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W opracowaniu przedstawiono możliwości wykorzystania różnych testów bakteryjnych i eukariotycznych do oceny genotoksyczności środowiskowych substancji chemicznych.
The mutagenic activity of captan and captafol was tested using Ames stzaiiis and strains showing an SOS response. Captafol was mutagenic in S. typhimurium strain TA102 (uvr+) and captan in strain TA104 (uvrB). Both captan and captafol elicit damages in DNA recognized by correndonuclease U, as shown by the repair teat, and induced the SOS repair system in E. coli PQ37 (uvrA) strain. Only captafol induced the SOS system in PQ35 (uvr+). The lack of induction of p-galactesidaac at nonpermissive temperature in E. coli MD332 (dnaCs uvrA) strain showed that neither chemical was able to produce DNA breaks. In V79 Chinese hamster fibroblasts higher induction of c-mitosis by captafol than by captan (22% and 15% over th* control, respectively) was accompanied by a higher decrease in nonprotein sulfhydryl groups, mainly GSH (41% and 77%, respectively). The content of protein sulfhydryl groups was decreased by either fungicide to a similar extent.
Looking for a mechanism of induction of chromosomal aberrations by thiram we checked its ability to damage bacterial DNA. Evaluation of the mutagenic effect of thiram was by: (i) the Salmonella!mammalian microsome Ames test with S. typhimurium TA102 strain, (ii) E. coli PQ37 strain to reveal an induction of the SOS response, (iii) test with E. coli BH20 fpg- and BH190 fpg-uvrA- mutants to check possible effect of modification of purines, (iv) test with E. coli AB1157 and AB1151 ada3 to detect DNA alkylation in 06 -position of guanine. The results of this work exclude DNA cross-links, oxidative damage of DNA, opening of imidazole ring in purines and events leading to activation of the SOS system as well as adaptive response as direct reasons of chromosomal abnormalities induced by thiram.
In the recent paper genotoxic effects of daminozide and its metabolites were tested. Evaluation of the mutagenic effect of daminozide was by: (i) the Salmonella/mammalian microsome Ames test with S.typhimurium TA97, TA98, TA100 and TA102. (ii) E. coli PQ37 strain to reveal an induction of the SOS response, (iii) S. typhimurium TA1538 (uvrB) and TA1978 (uvr+) to detect the chemicals bound covalently to DNA (repair test). Daminozide was not mutagenic in any of the S.typhimurium strains and did not induce damages in DNA recognized by correndonuclease II, as shown by the repair test. Only metabolites of daminozide induced the SOS system.
The Chinese hamster cell mutant V-C8 is defective in the Brca2 gene (Kraakman-van der Zwet et al., 2002, Cell Biol.-, 22: 669). Here we report that V-C8 cells were 10-fold more sensitive to camptothecin, an inhibitor of topoisomerase I, than the parental V79 cells. The level of the relaxation activity of topoisomerase I in nuclear extracts was also lower (4-fold) in V-C8 than V79 cells, in spite of the fact that the level of the topoisomerase I protein was the same in these cells. The survival of V-C8 cells in the presence of camptothecin, the sensitivity of V-C8 topoisomerase I to camptothecin, and the level of the relaxation activity in V-C8 nuclear extract were almost completely restored by transfection of V-C8 cells with the murine Brca2 gene or by the transfer of human chromosome 13 providing the BRCA2 gene. These results indicate that the ob­served changes in the topoisomerase I activity in V-C8 are due to the defective func­tion of the Brca2 gene.
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