Document bB6XwjkDLY8N8JBe2DkaJoea6

/ r February 1990 >V Appearing monthly Vol. 240. No. 2 [ ]K-17n-(100) 1990 MUREAV 240(2) 47-158(1990) R&S 024963 Editor-in-Chief: F.H. Sobels, Leiden Board of Managing Editors J. Ashby. Macclesfield: C. Franceschi. Modena (DNAging); S.M. Galloway, lies/ Point. PA (Mutation Research Letters); J.M. Gentile. Holland. Ml (Mutation Research Letters); B.W. Glickman. Toronto. On/.. P.C. Hanawalt, Stanford. CA (DNA Repair); P.H.M. Lohman. Leiden (DNA Repair); K. Sankaranaravanan, Leiden: F.J. Ue Serrcs. Research Triangle Park. A'C; R.B. Setlow. Upton. A ) (DNAging); M.D. Shelby Research Triangle Park. S'C: T. Sugimura. Tokyo (DNAging); II. Takehe. Ki oto I DNA Repair); J. Vijg. Rijswijk (DNAging); E. Vogel, Leiden: J.S. Wassom. Oak Ridge. 7A Genetic Toxicology Testing and Biomonitoring of Environmental or Occupational Exposure R&s 024964 i '> Mutation Research, 240 (1990) S3--85 i Kevier MUTGf'N 01507 Cytogenetic analysis of peripheral blood lymphocytes in workers exposed to vinyl chloride L. Hrivnak, Z. Rozinova, S. Korony and E. Fabianova Regional Institute of Hygiene, 975 5ft Banska Ihstnca tCzechosUn'iikia) (Received 21 Februjry 19X9) (Revision received 31 August 19X9) (Accepted 4 September 19X9) Keywords: Vinyl chloride-exposed workers; Chromosomes; Peripheral blood lymphocytes: Lymphocytes; Peripheral blood; Chro mosomal aberrations Summary In the present study, the method of cytogenetic analysis of peripheral blood lymphocytes was used to examine 43 workers exposed to vinyl chloride monomer (average exposure 11.2 years) and 22 subjects selected from the same locality (control group). A total number of 8650 metaphases were analysed. All cytogenetic parameters examined were increased in the exposed group as compared to the control group and 3 parameters, chromatid breaks, percentage of aberrant cells and breaks per cell, were significantly increased (P < 0.001). Vinyl chloride monomer (VCM) is a basic com ponent for the production of polyvinylchloride (PVC). PVC, with an estimated yearly production of 16 million tonnes in 1980 (Groth et al., 1981), is in daily contact with the human population. In exposed workers VCM leads to an increased inci dence of angiosarcomas of the liver, brain tumours, malignancies of the haema.opoctic system and cancer of the lungs (IARC, 1979; Buffer et ah, 1979; Nicholson et ah, 1975). Similar results were obtained in experimental animals (Suzuki, 1978, 1981). Increased frequencies of chromosomal aberra tions in exposed workers indicate that VCM has mutagenic effects in man (Ducatman et al., 1975; Correspondence: Dr. L. Hrivnak, Regional Institute of Hygienc. 975 56 Banska Bystrica (Czechoslovakia). Kueerova et ah, 1979). We analysed the chro mosomes of peripheral lymphocytes of workers exposed to VCM. The maximum allowable con centration for VCM in Czechoslovakia is 10 mg/m3. Materials and methods The exposed group consisted of 43 workers (average age 34.4 + 8.5 years), with an average VCM exposure of 11.2 7.2 years. According to measurements at the work place during the last 6 months before the blood was sampled the VCM concentrations varied from 2 to 40.9 mg/m3. The 22 subjects of the control group had similar social habits and living environments, but they were not exposed to VCM or other known mutagenic agents. All subjects who had been ill or taken medicines (including X-ray examinations) during the previ ous 3 months were excluded. 0165-1218/90/S03.50 O 1990 Elsevier Science Publishers B.V. (Biomedical Division) fff; .' yj. ' t :! > - 84 TABLE1 RESULTS OR CYTOGENETIC ANALYSIS OF PERIPHERAL FLOOD LYMPHOCYTES OF VCM-EXPOSED WORKERS Group N Age (years) Average exposure to VCM (years) Number of Chromosomal mctaphjses aberrations analysed B' B" E' G Percent of aberrant metaphascs (S AB,C) Breaks per inctaphasc (B/C) Exposed Control 43 34.4 8.5 22 40.3 8.2 11.2 7.2 - 4650 2200 119 * 24 52 18 19 184 3.30 * 5 55 1.64 0.037 0.019 B'. chromatid hrcaks: B", chromosome breaks: E/. chromatid exchanges: G. gaps *P <0.001. Peripheral blood lymphocytes were cultured according to Hungerford (1965). Heparinised blood (0.7 ml) was cultured for 48 h at 37 C in RPMI medium (USOL, Prague), supplemented with 20% inactivated calf serum (USOL, Prague), antibiotics (penicillin, streptomycin, neomycin) and stimulaicd with phytohacmagglutinin (PHA, USOL, Prague). Colchicine (0,7 g/ml, Fluka) was added to the culture for the last 2 h of incubation. Cytological preparations were made using a routine protocol and the slides were stained with Giemsa. At least 100 metaphascs with 46 centromeres were analysed per person for the presence of chromosomal aberrations. The following types of chromosomal aberra tions were recorded: chromatid breaks (B'), chro mosome breaks (B"), chromatid exchanges (E'), gaps (G). Gaps were not considered aberrations. Differences between groups were statistically evaluated using the /-test. Relative frequencies were transformed by inverse trigonometric func- TABLE 2 PERCENT ABERRANT METAPHASES ((6 AB.C) IN SMOKERS AND NON-SMOKERS Group Exposed N Range Control N Range % AB.C 3.3 1.09 43 1-6 1.64 0.9 22 0-3 e H AB.C smokers 3.45 1.04 * 15 2-5 1.73 0.76* 7 1-3 * P < 0,05: * P< 0.001. non-smokers 3.23 1.12 * 28 1-6 1.60 1.0 ** 15 0-3 lion (2 arc sin //) because of their small values (Reisenaucr. 1970; Draper and Smith. 1981). Results and discussion The results are summarized in Table 1. All aberration types were increased in the exposed group as compared to the control group. The differences were highly significant for chromatid breaks (B'), percentage of aberrant cells (% AB. C) and for breaks per cell (B/C) (P <0.001). Table 2 shows the percentages of aoerrant cells in the groups of exposed and control persons with respect to smoking habits. In both groups there was no difference between smokers and nonsmokers. The difference in aberrations given as % AB.C between non-smokers in the exposed and the control groups was highly significant {p < 0.001), with respect to the smokers the difference was significant (P< 0.05). These data indicate that the main reason for the increased aberration frequencies is the exposure to VCM. Our data agree xvith the results of Gcryk and Zudova (1986) who reported 4.1% AB.C in VCMexposed workers. Acknowledgements We are grateful to Dr. T, Geist for his kind support. We thank Dr. E. Horvathovi for her useful medical advice and D, Knoppova for her laboratory assistance. References Buffer, A.. 5. Wood. C. Eincr, L. Suarez and DJ. Kilian (1979) Mortality experience of workers in vinyl chloride monomer production plant. J. Occup. Med., 21. 195-202. ....... ........... ................... ..... ..... ;" >';/' '-: . m-:;|?;-v - 'V '' '?* . :W Draper, N.R., and H. Smith (1981) Applied Regression Analy sis, 2nd edn., John Wiley and Sons, New York. (Russian translation, Moscow, 1986). Ducatman, A., K. Hirshom and J.I. Selikoff (1975) Vinyl chloride exposure and human chromosome aberrations, Mutation Res., 31,163-165. Geryk, E,. and Z. ZudovA (1986) Vinyl chloride as occupa tional health hazard, Pracov. Lk., 38,1-8. Groth, D.H., D.W, Lynch, WJ, Moorman, L.E. Stealer, T.R, Lewis, W.D, Wagner and Ch. Kommincni (1981) Pneumo coniosis in animals exposed to poly(vinyl) chloride dust, Environ. Health Perspect., 41, 71-81. Hungerford, D.A. (1965) Leucocytes cultured from small inocula of whole blood and the preparation or metaphase chromosomes by treatment with hypotonic KC1, Stain Technol., 40, 333-338. 85 IARC (1979) Monographs SuppL, International Agency for Research on Cancer, Lyon. Kuierova, M,, Z. PolivkovA and J. BAtora (1979) Mutagenni a karcinogenni uiinky vinylchloridu. Prac. Lck.. 31. 249-252. Nicholson, W.J., E.C. Hammond, H. Seidman and IJ. Selikoff (1975) Mortality experience of a cohort of vinyl chloride workers, Ann. N.Y. Acad. Sci,, 246, 225-230. Reisenaucr. R. (1970) Methods of Mathematical Statistics and Their Application. SNTL, Prague, 239 pp. (in Czech). Suzuki, Y, (1978) Pulmonary tumors induced in mice by vinyl chloride monomer. Environ. Res.. 16, 285-301. Suzuki, Y. (1981) Neoplastic and nonneoplastic effects of vinyl chloride in mouse lung. Environ. 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