information on whether pre-existing fetalhypoxia alters hemodynamic responses and changes inT/QRS ratio and ST waveform shape during subsequentsevere asphyxia. Chronically instrumented near-termsheep fetuses (124±1 days) were identified as eithernormoxic PaO 2 > 17 mmHg (n=9) or hypoxic PaO 2 ≤ 17mmHg (n=5), and then received complete occlusion of theumbilical cord for 15-min. Umbilical cord occlusion lead tosustained bradycardia, severe acidosis and transienthypertension followed by profound hypotension in bothgroups. Pre-existing hypoxia did not affect changes inmean arterial blood pressure, but was associated with amore rapid initial fall in femoral blood flow and vascularconductance and with transiently higher fetal heart rate at2 minutes and from 9 to 11 minutes of occlusioncompared with previously normoxic fetuses. Occlusionwas associated with a significant but transient rise inT/QRS ratio; pre-existing hypoxia was associated with asignificant delay in this rise (maxima 3.7 ± 0.4 vs. 6.2 ± 0.5minutes), but a slower rate of fall. There was a similarelevation in troponin-T levels 6 hours after occlusion in thetwo groups (median (range) 0.43 (0.08, 1.32) vs 0.55(0.16, 2.32) µg/L, N.S.). In conclusion, mild pre-existinghypoxia in normally grown singleton fetal sheep isassociated with enhanced centralisation of circulationafter umbilical cord occlusion and delayed elevation of theST waveform and slower fall, suggesting that chronichypoxia improves myocardial dynamics during asphyxia.NotesPage 118

PosterFetal brain, liver and renal blood flow assessment using 3Dpower Doppler ultrasound12Fong-Ming Chang, Chiung-Hsin Chang, Chen-Hsiang Yu, Lin Kang,Pei-Ying Tsai and Huei-Chen Ko*Department of Obstetrics and Gynecology, and Research Institute of Behavioral Medicine*,National Cheng Kung University Hospital, Tainan 70428 TaiwanPurpose: To assess the blood flow of fetal brain, liver,kidney at second and third trimesters in normal pregnancyusing 3D power Doppler ultrasound.Materials and Methods: We underwent a series ofprospective, cross-sectional, consecutive and randomizedstudies to assess the fetal organ blood flow in normalsingleton fetuses from 20 to 40 weeks of gestation at theultrasound lab of a tertiary medical center. All the mothersand fetuses are free of any medical or obstetricalcomplications. We assessed the vascularization index (VI),flow index (FI) and vascularization-flow index (VFI) of fetalbrain, liver and kidney using the quantitative 3D powerDoppler ultrasound. All the fetuses were followed todelivery to ensure they were born normal.Results: All the VI, FI and VFI of fetal brain, liver andkidney increased significantly with gestational age (GA).Using GA as the independent variable, and the VI, FI andVFI as the dependent variable, the linear regressionequations as well as the correlation coefficients for VI, FIand VFI of fetal brain, liver and kidney versus GA were allsignificant (all P

Poster13Molecular Genetic Study on Angiotensin-Converting EnzymeGene and Preeclampsia in Taiwanese:Two polymorphismsTested and as HaplotypesFong-Ming Chang , Ming-Hui Chen, Chiung-Hsin Chang, Chia-Fu Li * ,Chen-Hsiang Yu, and Huei-Chen Ko **Department of Obstetrics and Gynecology, National Cheng Kung University Medical College,Tainan, Taiwan, *831 Army Hospital, Pei-Tou, Taipei, Taiwan, and ** Research Institute ofBehavioral Medicine, National Cheng Kung University Medical College, Tainan, TaiwanPurpose: To test whether the angiotensin-convertingenzyme (ACE) gene polymorphisms are associated withpreeclampsia-eclampsia (PE).Method: The study included 99 patients with PE and 146healthy controls. From genomic DNA, two polymorphismsin genes for ACE were typed. Allelic frequencies werecompared between PE patients and controls. Forstatistical analysis, chi-squared tests were used for twopolymorphic sites and as haplotypesResults : The genotype frequencies of the 287 bpinsertion-deletion (I/D) polymorphic site at the intron 16 ofthe ACE gene in PE patients were different with those incontrols (χ 2 = 6.68, p=0.035). However, the allelefrequency of the 287 bp I/D polymorphic site at the intron16 was not different with controls. In addition, thegenotype frequencies and the allele frequencies of Pst Isite at intron 7 in PE patients were not different withcontrols. Furthermore, the haplotype frequencies of twopolymorphic sites were not different in PE patients andcontrols.Conclusion:Although significant difference exists in thegenotype frequencies of the 287 bp I/D polymorphic siteat the intron 16 of the ACE gene between PE patients andcontrols, no significant differences can be detected for allthe other tests, either as single locus or as haplotype. Wespeculate the disease-gene association between PE andthe ACE gene in Taiwanese population. Further studiesare warranted.NotesPage 120

Clinical application of electromechanical delay time for indirectevaluation of fetal blood pressure during labor and delivery. Apreliminary study of 18 full-term infants.Yasuyuki Kawagoe, MD, Hiroshi Sameshima, MD, Tsuyomu Ikenoue, MD.Department of Obstetrics and Gynecology and Perinatal Center, Faculty of Medicine, Universityof Miyazaki, Japan.Poster14Objective: We previously showed a reciprocal relationshipbetween the electro-mechanical delay time and bloodpressure in fetal goat models. We applied this techniqueto clinical settings to determine whether the chronologicalchanges in delay time during labor and delivery correlatewith fetal heart rate monitoring patterns and acid-basestatus.Methods: Informed consent was obtained from 18 womenwith full-term, singleton pregnancy. Delay time wasobtained on a beat-by-beat basis from electrocardiogramto pulse oximeter waveform and averaged during eachbaseline period defined by the interpretation of fetal heartrate monitoring. According to the > 10% change ormore from the control value, chronological changes duringlabor and delivery were categorized into shortened,unchanged, and prolonged. Incidence of abnormal fetalheart rate monitoring patterns, hypoxemia, acidemia, lowApgar score was compared according to the chronologicalchanges in delay time.Results: Delay time was available in 82±11% of the time ofrecordings. Chronological changes were assessed in 15fetuses, 2 (13%) showed prolonged, 7 (47%) showedshortened, and 6 (40%) showed unchanged.Comparisons of the shortened and unchanged revealedthat severe variable deceleration was significantlyincreased and half or more fetuses showed mildlyhypoxemic in the shortened category.Conclusions: Shortening of delay time during delivery,theoretically indicating a hypertensive condition, wasmore likely associated with the presence of severevariable decelerations and fetal hypoxemia, suggestingthat the delay time may supplement the interpretation offetal heart rate monitoring.NotesPage 121

Poster15Temporal Changes in Mediators of Microvascular Tone areInfluenced by Sex and Glucocorticoid Exposure in PretermInfantsStark MJ, Clifton VL, Wright IMRMother and Babies Research Centre, Hunter Medical Research Institute, University ofNewcastle, NSW, AustraliaIan.Wright@hnehealth.nsw.gov.auAIM: Sexually dimorphic differences in microvascularfunction following preterm birth, with dysregulation ofvascular tone, may contribute to circulatory compromiseand a male excess of morbidity and mortality. The fetusexhibits sex-specific differences in glucocorticoidmetabolism with downstream pathways involved in thecontrol of vascular tone. We hypothesized that therewould be temporal changes in local mediators ofmicrovascular tone and that these would be influenced bysex and antenatal steroid exposure in extremely preterminfants.METHOD: Forty-four infants (male: n=29, female: n=15)of 24-28 weeks gestation were studied at 24, 72, and 120hours of age. Venous endothelin-1 and urinary nitric oxidemetabolites and normetanephrine were determined byEIA. Venous haemoglobin-bound carbon monoxide(COHb) was determined by spectrophotometry. Data wasanalysed by repeated measures ANOVA.RESULTS: Endothelin-1 altered with time [F(2,30)=2.65,p=0.048] but not sex or steroid exposure. No temporal orsex-specific differences in urinary nitric oxide metaboliteswereobserved but levels were lower in infants exposed toantenatal betamethasone [F(1,15)=4.75, p=0.046].Urinary normetanephrine was lower in male infants[F(1,10)=11.03, p=0.008] and in those infants exposed toantenatal glucocorticoids [F(1,30)=2.51, p=0.041] but notemporal change was observed. Venous COHbdecreased with time [F(2,76)=18.9, p

The effect of carperitide, human atrial natriuretic peptide, onacute heart failure caused by endocardial cushion defect withatrioventricular block in a premature infant: a case reportYuichiro Miura, Tadashi Matsuda, Masaki Sato, Shizuko Akiyama, RyutaKitanishi, Takushi Hanita, Tatsuya Watanabe, Hidenobu OhtaDivision of Perinatal Medicine, Tohoku University Hospital, JapanPoster16[Introduction]Systemic vasodilator therapy has proved useful in thetreatment of low cardiac output in adult and child patients.Both carperitide and nitroglycerin have vasodilatingactions, which are exhibited by elevation of cyclicguanosine monophosphate. In addition to the vasodilatingaction, carperitide has more pathophysiological rolesincluding increase in the glomerular filtration rate andinhibition of the renin-angiotensin-aldosterone (RAA)system.We describe a premature infant with acute heart failuredue to complete atrioventricular block and atrioventricularregurgitation. In this case, elevation of the mean bloodpressure (MBP) and adequate diuresis were achievedwith carperitide while nitroglycerin was of no effect even athigh dose, indicating that carperitide effectively inhibitedthe accelerated RAA system, which nitroglycerin did not.[Case report]A 22-year-old woman was pointed out fetal heart anomalyat 27weeks of gestation. The fetal echocardiographyshowed complete endocardial cushion defect and completeatrioventricular block; the fetal heart rate was 50-60 bpm,and the cardio-thoracic area rate was 40.8%. Cesareandelivery was performed at 31 weeks and 1day of gestationbecause fetal pleural effusion and ascites appeared.A female infant weighing 2,182g was delivered. Theheart rate was 40-60 bpm and the MBP was as low as 23mmHg. External pacing at 120-140 bpm was introducedimmediately after birth. However, the MBP remained lowand finally elevated to 31 mmHg after additionaladministrations of dopamine (10 µg/kg/min) andhydrocortisone (5 mg/kg), resulted in an increasing flow ofatrioventricular regurgitation. Nitroglycerin and diureticswere given for improvement of the regurgitation andoliguria but no significant effect was observed.After 30 hours of unsuccessful management forpersistent hypotension and hypoperfusion, carperitide(0.05 µg/kg/min) was administered. Immediately after theadministration, the decrease of the atrioventricularregurgitation and the increase of the diastolic flow velocityin the renal artery started to be detected by pulseddoppler echocardiography. By increasing carperitide up to0.20 µg/kg/min, appropriate MBP (>30 mmHg) andurination (>3 ml/kg/h) were achieved.[Comments]Carperitide has a potential effect not only as avasodilator but also as an inhibitor of RAA system in thetreatment for low cardiac output in premature infants.NotesPage 123

Poster17Effect of Maternal Stress on Fetal Heart Rate Assessed byVibroacoustic StimulationIkuko Makino 1) , Yoshio Matsuda 2) , Marie Yoneyama 2) , Kyoko Hirasawa 3) ,Koichiro Takagi 1) , Horoaki Ohta 2) , Yukuo Konishi 3)1) Depertment of Obstetrics and Gynecology Tokyo Medical University, Medical Center East2) Department of Obstetrics and Gynecology Tokyo Women’s Medical University3) Department of Infants’ Brain & Cognitive Development Tokyo Women’s Medical UniversityObjective: To determine whether maternal stress level,state and trait anxiety levels and stress hormones, affectfetal heart rate (FHR) pattern after vibroacousticstimulation (VAS) at 30 weeks of gestation.Materials and methods: Twenty-four healthy pregnantwomen with a singleton pregnancy were enrolled.Maternal anxiety was assessed using the SpielbergerState-Trait Anxiety Inventory (STAI). Women in the highanxiety groups had scores equal to or above the median.Corticotropin releasing hormone (CRH) andadrenocorticotrophic hormone (ACTH) in maternal plasmaand cortisol and chromogranin A in saliva were measuredas maternal stress hormone. FHR pattern after VAS weredivided into three response types; type I: a long period ofacceleration, or one acceleration lasting > 1 minute or atleast two accelerations lasting > 15 seconds’ duration,type II: a biphasic response with acceleration followed bydeceleration, type III: no response or prolongeddeceleration.Results: Only of these hormones, the value of CRH in thehigh trait anxiety group was significantly higher than in thelow trait anxiety group. In pregnant women with high traitanxiety, the FHR response after VAS showed mostly atype II pattern rather than normal response as type I(p=0.036) and the mean amplitude of FHR accelerationafter VAS was significantly lower in the high trait anxietythan in the low trait anxiety group (p=0.012).Conclusions: These findings suggest that stress inpregnant women with high anxiety may influence the FHRpatterns after VAS. The maternal environment exertsinfluence on the fetal autonomic nervous system.NotesPage 124

PosterNon-Invasive long-term fetal ECG monitoring 18E.M. Graatsma, J.B.Derks, E. Mulder, G. VisserDept of Perinatology, University Medical Center Utrecht,The NetherlandsNon-invasive fetal heart rate (FHR) monitoring using thefetal ECG (fECG) signal as obtained from the maternalabdomen has several advantages compared to FHRmonitoring using ultrasound. Main advantage is itspotential of obtaining prolonged recordings. Previously,antepartum fECG-monitoring has been hampered bytechnical difficulties (poor signal pickup, and a decline insignal quality around the formation of the vernix caseosabetween GA 28-32 wks). Most technical problems appearto have been solved now.We report on our first experience with an updatedfECG-monitor that is currently being tested for clinical use.We hypothesized that recording quality would be betterduring the night.abdominal electrodes. Recordings were consideredsuccessful if data loss was limited to a maximum of 60%).When total recording time (5pm-8am) was considered,62/75 (83%) recordings were successful. The number ofsuccessful recordings increased to 68/75 (91%) when thenight part (11pm-7am) was analyzed separately. Mean SRfor total recording time was 75%, but was higher overnight(85%).SR declined transiently between 26-34 wks, which may bedue to the vernix formation. Fortunately, despite theoverall decline in SR at this age range, the overnight SRwas sufficient to perform numerical FHR analysis.fECG recordings were made in 75 women at GA 20-39wks with singleton pregnancies between 5pm and 8am(total duration 15h) using a fECG-recorder (MonicaHealthcare). Patients used the recorder at home (n=40) orin hospital (n=35). fECG data was obtained from fiveThe newly developed fECG-recorder appeared anaccurate tool for long-term antepartum FHR monitoring,especially during the night. In the future the fECG will beof particular interest for fetal monitoring in at riskpregnancies.NotesPage 125

Poster19 The Relationship of Microchimerism to AbortionTomoko Sato 1) , Keiya Fujimori 2) , Akira Sato 2) , Hitoshi Ohto 3) , KenichiHata 4) , Hiroko Sasaki 4) , Mihoko Yazawa 4) , Shizuka Honda 5) , MayumiNoguchi 5)1) Ohara General Hospital, Fukushima, Japan 2) Fukushima Medical University OB/GYN,Fukushima, Japan 3) Fukushima Medical University Transfusion/Immunology, Fukushima, Japan4) Meiji Hospital, Fukushima, Japan 5) Nishiguchi Clinic, Fukushima, JapanObjective: Feto-maternal transfusion occurs in mostpregnancies. Researchers have recently reported anassociation between fetal cell microchimerism andautoimmune diseases; the phenomenon was related tothe persistence of fetal DNA postpartum. The purpose ofthis study was to determine the perseverance of fetal cellmicrochimerism after artificial or spontaneous abortion inthe first trimester.Methods: A total of 84 women who had never had anabortion or a male delivery were registered. Peripheralblood samples were obtained from 67 women whounderwent a dilatation and curettage in the first trimester.Samples were collected from each woman at three points:before, 7 days after, and 30 days after the abortion. Ychromosome-specific, nested PCR targeting SRY wasused to test DNA extracted from lymphocytes in the buffycoat. DNA was also extracted from the chorion todetermine sex. The sensitivity of our assay was thedetection of approximately one male cell in 100,000female cells.Results: A total of 31 male (48%) and 36 female (52%)chorions were obtained. Male DNA was found in 54.8%of women who had a male chorion before the abortion,and decreased to 6.5% at 7 days after the abortion. At 30days after the abortion, the DNA had completelydisappeared. Male DNA was never detected at any pointfrom women who had a female chorion. No differencewas found between artificial and spontaneous abortion.Conclusion: Fetal cell microchimerism was not detectedafter any abortion. Male cellular DNA disappeared frommaternal circulation 30 days after the abortion.NotesPage 126

PosterThe effect of cerebral hypothermia on cortisol and ACTHresponses after umbilical cord occlusion in preterm fetal sheep.20Joanne Davidson, Mhoyra Fraser, Andrew S. Naylor, Vincent Roelfsema,Alistair J. Gunn, Laura Bennet.Fetal Physiology and Neuroscience Group, Dept of Physiology, The University of Auckland,Auckland, New ZealandThe hypothalamus-pituitary-adrenal (HPA) axis isessential for adaptation to stress. It is unknown whetherthese endocrine responses are impaired or exaggeratedby mild systemic hypothermia induced as part of headcooling. We therefore examined HPA responses inchronically instrumented preterm fetal sheep (104 d ofgestation, term is 147 d), allocated to sham occlusion(n=7), 25 min of complete umbilical cord occlusion (n=7)or occlusion and head cooling with mild systemichypothermia (n=7, mean±SEM esophageal temperature37.6±0.3°C vs. 39.0±0.2°C; P

Poster21The prevention of fetal brain and lung injuries based oninterleukin-6 levels in amniotic fluid before birthTakushi Hanita, Tadashi Matsuda, Masaki Sato, Shizuko Akiyama, RyutaKitanishi, Yuichiro Miura, Tatsuya Watanabe, Hidenobu OhtaDivision of Perinatal Medicine, Tohoku University Hospital, Japan.Objective:To determine whether interleukin-6 (IL-6) levels inamniotic fluid before birth could predict postnataldevelopment of periventricular leukomalacia and chroniclung disease in premature infants.Study design:A total of 16 pregnant women in cases of clinicalchorioamnionitis (CAM) were underwent transabdominalamniocentesis and divided into two groups on the basis ofIL-6 levels in amniotic fluid, the high IL-6 group (≥17 ng/ml,n=8) and the low IL-6 group (

Effect of intrauterine inflammation on induction of antenatalperiventricular leukomalacia in chronically instrumented fetalsheepMasatoshi SaitoDepartment of Obstetrics & Gynecology, Tohoku University Graduate School of Medicine,Sendai, JapanPoster22Objective:Our purpose was to determine effects of intrauterineinflammation on induction of antenatal periventricularleukomalacia (PVL) in premature fetal sheep.Methods:Fetuses were infused with 40 µg/day ofgranulocyte-stimulating factor intravenously at 105-109days gestation and 20 mg of lipopolysaccharide (LPS) intothe amniotic cavity at 107 days gestation. At 24 hoursafter the LPS infusion, the hemorrhage group received anacute withdrawal of 40% of the fetoplacental blood volume(n=5), whereas an isovolemic exchange transfusion wasperformed in the control group (n=5). Changes in the totalhemoglobin (Hb), oxy-Hb, and deoxy-Hb levels in thecerebral tissue were assessed using near-infraredspectroscopy throughout the study period and comparedstatistically (ANOVA). Five days after the insult, fetuseswere processed for histologic analysis.Results:Four of 5 fetuses in the hemorrhage group and four of 5fetuses in the control group exhibited PVL. In all fetuses inthe both groups, necrotizing membranitis and funisitiswere observed and both the brain total-Hb and deoxy-Hbwere continuously increased from 6 hours after the LPSinfusion. After the insult, the brain total-Hb, oxy-Hb anddeoxy-Hb in the both groups decreased seriously (p

Poster23Maternal Low Protein Diet Aggravates Fetal White MatterDamage Caused by InfectionTakuya Ito *1 , Kaori Uchida *2 , Michiyo Nakamura *2 , Hiroshi Chisaka *2 ,Yoshitaka Kimura *1 and Kunihiro Okamura *2*1 Tohoku University Biomedical Engineering Research Organization*2 Department of Gynecology and Obstetrics, Tohoku UniversityGraduate School of MedicineMaternal infection causes release of cytokines such aschorioamnionitis. Such cytokines are known to damageoligodendrocytes or oligodendrocyte precursor cells(OL/OPC) and affect the white matter in fetal brain. Lossof white matter has been related to various mentaldisorders. However, white matter damage in fetus doesnot necessarily occur in all cases of maternal infection.Therefore, there may be a critical factor that determinesthe occurrence of white matter damage in fetal brainduring infection. In this study we have focused on therelationship between maternal nutritional and fetal braindamage. Our hypothesis about undernutrition is based onthe well-known results from epidemiological survey of theDutch Hunger Winter in 1944.We fed low protein diet (L) or normal diet (N) to femaleC57BL/6N mice and performed transvaginaladministration of LPS (0.1mg/ml 30µl) or PBS (30µl) fromthe 14 th day of pregnancy. In total, we investigated thefollowing four groups (L_LPS, L_PBS, N_LPS, N_PBS).We collected brain samples from E17 fetuses and P7neonates, and made cross-sections for immunohistochemistry.We performed immunostaining with olig2, PDGFR alpha,MBP, GFAP and nestin antibodies and examineddifferentiation to Glia. We also investigated activecaspase3 and performed TUNEL assay for measurementof apoptosis. Upon performing the above studies, wephotographed periventricular white matter area in thebrain, measured the number of positive cells and madestatistical analyses.We detected no difference in E17 fetus brain uponadministration of LPS with/ without undernutrition.However, in P7 neonatal mice GFAP (+) and nestin (+)cells / GFAP (+) cells increased upon administration ofLPS both with normal diet or undernutrition diet. Decreaseof MBP (+) cells was observed in L_LPS mice only.Transvaginal administration of LPS caused increase ofGFAP (+) and nestin (+) cells / GFAP (+) cells. Theseresults indicate that maternal infection may affect fetalbrain in an adverse way. Brain damage observed in onlyL_LPS mice suggests maternal under nutrition mayinduce hyperactive response for infection in fetal brain.NotesPage 130

PosterPrenatal diagnosis of bradycardia using fetal electrocardiogramvia maternal abdomen24Yoshitaka Kimura *1 , Michiyo Nakamura *2 , Kaori Uchida *2 , Takuya Ito *1 ,Hiroshi Chisaka *2 , and Kunihiro Okamura *2*1 Tohoku University Biomedical Engineering Research Organization*2 Department of Gynecology and Obstetrics, Tohoku UniversityGraduate School of MedicineFetal bradycardia is often associated with severediseases such as fetal distress, sick sinus syndrome, andatrioventricular block. Generally, it is difficult to make adiagnosis before birth because we cannot perform directfetal electrocardiogram. Here we report that we havecarried out fetal electrocardiogram via maternal abdomento patients showing severe bradycardia, and successfullyperformed appropriate prenatal diagnoses of arrhythmia.All the patients subjected to this study were referred to ourhospital for severe bradycardia. We performed fetalelectrocardiogram via maternal abdomen. One patientwas diagnosed with sick sinus syndrome. After vaginaldelivery, the baby underwent pacemaker implantation.Another patient was diagnosed with premature atrialcontraction. After one week of vaginal delivery, thearrhythmia disappeared. The last patient was diagnosedwith atrioventricular block. The baby underwentpacemaker implantation after vaginal delivery.Prenatal diagnosis of bradycardia using fetalelectrocardiogram can help us to determine the mode andthe timing of the delivery, leading to appropriate therapyafter birth.NotesPage 131

Poster25Endothelin-1 inhibits both L-type Ca 2+ current and ATPsensitiveK + current in neonatal rat ventricular myocytesYasuhiro Katsube, Nobuko Suzuki, Makoto Watanabe, Masanori Abe,Miharu Hajikano, Mitsuhiro Kamisago, Ryuji Fukazawa, Shunichi OgawaDepartment of Pediatrics, Nippon Medical SchoolBackground: Endothelin-1 (ET-1), a potent vasoactivepeptide, has a wide range of biological effects in variousorgans and tissues. In the heart, ET-1 is released byendothelial, vascular smooth muscle and myocardial cells,particularly in cardiovascular disorders such ascongestive heart failure and ischemic heart disease. Inthese pathophysiological conditions, intracellular ATPcontents decrease, followed by the simultaneous releaseof catecholamines and ET-1, which may contribute to theregulation of cardiac function. We previously presentedthat ET-1 inhibits voltage-gated and ATP-sensitive K +channel in resistance pulmonary artery from neonatalrabbit, suggesting that ET-1 may act as a mediator ofvasospasm and hypertension even in immature tissue.Objective: In this study, we investigated the effects ofET-1 on L-type Ca 2+ current (I Ca,L ) and ATP-sensitive K +current (I K,ATP ) in immature ventricular myocytes.Methods: Cells were enzymatically isolated from theventricles of neonatal rats (aged 3 to 10 days). Whole-cellwas measured as the peak inward current at a testpotential of +10 mV from a holding potential of -40 mV.I K,ATP was measured at a test potential of +60 mV from aholding potential of -70 mV. All experiments wereperformed at 37°C.Results: ET-1 (10-100 nM) decreased not only basal I Ca,L ,but also forskolin (a direct activator of adenylate cyclase)-stimulated and isoproterenol (β-adrenoceptor agonist)-stimulated I Ca,L . On the other hand, ET-1 (10 nM) almostcompletely inhibited the ATP-sensitive K + channel opener,pinacidil-induced outward current (i.e., I K,ATP ). Furthermore,ET-1 inhibited the shortening of cardiomyocyte actionpotential induced by pinacidil. These effects were reducedin the presence of BQ-123, an ET A -receptor antagonist,which suggests that the action of ET-1 is mediated by theET A receptor.Conclusions: In the neonatal heart, ET-1 inhibits not onlycontractility, but also cardioprotecive action by inhibitingI Ca,L and I K,ATP .voltage and current clamp techniques were applied. I Ca,LNotesPage 132

PosterPlacenta accreta/increta in unscarred uterus 26Akagi K, Shima T, Ishigaki N, Oota S, Hayasaka A, Fujita N, Asano K andWada YNational Health Organization Sendai Medical Center, JapanWhen the obstetrician encounter placenta previa after aprior cesarean section, careful preoperative managementwill take place because of the significant possibility ofplacenta accreta/increta. However, it is difficult toanticipate placenta accreta/increta in patients without prioruterine surgery or cesarean delivery. When placentaaccreta/increta do occur in these patients, promptintervention will be needed on site.Methods: We retrospectively reviewed our hospitalrecords for patient with placenta accreta/increta caseswithout prior uterine surgery including cesarean delivery.Clinically diagnosed cases and pathologically confirmedcases were included.Results: There were 8 cases of placenta accreta/increta(0.06% of all deliveries) without prior cesarean birth oruterine surgery between July 1992 and June 2007. 5cases (62.5%) were occurred after 2004. In 4 cases,postpartum bleeding was controlled and retained placentawas successfully treated by methotrexate. In two cases,massive bleeding after vaginal delivery was onlycontrolled by hysterectomy on the same day. In twocases, cesarean section was performed because ofposterior wall low lying placenta. In one of these twocases, placenta was firmly adherent to the uterus andhysterectomy was done without attempting to remove theplacenta. Although bleeding was controlled by packing theuterus on surgery in other case, hysterectomy was donebecause of massive bleeding on the next day. At leastone uterine curettage was done in 4 cases (50%) butmaternal age was over 35 only in 2 cases (25%).Conclusions: The most important risk factors for placentaaccreta/increta is prior uterine surgery and the mostcommon setting is placenta previa after a prior cesareandelivery. In our study, there were 0.06% incidence ofplacenta accrete/increta without previous uterine surgery.Therefore, the obstetrician has to be cautious of thepossibility of placenta accrete/increta even in the absenceof previous uterine surgery.NotesPage 133

Poster27 QT interval during pregnancy in Long QT syndromeYuri Tokito, Akiko Omoto, Naoshi Yamada, Naoko Iwanaga, KaoruYamanaka, Keiko Ueda, Masayo Nozawa, Reiko Neki, Wataru Shimizu,Tomoaki IkedaNational Cardiovascular Center, Suita Osaka, JapanObjectivesThe hereditary long QT syndrome (LQTS) ischaracterized by prolonged ventricular repolarization,manifesting arrhythmia-related recurrent syncope,aborted cardiac arrest, and sudden death. Pregnancy isassociated with frequent cardiac events in the women withLQTS. In the animal study, QT interval was significantlyprolonged in pregnanct mice compared withnon-pregnancy mice. (Mansoureh Eghbali et al,Circulation Research 2005) We retrospectivery analizedelectro cardiogram(ECG) of pregnant women with LQTSin perinatal period.Methods17 pregnancies in 7 LQTS cases were treated from 1992to 2006 in National Cardio Vascular Center, Japan. 15pregnancies succesfuly resated in live infant, 10 of whichwas diagnosed as LQTS subsequently. 2 pregnancieswas terminated as intentional abortion. In 8 pregnanciesfrom 6 cases, ECG was available both in pre- andpostpartum periods.We measured the QT interval corrected the heart rate(cQT) and were compared cQTs between pre- andpostpartum periods.ResultThe averages of cQT interval in pre-partum periods were568±82msec, longer than that in post-partum periods:504±55msec, in 8 pregnancies. There were no differentsin cQT interval among three different trimester ofpregnancy. In the LQT type2 case, which were seriallymeasured cQT interval, significant decrease in cQTinterval was observer just after the delivery, remaining upto postpartum day 150.ConclusionsQT interval was prolonged during pregnancy in the LQTS,consistant with animal study of pregnant mice. Thisprolongation may be associated with frequent cardiacevents during pregnancy of LQTS.NotesPage 134

High altitude chronic hypoxia modifies cardiovascularresponses in newborn sheep depending on the time andduration of the hypoxic insult during gestation.Poster28a,d EA Herrera, a G Ebensperger, b RA Riquelme, a C Torres-Farfan, a VRReyes, c JT Parer, d DA Giussani, a AJ Llanos.a Facultad de Medicina, b Facultad Ciencias Químicas y Farmacéuticas, Universidad de Chile,Chile; c University of California San Francisco, USA; d University of Cambridge, UK.Chronic hypoxia is a common insult during fetaldevelopment inducing structural and functional changes inpulmonary and systemic circulations. Hypothesis:Maternal hypoxia induces neonatal cardiovascularchanges depending on the time and duration of thehypoxic insult during fetal life. Methods: Pregnant eweswere divided into three groups: conception, pregnancyand delivery in lowland (500m, LLL); conception inlowland and at 30% gestation taken to highland hypoxia(3,600m) until delivery and then taken back to lowland(LHL); and conception, pregnancy and delivery inhighland hypoxia (3,600m, HHH). LLL and LHL werestudied at lowland and HHH at highland. Newborn lambswere catheterised under general anesthesia andsubmitted to acute hypoxic episodes. Results: LHL grouppresent a high abortion (24%) and stillbirth (11%) rate, notseen in the other groups. Under basal conditions, LHL andHHH had higher pulmonary arterial pressures (PAP,20.8+1.2 and 20.2+2.4 vs. 13.6+0.5 mmHg, p

NotesPage 136

List of delegatesYuka AbeDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanabe-9.27.-yuka@s3.dion.ne.jpSamana AliDepartment of Pediatrics.Aga Khan UniversityStadium Road, P.O. BOX 3500Karachi 74800, Pakistansamana.ali@aku.eduKozo AkagiNational Health Organization,Sendai Medical Center2-8-8 Miyagino Miyagino-kuSendai 983-8520Japanakagik@snh.go.jpMaiko AraiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmaiko-k@mail.tains.tohoku.ac.jpKazunori BabaCenter for Maternal, Fetal andNeonatal Medicine, SaitamaMedical Center,Saitama Medical University1981 Kamoda, Kawagoe,Saitama Japan 350-8550baba-tokyo@umin.netLaura BennetDepartment of Physiology,The University of Auckland85 Park Road, Grafton,Auckland 1023New Zealandl.bennet@auckland.ac.nzAlan D BockingUniversity of TrontCanadaabocking@mtsinai.on.caFong-Ming ChangOb/Gyn, Natl Cheng Kung Univ MedCollege & Hospital138 Victory Road 70428Taiwanfchang@mail.ncku.edu.twHiroshi ChisakaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanchisaka@mail.tains.tohoku.ac.jpJason H Collins MdPregnancy InstituteP.O. Box 806 New Roads LA 70760U.S.A.haydel1@bellsouth.netJoanne DavidsonDepartment of Physiology,The University of Auckland85 Park Road, Grafton,Auckland 1023New Zealandjoanne.davidson@auckland.ac.nzRobert De MatteoMonash UniversityDepartment of Anatomy and CellBiologyWellington Road, Clayton 3168Australiarobert.dematteo@med.monash.edu.auJan DerksUniversity Medical Center UtrechtKerkstraat 6 Utrecht 3581rdNetherlandsjbderks@hotmail.comJ.B.Derks@umcutrecht.nlAlison ForheadUniversity of CambridgeDepartment of Physiology,Development and NeuroscienceDowning Street,Cambridge CB2 3EGU.K.ajf1005@cam.ac.ukMhoyra FraserLiggins Institute, University of AucklandLiggins Institute, Faculty of Medicaland Health Sciences Universityof Auckland,Private Bag 92019, Auckland 1New Zealandm.fraser@auckland.ac.nzKeiya FujimoriDept. Ob & Gyn. Fukushima MedicalUniversity1-Hikarigaoka Fukushima 960-1295Japanfujimori@fmu.ac.jpHideoki FukuokaWaseda University1-1-1, Nishiwaseda, Shinjuku-KuTokyo 169-0051Japanhideoki.fukuoka@gmail.comHelena M GardinerImperial College52 Valiant House, VicarageCrescent London SW11 3LUU.K.helena.gardiner@imperial.ac.ukSherly GeorgeDepartment of Physiology,The University of Auckland85 Park Road, Grafton,Auckland 1023New Zealandsa.george@auckland.ac.nzCaterina GuiotUniversity of TorinoDept Neuroscience C. Raffaello30 10125Italycaterina.guiot@unito.itAlistair J GunnDepartment of Physiology,The University of Auckland85 Park Road, Grafton,Auckland 1023New Zealandaj.gunn@auckland.ac.nzTakushi HanitaDivision of Perinatal Medicine,Tohoku University Hospital,Japan1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanpunita@olive.ocn.ne.jpMark HansonUniversity of SouthamptonInstitute of Developmental Sciences,Mailpoint 887, SouthamptonGeneral Hospital, Tremona Road,Southampton, Hants SO16 6YDU.K.m.hanson@soton.ac.ukRichard HardingDept of Anatomy and Cell BiologyMonash University 3800Australiarichard.harding@med.monash.edu.auPage 138

Steve HarveyUniversity of AlbertaDepartment of Physiology Universityof Alberta T6G2H7Canadasteve.harvey@ualberta.caShinichi HayasakaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japansiniti-hayasaka@pref.iwate.jpChika HayashiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanchayashi@mail.tains.tohoku.ac.jpEri HirokiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanerimer@zg7.so-net.ne.jpYoshiyasu Hombo1-3-23, Izumino-Demachi,Kanazawashi, Ishikawaken921-8116Japanyoshiyasu@spacelan.ne.jpJeng-Hsiu HungDepartment of Obstetrics andGynecology,Taipei Veterans General Hospital,201, Section 2, Shih-Pai Rd,Taipei 112,Taiwanjhhung@vghtpe.gov.twTomoaki IkedaNational Cardiovascular Center5-7-1 Fujishiro-Dai 565-8565Japantikeda44@hotmail.comtikeda@hsp.ncvc.go.jpTsuyomu IkenoueUniversity of Miyazaki5200, Kihara, Kiyotake,Miyazaki 889-1692Japansatomako@fc.miyazaki-u.ac.jpTakuya ItoTohoku University BiomedicalEngineering ResearchOrganizationTetsuya ItoNagoya City University,Department of PediatricsMizuho-Ku, Mizuho-Cho,Kawasumi 1 Nagoya 467-8601Japanitotetsu@med.nagoya-cu.ac.jpKiyoshi ItoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japankito@mail.tains.tohoku.ac.jpHiroaki ItohOsaka National Hospital2-1-14 Houenzaka, Chuou-Ku,Osaka, 540-0006Japanhitou-endo@umin.ac.jpNoriyuki IwamaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japannoribou@mail.tains.tohoku.ac.jpSharif JafarDepartment of Obstetrics andGynecology, Tohoku UniversitySchool of Medicine1-1 Seiryo-machi, Aoba-ku,Sendai-shi, Miyagi 980-8574, Japanjafarsharif@tubero.tohoku..ac.jpGraham JenkinMiscl, Bldg 75, Strip 1, 3rd Floor,Monash UniversityClayton Victoria 3800Australiagraham.jenkin@med.monash.edu.auNaru KageyamaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japannarukageyama@kcd.biglobe.ne.jpYukika KakutaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japansnowsmell@mail.tains.tohoku.ac.jpIneko KatoDepartment of Pediatrics,Nagoya City University1 Kawasumi, Mizuho,Nagoya 467-8601Japanikato@med.nagoya-cu.ac.jpYasuhiro KatsubeNippon Medical SchoolMusashikosugi Hopital1-396 Kosugi-Cho Nakahara-KuKawasaki 211-8533Japankatsube@nms.ac.jpIchiro KawabataDirector of the Department of Fetaland Maternal MedicineNational Nagara Medical Center1300-7Nagara Gifu city,Gifu,Japan502-8558kawabata@nagara-lan.hosp.go.jpYasuyuki KawagoeDep. of Ob & Gyn, University ofMiyazaki5200 Kihara, Kiyotake-Cho,Miyzaki 880-1692Japanyasuk@fc.miyazaki-u.ac.jpMie KawaiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanPage 139

Kumiko KawanoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japank-kawano@mail.tains.tohoku.ac.jpKei KawashimaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanYoshitaka KimuraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanykimura@ob-gy.med.tohoku.ac.jpEllen KnappDepartment of Physiology,The University of Auckland85 Park Road, Grafton,Auckland 1023New Zealande.knapp@auckland.ac.nzAnna Hauntoft KongstedUniversity of CopenhagenGr Negaardsvej 7Frederiksberg 1870Denmarkahtp@life.dkBrian John KoosUcla22-139 Chs 10833 Le Conte Avenue,Los Angeles 90095-1740U.S.A.bkoos@mednet.ucla.eduShiro KozumaUniversity of Tokyo7-3-1 Hongo Bunkyo-Ku Tokyo113-8655Japankozuma-tky@umin.ac.jpTomoyuki KuwataJichi Medical University3311-1, Yakushiji, Shimotsuke,Tochigi 329-0433Japankuwata@jichi.ac.jpIrina G. MakarenkoInstitute of Developmental BiologyRas Vavilov St. 26 Moscow 119334Russiaimakarenk@mail.ruIkuko MakinoTokyo Women’s Medical University2-1-10, Nishiogu,Arakawa-Ku 116-8567Japanmakinoog@dnh.twmu.ac.jpTadashi MatsudaCenter for Perinatal Medicine,Tohoku University Hospital1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanchoku@mail.tains.tohoku.ac.jpYoshio MatsudaTokyo Women’s Medical University2-1-10, Nishiogu,Arakawa-Ku 116-8567Japanym0709@obgy.twmu.ac.jpNaomi MatsudaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japannaomim@mail.tains.tohoku.ac.jpSuzanne Lee MillerMonash UniversityDepartment of Physiology,Building 13f Monash University 3800Australiasuzie.miller@med.monash.edu.auTakahiro MinatoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japant-minato@mail.tains.tohoku.ac.jpMariko MinouraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmari-m@mail.tains.tohoku.ac.jpJunko MinouraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanjminoura@zb.cyberhome.ne.jpKohzoh MitsuyaBiofunctional Science, TohokuUniversity BiomedicalEngineering ResearchOrganization (TUBERO)2-1 Seiryomachi, Aobaku,Sendai 980-8575, Japankmitsuya@tubero.tohoku.ac.jpYuichiro MiuraPerinatal Medicine, TohokuUniversity Hospital1-1, Seiryo-cho, Aoba-ku,Sendai, Miyagi, 980-8574, Japany-miu@mail.tains.tohoku.ac.jpLeon MuldersMaxima Medical Center VeldhovenMarie Curielaan 3Eindhoven 5644 DrNetherlandslgm.mulders@chello.nlTakashi MurakamiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmurakami@mail.tains.tohoku.ac.jpYuji MurataAizenbashi Hospital5-16-15 Nipponbashi Naniwa-kuOsaka-shi Osaka 556-0005JapanJun MurotsukiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmurotsuki@mail.tains.tohoku.ac.jpPage 140

Tomoyuki NagaiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanSatoru NagaseDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japannagases@mail.tains.tohoku.ac.jpMichiyo NakamuraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanminakamura@mail.tains.tohoku.ac.jpMichiya NatoriNational Center For Child Health andDevelopment Ohkura2-10-1 Setagaya, Tokyo 157-8535Japannatori-m@ncchd.go.jpMarina V NechaevaInstitute of Developmental BiologyRas Vavilov St. 26 Moscow 119-334Russiamnechaeva2003@yahoo.comReiko NekiNational Cardiovascular Center5-7-1, Fujishiro-Dai SuitaOsaka 565-8565Japanrneki@hsp.ncvc.go.jpHitoshi NiikuraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanniikura@mail.tains.tohoku.ac.jpHidekazu NishigoriDept. of Ob & GynIwate Medical University19-1 Uchimaru MoriokaIwate 020-8505Japanhknishig@iwate-med.ac.jpYasuhisa NomuraFukushima Nartional Hospital13 AshidadukaSukagawa City 962-8507Japannomuyasu1969@yahoo.co.jpTakashi OkaiDept. of Obst and Gyne,Showa University1-5-8 Hatanodai Shinagawa-KuTokyo 142-8666Japanokai.t@med.showa-u.ac.jpKunihiro OkamuraDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanokamura@mail.tains.tohoku.ac.jpTeruo OkanoInstitute of Advanced BiomedicalEngineering and ScienceTokyo Women’s Medical University8-1kawada-cho, Shinjuku-ku,Tokyo,162-8666 JapanHitoshi OshitaniDepartment of VirologyTohoku University, Graduate Schoolof Medicine1-1Seiryo-Machi Aoba-KuSendai 980-8574JapanHidenobu OtaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanhideohta@mail.tains.tohoku.ac.jpJulian T. ParerUniversity of California SanFrancisco505 Parnassus AvenueSan Francisco CA 94143-0132U.S.A.pareb@obgyn.ucsf.eduUsha RajLos Angeles Biomedical ResearchInstitute1124 West Carson St.,Rb-1 Torrance, CA 90502U.S.A.raj@labiomed.orgKusol Russameecharoen4-25-5, Mitsukoshi Higashi NakanoMansion, Room No. 602,Nakano-Ku, Tokyo 164-0003Japanrkusol@yahoo.comBurcu SaglamMonash University Dept ofObstetrics and Gynaecology,Level 5 Monash Medical Centre246 Clayton Road, Clayton,Victoria 3168AustraliaBurcu.Saglam@med.monash.edu.auMasatoshi SaitoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmasa-saito@mail.tains.tohoku.ac.jpTsukuru SaitoSuzuki Memorial Hospital3-5-5 Satonomori Iwanuma-shiMiyagi 989-2481Japant.saito@suzukihp.or.jpHiroshi SameshimaUniversity of Miyazaki5200 Kihara, KiyotakeMiyazaki 889-1692Japanhsameshima@fc.miyazaki-u.ac.jpAkira SatoDept. Ob & Gyn.Fukushima Medical University1-Hikarigaoka Fukushima 960-1295Japanakira-s@fmu.ac.jpPage 141

Tomoko SatoOhara General Hospital, Fukushima6-11 Omachi Fukushima-City,Fukushima 960-8611Japantomoko-s@ohara-hp.or.jpKazuyo SatoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japankazusato@mail.tains.tohoku.ac.jpNaoaki SatoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanny-satou@cb3.so-net.ne.jpTakenobu ShimizuDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanNorio Shinozuka4-12 Mitsuka Hiratsuka,Kanagawa 254-0045Japannorio@shinozuka.comJunichi SugawaraKyono Art Clinic1-1 Honcyo Aoba-ku Sendai1-2 Miyagi 980-0014Japansugawara@ivf-kyono.or.jpVeena Gayathri SupramaniamMonash UniversityMonash Immunology & Stem CellCentre Strip 1,Monash University 3800Australiaveena.supramaniam@med.monash.edu.auKazunao SuzukiHamamatsu University School ofMedicine Handayama1-20-1 Higashiku Hamamatsu CityJapanks101865@f8.dion.ne.jpKeiji SuzukiSaitama Medical Center1981 Tsujido-Machi, Kamoda,Kawagoe, Saitama 350-8550Japandks@saitama-med.ac.jpSatoshi SuzukiDepartment of Pediatrics, NagoyaCity University1 Kawasumi, Mizuho,Nagoya 467-8601Japans.suzu@med.nagoya-cu.ac.jpKichiya SuzukiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japankichiyasuzuki@mail.tains.tohoku.ac.jpHidenori TakahashiDepartment of Obstetrics andGynecology Fukushima MedicalUniversity School of Medicine,Fukushima1 Hikarigaoka Fukushima city, Japanhide-t@fmu.ac.jpToshifumi TakabayashiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanYuha TakadaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japantakadayh@mail.tains.tohoku.ac.jpNaomi TakahashiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574JapanTadao TakanoDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanttakano@mail.tains.tohoku.ac.jpKojiro TanabeDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanmar22tana@aol.comSouta TanakaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japantanakasouta@hotmail.co.jpShingo TanigawaraJapanese Red Cross SendaiHospitalDepartment of Obstetrics andGynecology2-43-3 Yagiyamahoncyo Taihaku-kuSendai Miyagi 982-8501Japantanigawara@miyagi.med.or.jpYukihiro TeradaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanterada@mail.tains.tohoku.ac.jpHajime TogariDepartment of Pediatrics,Nagoya City University1 Kawasumi, Mizuho,Nagoya 467-8601Japantogari@med.nagoya-cu.ac.jpYuri TokiouNational Cardio Vasuculer CenterFujishirodai 5-7-1 SuitaOsaka 565-8565Japanyuri_tokitou@hotmail.comPage 142

Claudia Torres FarfanUniversidad De ChileSalvador 486, Providencia, SantiagoChilectorresf@med.uchile.clMasafumi ToyoshimaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanm-toyo@mail.tains.tohoku.ac.jpShinya TsuchidaThe University of Tokyo7-3-1 Hongo Bunkyo-KuTokyo 113-8655Japanshinyatsuchida556@hotmail.comKaori UchidaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japankao-u@mwe.biglobe.ne.jpTomohisa UgajinDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japantugajin@mail.tains.tohoku.ac.jpNobuya UnnoKitasato University, School ofMedicine1-15-1, Kitasato, Sagamihara City,Kanagawa 228-8555Japanunno@med.kitasato-u.ac.jpHiroki UtsunomiyaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanuskichi@mail.tains.tohoku.ac.jpGuido WassinkDepartment of PhysiologyThe University of Auckland,Auckland 1003New Zealandg.wassink@auckland.ac.nzFlora Y WongRitchie Centre For Bhr,Monash UniversityLevel 5, 246 Clayton RoadClayton 3168Australiaflora.wong@med.monash.edu.auTze-Fang WongDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanwongtzefang@mail.tains.tohoku.ac.jpIan M WrightMother And Babies Research CentreHunter Medical Research Institute,Lookout Road, New Lambton Hts,Newcastle 2287AustraliaIan.Wright@hnehealth.nsw.gov.auShanhai XuDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanshanhaix@mail.tains.tohoku.ac.jpHuixia YangDepartment of Obstetrics andGynecology, Peking UniversityFirst Hospital,Beijing, 100034 Chinayanghuixia688@sina.comNobuo YaegashiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanyaegashi@mail.tains.tohoku.ac.jpShun YasudaTakeda General HospitalJapanshunyasuda335@yahoo.co.jpKosuke YoshinagaDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japany-kou-m@mtb.biglobe.ne.jpHiromitsu YukiDepartment of Obstetrics &Gynecology,Tohoku University Graduate Schoolof Medicine1-1 Seiryo-Machi Aoba-KuSendai 980-8574Japanhiromitsu@mail.tains.tohoku.ac.jpTakanori WatanabeSendai City Hospital3-1 Shimizukouji Wakabayashi-kuSendai-shi Miyagi 984-8501JapanPage 143

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Author IndexAkagi, Kozo··························· P26Ali, Samana··························· Abstract13,Abstract14Bennet, Laura ······················· Plenary lecture 5,Abstract35, P11Bocking, Alan ························ Abstract38Chang, Fong–Ming ··············· P12,P13Collins, HJason ····················· Abstract17Davidson, Joanne ················· Abstract24,P20Derks, B Jan ························· Abstract34,Abstract46,P18Forhead, J Alison ·················· Abstract40Fraser, Mhoyra······················ Abstract5Fukuoka, Hideoki ·················· Abstract36Gardiner, Helena··················· Abstract18,Abstract20George, Sherly······················ Abstract31Guiot, Caterina······················ P3,P4Gunn, J Alistair······················ Abstract10,Abstract25Hanita, Takushi ····················· P21Hanson, Mark························ Plenary lecture 2Harding, Richard ··················· Abstract1,Abstract41Harvey, Steve ······················· Abstract23Hombo, Yoshiyasu················ Abstract16Hung, Jeng-Hsiu ··················· Abstract15Ishino, Fumitoshi··················· Invited lectureIto, Takuya ···························· P23Itoh, Hiroaki··························· Abstract3Jafar, Sharif··························· P2Jensen, Ellen ························ Abstract4,P1Katsube, Yasuhiro················· P25Kawagoe, Yasuyuki··············· P14Kimura, Yoshitaka················· Abstract45,P24Kongsted, Hauntoft Anna ······ Abstract2Levine, Richard ····················· Plenary lecture 3Makarenko, G. Irina·············· Abstract29Makino, Ikuko ······················· P17Matsuda, Naomi ··················· P10Matteo, De Robert················ Abstract26Miller, L. Suzanne ················ Abstract28,Abstract30Mitsuya, Kohzoh··················· Abstract37Miura, Yuichiro ····················· P16Murotsuki, Jun······················ Abstract8Nakamura, Michiyo ·············· Abstract9Nechaeva, V. M.··················· Abstract33Neki, Reiko··························· Abstract21Ohta, Hidenobu ···················· Abstract7Okano, Teruo ······················· Invited lectureOshitani, Hitoshi ··················· Geoffrey Dawes LectureParer, T Julian······················ Plenary lecture 1Raj, Usha J··························· Abstract42Saglam, Burcu······················ Abstract27,Abstract47Saito, Masatoshi··················· P22Sato, Kazuyo························ Abstract39Sato, Tomoko······················· P19Shinozuka, Norio·················· Abstract19,Abstract43Supramaniam, Veena ·········· Abstract44Suzuki, Keiji·························· Abstract12Suzuki, Kazunao ·················· Abstract22Takahashi, Hidenori ············· P5,P6,P7,P8,P9Tokitou, Yuri ························· P27Torres-Farfan, C··················· Abstract6,P28Wassink, Guido ···················· Abstract32Wong, Flora·························· Abstract11Wright, Ian···························· P15Yang, Huixia························· Plenary lecture 4Page 146