Cigarette smokers have been reported to void urine which is more mutagenic, as measured in the Ames assay, than urine voided by non-smokers. Condensate from the mainstream smoke of a cigarette which primarily heats tobacco (test cigarette) has shown significantly reduced mutagenicity in a battery of in vitro genotoxicity assays compared with tobacco-burning cigarettes. The objective of this study was to determine whether the reduction in mutagenic activity observed in the in vitro assays would be reflected in the urine of smokers of the test cigarette. Twenty smokers were enrolled in a 4-week crossover study, with each smoker consuming test cigarettes ad libitum for a week and their usual brand of tobacco-burning cigarettes the other 3 weeks. Diet was strictly controlled throughout the study, and broiled and pan-fried meat was not served to minimize ingestion of mutagenic protein pyrolysis products. There was no statistically significant difference (p = 0.06) in consumption of tobacco-heating and tobacco-burning cigarettes. There were no statistically significant differences (p = 0.22) in salivary cotinine concentrations for smokers when smoking either tobacco-burning or tobacco-heating cigarettes. Urinary nicotine (ng/mg creatinine) was not different (p = 0.31) for smokers when smoking either tobacco-burning or tobacco-heating cigarettes. Urinary cotinine (ng/mg creatinine) was 32% lower (p = 0.0004) when smoking tobacco-heating cigarettes as compared with smoking tobacco-burning cigarettes. Twenty-four-hour urine samples were collected twice weekly, concentrated using XAD-2 resin and tested in Ames strains TA98 and YG1024 with metabolic activation. Tobacco-burning cigarette smokers experienced a 79% reduction in urinary mutagenicity as measured in strain YG1024 and a 72% reduction as measured in strain TA98 during the week that they smoked the tobacco-heating cigarette while maintaining a fixed dietary regimen. The results of this study indicate that smokers of tobacco-heating cigarettes void urine which is significantly less mutagenic than urine voided by smokers of tobacco-burning cigarettes.
Increased consumption of green vegetables in the diet has been associated with protection against carcinogenic effects and related mutagenic and clastogenic (chromosome breaking) activity of genotoxic agents. Chlorophyll, present in all green plant parts, has been suggested to be a major protective factor in the process. We have, however, observed that while a crude aqueous extract of Indian spinach leaf significantly reduced genotoxic effects, chlorophyll alone was ineffective. On the other hand, chlorophyll, both as an aqueous extract from the leaf and in a purified commercial form, induced a significantly high frequency of chromosome breaks in bone marrow cells of mice on oral administration. The crude aqueous extract of the leaf was non-toxic. The protective activity of the crude leaf extract may be attributed to the total effect of the interaction between different components, in which the clastogenicity of chlorophyll has been neutralized.
Involvement of human CYP2E1 expressed in genetically engineered cells in the metabolic activation of promutagens and procarcinogens was studied, An expression plasmid containing an insert of CYP2E1 cDNA and SR alpha promoter was constructed and transfected into the cultured cell line CR-119 which had previously been established by introducing a cDNA coding for NADPH-cytochrome P450 reductase. Among newly established cell lines, ER-181 showed the highest expression of CYP2E1 mRNA. Production of the CYP2E1 protein was confirmed by Western blot analysis using anti-rat CYP2E1 antibodies. Assay of 7-ethoxycoumarin O-deethylase activity demonstrated that ER-181 cells acquired the catalytic function of CYP2E1. ER-181 cells showed higher sensitivity to N,N-dimethylnitorosamine (DMN) in cytotoxicity assays as compared to parental CR-119 cells. Hypersensitivity to DMN of ER-181 cells was completely suppressed by 3-amino-1,2,4-triazole, a known inhibitor of CYP2E1. These results indicate that ER-181 cells which express human CYP2E1 are a useful tool to investigate toxicological functions of the cytochrome.
The possible mutagenicity of the water-soluble contents of cigarette smoke (WSCS) was evaluated by using the Vicia faba root tip micronucleus test. The results showed significant changes in micronucleus frequency which were caused by each different concentration of WSCS. This indicates that the Vicia faba root tip micronucleus test might be used as one kind of mutagenic detection method for cigarette smoke. A comparative evaluation on the mutagenicity of 10 brands of cigarettes was carried out. Results confirmed that various degrees of mutagenicity were found for all of the brand cigarettes, among them, Huaihai was the highest, while Camellia was the lowest. The micronucleus frequencies were reduced by adding tea polyphenol, nicotinamide adenine, vitamin C and sodium selenite to the WSCS. The results suggest that these added substances might reduce the genetic injury induced by cigarette smoke.
