Metabolism of aflatoxins B1 and G1 byAspergillus parasiticus

Résumé Un examen préliminaire fait avec une préparation d'Aspergillus parasiticus est rendu évidente la conversion de l'aflatoxine B en G. Il est probable que l'aflatoxines B₁ est le précurseur biogénétique d'autres aflatoxines produite par le champignon.

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K. K. M. is grateful to the Council of Scientific and Industrial Research, New Delhi, India, for the award of a Senior Research Fellowship.     

Binding of aflatoxins B1 and G1 to human serum proteins

Zusammenfassung In vitro Experimente mit¹⁴C-markiertem, gereinigtem Aflatoxin zur Untersuchung der Bindung von Aflatoxin B₁ und G₁ an verschiedene Serumproteine ergaben, dass Aflatoxin B₁ hauptsächlich mit-Globulin, G₁ dagegen vorwiegend mit Albumin bindet.

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This work was supported by part by a grant from the China Medical Board of New York, Inc., and was performed during one of us (S.S.L.) received a class C. research award from the National Science Council, Republic of China.     

Biosynthesis of aflatoxins by cell-free preparations fromAspergillus flavus

Résumé Incorporation d'acétate-1-¹⁴C, de leucine-U-¹⁴C et de mévalonate-2-¹⁴C sous forme d'aflatoxine dans les extraits libres des cellulesd'Aspergillus flavus. Dans le cas du mévalonate, l'incorporation de l'ordre de 0,4 pour 100 et les activités spécifiques sont 13 000 à 40 000 cpm/mg d'aflatoxine. La capacité d'incorporation des substrats en aflatoxine s'observe, en particulier, dans les mitochondries.     

Resolution of small G1 and G2 populations of L1210 leukemia by flow microfluorometric analysis aided by velocity sedimentation

L1210 leukemic cells grown in vitro were subjected to kinetic analysis using a flow microfluorometer. A single broad peak was found for the DNA content distribution if unfractionated cells were used; prior fractionation using lg velocity sedimentation allowed the separation of small peaks with smaller (G1) and larger (G2) DNA contents from the dominant S phase peak with intermediate DNA content.     

Resolution of small G1 and G2 populations of L1210 leukemia by flow microfluorometric analysis aided by velocity sedimentation

Summary L1210 leukemic cells grown in vitro were subjected to kinetic analysis using a flow microfluorometer. A single broad peak was found for the DNA content distribution if unfractionated cells were used; prior fractionation using lg velocity sedimentation allowed the separation of small peaks with smaller (G₁) and larger (G₂) DNA contents from the dominant S phase peak with intermediate DNA content.This work was supported by grant number 5P01CA13053 awarded by the National Cancer Institute, DHEW USA, and by grant num ber RBI 76-1 from the Queen Wilhelmina Fund, The Netherlands.     

Uptake of aflatoxin B1 by plastic materials

Zusammenfassung Aflatoxin wird von einigen gebräuchlichen Kunststoffen innerhalb kurzer Zeit in verhältnismässig grosser Menge absorbiert. Die vorliegenden Ergebnisse erlauben es, analytische Fehler bei Verwendung ungeeigneten Materials zur Zirkulation oder Filtration von Aflatoxinlösungen zu vermeiden.

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This is contribution No. 660 of the Euratom Biology Division.     

Regulation of G1 phase progression by growth factors and the extracellular matrix

Cell cycle progression is regulated by both intracellular and extracellular control mechanisms. Intracellular controls ensure that cell cycle progression is stopped in response to irregularities such as DNA damage or faulty spindle assembly, whereas extracellular factors may determine cell fate such as differentiation, proliferation or programmed cell death (apoptosis). When extracellular factors bind to receptors at the outside of the cell, signal transduction cascades are activated inside the cell that eventually lead to cellular responses. We have shown previously that MAP kinase (MAPK), one of the proteins involved in several signal transduction processes, is phosphorylated early after mitosis and translocates to the nucleus around the restriction point. The activation of MAPK is independent of cell attachment, but does require the presence of growth factors. Moreover, it appears that in Chinese hamster ovary cells, a transformed cell line, growth factors must be present early in the G1 phase for a nuclear translocation of MAPK and subsequent DNA replication to occur. When growth factors are withdrawn from the medium immediately after mitosis, MAPK is not phosphorylated, cell cycle progression is stopped and cells appear to enter a quiescent state, which may lead to apoptosis. Furthermore, in addition to this growth-factor-regulated decision point in early G1 phase, another growth-factor-sensitive period can be distinguished at the end of the G1 phase. This period is suggested to correlate with the classical restriction point (R) and may be related to cell differentiation.     

Metabolism of the normal cardiovascular wall. 1. Utilization of glucose by arteries and veins

Riassunto Il destino del glucosio generalmente mercato¹⁴C in diverse arterie e vene del cane é stato studiato con l'uso di una tecnica quantitativa di radiocromatografia su carta.I risultati non solo mostrano che nella parete arteriosa ha luogo una notevole glicolisi aerobica ed anaerobica, ma anche chiaramente indicano che una percentuale relativamente alta di glucosio viene trasformata in amino acidi (probabilmente attraverso il ciclo degli tricarbossilici) ed in grassi.Quantitativamente è stato osservato che il tessuto arterioso utilizza maggiore quantità di glucosio di quello venoso e che tale differenza è tanto maggiore quanto più i vasi sono periferici.

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Investigation was conducted during tenure of Special Research Fellowship (No. 11091) U.S. Public Health Service.     

Production and characterization of antibodies cross-reactive with major aflatoxins

Antibodies cross-reactive with 4 major aflatoxins were demonstrated three weeks after immunization of rabbits with an immunogen which was prepared by conjugating aflatoxin B3 to bovine serum albumin. Aflatoxin B3 was first converted to its hemisuccinate before conjugation to the protein. Tritiated aflatoxin B1 (AFB1) was used as the marker ligand both for antibody titer determination as well as for analysis of antibody specificity. Competitive RIA revealed that the antibodies have good cross-reactivity with aflatoxins B1, B2, G1, and G2 when tritiated AFB1 was used as the marker ligand. The concentrations causing 50% inhibition of binding of 3H-AFB1 to the antibodies by unlabeled aflatoxins B1, B2, G1, G2 and B3 were found to be 0.25, 3.34, 0.32, 4.0 and 0.53 ng/assay, respectively. The antibodies could be used for simultaneous analysis of aflatoxins B1 and G1, two of the most important toxic metabolites produced by Aspergillus flavus and A. parasiticus.     

Production and characterization of antibodies cross-reactive with major aflatoxins

Antibodies cross-reactive with 4 major aflatoxins were demonstrated three weeks after immunization of rabbits with an immunogen which was prepared by conjugating aflatoxin B₃ to bovine serum albumin. Aflatoxin B₃ was first converted to its hemisuccinate before conjugation to the protein. Tritiated aflatoxin B₁ (AFB₁) was used as the marker ligand both for antibody titer determination as well as for analysis of antibody specificity. Competitive RIA revealed that the antibodies have good cross-reactivity with aflatoxins B₁, B₂, G₁, and G₂ when tritiated AFB₁ was used as the marker ligand. The concentrations causing 50% inhibition of binding of³H-AFB₁ to the antibodies by unlabeled aflatoxins B₁, B₂, G₁, G₂ and B₃ were found to be 0.25, 3.34, 0.32, 4.0 and 0.53 ng/assay, respectively. The antibodies could be used for simultaneous analysis of aflatoxins B₁ and G₁, two of the most important toxic metabolites produced byAspergillus flavus andA. parasiticus.     

The desulphation of hexosamine sulphates by arylsulphatase B1

Summary The sulphation of Carbobenzoxyglucosamine by chlorosulphonic acid resulted in formation ofN-carbobenzoxyglucosamine-4,6-disulphate. UDP-galactosamine 4-sulphate and glucosamine 4,6-disulphate were the competitive inhibitors of arylsulphatase B. Arylsulphatase B can hydrolyze UDP-galactosamine 4-sulphate and glucosamine 4,6-disulphate but not galactosamine 6-sulphate.The work was done at the Australian National University, Canberra, Australia, and the Aligarh Muslim University, Aligarh, India.     

Uptake of aflatoxin B1 by the skin of rats

Zusammenfassung Nachweis der Absorption von Aflatoxin B₁ durch die Rattenhaut. Die toxische Leberwirkung nach der Hautbepinselung entsprach den von anderen Autoren gefundenen Verhältnissen bei peroraler oder intraperitonealer Verabreichung.     

Metabolism of dimethylnitrosamine by amphibians and fish in vitro

Résumé On a examiné le métabolism de la (C¹⁴)-DMN par des coupes de foie de truite, de poisson rouge et de trois espèces d'amphibiens, ainsi que l'excretion du cancérogène après injection par voie intrapéritonéale chez le triton et le poisson rouge. La relation entre le métabolisme et l'activité toxique et/ou cancérogène est discutée.     

Metabolism of hydrazines and hydrazides by the intestinal microflora

Summary Intestinal microorganisms are able to effect the metabolic reductive fission of hydrazines but not hydrazides during incubation in vitro.We thank the Medical Research Council for a studentship (to GCB).