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C Ramel

Publications and source records attributed to C Ramel.

At least 55 records · Page 3Linked to original sources

Advantages of and problems with short-term mutagenicity tests for the assessment of mutagenic and carcinogenic risk.

The Salmonella microsomal assay has become an indispensible tool for the screening of mutagens and carcinogens, particularly when a large number of samples have to be tested, as in the present context for the screening of air pollution. However, for a more definite identification of potential carcinogens, a verification of the results from bacterial tests has to be performed with a battery of other tests, including point mutations and chromosomal aberrations in eukoaryotic systems. While there is a close qualitative correlation between the mutagenic and carcinogenic property of chemicals, a corresponding quantitative correlation between the mutagenic and carcinogenic potency is not always found. One reason for this lack of quantitative correlation presumably depends on the fact that cancer is induced in two steps, of which only the initiating, but not the promoting, step constitutes a mutational event, which is reflected by mutagenicity tests. Present mutagenicity tests have concentrated on discrete major mutations, while mutations of polygenes, acting on quantitative characters, have largely been omitted. Mutational data from Drosophila indicate, however, that polygenes mutate at a considerably higher rate than major genes and that they have a comparatively strong effect in heterozygous condition. It seems of great importance to develop appropriate methods to study induced mutations of polygenic systems and to get a better understanding of the properties of these genetic systems and an evaluation of the risk connected with induced mutations in polygenes.

Air Pollutants↗

Relation between the somatic toxicity of dimethylnitrosamine and a genetically determined variation in the level and induction of cytochrome P450 in Drosophila melanogaster.

The insecticide-resistant Drosophila strain Hikone R differs genetically from sensitive strains by having a higher activity of cytochrome P450 and a lack of inducibility with phenobarbital. Toxicity tests with dimethylnitrosamine (DMN) showed that: (a) the toxic effect of DMN was higher in the insecticide-resistant strain than in sensitive strains in accord with data by Vogel (1980); and (b) induction with phenobarbital increased the toxicity of DMN in the sensitive strains but not in the resistant strain. A dominant gene, responsible for the metabolic pattern of Hikone R and the non-inducibility, was located to 66 cM, near the gene vg in chromosome 2. This gene is most likely identical with that causing insecticide resistance, described by Kikkawa (1961).

Animals↗

The metabolism of drugs and carcinogens in isolated subcellular fractions of Drosophila melanogaster. I. Activation of vinyl chloride, 2-aminoanthracene and benzo[a]pyrene as measured by mutagenic effects in Salmonella typhimurium.

The capacity of microsomal fractions from different Drosophila strains to activate three premutagens, 2-aminoanthracene (2-AA), vinyl chloride (VCM) and benzo[a]pyrene (BP) was investigated, using Salmonella typhimurium as the indicator organism. A significant increase in the mutation response in the Salmonella test system was obtained with all three substances in the presence of a metabolizing system (S9) from Drosophila larvae. 2-AA was converted to highly mutagenic metabolite(s) by the Drosophila S9 and the mutagenic effect was further increased after pretreatment with Aroclor 1254 (PCB) or beta-naphthoflavone (BNF). BP had only marginal mutagenic effects, causing less than a 2-fold increase in the number of mutants over the control. The data indicate that the metabolic conversion of BP is different in the Drosophila as compared to the rat liver microsomal fraction. In accordance with mutagenic data on Drosophila in vivo, vinyl chloride was a fairly weak mutagen in this Drosophila/Salmonella in vitro system.

Animals↗

Mutagenic effects of effluents from chlorine bleaching of pulp.

Effluents from the bleaching of kraft pulp were tested for mutagenicity. Samples from different mills in which softwood kraft pulp is bleached in a conventional sequence of stages were spot-tested with the Escherichia coli pol A-/pol A+ system. All samples were nontoxic and therefore no difference could be noted between the repair-proficient and the repair-deficient strain. Also no toxic mutagenic effects were seen in spot tests with Salmonella. In a quantitative test with Salmonella, using the plate incorporation assay, effluents from the chlorination (C) stage induced point mutations of the base-substitution type. No frameshift mutations were noted. In the presence of a metabolizing system the direct mutagenic effect was reduced. This reduction was essentially NADPH-independent. Tests on effluents concentrated by freeze-drying indicated, in addition to the base-substitution effect of the C-stage effluent, the presence of frameshift mutagen(s) in this effluent and a base-substitution effect of the hypochlorite stage effluent. Fractionation of the C-stage effluent showed that the mutagenic components are of low molecular weight, extractable with diethyl ether, and sensitive to alkali treatment. Most of the mutagenicity is found in the neutral ether fraction. Induction of 6-thioguanine-resistant mutants in Chinese hamster V79 cells confirmed the mutagenic properties of the C-stage effluent.

Animals↗

Relationship between chemical damage of DNA and mutations in mammalian cells. I. Dose-response curves for the induction of 6-thioguanine-resistant mutants by low doses of monofunctional alkylating agents, X-rays and UV radiation in V79 Chinese hamster cells.

The shape of the dose-response curve for mutations induced at low doses of mutagenic agents in mammalian cells was studied. With the exception of X-rays and MMS, which are very toxic in relation to their mutagenic potency, dose-response studies with EMS, MNU, ENU and UV radiation were performed at doses giving about 100% survival. The results from several experiments were pooled for each agent to get higher resolution power at low doses, and the result was compared with what could be expected when linear interpolation is performed from higher doses. The dose response for induction of mutations by UV- and X-irradiation did not deviate from linearity at low doses. In contrast with irradiation and the ethylating agents EMS and ENU, the methylating agents MMS and MNU showed a significantly lower effect at low doses as compared with estimation from a dose 5 times as high. The extent of alkylation of DNA by various doses of MMS was linear in the same dose intervals. It is suggested that the decreased response at low doses of the methylating agents found here may be connected with changes in the mechanism of repair of the lesions induced at different dose levels.

Animals↗

The micronucleus test as part of a short-term mutagenicity test program for the prediction of carcinogenicity evaluated by 143 agents tested.

To evaluate the usefulness of the micronucleus test as a short-term assay for the detection of carcinogens, the correlation between micronucleus test data for 143 chemicals and corresponding cancer data, has been analyzed. For comparison, analogous data from Ames's test have also been collected for the same chemicals. In a comparison of the micronucleus test and Ames's test it was found that they had about the same specificity (around 80%) and predictive value (around 90%), while there was a significant difference in sensitivity in favor of Ames's test. The difference in sensitivity could be partly explained by differences in metabolizing capacities of these two test systems. It is concluded that a more elaborate test procedure for the micronucleus test would increase that sensitivity of this test. The principal value of the micronucleus test lies in the fact that it is an in vivo method, which may pick up effects at the chromosomal level not covered by bacterial assays. This is emphasized by the finding that the combination of Ames's test and the micronucleus test did increase the sensitivity of the screening procedure for the prediction of carcinogenic effects.

Animals↗

Studies on metabolic activation of vinyl chloride in Drosophila melanogaster after pretreatment with phenobarbital and polychlorinated biphenyls.

It is known that vinyl chloride is metabolized by the mixed function oxygenase system in the liver to reactive mutagenic and carcinogenic metabolites. This metabolic activation was studied in Drosophila melanogaster by measuring the uptake of 14C from labelled vinyl chloride in different strains and with different pretreatments with phenobarbital and polychlorinated biphenyl (PCB) Clophen A50), well known inducers of cytochrome P-450. In accordance with previously obtained data on vinyl chloride induced sex linked recessive lethals, it was shown that pretreatment with inducers increased the uptake of labelled compound up to ten times. There was, however, a marked difference in response between the five strains used. In particular, the strain Hikone, known to be resistant to insecticides, had a comparatively high initial radioactivity from vinyl chloride without any pretreatment, but it was not or insignificantly inducible with phenobarbital or PCB. Crosses between Hikone and an inducible strain indicated essentially a dominance for the Hikone genotype. Tests on inducible strains showed the same response to phenobarbital by 2 h old larvae and adult male and females. Dimethylsulphoxide (DMSO) used as a solvent decreased both the initial uptake of 14C and particularly the induction by PCB. The use of Tween 80 as an emulsifier did not have such an effect. It is emphasized that the interstrain variation in metabolic activation and inducability has to be taken into consideration in order to optimize the use of Drosophila for mutagenicity testing. This variation also opens up new possibilities of analyzing the mixed function oxygenase system biochemically and genetically.

Animals↗

Mutagenicity testing on chinese hamster V79 cells treated in the in vitro liver perfusion system. Comparative investigation of different in vitro metabolising systems with dimethylnitrosamine and benzo[a]pyrene.

A comparative study of three in vitro metabolising systems was performed in combination with Chinese hamster V79 cells, at which point mutation to 6-thioguanine resistance was scored. The three metabolising systems used were: (1) rat liver microsomal fraction (S9-mix); (2) feeder layer of primary embryonic golden hamster cells, according to Hubermann's system; (3) in vitro perfusion of rat liver according to the system of Beije et al. As model substances dimethylnitrosamine (DMN) and benzo[a]pyrene (BP) was used. The liver perfusion was more efficient than S9-mix as an activating system of DMN, while the feeder layer of embryonic cells was unable to activate this compound. The activation of DMN with S9-mix was dependent on the presence of NADP. By exposing the target cells in the liver perfusion at different distances from the liver the biological half life of the active metabolite of DMN could be estimated to less than 5 s. With BP the three metabolising systems showed reversed results as compared with DMN--both the feeder layer cells and S9-mix activated BP, the feeder layer cells being most efficient. With liver perfusion, the perfusate itself was totally negative. Only the bile showed a week mutagenic effect. These results are in accordance with the notion that intact liver cells perform both an activation and a subsequent deactivation of BP. Because of the importance of hepatic bio-transformation in chemical mutagenesis and carcinogenesis it is emphasied that a liver perfusion system could be used in a testing protocol for genotoxic effects as a valuable tool in order to analyse the mechanism of action of mutagenic and carcinogenic compounds detected in other test systems, for instance bacterial/microsomal tests.

Animals↗

Mutagenicity and metabolism studies on 12 thiuram and dithiocarbamate compounds used as accelerators in the Swedish rubber industry.

12 thiuram and dithiocarbamate compounds used in the rubber industry as accelerators, and to some extent as sources of sulfur, were tested, as well as carbon disulfide, a metabolite found in vivo after dithiocarbamate treatment, for mutagenicity in Salmonella typhimurium. A mutagenic effect on the base-substitution-sensitive strains TA1535 and TA100 was found for 7 compounds. The most potent directly acting mutagens were: tetramethylthiuram disulfide (TMTD), zinc dimethyldithiocarbamate (ziram), cadmium diethyldithiocarbamate and zinc diethyldithiocarbamate. Tetraethylthiuram disulfide (TETD), also known as Antabus, and carbon disulfide were non-mutagenic. The relatively low direct mutagenic effect of tetramethylthiuram monosulfide (TMTM) was enhanced in the presence of a metabolizing system (S9 mix). A hypothesis is given regarding the activation process of the monosulfide TMTM.

Animals↗

Some aspects on the organization of microfilaments and microtubules in relation to nondisjunction.

One possible mechanism behind nondisjunction is a malfunctioning spindle. A defect or lack of spindle is a criterion for c-mitosis. In chemical mutagenesis research the c-mitotic effect is a well known cytological phenomemon, which can be induced by many different compounds. Pioneer work was performed during the 1940's by Ostergren and Levan, and their results and conclusions are briefly discussed. Since colchicine can induce c-mitosis and c-meiosis, by definition, and nondisjunction, a correlation between these phenomena is logical. The general importance of the spindle protein tubulin is considered and some new data from the cell biology literature on spindle formation and function as well as chromosome structure are briefly summarized. This knowledge can be used to correlate cytological and biochemical parameters among which c-mitosis and changes in sulfhydryl group metabolism after chemical treatment are the most obvious ones.

Animals↗

Chemical induction of nondisjunction in drosophila.

Tests for chemically induced nondisjunction and loss of the sex chromosomes in Drosophila were performed. Of 31 compounds tested four gave rise only to an increase of XO exceptions, indicating the induction of chromosome loss. Six compounds, all known spindle inhibitors (colchicine, organic mercury, lead, and tin compounds) gave rise to an increase both of XXY and XO or of only XXY. The effect by metalloorganic compounds of which methylmercury was studied particularly closely, follows a peculiar pattern. In females with structurally normal X chromosomes only an increase of XX gametes is obtained, while with X chromosomes heterozygous for long inversions only O gametes are increased. The data indicates that the effect of the metal compounds occurs at first meiosis and that the process is connected with a meiotic drive, giving rise to a preferential segregation of the two X chromosomes to the functioning pole. The increase only of O gametes with structurally heterozygous X chromosomes can tentatively be explained by a loss due to crossing over within the inversion. An increase of the effect of methyl mercury was obtained where the normal pairing of the X chromosomes was interfered with by means of autosomal inversions. Likewise a synergistic increase of nondisjunction was obtained when a temperature chock of 10 degrees C was applied together with treatment with methylmercury. It is concluded that chemical induction of nondisjunction can be studied in Drosophila, but the sensitivity of the test is rather low and large amount of material is required.

Animals↗

The mutagenic effect of 1,2-dichloroethane on Salmonella typhimurium I. Activation through conjugation with glutathion in vitro.

One of the main components in the waste products from vinyl chloride industries (EDC-tar), is ethylene dichloride (1,2-dichloroethane). This compound has been tested for mutagenicity on Salmonella typhimurium TA 1535. It is concluded that 1,2-dichloroethane gives a weak direct mutagenic effect, which is enhanced by addition of the postmitochondrial liver fraction (S-9). This activation is NADPH-independent and non microsomal. It is caused by a factor in the soluble fraction (115 000 g supernatant). This activation was further enhanced by the addition of glutathione but not by the addition of L-cysteine, N-acetyl-L-cysteine or 2-mercaptoethanol. No activation was observed when glutathione was added in the presence of a totally denaturated S-9 fraction or in the absence of this fraction. Activation of 1,2-dichloroethane was also found in the presence of glutathione and glutathione S-transferase A and C but not with glutathione S-tranferase B. A synthetic conjugate S-(2-chloroethyl)-L-cysteine gave a strong direct mutagenic effect at concentrations where no effects were seen with 1,2-dichloroethane. It is thus concluded that 1,2-dichloroethane is activated by conjugation to glutathione. Another main component in EDC-tar, 1,1,2-trichloroethane, was not mutagenic under any of our experimental conditions. For comparison 1,2-dibromoethane was also tested and gave a stronger direct mutagenic effect than 1,2-dichloroethane. Like the latter 1,2-dibromoethane was also activated by a NADPH-independent process.

Animals↗