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CHARACTERIZATION Of IDIOTOPES ON MOPC-315 IGA USING MONOCLONAL ANTIIDIOTYPIC ANTIBODIES

MOLECULAR IMMUNOLOGY 1983年第5期20卷 537-547页

作者： NUSAIR, T BAUMAL, R ROSENSTEIN, R JORGENSEN, T MARKS, A UNIV TORONTO CHARLES H BEST INST BANTING & BEST DEPT MED RES TORONTO M5G 1L6 ONTARIO CANADA HOSP SICK CHILDREN DEPT PATHOL TORONTO M5G 1X8 ONTARIO CANADA YALE UNIV SCH MED DEPT PATHOL NEW HAVEN CT 06510 USA UNIV TROMSO SCH MED INST MED BIOL N-9001 TROMSO NORWAY

The isologous antiidiotypic response in BALB/c mice to immunization with the DNP[dinitrophenyl]-binding IgA myeloma protein, MOPC 315, alters the expression of the anti-DNP antibody repertoire and confers immunity against MOPC 315 myeloma tumors. To characterize the idiotopes on MOPC 315 IgA which elicit this response 4 monoclonal antiidiotypic antibodies (AIA), D10 (IgG2a), A2 (IgG1), G3 (IgG2b) and f1 (IgG2a), produced by splenocytes of BALB/c mice immunized with MOPC 315 IgA were isolated in 3 independent fusion experiments. These AIA react with MOPC 315 IgA, reassociated H315 L315 and .**GRAPHIC**. but not with free H315, L315, .**GRAPHIC**. or .**GRAPHIC**. The AIA do not react with the closely related DNP-binding IgA myeloma protein, MOPC 460, suggesting that they are directed against private idiotypes on MOPC 315 IgA. These idiotopes can be divided into 2 groups. Group I, defined by D10, A2 and G3, consists of 2 overlapping idiotopes, 1 of which is related to the hapten-binding site. The 2 idiotopes are formed by an interaction of amino acids in H315 and L315. Group II, defined by f1, consists of 1 idiotope which is related to the hapten-binding site. This idiotope is composed of an amino acid sequence on H315 which requires an interaction with either L315 or L460 for expression. A2 and G3 react identically with the same idiotope but were derived from 2 independent fusion experiments. An identity of AIA clonotypes among individual mice was indicated and the isologous AIA response to MOPC 315 IgA may be restricted.

关键词： AIA antiidiotypic antibody CfA complete freund's adjuvant DNP 2,4 dinitrophenyl DNP-cap N -2,4-DNP-ϵ-amino-caproic acid fCS fetal calf serum KLH keyhole limpet hemocyanin PBS phosphate buffered saline (140 m M NaCl, 3 m M KCl, 0.15 m M KH 2 PO 4 , 8 m M Na 2 HPO 4 ·7H 2 O, pH 7.4) Ig immunoglobulin H heavy L light L 315 L 460 free L chains of MOPC 315 460 IgA respectively H 315 H 460 free H chains of MOPC 315 460 IgA respectively f 315 V fragment consisting of V domains of MOPC 315 IgA V 315 2 fragment consisting of V domains of L 315 V 315 H fragment consisting of V domains of H 315

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DELAYED DIMETHYLfORMAMIDE BIOTRANSfORMATION AfTER HIGH EXPOSURES IN RATS

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TOXICOLOGY LETTERS 1983年第1-2期17卷 29-34页

作者： LUNDBERG, I PEHRSSON, A LUNDBERG, S KRONEVI, T LIDUMS, V NATL BOARD OCCUPAT SAFETY & HLTH S-17184 SOLNA SWEDEN

Rats were exposed to 2 dimethylformamide (DMf) air concentrations (2250 and 565 ppm for 4 h). Concentrations of DMf and the biotransformation product monomethylformamide (MMf) were measured in blood and some tissues at different times after the end of exposure. MMf concentrations 0 and 3 h after the end of the high exposure were generally lower than MMf concentrations at the same time after the low exposure. DMf biotransformation to MMf was delayed after the high exposure. As the hepatotoxic effect of DMf was correlated with MMf, this may have contributed to the previously observed slower appearance of hepatotoxicity after a high DME dose.

关键词： DMf dimethylformamide MMf monomethylformamide f formamide SDH sorbitol deny drogenase

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A HOMOPOLYMER Of THE POTENT ANTI-TUMOR DRUG 2'-DEOXY-5-fLUOROURIDYLATE

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BIOCHIMICA ET BIOPHYSICA ACTA 1983年第2期741卷 224-229页

作者： AMARNATH, V MILLER, TL BROOM, AD UNIV UTAH COLL PHARM DEPT MED CHEM SALT LAKE CITY UT 84112 USA

Poly(5-fluoro-2''-deoxyuridylic acid) was synthesized and its properties were compared with those of poly(dT) and poly(dU). It readily complexed with poly(dA). The 1:1 complex melted at about ***. C lower than poly(dA).***(dT). A triple-stranded helix, poly(dA).cntdot.2 poly(df5U) was formed only in high salt (2.0 M NaCl).

关键词： Antitumor drug Deoxyfluorouridylate Polynucleotide complex formation f 5 Ura 5-fluorouracil, df 5 Urd, 2′-deoxy-5-fluorouridine df 5 UMP 2′-deoxy-5-fluorouridylic acid poly(f 5 U) poly-(5-fluorouridylic acid) poly(df 5 U) poly(2′-deoxy-5-fluorouridylic acid) Mops 3-( N -morpholino)propanesulfonic acid Pipes piperazine- N , N ′-bis(2-ethanesulfonic acid)

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SERUM-PROTEINS Of WILD TURKEY VULTURES (CATHARTES-AURA)

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COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY 1983年第4期76卷 907-913页

作者： APANIUS, V TEMPLE, SA BALE, M UNIV WISCONSIN DEPT WILDLIFE ECOL MADISON WI 53706 USA UNIV WISCONSIN DEPT CHEM MADISON WI 53706 USA

The major serum proteins of turkey vultures (C. aura) were separated and identified: albumin, HDL [high density lipoproteins], LDL [low density lipoproteins], IgG and M, transferrin, .alpha.2M [.alpha.2 macroglobulin] and, putatively, IgA, ceruloplasmin and fibronectin. Turkey vulture HDL and LDL had similar electrophoretic mobility and solution properties as those from gallinaceous birds. Turkey vulture IgG and M, and their subunits, had MW comparable to other birds. Serum IgG and M levels in wild turkey vultures were within the range of values reported for domestic avian species.

关键词： α 2 M, α 2 macroglobulin Ig immunoglobulin BSA bovine serum albumin SDS sodium dodecyl sulfate M r apparent molecular weight (in kilodaltons) as determined by SDS-polyacrylamide electrophoresis R f relative mobility HDL high density lipoprotein LDL low density lipoprotein VLDL, very low density lipoprotein

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INTERACTIONS Of ACRIDINE-ORANGE WITH NUCLEIC-ACIDS - PROPERTIES Of COMPLEXES Of ACRIDINE-ORANGE WITH SINGLE STRANDED RIBONUCLEIC-ACID

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BIOCHEMICAL PHARMACOLOGY 1983年第24期32卷 3679-3694页

作者： KAPUSCINSKI, J DARZYNKIEWICZ, Z MELAMED, MR Memorial Sloan-Kettering Cancer Center New York NY 10021 United States

Interactions between acridine orange (AO) and nucleic acids (calf thymus DNA, and homoribopolynucleotides and homodeoxyribopolynucleotides) were studied in solutions containing ethanol as a cosolvent. Light absorption, scattering and luminescence were measured as a function of AO concentration at different dye/phosphate (D/P) ratios, and the data were analyzed using the McGhee-von Hippel probabilistic model of the polymer-ligand interactions. The absorption spectra of AO complexes with 4 homoribopolymers are presented. The intrinsic association constants and cooperativity coefficients of the formation of the complexes were calculated. The effects of ethanol (up to 35%, vol/vol) on these interactions were concentration dependent and may be extrapolated to 0 concentration of this cosolvent. The possibility of destabilization of the double helix of nucleic acids by AO at high D/P ratios is discussed in light of the available thermodynamic data.

关键词： AO 3,6-bis (dimethylamino)acridine hydrochloride = acridine orange ss single stranded ds, double stranded D/P, dye/phosphate molar ratio r , binding density n , binding site size ω, cooperativity coefficient K , association constant C f C B , free and bound dye concentrations E λ , molar extinction coefficient Δ G ° =- RTlnK, Gibbs free energy change and Hepes, 4- (2-hydroxyethyl)-1-piperazine-ethanesulfonic acid

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CHRONIC LYMPHOCYTIC-LEUKEMIA - A PROLIfERATIVE OR ACCUMULATIVE DISORDER

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LEUKEMIA RESEARCH 1983年第1期7卷 1-&页

作者： DORMER, P THEML, H LAU, B STADT KRANKENHAUS MUNCHEN MUNCHENFED REP GER

In two untreated patients with progressive CLL, quantitative 14C autoradiography of lymph nodes and, subsequently, continuous infusion of [3H] thymidine over eight and nine days, respectively, were performed in order to analyse the lymph node cell kinetics. Simultaneously, the turnover of labelled lymphocytes in the peripheral blood was evaluated. from another CLL patient a regional lymph node was removed 6 h after an intralymphatic flash injection of [3H] thymidine and sectioned for autoradiographic study of the distribution of labelled cells within the lymph node tissue. While the durations of DNA synthesis were found to be normal, the labelling indices were reduced. The relative cell production rate was far lower than normal. Very small growth fractions were calculated, amounting to less than 1% in one patient, and to 2.4% in the other. The distribution of labelled cells in the lymph nodes was focal, which supports the finding of low growth fractions. According to the present data, CLL is a disorder in which a very small number of cells cycle at a roughly normal rate. A kinetic definition of accumulative and proliferative tumour growth is introduced. Tumour growth is termed accumulative if growth appears to result from a decreased relative cell loss rate rather than an increased relative cell production rate. According to this definition, the kinetics in CLL may be classified as accumulative. In absolute terms, however, the number of lymphoid cells produced per unit of time was found to be far higher in CLL than in the healthy state.

关键词： Chronic lymphocytic leukemia proliferation kinetics growth fraction tumour growth CLL , chronic lymphocytic leukemia, TdR , thymidine WBC , white blood cells BM , bone marrow l s , DNA synthesis time K p , relative cell production rate (per 100 cells in the compartment), N S , relative number of cells in DNA synthesis (per 100 cells in the compartment) K L , relative rate of cell loss (per 100 cells in the compartment) K R , relative rate of lymphocyte release from lymph nodes (per 100 lymph node cells) K D , relative rate of cell death (per 100 lymph node cells) K l , relative rate of lymphocyte influx into the lymph node (per 100 lymph node cells) K f , relative rate of disappearance of lymphocytes from the peripheral blood (per 100 peripheral blood lymphocytes)

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MUTAGENIC ACTIVITY Of HETEROCYCLIC AMINES IN CHINESE-HAMSTER LUNG-CELLS WITH DIPHTHERIA-TOXIN RESISTANCE AS A MARKER

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MUTATION RESEARCH 1983年第1-2期118卷 91-102页

作者： NAKAYASU, M NAKASATO, f SAKAMOTO, H TERADA, M SUGIMURA, T National Cancer Center Research Institute I-I Tsukiji 5-chome Chuo-ku Tokyo 104 Japan

The mutagenicities of 8 heterocyclic amines, which were purified from pyrolysates of amino acids and proteins, broiled fish and fried beef, were assayed in Chinese hamster lung cells (CHL cells) in culture in the presence and absence of a metabolic activation system, with diphtheria toxin resistance as a selective marker. 3-Amino-1-methyl-5H-pyrido-[4,3-b]-indole (Trp-P-2) was the most mutagenic of the heterocyclic amines tested. 3-Amino-1,4-dimethyl-5H-pyrido-[4,3-b]-indole (Trp-P-1), 2-amino-3,4-dimethylimidazo-[4,5-f]-quinoline (MeIQ), 2-amino-3-methylimidazo-[4,5-f]-quinoline (IQ), 2-amino-9H-pyrido-[2,3-b]-indole (***.C), 2-amino-3,8-dimethylimidazo-[4,5-f]-quinoxaline (MeIQx), 2-amino-6-methyldipyrido-[1,2-a:3'',2''-d]-imidazole (Glu-P-1) and 2-aminodipyrido-[1,2-a:3'',2''-d]-imidazole (Glu-P-2) showed mutagenic activity on CHL cells, their specific activities being in the order shown above. All the compounds except Trp-P-1 required the presence of a metabolic activation system for mutagenicity on CHL cells. [This study may be applicable to carcinogenesis.].

关键词： Trp-P-1 3-amino-1,4-dimethyl-5 H -pyrido[4,3- b ]indole Trp-P-2 3-amino-1-methyl-5 H -pyrido[4,3- b ]indole Glu-P-1 2-amino-6-methyldipyrido[1,2- a :3′,2′- d ]imidazole Glu-P-2 2-aminodipyrido[1,2- a :3′,2′- d ]imidazole αC 2-Mamino-9 H -pyrido[2,3-b]indole IQ 2-amino-3-methylimidazo[4,5- f ]quinoline MeIQ 2-amino-3,4-dimethylimidazo[4,5- f ]quinoline MeIQx 2-amino-3,8-dimethylimidazo[4,5- f ]quinoxaline DMNA dimethylnitrosamine B[ a ]P, benzo[ a ]pyrene [ 14 C]NAD nicotinamide[U- 14 C]adenine dinucleotide DT r cells diphtheria toxin-resistant cells Ef-2 elongation factor 2 VAMEM Eagle's minimum essential medium supplemented with double concentrations of vitamins and amino acids fBS, fetal bovine serum

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STUDIES ON THE ANTIGENICITY AND CARBOHYDRATES Of HUMAN LOW-MOLECULAR WEIGHT KININOGEN

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MOLECULAR IMMUNOLOGY 1983年第12期20卷 1411-1418页

作者： TURPEINEN, U Department of Biochemistry Univeristy of Helsinki SF-00170 Helsinki 17 FinlandReceived 27 May 1983 Accepted 8 July 1983 Available online 12 November 2002

Native MW (LMr) kininogen from human plasma and a kinin-free kininogen from Cohn''s plasma fraction IV (HC-antigen) were isolated and studied with regard to their immunological reactivity and carbohydrate heterogeneity. Antisera were prepared against the conformational and sequence-dependence determinants of the H chain which is the common denominator of plasma kininogens. The molecules were characterized by single radial immunodiffusion and SDS [sodium dodecyl sulfate] gel electrophoresis. The effect of exoglycosidase treatment was investigated by a radioimmunoassay. The antigenic combining sites were not influenced by the partial removal of carbohydrates. These results suggest that the determinant structures of LMr kininogen reside in the peptide backbone of the protein. Carbohydrate heterogeneity was shown by different binding to concanavalin A and wheat germ lectin. With concanavalin A 52% of the immunoreactive HC-antigen was reactive compared to 76% of the native LMr kininogen. The weak reactivity (11%) of the HC-antigen towards the wheat germ lectin indicates that the N-acetylglucosamine residues are apparently blocked or missing. Immunoreactive kininogen was in all cases measured by single radial immunodiffusion or an RIA [radioimmunoassay]. The molecular nature of the heterogeneity is discussed in relation to the known isoelectric variations of kininogen.

关键词： Con A concanavalin A WGA wheat germ agglutinin HM λ high mol. wt LM λ low mol. wt EACA 6-aminocaproic acid H c -antigen kinin-free LM λ kininogen isolated from Cohn's plasma fraction IV SDS-PAGE sodium dodecyl sulfate-polycrylamide gel electrophoresis RCM reduced and carboxymethylated RIA radioimmunoassay PhMeSO 2 f phenylmethysulfonyl fluoride MeMan α-methyl-mannoside GlcNAc N -acetyglucosamine

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ULTRAVIOLET LIGHT-INDUCED MUTATION Of DIPLOID HUMAN-LYMPHOBLASTS

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MUTATION RESEARCH 1983年第2期107卷 347-370页

作者： DELUCA, JG WEINSTEIN, L THILLY, WG MIT DEPT NUTR & FOOD SCI TOXICOL GRP CAMBRIDGE MA 02139 USA

UV irradiation (254 nm) of immortal diploid human lymphoblasts killed cells, caused mutation at 3 genetic loci studied, and transiently inhibited 3H-TdR [H3-trifluorothymidine] uptake into DNA. A shoulder of about 6 J/m2 and a Do [mean lethal dose] of 6 J/m2 were observed for survival. Mutation rose in a monotonic nonlinear fashion through 6 J/m2;above 6 J/m2, complex behavior approximating a plateau in induced mutation was observed. Irradiation at 4.4 J/m2 caused a transient increase in the number of cells synthesizing DNA and a decrease in the rate of DNA synthesis relative to mock-irradiated controls. The parameter of rate of DNA synthesis per cell in DNA synthetic phase showed a rapid recovery toward control values between 2 and 4 h after irradiation and a slower recovery to control values by 22 h postirradiation. fractionated dose schedules were used to measure the effects of allowing a time interval between doses at nontoxic fluences (2.2 J/m2), moderately toxic fluences (8.8 J/m2) and toxic fluences (17.6 J/m2). These measurements indicate that, in the nontoxic range of fluences common to human exposure, mutational response is mediated by a postirradiation process which seems to show an enhanced ability to protect against mutation induced by subsequent irradiation. At moderatley toxic fluences there was little effect of dose fractionation, and, at toxic fluences, a time-dependent increase in mutation fraction was observed at separation times > 7 h. These latter observations arise primarily from cell cycle heterogeneity with regard to sensitivity to UV killing and mutation.

关键词： CfA clone-froming ability f 3 TdR trifluorothymidine fUdR fluorodexyuridine GCE growth curve extrapolation HGPRT hypoxanthine-guanine phosphoribosyl transferase IMf induced mutant fraction OUA ouabain PE plating efficiency S.E.M. standard error of mean TCA Trichloroacetic acid 6TG 6-thioguanine TK thymidine kinase UV 254 nm ultraviolet light XP xeroderma pigmentosum

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ISOLATION AND PARTIAL CHARACTERIZATION Of A ALPHA-2-BETA-1-GLYCOPROTEIN fROM HORSE PLASMA

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INTERNATIONAL JOURNAL Of BIOCHEMISTRY 1983年第8期15卷 1013-1018页

作者： PELLEGRINI, A ZWEIfEL, HR VONfELLENBERG, R Department of Veterinary Physiology University of Zurich Winterthurerstr 260 8057 Zürich Switzerland

A .alpha.2-.beta.1-glycoprotein was isolated from horse plasma by classical methods. The final product appeared homogeneous by agarose gel and pore limit SDS[sodium dodecyl sulfate] polyacrylamide gel electrophoresis, immunoelectrophoresis and crossed immunoelectrophoresis. The protein moved in agarose gel electrophoresis just above the .beta.1 region and seemed composed of a single polypeptide chain. A highly heterogenic banding pattern, focused between pH 5.1 and 6.5 was revealed by isoelectric focusing. The MW determined by gel filtration on Sephadex G100 and by a pore limit polyacrylamide gel electrophoresis in presence of SDS were 65,000 and 82,300 dalton, respectively. No serological relation was found between the horse .alpha.2-.beta.1-glycoprotein and human and bovine plasma proteins.

关键词： SDS-sodium dodecyl sulphate BIS—methylenebisacrylamide TEMED- N , N ′, N ′, N ′-tetramethy-lenediamine K 2 EDTA-ethylenediaminetetraacetic acid dipotassium salt Affi Gel Blue-Cibacron Blue f3GA crossed-linked to Bio Gel® A5-M 2-ME-2 mercaptoethanol f α 1 -PI-fast α 1 -protease inhibitor T -Total acrylamide concentration C -methylenebisacrylamide concentration

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