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and without ~wever, in a ion of TA98 ,utable to the is, including tnd 7,12-di- J to express ~genicity to TA98 strain :olic activa- • e the domi- )-dependent tose-depen- ~ individual . other than In addition dr, nitrated PAHs) are as a result is !25]. It is yes could autagenie- bacterial :vation by ~resent in ~ cells for uction of ~ the YG ~ransform tes. Re- ' metabo- ependent O-acety- urinary $9 and • rease of ites ob- etion of urinary ume of ~ns ex- e about Ill Ill Ill • Ill M. ~ernd et al. / Mutation Research 391 (1997) 99-1 I0 109 [8] and needs substantially lower volumes of urine extracts as well as of $9 mix. Therefore, both stan- dard incorporation and microsuspension bioassays were used in this study. In comparison with the standard plate test results, indeed, the number of obtained revertants was more than 10 times higher. The urine from most individuals was found to be mutagenic in the Kado assay based on the signifi- cance in the Bernstein model. However, substantial interindividual variations with overlapping ranges of the number of induced revertants in both groups resulted in statistically insignificant differences be- tween the TP and PT groups. A positive correlation between some of the urinary PAH/metabolites ana- lyzed and the YGI041 revertants without external metabolic activation may suggest that, in contrast to the plate incorporation test, the differences in treat- ment conditions, such as concentrated bacteria and preincubation with longer direct contact of indicator strains with urine extracts and enzymes could affect the metabolic transformation of excreted direct- and indirect-acting mutagens [28]. A high interindividual variability in urine muta- genicity responses seemed to be the main disadvan- tage of this biomarker. These differences may be explained by genetic, environmental and lifestyle factors. The metabolic activation and detoxification processes are crucial for individual susceptibility to the exposure of genotoxic carcinogens and influence the excretion of mutagens in urine. The dependence of urine mutagenicity for smokers on glutathione-S- transferase genotype has recently been presented [29]. The glutathione S-transferase M1 genotype also had a significant effect on urine mutagenicity and urinary PAH metabolites, but the effect of personal expo- sures to PAHs on the variability of biomarkers might be even higher [30]. The present study has shown that the use of two biomarkers - urinary mutagenicity and urinary PAH/metabolites analysis - may reflect the expo- sure of the population to genotoxic contaminants in ambient air. The sensitivity of the TA98 strain seemed to be insufficient in detecting the differences in ambient air exposure. On the contrary, the use of YG1041 tester strain could facilitate the detection of mutagens excreted in the urine arid it is to be recom- mended to the biological monitoring. The microsus- pension modification did not show any substantial advantage over standard plate incorporation assay in this study and its use in urinary mutagenicity assays needs to be validated in further comparative studies. Acknowledgements This study was supported by a grant from the Czech Ministry of the Environment (Teplice Pro- gram), the US. Environmental Protection Agency/the US Agency for International Development and CEC (PHARE II, EC/HEA/18/CZ). The authors ac- knowledge the support of the Directors of the Dis- trict Institutes of Hygiene in Teplice (Dr. F. Kot~ovec) and Prachatice (Dr. J. No~i~ka) and the technical support of the staff of these institutes. They acknowledge the technical support of A. Truhl~ovfi and H. ~muhat~ov~ of NIPH, Prague, technical ad- vice of J. ~mld and statistical advice of Dr. E. ,~vandovfi. References [1] EHC (Environmental Health Criteria) 155 (1993) Biomarkers and Risk Assessment: Concepts and Principles, WHO, Geneva, 82 pp. [2] B.N. Ames. J. McCann, E. Yamasaki, Methods for detecting carcinogens and mutagens with the Salmonella/ mammalian-microsome mutagenicity test, Mutation Res. 31 (1975) 347-364. [3] E. Yamasaki, B.N. Ames, The concentration of mutagens from urine with the nonpolar resin XAD-2: cigarette smokers have mutagenic urine, Proc. Natl. Acad. Sci. USA 74 (1977) 3555-3559. [4] V. Minnich, M.E. Smith, D. Thompson, S. Komfeld. Detec- tion of mutagenic activity in human urine using mutant strains of Salmonella typhimurium, Cancer 37 (1976) 665- 670. [5] M.S. Legator, E. Bueding, R. Batzinger, T.H. Connor, E. Eisenstadt. M.G. Farrow, G. Fiscor, A. Hsie. J. Seed, R.S. Stafford, An evaluation of the host-mediated assay and body fluid analysis. A report of the U.S. Environmental Protection Agency Gene-Tox Program, Mutafiot~ Res, 98 (1982) 3 t9- 374. [6] R.B. Everson, Detection of occupational and environmental exposures by bacterial mutagenesis assays of human body fluids. J. Occup, Med. 28 (1986) 647-655. [7] R.P. Bos, J.L.G. Theuws, P.T. Henderson, Excretion of mutagens in human urine after passive smoking, Cancer Lett. 19 (1983) 85-90.

llO [81 M. ~ern6 et al. / Mutation Research N.Y. Kado, D. Langley, E. Eisenstadt, A simple modification of the Salmonella liquid-incubation assay. Increased sensitiv- ity for detecting mutagens in human urine, Mutation Res. 121 (1983) 25-32. [9] M. Watanabe, T. Nohmi, M. lshidate Jr., New tester strains of Salmonella typhimurium highly sensitive to mutagenic nitroarenes, Biochem. Biophys. Res. Commun. 147 (1987) 974-979. [10] RA. ~r'~rn, I. Bene.~, B. BinkovL J. Dejmek, D. Horstman, F. Kot~]ovec, D. Otto, S.D. Perreault, J. Rube], S.G. Selevan, I. Skal~, R.K. Stevens, J. Lewtas, Teplice Program - the impact of air pollution on human health, Environ. Health Perspect. 104 (Suppl. 4) (1996) 699-714. [11] D. Szadkovski, A. J~Srgensen, H.G. Essing, K.H. Schaller, Die kreatinineliminationsrate als Bezugsgr~Sss for Analysen aus Hamproben, Z. Klin. Chem. U. Klin. Biochem. 8 (1970) 529-533. [12] R.W. Williams, T. Pasley, R. Watts, J. Inmon, J. Fitzgerald, L. Claxton, Comparative yields of mutagens from cigarette smokers urine obtained by using solid-phase extraction tech- niques, Environ. Mol. Mutagen. 14 (1989) 20-26. [13] D.M. Maron, B.N. Ames, Revised methods for the Salmonella mutagenicity test, Mutation Res. t 13 (1983) 173-215. [14] Y. Hagiwara, M. Watanabe, Y. Oda, T. Sofuni, T. Nohmi, Specificity and sensitivity of Salmonella typhimurium YGI041 and YG1042 strains possessing elevated levels of both nitroreductase and acetyltransferase activity, Mutation Res. 291 (1993) 171-180. [15] O.H. Lowry, N.J. Rosebrough, A.L. Fan', R.J. Randall, Pro- tein measurement with the Folin phenol reagent, J. Biol. Chem. 193 (1951) 265-275. [16] D. DeMarini, M.M. Dallas, J. Lewtas, Cytotoxicity and effect on mutagenicity of buffers in a microsuspension assay, Teratogen. Carcinogen. Mutagen. 9 (1989) 287-295. [17] L.D. Claxton, J. Creason, J.A. Nader, W. Poteat, J. Orr, GeneTox manager for bacterial mutagenicity assays: a per- sonal computer and minicomputer system, Mutation Res. 342 (1995) 87-94. [18] L. Bernstein, J. Kaldor, J. McCann, M.C. Pike, An empirical approach to the statistical analysis of mutagenesis data from the Salmonella test, Mutation Res. 97 (1982) 267-281. [19] H. Vainio, M. Sorsa, K. Falck, Bacterial urinary assay in monitorir!g exposure to mutagens and carcinogens, in: A. Berlin, M. Draper, K. Hemminki, H. Vainio (Eds.), Monitor- ing Human Exposure to Carcinogenic and Mutagenic Agents, 391 (1997) 99-110 International Agency tbr Research on Cancer, Lyon, 1984, pp. 247-258. [20] R. Pasquini, S. Monarca, G. Scasellati Sforzolini, R. Conti, F. Fagioli, Mutagens in urine of carbon electrode workers, Int. Arch. Occup. Environ. Health 50 (1982) 387-395. [21] B. Binkov/i, J. Lewtas, I. M~kov~, J. Len~ek, R.J. ~r-,im, DNA adducts and personal monitoring of carcinogenic poly- cyclic aromatic hydrocarbons in an environmentally exposed population, Carcinogenesis 16 (1995) 1037-1046. [22] M. ~erml, V. Hfijek, L. Dobifi], R.J. ~rfim, Mutagenic activ- ity of the urine of workers employed in the processing of coal tar, Prac. L6k. 36 (1984) 41-44, in Czech. [23] R. Pasquini, S. Monarca. G. Scasellati Sforzolini, A. Savino, C. Fatigoni, P. Pucetti, Urine mutagenicity and biochemical parameters as markers of exposure to petroleum pitch using a rat model, Environ. Mol. Mutagen. 15 (1990) 56-66. [24] P. Einist~5, T. Nohmi, M. Watanabe. M. Ishidate Jr. Sensitiv- ity of Salmonella typhimurium YGI024 to urine mutagenic- ity caused by cigarette smoking, Mutation Res. 245 (1990) 87-92. [25] R. Watts, J. Lewtas, R. Stevens, T. Hartlage, J. Pinto, R. Williams, K. Hattaway, I. Mi'~kowl, I. BeneL F. Kot~ovec, R.J. ~rfim, Czech-U.S. EPA health study: assessment of personal and ambient air exposures to PAH and organic mutagens in the Teplice district of Northern Bohemia, Int. J. Environ. Anal. Chem. 56 (1994) 271-287. [26] H.S. Rosenkranz, R. Mermelstein, Mutagenicity and geno- toxicity of nitroarenes. All nitro-containing chemicals were not created equal, Mutation Res. 114 (1983) 217-267. [27] P.F. Rosser, P. Ramachandran, R. Sangaiah, R.N. Austin, A. Gold, L.M. Ball, Role of O-acetyltransferase in activation of oxidised metabolites of the genotoxic environmental pollu- tant l-nitropyrene, Mutation Res. 369 (1996) 209-220. [28] T. Watanabe, M.J. Kohan, D. Walsh, L.M. Ball, D.M. DeMarini, J. Lewtas, Mutagenicity of nitrodibenzopyranones in the Salmonella plate-incorporation and microsuspension assays, Mutation Res. 345 (1995) 1-9. [29] A. Hirvonen, L. Nylund, P. Kociba, K. Husgavfel-Pursiainen, H. Vainio, Modulation of urinary mutagenicity by genetically determined carcinogen metabolism in smokers. Carcinogene- sis 15 (1994) 813-815. [30] B. Binkov,5., J. Lewtas, I. M~kovL P. R~ssner, M. ~ern.4, G. Mrfi~kovfi, K. Peterkov~, J. Mumford, S. Myers, R. ~rfim, Biomarker studies in Northern Bohemia, Environ. Health Perspect. 104 (Suppl. 3) (1996) 591-597. ~ Dicisi . /stra fibers. il24-h e (48E). only u~ signifit - --I~ groups silica. -~ under _ I~] treatml Keywo~ 1. Int Sil COrlSt; monL tries.