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32P-POSTLABELING ANALYSIS OF DNA ADDUCTS IN HUMAN SPERM CELLS FROM SMOKERS AND NON-SMOKERS
Citation:
Gallagher, J., M. Vine, M. Schramm, J. Lewtas, M. George, B. Hulka, AND R. Everson. 32P-POSTLABELING ANALYSIS OF DNA ADDUCTS IN HUMAN SPERM CELLS FROM SMOKERS AND NON-SMOKERS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-94/048 (NTIS PB94137064), 1993.
Description:
To determine the feasibility of using human sperm cells for DNA 32postlabeling 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 non-smokers. he mean adduct level for the three exposure groups was not related to smoking status. ased on the level of radioactivity associated with various dilutions of a Benzo(a)pyrene (B(a)P)-derived adduct our limit of sensitivity was at least 1.2 adducts/109 nucleotides. utative adducts/nucleotide was not related to smoking status. ne major discrete DNA adduct of unknown chemical structure was detected in three of the 36 samples analyzed (one nonsmoker and two smokers). ur 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. his point is particularly relevant given that the 32p-postlabeling procedures rely, in part, on visual verification of the presence of DNA adducts.