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Ethyl carbamate is a known carcinogen occurring in fermented food and beverages and is therefore of interest for food safety assurance. We studied the genotoxicity of ethyl carbamate in Salmonella typhimurium, in Saccharomyces cerevisiae and in human lymphoblastoid TK6 cells. In absence of cytochrome P450 enzymes, no ethyl carbamate-mediated genotoxicity was observed in any of the three test systems...
Two assays, the alkaline single cell gel electrophoresis (Comet) assay and the fluorescence in situ hybridisation (FISH) of a whole chromosome or `chromosome painting' assay have gained importance in recent years as witnessed by the increasing yield of scientific literature using these techniques. Thus, it would be useful to have one system to measure both endpoints. In the present communication,...
Mediterranean mussels were exposed to benzo[a]pyrene for 2 days at doses which had previously caused the formation of specific adducts in gill DNA. Micronuclei and other nuclear abnormalities were detected in gill cells and haemocytes in order to ascertain the induction of cytogenetic damage in two different target cells in parallel. A number of procedural details were examined initially to improve...
Genotoxic effects of benzo[a]pyrene (BP) and its reactive metabolites (±)-anti-benzo[a]pyrene-7,8-diol 9,10-oxide ((±)-anti-BPDE) were comparatively investigated in vitro with the permanent human fibroblast cell line MRC5CV1. Induced DNA adducts were measured by 32 P-postlabeling, DNA strand breakage was determined by the comet assay and the HPRT gene mutation test was used to detect cytotoxicity...
The effects of the flavonoids quercetin, myricetin and silymarin on DNA damage and cytotoxicity in human cells were investigated. DNA strand breaks and oxidised pyrimidines were determined using alkaline single cell gel electrophoresis (the comet assay). Inhibition of cell growth was also measured. Caco-2 (colon), HepG2 (liver), HeLa (epithelial) cells and normal human lymphocytes showed different,...
Mediterranean mussels were exposed to benzo[a]pyrene for 2 days at doses which had previously caused the formation of specific adducts in gill DNA. Micronuclei and other nuclear abnormalities were detected in gill cells and haemocytes in order to ascertain the induction of cytogenetic damage in two different target cells in parallel. A number of procedural details were examined initially to improve...
DNA adducts in human placenta have been studied in relation to metabolic genotype for glutathione S-transferase M1 (GSTM1) in 98 mothers living in two regions with a different annual average air pollution levels: Northern Bohemia – the district of Teplice as polluted industrial area (mines, brown coal power plants) and Southern Bohemia – the district of Prachatice as agricultural area without heavy...
The Allium anaphase–telophase assay was used to show genotoxicity of N-methyl-N-nitrosourea (MNU), maleic hydrazide (MH), sodium azide (NaN 3 ) and ethyl methanesulfonate (EMS). All agents induced chromosome aberrations at statistically significant levels. The rank of the lowest doses with positive effect was as follows: NaN 3 0.3 mg/l<MH 1 mg/l<MNU 41 mg/l<EMS 100 mg/l. The...
Despite the remarkable resistance to desiccation, Bacillus subtilis spores manifest indications of DNA damage when being kept in an extremely dry environment made by high vacuum. Spores of strain TKJ3422 (uvrA10 spl-1 recA4) with triple repair defects lost colony-forming capacity dependent on the duration and strength of the exposure. Mutations to rifampicin resistance were induced in the spores of...
The mouse micronucleus test was conducted to investigate the effect of high body temperature on micronucleus induction. Groups of 10 male ddY mice were exposed to 30°C for 1, 3 or 6 h, 37°C for 0.5, 1, 2, 3 or 4 h, and 40°C for 1 or 2 h. Bone marrow cells were sampled 24 h after heat exposure. Exposure of mice to 37°C for 3 or 4 h and 40°C for 1 or 2 h raised body temperature to approximately 40.5°C...
A micronucleus assay in vivo has been developed that is based on the use of freshly isolated kidney cells from mononephrectomized rats. In this validation study, a statistically significant increase in the frequency of micronucleated cells was detected in rats given i.p. a single dose of four kidney carcinogens, N-nitrosodimethylamine, N-nitrosodiethylamine, N-ethyl-N-hydroxyethylnitrosamine and N-nitroso-N-methylurea...
The genotoxicity of effluents collected from a conventional 5-stage softwood kraft pulp bleaching process was studied in Chinese hamster ovary (CHO) cells in vitro. Spent liquor from the first chlorination stage (C/D), where elemental chlorine and chlorine dioxide had been used in equal proportions, was shown to induce a dose-dependent increase in sister chromatid exchanges (SCEs) without metabolic...
We earlier developed the Chinese hamster ovary UV5P3 cell line that expresses cytochrome P4501A2 and lacks nucleotide excision repair for studying metabolism and mutagenicity of heterocyclic amines. The Chinese hamster ovary UV5P3 cells are approximately 50-fold more sensitive to the cooked food mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) than 2-amino-3-methylimidazo[4,5-f]quinoline...
Genotoxic effects of benzo[a]pyrene (BP) and its reactive metabolites (±)-anti-benzo[a]pyrene-7,8-diol 9,10-oxide ((±)-anti-BPDE) were comparatively investigated in vitro with the permanent human fibroblast cell line MRC5CV1. Induced DNA adducts were measured by 32 P-postlabeling, DNA strand breakage was determined by the comet assay and the HPRT gene mutation test was used to detect cytotoxicity...
In 7 non-smoking healthy volunteers, the number of micronuclei (MN) was determined in exfoliated buccal mucosa cells before and after rinsing the mouth with an aqueous 10 ppm solution of 2-trans-hexenal during 3 consecutive days. All individuals showed at least a doubling of the MN frequency during one of the next 4 days. An increase of the mean group MN frequency was observed on the fourth day, becoming...
We have demonstrated that free radicals generated by hydrogen peroxide (H 2 O 2 ), in the presence of divalent iron (Fe 2+ ) and a chelator (EDTA), oxidize 2′-deoxyguanosine (dG) to 8-hydroxy-2′-deoxyguanosine (8-OHdG). The 8-OHdG formed by this reaction was isolated and quantitated using reverse-phase HPLC with UV and electrochemical detection. A 1-h incubation of dG with...
The genotoxicity of effluents collected from a conventional 5-stage softwood kraft pulp bleaching process was studied in Chinese hamster ovary (CHO) cells in vitro. Spent liquor from the first chlorination stage (C/D), where elemental chlorine and chlorine dioxide had been used in equal proportions, was shown to induce a dose-dependent increase in sister chromatid exchanges (SCEs) without metabolic...
Ethyl carbamate is a known carcinogen occurring in fermented food and beverages and is therefore of interest for food safety assurance. We studied the genotoxicity of ethyl carbamate in Salmonella typhimurium, in Saccharomyces cerevisiae and in human lymphoblastoid TK6 cells. In absence of cytochrome P450 enzymes, no ethyl carbamate-mediated genotoxicity was observed in any of the three test systems...
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