||32P-Postlabeling Analysis of DNA Adducts in Human Sperm Cells from Smokers and Nonsmokers.
Gallagher, J. E. ;
Vine, M. F. ;
Schramm, M. M. ;
Lewtas, J. ;
George, M. H. ;
||Health Effects Research Lab., Research Triangle Park, NC. Genetic Bioassay Branch. ;North Carolina Univ. at Chapel Hill. Dept. of Epidemiology. ;SRA Technologies, Inc., Research Triangle Park, NC. ;Environmental Health Research and Testing, Inc., Research Triangle Park, NC.
DNA adducts ;
Phosphorus 32 ;
Feasibility studies ;
Thin layer chromatography ;
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||To determine the feasibility of using human sperm cells for DNA (32)P-postlabeling analyses, and to evaluate the baseline level and the possible presence of smoking-related DNA adducts in these cells, sperm DNA was isolated from 12 heavy smokers, 12 light smokers and 12 nonsmokers. The mean adduct level for the three exposure groups was not related to smoking status. Based on the level of radioactivity associated with various dilutions of a benzo(a)pyrene-derived adduct our limit of sensitivity was at least 1.2 adducts/10 to the 9th power nucleotides. The level of radioactivity expressed as putative adducts/nucleotide was not related to smoking status. One major discrete DNA adduct of unknown chemical structure was detected in three of the 36 samples analyzed (one nonsmoker and two smokers). Our study emphasizes the need to more clearly define the significance of background radioactivity associated with DNA adduct maps where the measured adduct levels approximate detection limits defined by visual observance of adduct spots. This point is particularly relevant given that the (32)P-postlabeling procedures rely, in part, on visual verification of the presence of DNA adducts.
||Pub. in Cancer Epidemiology, Biomarkers and Prevention, v2 p581-585 Nov/Dec 93. Prepared in cooperation with North Carolina Univ. at Chapel Hill. Dept. of Epidemiology, SRA Technologies, Inc., Research Triangle Park, NC., and Environmental Health Research and Testing, Inc., Research Triangle Park, NC.
|NTIS Title Notes
||Reprint: 32P-Postlabeling Analysis of DNA Adducts in Human Sperm Cells from Smokers and Nonsmokers.
||PC A02/MF A01