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- - - A B c D E F G -110 66 - 40 - 27 ~ 15 19 Fig. 5. Label of transformed cells in vivo by 32P-orthophosphate and pIS c1eavage. RSV -infected chicken fibroblasts (A-C), MC 29 nonproductively infected quail cells (D-F), and SSV(SSAV)-infected marmoset fibroblasts (G-I) were labeled with 1 uCi/ml 32P-orthophosphate for 2 h, lysed, immunoprecipitated (B, D, H), or subjected to an overnight incubation at 3rC in presence of pIS (A, E, G) or without pl5-protease (C, F, I). The sampies were analysed by SDS-PAGE followed by autoradiography Acknowledgments We thank Dr. F. Deinhardt very much for his interest and support of this work. The technical assistance of Miss D. Rose is gratefully acknowledged. Preparations of AMV have been provided by Dr. J. Beard. This work was supported by the Deutsche Forsch ungsgemeinschaft. References Arcement U, Karshin OL, Naso RB, Jamjoom G, Arlinghaus RB (1976) Biosynthesis of Rauscher Leukemia viral proteins : Presence of p30 and envelope pIS sequences in precursor polypeptides. Virology 69:763-774 - Bister K, Hayman MJ, Vogt PK (1977) Defectiveness of avian myelocytomatosis virus MC 29: Isolation of long term non-producer cultures and analysis of viral speeific polypeptide synthesis. Virology 82:431-448 - Brugge JS, Erikson RL (1977) Identification of a transformation specific antigen induced by an avian sarcoma virus. Nature 269:346-348 - Deinhardt F, Bergholz CM, Hunsmann G, Schneider J, Thiel HJ, Beug H, Schäfer W (1978) Studies of simian sarcoma and simian sarcoma associated virus. 1. Analysis of viral structural proteins, and preparation and characterization of antiserum specific for viral envelope components. Z Naturforsch [C) 33: 969-980 - Dittmar Kl, Moelling K (1978) Biochemical properties of pl5-associated protease in avian RNA tumor virus. J Virol28: 106-118 - Eisenman RN, Vogt VM (1978) The biosynthesis of oncornavirus proteins. Biochim Biophys Acta 473:187-239 - von der Helm K (1977) Cleavage of Rous sarcoma viral polypeptide precursor into internal structural proteins in vitro involves viral pro tein p15. Proc Natl Acad Sci USA 74:911-915 - von der Helm K, Konze-Thomas B (1980) Avian oncornaviruses contain a virus protease (pI5) which processes its own gag protein precursor and the 110 kd polypeptide of MC 29. Arch Geschwulstforsch 50:299-305 - Konze-Thomas B, von der Helm K (to be published) Processing of viral protein precursors by the avian myeloblastosis viral (AMV) protein p15 - Langlois AJ, Sankaran S, Hsiung PHL, Beard JW (1967) Massive direct conversion of chick embryo cells by strain MC 29 avian leukosis virus. J Virol 1: 1082-1084 - Mellon P, Pawson A, Bister K, Martin GS, Duesberg PH (1978) Specific RNA sequences and gene products of MC 29 avian acute leukemia virus. Proc Natl Acad Sei USA 75 :5874-5878 - Okasinsky GF, Velicer LF (1976) Analysis of intracellular feHne leukemia virus proteins. 1. Identification of a 60,000 dalton precursor of FeLV p30. J Virol 120:98-106 - Vogt VM, Eisenman R, Diggelmann H (1975) Generation of avian myeloblastosis virus structural pro teins by proteolytic c1eavage of aprecursor polypeptide. J Mol Biol 96:471-493 - Witte ON, Dasgupta A, Baltimore D (1980) Abelson murine leukemia virus protein is phosphorylated in vitro to form phosphotyrosine. Nature 283: 826-831 - Wolfe LG, Deinhardt F, Theilen GH, Rabin H, Kawakami T, Bustad LK (1971) Induction of tumors in marmoset monkeys by simian sarcoma virus, type I (Lagothrix): A preliminary report. J Natl Cancer Inst 47 : 1115-1120 413
Haematology and Blood Transfusion Vol. 26 <strong>Modern</strong> Trends in Human Leukemia IV Edited by Neth, Gallo, Graf, Mannweiler, Winkler © Springer-Verlag Berlin Heidelberg 1981 Control 01 Protein Synthesis. Phosphorylation and Dephosphorylation 01 Eukaryotic Peptide Initiation Factor 2 G. Kramer, N. Grankowski, and B. Hardesty Differentiation, virus infection, or certain pathologieal conditions are examples of physiologie events that cause dramatic qualitative and quantitative changes in protein synthesis. lt has been assumed and directly shown in several model systems that these changes involve a block in peptide initiation. Systems in which some of the underlying molecular mechanisms have been studied include dimethyl sulfoxide (DMSO) induced differentiation in Friend leukemia cells (FLC) , iron or heme deficiency in retieulocytes or their lysates, and interferon-treated cells that are sensitive to double-stranded RNA (dsRNA). It has been shown with FLC that DMSO and other inducers of differentiation cause a rat her rapid inhibition of peptide initiation that precedes the induction of hemoglobin synthesis (Bilello et al. 1979). These authors also demonstrated that inhibitors of peptide initiation, but not inhibitors of elongation, may be able to induce these cells to differentiate. Cell-free systems have been used to study the mechanism by which peptide initiation is blocked during heme deficiency and in the presence of dsRNA. lt was found that the peptide initiation factor elF -2 plays a key role in the regulation of eukaryotic pro tein synthesis. The activity of e1F-2 in the reaction by which Met-tRNA f is bound to 40S ribosomal subunits is controlled by phosphorylation/dephosphorylation of its smallest subunit, elF-2a (reviewed by Kramer et al. 1980). Binding of Met-tRNA f to 40S ribosomal subunits that are dependent on eIF-2, GTP, and otherinitiation factor(s) is inhibited if elF-2a has been phosphorylated by the cAMP-independent, hemecontrolled reticulocyte protein kinase that is specifie for this substrate (Pinphanichakarn et al. 1976). Conversely, aphosphoproteinphos- phatase isolated from retieulocytes will reverse this inhibition (Grankowski et al. 1980a). The elF-2a protein kinase and the counteracting phosphatase thus form a pair of regulatory components in the control of eukaryotic peptide initiation. The activity of the regulatory proteins is controlled by other factors. Here we consider some aspects of the regulation of the phosphoprotein phosphatase. A. Materials and Methods Preparation of the following components from rabbit reticulocytes has been described in detail: eIF-2 (Odom et al. 1978), phosphat ase (Grankowski et al., 1980a), and phosphatase activators (Grankowski et al. 1980b). eIF-2 was phosphorylated in its asubunit by the heme-controlled protein kinase from reticulocytes, reisolated by chromatography on phosphocellulose, and used as substrate in the phosphatase assay as reported previously (Grankowski et al. 1980a). B. Results and Discussion In contrast to the highly specific protein kinase that phosphorylates eIF-2a, phosphoprotein phosphatase activity isolated from rabbit reticulocytes has been found to have a rather broad substrate range (Grankowski et al. 1980a). Proteins phosphorylated by both cAMP-independent and cAMP-dependent protein kinases are readily dephosphorylated. However, stimulation and a certain degree of substrate specificity may be imposed on phosphatase activity by specific low molecular weight, heat-stable peptides. We have purified two such peptides from retieulocytes to homogeneity (Grankowski et al. 1980b). 414
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Haematology and Blood Transfusion 2
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Dr. Rolf Neth, Universitäts-Kinder
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Participants of the Meeting Aaronso
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Kurth, R, Friedrich-Miescher-Labora
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Wilsede Scholarship Holders (Grante
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Preface Gut ist eine Lehrart, wo ma
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Personal and scientific discussion
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Acknowledgement We should like to t
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Wilsede, June 21 1978 Memorial Trib
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limiting benign lymphoproliferative
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this most unlikely, however (for re
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longer subject to the same control
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Bregula U, Wiener F, Harris H (1971
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fresh lymph node biopsies from ten
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Fig. 3. Binucleate mitosis in an ob
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y Fig. 4. Schematic diagram of post
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than that among patients with Stage
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Tests of binding affinity, agglutin
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N Engl J Med 297: 963-968 - Green I
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Clinical Aspects
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"ring chromosome". "Mar" is marker,
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191 chromosomally abnormal patients
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tumors of both African and non-Afri
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Haematology and Blood Transfusion V
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Retrospective group 26/93 evolved l
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Treatment
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advantage in terms of duration of f
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leukaemia. An analysis of 168 patie
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11. Treatment Program Chemotherapy
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efractory congestive heart failure
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period (Sequence II) was gene rally
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children with a multiple drug proto
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DAYS 1 5 10 15 20 25 30 35 , I I I
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Biologically Drug Outcome fit" sens
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may counterbalance the potential be
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Ht (0--0) 50 40 30 20 10 0 Platelet
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10 7 ,-----------------------------
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11. Haematological and Biochemical
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0\ 0\ INTERFERON ... 10 3 rl E "'
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("pre-B" phenotype) has been descri
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human bone marrow cells exhibit the
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10 9.,0 MBKCC .. .,4 B prot:oool 1'
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demann W, Büehner T, Arlin Z, Gee
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-...l 00 a-tLDRENS CAt«:ER S TU>Y
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prognostic characteristics under in
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significantly higher in HR} (14.5%,
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normal or elevated immunoglobulin l
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References Andreeff M, Darzynkiewic
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R GROUP 1 ( 1970 - 1971 ) 17 PTS. R
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Table 3. Influence of initial featu
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LIJ (f) a.. < -1 LIJ Q: u. 0 >- I-
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me nt indicated that additional the
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42:2123-2134 - Chessells JM, Cornbl
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Baum et al. 1979). This study was b
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1. 0 '--'1--" 0.9 e: o .- VI VI E C
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e: o e: 1. 0 ,.--...-- 1'"'\ ......
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In conclusion, we can state that th
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Haematology and Blood Transfnsion V
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INDUCTION VCR + PRED + ASNASE (wk 3
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SCHED.INT 1 MTX • ! Adria MTX •
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~ looh---------ooooc>----o---c>----
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Consoll- Irra .tlon t) .tlon Indu
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Highrisk Standard risk Table 1. Cli
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Cumulo/ive comp/el9 remission 1.0 .
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10° 8 = 6 ö 4 Non T Cell .~ :ö
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% SURVIVAL 100 ~ ~II 0 80 60 40 - -
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a) Percentage of donor cell mitoses
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Table 5. Results of bone marrow tra
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...... w N Table 7. Clinical result
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agent had to be added and that in t
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C. Chronic Myelogenous Leukemia The
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Patients with a suitable donor rece
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No. Recurrent Leukaemia Other death
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followed by dialysis against isoton
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HS, HL- Heme >_ 40 0 HS I ;:. =====
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knowing at present. These genes may
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esistance to antifolates. In Vitro
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a DNA probe specific for the gene t
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translocation between chromosomes 9
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Table 1. Subdivision of 1827 Myelo-
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NORMAL CELL ®
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some change of the primary type, in
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F. Transformation of Mouse Bone Mar
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were carried out with 3H-MTX in the
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the lysozyme cDNA prepared from ovi
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et al. 1976). The BJAB cellline in
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dalton bands corresponding to light
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1979). The EBV-carrying cell lines
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producer lines (DAUDI and P3HR-1),
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G, Giovanella B, Westman A, Stehlin
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Dc;bt CT + i J.I9 Raji; 1. i .. 100
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c. Discussion During infectious mon
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C. Nonepisomal Viral DNA We have us
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A 260 3.0 A B 1670 70 N2 2.0 c: E .
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vivo. Nature 260:302-306 - Kirk JTO
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Fig. 1. a Polyacrylamide gelelectro
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1979). The lymphocyte subpopulation
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lysin 0 antigens are found, and mix
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Pope JH (1978) Long-term T cell-med
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Haematology and Blood Transfnsion V
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Charaeteristie Morphologie maturati
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after TPA-induction (Table 3). The
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PA (1978) Reactivity of a rabbit an
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Blood Monocyte. B lood Monocytes 1-
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and nonhistone proteins can serve a
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The findings of the evaluation of s
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Fig. 3. Tumor grawth of L 428 cells
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Cell biological and Immunological A
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consideration. Sanel (1973) obtaine
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endotoxin serum, were used. A parti
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monocytic leukemia cell line in cul
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B. Isolation, Characterization and
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don al hemopathies generally displa
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nitors. Blood Cells 6:595-607 - Min
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\I) ....I ....I IU Jl 200 r 100 90
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their haemopoietic cells' ability t
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normal normal ALL ALL ALL persons p
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64.5 { V)
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References Biermann E, Neth R, Gros
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ehambers, the growth pattern observ
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References Anderssen LC, 10kinen M,
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REH CELL UNE TO CALLb~Ö ~q~ V.M. c
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the macrophage series in which the
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Table 1. Production of factor depen
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Table 1. Characteristics of the adh
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Fig. 3. A human marrow culture at 6
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300 250 • HYPERPROLIFERATIVE PATT
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Fig. 9. Nonadherent population from
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The cobblestone areas were often no
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L (1976) Induction of colony format
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B. Materials and Methods J. Tissue
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Fig. 2. Ultrastructural appearance
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Page 307 and 308:
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Table 1. Monoclonal antibody reacti
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al. 1979; Janossy et al. 1979) is e
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to which this is an exact replica o
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inger JL (1976) Distribution of la-
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I region antigens were found to be
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have been reported to be present on
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V. FunctionalAssays Assays for PHA
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Functional studies of UCHT1 separat
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40 ,...... (5 L.. C o u 30 .9 ~ .~
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with immunohistochemieal methods (L
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can be dissected by cloning. Utiliz
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Table 1. Origin and marker profile
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complete identity of gp 100 from no
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Table 1. Reaction of single anti se
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media. Following 24-h incubation, c
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nonleukemic eells (Greaves 1979). L
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Fig. 2. Cytocentrifuged smears of b
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Modification of amino-groups of hum
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disturb this balance in the directi
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The majority of human blood lymphoc
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of the EBV infected B cells. Howeve
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and immunoglobulins. J Immunol 120:
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" K 10 • j ~ ALL RNlL AL wtfh les
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tion akuter Leukämiezellen in "spo
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D. Results and Discussion J. Acute
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selectively responsive to different
Page 371 and 372: Haematology and Blood Transfusion V
Page 373 and 374: l1J ~ 100 ~ Q. ::) l1J Z 0 ~ z >- .
Page 375 and 376: Table 2. Growth inhibition of S49 e
Page 377 and 378: Reverse Infectious ASC/2X1Q6 transc
Page 381 and 382: munized animals to 1316 tumor cells
Page 383 and 384: Haematology and Blood Transfnsion V
Page 385 and 386: Fig. 3. Expression of retroviral gp
Page 387 and 388: pg70. J Exp Med 143: 151-166 - McGr
Page 389 and 390: Table 1. Expression of tumor antige
Page 391 and 392: Virological and Molecularbiological
Page 393 and 394: Table 1. Oncogenic properties of re
Page 395 and 396: also Fig. 1). It followed that the
Page 397 and 398: whether this sequence is related to
Page 399 and 400: zed in infected cells argue that th
Page 401 and 402: a specific viral transforming prote
Page 403 and 404: detect viral RNA as the only charac
Page 405 and 406: Eisen H (1980) Myeloproliferate vir
Page 407 and 408: p,60- -34K - 1 2 3 4 Fig. 1. In vit
Page 409 and 410: .. p - • a'T ... I .t'l Fig. 4. D
Page 413 and 414: Fig. 2. Photomicrographs of NY68 in
Page 415 and 416: DEAE (o(lJmn 'liI GI -2 2 o e s 1
Page 417 and 418: vity was deterrnined. For endogenou
Page 419 and 420: pr 180 - - - P 12/15 - A B c D E F
Page 421: 85,- 66- -27 - -66 40- 27- -1'9 19-
Page 425 and 426: = 0.5 o E Q. o ILI 0.4 (J) « ILI .
Page 427 and 428: Table 1. Oncogenic potential of avi
Page 429 and 430: 100 III GI c: o "8 80 > w
Page 431 and 432: Expt. No. Infecting virus Proportio
Page 433 and 434: Haematology and Blood 'fransfusion
Page 435 and 436: sequences, provided its gag portion
Page 437 and 438: 77:3009-3013 - Hu SSF, Moscoviei C,
Page 439 and 440: RI 2.2 md and Hind III 2.6 md leuke
Page 443 and 444: RELATIONSHIP OF ENDOGENOUS AND EXOG
Page 445 and 446: E ........ E Cl. U E :::l .... Q) (
Page 447 and 448: References Astrin S (1978) Endogeno
Page 449 and 450: the same animal were examined (e.g.
Page 451 and 452: I) 'IQ a \0 - CL -- Fig. 2. Restrie
Page 453 and 454: ~ ..................... ~ Cell 3' 5
Page 455 and 456: a :Sa,C! + . ~ 111 i' ( " '~ .J. b
Page 457 and 458: Table 1 Sampie Tissue TS fragment"
Page 459 and 460: a b Sac I / rep ._-_.-_ K J nl rep
Page 461 and 462: Haematology and Blood 'fiansfusion
Page 463 and 464: Osteopetrosis and osteosarcomas occ
Page 465 and 466: Lymphoma cellline Table 1. T and B
Page 467 and 468: Table 2. In vivo growth and oneogen
Page 471 and 472: Table 1. Transforming aetivity of c
Page 473 and 474:
Table 3. Transformation aetivity of
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- Shoemaker C, Goff S, Gilboa E, Pa
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indieate a shifting of target eell
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Table 2. Expression of Jl and Sourc
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~ i ,..1---------,,':::::::::::::::
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fragment of pBR322 DNA, a 3.0-kbp S
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Mak TW (1979) Proc Natl Acad Sci US
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Table 1. Transformation of 3T3 cell
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inserts in the genome of rapidly tr
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pulations, it seemed likely that th
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contains antigens which react with
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le of reacting in sensitive radioim
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gic approach. In: Hiatt HH, Watson
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12 Eco R [ Fig. 1. Hybridization pa
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10 - '1~ )( -~ S:! E A U I o 1 2
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Haematology and Blood 'fransfusion
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cultures. These cells did not conta
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indication of malignant T-cells in
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Fig. 3. Thin-section electron micro
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Table 3. Lack of detectable related
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J. HTLV Was Present in the Primary
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specific signals into nonspecific s
Page 525 and 526:
Table 2. Distribution of HTLV-relat
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u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
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Page 531 and 532:
SSV TrS-gp labeled effectively with
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Page 535 and 536:
60 HFF/SSV cDNA CELL RNAs: 0 HF/SSV
Page 537 and 538:
Antisera a Table 1. Immunofluoresce
Page 539 and 540:
Haematology and Blood 'ftansfusion
Page 541 and 542:
y competition RIA yielded virtually
Page 543 and 544:
SiS'V' gp70: ANTIGEN, ANTI BODY, IM
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type-C virus p30 antigen in cells f
Page 547 and 548:
c. Results The fractionated whole-b
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induced antibodies as well as exper
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- Fig. la-f. Retravirus particles p
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eplication of infecting murine leuk
Page 555 and 556:
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immunodeficiency, multic10nal lymph
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Bovine leukemia --, proteins of 499
Page 561 and 562:
Human T- cell - -, characterization
Page 563 and 564:
RNA -, of tumor virus 439 Rous sarc
Page 565 and 566:
Haematology andBlood Transfusion Su
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