Document omBzOVBpOVg3py9N2GNw5n3oR

. 1 ' ---uy ' , . ` f, / ,,L `r~(t1$o) Appearing monthly Volume 77, No. 2, February 1980 ISSN 01G5-1218 . . /, .... MUHEAV 77 (2199-19G (10801 f ' V.l * ' _ * \ <\t -> ;*-*]*-' hr / 'j'rS -S3 - A /T \ J ] j :Y, A ' ( ' ; O' T r- ] i 7` . .. /, '! . /, -Jl -* *-+ V . v,, Genetic Toxicology Testing MANAGING KDITQIt: ASSOGIATK KOITOUS: F.M. Sobol'*, l.citlcn F J, tie Serrrs. fteseareft Triuwile Turk, S( K. Vojjel, l.i'uU'n KDITOKIAI. HOAKU: A. Abbondandolo, ftsu S, Abrahamson, Madison, SJ Mb Adler, Scuhcrbcrc K, J. Alberlini, liurltnatun, Vermont II. Hartsoh, l.ytm M.A Uender, t'ptnn ,N.K, UltHitn* Ithmui, SY |l,K. llmekman, Sonnet, tU l). Ilrusiek. Kensington L. Husk. ('ppsnla A.V. Garrumi, /.n-mure, (\\ I.K. ni'uu'r, .S\i/i Francisco, (\\ H..I. Dean, Niffuu'bomrie U.H. Khlim*, Scuherherj L. Khtealierc* Sttickludm 4.U. Kpirr, Oak Ku/tfA TS W.M, IIimuuw, lUdne, 7'V U. Graf, cc/<it cncnhoch \V,F. Grant, Sic, Anna tic HeHci'ue I. Mansmann. (it'd t mum .1 A. Ibitidle. itou-nteteu- (bit. Hoffmann, Washington, DC A.W. Hsi\Ou/{ HuUc, TS J. Md'.inn. Itcrttvlcv, ('A H K Manor. liasel K. MontcNiitu, l.yon K. MouMin't In, Ur^uy /\. Nasun. ttttau't* N,K, Notani. Tromhay. itomhay G, t )be, Hcritn Olivieri, Home Kamel, Sfti(d:hfJm Kay. Croton , Knhrhftrn, .S. K>k.*nkr.n/, Voii' York, S'* Scln *noieh. GY/er-deG'H l'\ Simmon. A/*n/' i'tirk, ( .1 Suak. t'anthruljr, AM , Sors.i, //e/*iri^.*l F. Sludi. 1`ufirout crt }i(` Sui'imura. Tokyo ,.J, Sram, t'ra^ue G. Tlully, f'i:nihrn!^c, St A ,S. W.issoin. Oak ,K Wurcb-r. /.urtch /amier. Washington. DC i l V` t- 'KC : t` }' 'AX - e-i.i.iif... *s^>i i 4 ' . ` N1 - 1 Mutation Itrxmrcli, 77 (10X0) Hin-lUO OK iM-vier/N'ortli-Holland liioniftijr.il I'rrst A /'/// ) 103 -Short Communication LACK OF MUTAGENIC EFFECT OF VIN YL CHLORIDE MONOMF.U IN THE MAMMALIAN SPOT TEST SAN'DOlt I'KTKK uml (JYOW.Y l.'S>iV.MI Y ' 1 Stair Institute of Hygiene, l.uftor.1tor', of Human tienelirs. rliiiuun Chmnieal am! t'hunnurvulwal tl Drits Ltd., and * Stair Institute of (lerufiational Itrallli, Iteinirtntent of t'.xtirrimrntul /uihuiuyy, ISudapesl tHungaryi (ICcivivi-il 2(i .liinr 1070) (Arerpn-tl 7 r \`.17`.1) Evaluation of genetic hazards of exposure to vinyl chloride monomer (VCM) is one of the most serious current problems in occupational medicine. Numer ous tests have been done to check the mutagenicity of VCM. Many of these were carried out on sttb-mammalum systems |2 -l,7,S.lf>.22.2r>.2<i.29--31 J, others on mammals m vim 11,12.27.2U.3.'J|, and on human lymphocytes in vitro [f>,0,12--1 1,10--18,20.21,27.2S.32]. The sub-mammalian examinations sited light on the importance of the metabolism of VCM in the relation to the mutagenicity [2--l,7,15,lf),22,2ib-31 ]. Chromosome analysis of human lymphocytes demonstrated dose-dependent (12,17,20| and time-dependent |.'12) mutagenic effeets, showing; the importance of the chronic exposure to VCM (0,1.2.11,lfi,27,28] as opposed to the aeute administration (SJ. Inducibility of reei'ssive gene mutations have been studied in Drosophila meUmogaster. too. These positive findings 123,2-1,29..Tl) have shown that it might be possible to induee somatic gene mutation in mammals in vivo, too, an assumption that we wanted to verify by the mammalian spot test. The net hod -,-e i-r;| was described by Fahring (10 j. Female mice of the inbred C57BI..G-I1 inn (a/a: wild-type) were mated to the Han rotated-bred males of the T-stock strain (ala. non-agouti; b/h. brown;cchp/ cchp, chineliilla, and pinked eye diluturn: d.sv/dse, dilute and short ear; s.'s, piebald spotting). On the morning of th** 10th day after the appearanee of the vaginal plugs, the C571il./r>Jllan females were exposed to 12 000 ing/nt4 t-KJOO ppm) atmospheric concentration of VCM for 5 h. The VCM concentration in the inhalation chamber was determined by a type 5S-10-A Hewlett l'ackard digital gas chromatograph. The F, offspring were examined at 3--5 weeks post partum for the appearance of mosaic coat colours. To check the ability to induce coloured spots in F, hybrids of ottr parent mice, we examined the muta genic effects of cyclophosphamide (.Jenapharm, CHIt) (Cl1), too. This experi ment served as positive control. The statistical evaluation of the da'a was done by the chi-square test. * refill aUilwvi: Drp*runrnt if Itetuviuur ci^n. -.<- t.. K..jv. Cnivvmtv. 11-2131 u. I 4 (HunfUfv). J.it.fkw S. R&S 024002 mmyStTi'J :% . ft '.U:J\ %%{.' ...' /-.v^T , V^.i 174W0&& V^l'"K' J.' ' ',; Yr^j ' ^ i: *. .-4-:i,:lc* :>r7:mm - `-;;it.J^i i'fi ?' :l'ls<?$ suit VIVA!. AM) smr tNl'HIICNCI-: IX OIKSI'III.Vi OK C57 HI./CJIIjii >: T-STOCK KOI.LOWING tkkatmkxt or VC"M AM) cr OS T11K Kill! DAY OK I'KKG NANCY I ^ ooo men 1 .11 HUI I1ic/k|! ill 0 'll 11 lluljii<m. i*,,. U iihoul i; I ri !7,2 lit* jhj.u *J17 UH.K W nh ColiUtT %|>cil Whtl** Itrou nisli (ri t'rl d ('.) 0 3 *. no coofr^o S9y _3"5r 3r?* *< ^Orj>. O^r* # p fe 3 5-S 3*g lxfii^iinii'-,?isig^iiiH v0^>Cr0JO :-aiaaft :?., aS8 The data and the results of tlx- experiment are summarized in Table I. Alto gether. 210 K, offspring of-H treated parent-, were observed in the VCM-t-xpusure group. There wti.s no difference in the average litter size at birth oral the lime of observation 0--o weeks after birth. No offspring with abnormal mor phology could be found among the animals. Mid-ventral white spots occurred at a frequency of 2.Swhich does not -ignificantly differ from the relative control value (p > O.Oo). Treatment with 10 mg CT/kg did not induce a marked difference in the number of offspring in comparison with the control or the V C.M-tri aled group. There .were differences, however, in the number of white and coloured spots; both were significant. All coloured spots were brownish. The distribution of these spot;, was random among litters. No data on the mutagenic effect of VCM on genes, studied on mammals in vivo, have yet been published. Studies on sub-mammalian systems show that VCM can cause point mutation, especially base-pair substitution after mcabolic activation, in vitro 12-1,1 n..10| and in vivo |u|, although VCM itself does not produce such effect without the presence of liver microsomes |H|. There fore it was assumed that VCM would exert a mutagenic effect in the somatic coat-colour (spot) test. The dose used m our test was based on the results ob tained in previous acute experiments. This toxicity study showed that this dose was just tolerable, hut not yet toxic for the animals. The* observed survival rate of the offspring in our work confirmed this conclusion. This work provides no evidence that VCM induces somatic gene mutation. The experiment with Cl* showed that this lack of inducihility of emit-colour mosaic in K, could not have been due to some inherent deficiency in the'test system used. Nevertheless there is no evidence that either the application of fractionated doses during the 10th day (pel or single do-eisi of VCM, and/or continual exposure on the S--10th days would not have led to a positive result. However, there are no data about chemical agents, which, though they have produced somatic-colour mosaics on the Sth or 9th day. fail to do so on the 10th. The reason for this may be that there are only two types of real effect chemical agents have in the mammalian spot test; < 11 an agent can produce a maximal response after treatment on one day between the Sth anti 10th days <pe),e.g. ethyl methanesulphonate (EMS) 1111: (2) the other type of eff. ct shows equal response at any lime from the Sth to 10th days in the spot test system. This type of effect can be' seen with benzo[i;]nyrcne (H|o|l`i |0|. Naturally, a negative result may be regarded as the ,`ird type of effect, because of the negative result on the 10th day, VCM does not have any of the,real effects, and the results provide no evidence that VCM causes gem- mutation. Acknowledgement We thank Miss M. Michels for excellent technical assistance. References 1. AnUrk>m, J)., Hotljie 4nd I.K.H. Curi'h^w1, CI>*| nucr, Muijimn Kr|., 40 'i-Vf '-.HTo, chlond**; dominant 41 in iti4]e 2 iUrtun. A.* II* a. t I*. J4r<|Uiiion, C, Miljtcille, It. M<nt**wno *n<i II. 1-n`rr miCfi,Honii`-ineiit4tt><| formation of 4Jkt Huiptit v He*, f oniniun,, ? <1 **'!*} .Viti - fr*rn \mv| chlundr *nd wnvj limntid*1, J)io< firm, 3 ilartsrh, II,. ('. Malairilh* and K. Montesaim. Hiifn^n. rat and mouse hver mediated mutagenieitv of vmyl chloride in >\ i\phltnurium strain*. Ini. J. Ca/icer, 15 i]*#TT*> 42*1-437. I Ifartych, Jl., and It. Monies.inn, MutaiPunc and carcinogenic effect* of vmvi < htonde, Mutation lies,, 32(1975)9.1 II I. 3 llarttch, II.. I'rt'chfliW' >jluc <il muiagemcitv tests in the chemical ru/i ino^cn^M^, Mul^iion Kes.. 38 < I li7<;> | |7--| <10, fi OdVidKMi, U.K., and I),tuon, The in>luc(tn of vimatir muidUiu in nituv* embryos by brn/n|u}pyrenr, Arch. ToxirnL, 3K (1977) 99 --103. 7 IlarUch. Jl., and S'. Lnprjrnn, Vmil chloride example lor evaluating adverse hiologj.'al effects in yhort-term tests, Mutation he*.. -10 (1977) 200 -201. V Drovdouitc/, H./., and I'.C. Huang, Lack of mutagenicity of \md chlonde in two strains of AVuro* sporo croua. Mutation lie*,. 48 (Will) 4.1- r>0. D Ducatmun, A., K, llir&chhorn and l.L Srlikofl. Vmvl chloride )`\poiurr and human rtironutvom*' aber* rations. Mutation Hr*,, 3 | (1075) 163'li.K, 10 F)hng, It ,, A mammalian spot text: indue ton o( genetic alterations in pigmcm cells of mnuw embryos with X-raV*. and chemical rnul.iiyiis, Mol. Gen. OciKl.. I 3* < 1 975) .It)1* 31 J, 1 I Kihng, It., The mammalian spot test: a sensible in mo method for the detection of genetic alt**ra* turns in somatic cells of mire, m: A. Iloil.ondcr ami I-'. ls- .xerres < I d*.). Chemical Mutagens, Vol. 5. I'lrnum, Nt-w Vork. I978.pp, 151 -176, 12 Mrg, 1.1 and A.M. Thiesx, Kxiernal chromosome studies undertaken on person and animals with VP Illness, Mutation Ilex.. 53 (|*J?K) 1*7. 13 Fh-ig, I., and A.M. Thiexs, KMern.il ehfotnoxome imestig itions <>f persons with .iiioostroaiij due to VC exposure. Mutation lie*,.53 (1978) 187 -IK*. 14 Funci*Craviotn. F,. H, Lambert, J. I.insten. U. Khrenberg. A.T. Nauruan and S. Ostermaii'Golkar. Chromosome aberrations in workers exposed to vmvl chloride, l.ancet, i <1975) 439. 15 Clarm, A.J., J.IL Ciuttcnpian and I*. MiLv. Vinyl chlonde dependent mutagenesis: effects of hser rs tracts and free radicals. Mutation hr*., 3K (1976) M - kk. Id llansteen, 1.1,., 1,, HiUextad ami K, Thus-Fsenxen, Chromosome studies in workers exposed to vmvl chlonde, Mutation lies,, 3H (1976) I ) 2. -7 Kansteen, 1,1,,, 1,, lldlestad. K. ThnvKien*en and S.S. Fffect*, of un>l chloride in n.an. A cytogenetical follow*up study, Mutation lies., 51 (1978) 271 JTK. Id iiaitstren, 1.1,., Acute exposure to a cs togeiictic studv of two workers. Mutation lies.. 53 (1*J7K) Hid. 19 Ivanrtich, K.M., I, Debra Ketr and I. Aronson, 1'he m-t.iindic actuation of cmvl chloride .n sitro. Mutation lies.. 53 (11)78) 204. 20 Kuccrava, M., C) vitals six t( human rhninunm*s and its s alue fer the estimation *f genetic risk. Mutation he*.. 41 (l'J?d) 123-130. 21 Leonard, A., CL Decal and K.D, Leonard, ("vwnienoiu' inv<*sto:aiions <>n )vmph<cvt*`S fr*m wock**/* exposed to vinyl chloride. Mutation he*., 53 (ll*7M 21b. 22 Lopncim, N.t H. Harelc, S. ftaroncelh, C. hauer, f. ('.iiiiindim, C. t'ercntnaim and C. (`orst, Kvaluation nt the ttenetic effeets in<luce<l l>> smsl clilond** monomer umier mammalian uieiahohr activation' studies in sitro and in w\o. Mutation Ht s.. 40 (IbTd) K5--*o*. 23 Maitnusson, L,, Mutam-nic effects f vmvl chloride in /Jron,/dwi rn'.*uoo*;ii.r*r. Mutation lies.. 3K I 15. 24 NiaKiiussoii, J.. and (T. Kamel. Mutagenic effects of miisI chjtnde >n 1/ritntptuIti nuduooft'usr.T with ami without prelre.itmenl with sodium phenoh^ftiituraie. Mutation Kes., 57 tlhTK) 307--312. 25 MalavedJe, C,, (I, Itartsch, A, Itartun, A.M. (`amu* and It. Montesano. Mutagenicity of vmvl chloride, thloroethvl oxide, ehlorOacetahlehyUe, ami chloroethanoj, Hiochem. Huiphys. Ites, Cormnun,. 03 (lb*5) .hi3 370. 2<> Matiern, I.I.., anti W,l$. van tier /..ujii, Muta^emcits testmu of unne from wis*l chloruie (VCM) treated rats usinK tile Salmonella u *i svtum, Mutation Kes.. ! (l!`77) 230 t231, 27 Purchase, O.K.. Ph.lt. Hichardsioi and I). Anderson, Chromosomal and dominant lethal effects >f sins 1 t hlonde. Lancet, ii (1075) 4 10. 2H I'urehave. LK.IL. Ch,K, hichardson. D. Antlersou, t*.M. Paddle and \V.<J.K. Ailams, ('tuorn>s<tm^l analysis in vinyl chlonde exposed wmkers. Mutation Key., 57 (lb7K) 323--334. 20 haincl. C.. Mutaib'nicitv research and tetm in Sweden, %!utation he*,, 33 (Jh75) 7h -- H*\. 30 hatmui.. t;., A. Jnhatisnn, C. Kamel and C,A. Wachtmeistef, The mut i^rmcity of vinsl chloruie after metah-die actisaiiou. Amliio. 3 (li7 1) HM--lb7. 31 Shahtn, M.M., The nim-mutaiienimy and recornhinitfieniciiy of sins I chloride in the alrvence of metu* hohe actuation. Mutation Kes,. 10 (|(.I7*D 2f.*J--272. 12 S/entesi, L. K, Ilomvak, U, t'n^v'arv, . C/ei/el and 31. Timar, llixh fate of chromosomal aberrations in l*VC-wofWer<, Mutation lies., 37 (IH78) 313--316. 73 Verbor^i. F.i;., and L, Vogel. Vinvl chlonde mutagenesis in tinurtphth m Unujatirr. Mutation Ilex.. 48 |ti*77) 327-336.