Document gaMpNwMjr5GKYEJ8g5Dzzn5XV

Seminar in Occupational Medicine Mutagenicity of Vinyl Chloride External Chromosome Studies on Persons With and Without VC Illness, and on VC Exposed Animals I. Fleig, Dr. rer. nat., and A. M. Thiess, Dr. Med. /?-7f oooom R&S 107199 Review of the Literature According to our knowledge, nine studies1'0 " relating to vinyl chloride (VO problems have been undertaken. However, no conformity could be reached from these results. As Table 1 shows, five studies had a higher rate of chromosome aberration in the VC exposed group than in the control group. On the other hand, four further studies produced negative results. All those investigations made on animal and sub-mammalian species' are shown in Fig 1. Personal Investigations Investigations on persons concerned with the manufacture of |VC/PVC with estimated exposure have been undertaken in our From the Oefwtmem (or Occupational Medicine and Health Protection. BASF AktiengeseNschaft. 6700 Ludwigshafefl, W. Germany (Corporate Medical Director: Professor Dr. Med- A. M. Thiess), Paper presented at the Second International Conference on Environmental Mutagenv July 11-15, 1977. in Edinburgh. medical department.' In this group we cannot make any definite statements regarding the rate of exposure, but it is most probable that the assumed values taken from abroad are also applicable to the Federal Republic of Cermany." These values are: 194S-19S0 -- 1,000 ppm: 1950-1960 -- 400-500 ppm: 1960-1970 -- 200400 ppm: and 1971-1973 -- below 150 ppm. In addition, investigations were undertaken on persons in the VC/PVC processing plants who have undergone monitored VC exposure (Fig 2).' Both groups were combined as "Exposed per sons showing no symptoms of VC illness" (Table 3 and Fig 3). Before commencing the study, anamnestic data were collected regarding sex, age, occupational exposure(s). medical x-ray treat ment either for diagnostic or therapeutic purposes, recent viral diseases and vaccinations, consumption of drugs, and smoking habits, in order to discard those subjects who have multiple ex posures. We matched these subjects with healthy controls of the same sex (in this case male) and approximately the same age who Authors Flei*. Thiess (1974) OuGttmin. Hirsdihom Setihoff (1975) FtmnOhitfo et at (1975) Hansteen, Hillestad Thns-Evensoo (1975) Ktlian. Ptooaoo (1975) Lange Schwinger. Veftmjn (19751 Purchase. Richardson Anderson (1975) FUij (19771 Itwwd it il (1977) Tibi* 1. -- A Survey el Results tram VC Exposed Persons. No. of Proband* 10 Ortwt 11 worsen 10 ConfroH 7 ortvs 3 controls 39 worths 16 Confront 203 workers 108 controls 12 workers LMfte of Etoosw* Cfears) 4 26 m 929 -- Up to 30 32521 56 workers 24 controls 20 workers 20 Controls -- 4 30 18 workers 10 controls 2(7 % Chromosomo Aberrations VC Eipau4 Control* (A**r>|.) 73 55 Mutagen* Effect Negative 9 82 9.52 64 Positive 194 Positive Pathological Remits (VC Syndrome! No pattotogioJ results -- 1 tiwomourinoptin 3 41 39 -- 813 112 19 <esp 28 179 Positive 55 Negative -- NegOn* -- TtoomOocytoptmo. ipWwmfU, acroosteofysis. Raynaud's Syndrom* sderodermia 418 Positive -- 55 Positive Tlwombocytopenu splenomegaly sderodermia Raynaud's syndrome liver ftoovi. esophagus, and stomach var<es of the farita. acroosfeciysis 26 More severe -- tfeome some anomalies j-- due to redtograpn* Journal of Occupational Medicine/Vol. 20, No. 8/August 1978 557 R&S 107200 B..M..-M--f-t - (17S> hAaMHBM town i > ftMMl row fMnw 0tr4> Citrn IMKMW TAtUOlTA 11U.04I IA 31 TA1S37, TAISM HATUAEI1l>3T1M MAMCS-T** TTAA1tS13MI TAtur. M AMCVTnt 0 HBOfrStTtw<MwOirliAnTitf0m4SrSirArYti (MOUMf 00MMMir4CTHM*T*M (MnuiM BONEUAAAO* a*0*U* 2,000.20.000. iMMOOOOpfw AantfAOh 200,000 90. CO, >0 13,000 ppm b* 1.000 CKM. d TOQmgAio fl gf 9.n0d02001,000,000p0om s,ooo ppm bMig--c ENM alpppmu^ M mgMm dpopfcv* 0 AtglPW Ipbmm UPOMM PPWPH* MOBbH pOMUi UbiwlMA Type PvdnuMwt WM wiMtaiiowi m iipm umpiwi MO domininf Imm wwmtoaa bWNBM wet not exposed to VC or to any other known chromosome damaging agent but worked in the same factory in different departments (e.g., clerical staff from the sales or technical department). The same questionnaire used for obtaining data on experimental subjects was also used for controls, and the same criteria for exclu sion from the study was applied. Lymphocyte cultures were prepared from both groups and finally processed using the so-called "micro-method." One hundred metaphases per person were in vestigated, making a total of 3,000. which were analyzed for structural chromosome anomalies. The frequency of aberrant metaphases totaled 3.1% excluding gaps and 7.5% including gaps. The correspond fig 1. -- A ur**y ol th* rnullt using VC In virion tnti. ing values in the control group were 2.1% Recording Point Fluid Mixer Fluid Mixer Exit Mixing-mill Mixing-mill directly above the nip Colander feeding point Colander take-off VC content in ppm <1 <1 <1 12 <1 <1 Remarks At headlevei in the position occupied by the operator At headlevei In the position occupied by the operator (after the mixing process.) At headlevei in the position occupied by the operator Position not occupied by personnel At headlevei in the position occupied by the operator . At headlevei in the position occupied by the operator and 5.5%, respectively. The individual datas are depicted in Tables 2 and 3. There was no significant difference in the rate of chromosome aberrations compared with the control group (Fig 3). Investigations were also made on workers not employed in the BASF but with VC symptoms after an unknown rate f ex posure. Within the framework of a re search program from the insurance associa tion of (he chemical industry and the Verband Kunststofferzeugende Industrie (VKE). 20 workers all showing symptoms of VC ill ness (Fig 4) were studied. Many of the workers examined showed signs of more Extruder jet Extruder feed-hopper < 1-4 < 1-28 Only temporarily occupied by the operator Only temporarily occupied by the operator than one symptom of VC illness. The statis tical evaluation revealed a significant differ ence between the rate of chromosome f]| 2.__ Rneordinp of VC concwitratiom in th* almosphtr* in th* region ol th* KAE Tochnieil D*p*rtm*nt. aberrations in these persons and those in the control group. The frequency of aberrant metaphases in the exposed group was 5.2% excluding gaps and 11.2% including gaps, compared with 2.1% and 5.5%. T*bl* 2. -- Chratnnom* Aberration Rat* of Control*. respectively, in the control group (Fig 5). The control group was A*atyr4 to. *6 PflSftWl SSS Inel G*p Eiel G*pt 18 2 100 7 3 22 SO too 7 3 26 33 100 t -- 29 39 too 6 2 132 37 too 3 -- 149 S6 100 2 153 46 100 3 -- 70S 54 100 3 1 207 44 100 5 1 206 64 too 9 7 209 37 100 3 210 62 100 9 4 2M 38 100 2 212 36 100 5 1 213 57 too 2 1 214 42 too 9 4 216 60 100 7 2 217 33 too 2 216 52 too 3 2 219 60 too 7 2 the same one mentioned earlier. The individual datas are depicted in Tables 2 and 4. External investigations were done on a patient with an angiosar coma due to VC exposure. According to information received Tabl* 3. -- Chramosom* Atwmtion Rat* ol Eipowd Parson* Showing No Symplomi of VC llln***. Dviatw* f ElpOtw* An*hfV*d to. Ac* (Y*an) 5 51 6 39 14 57 76 52 82 43 83 52 84 54 86 46 7 55 101 34 11 7 20 25 15 20 20 4 26 6 too 100 100 100 100 100 too 100 too too % Ab*rr<*t H*E*ph*M* Ind. G*pt End. Cip* 71 2 10 2 93 3 85 64 84 15 42 558 Mutagenicity of Vinyl Chloride/Fleig and Thiess V 0( 10 " ea> K T3 c o 5s a ts g 2 it 7.5 5.5 Control ind. gaps Control exd. gaps Exposed ind. gaps Exposed exd. gaps Til 3. --ChramoMnw atMcratlM ral* f upsud workn ihowing no tymptomi of VC IIIAM*. from the Berufsgenossenschaft, 11 cases from a total of 57 cases of hemangioendothelial sarcoma, attributable to VC/PVC exposure, were discovered in the Federal Republic of Cermany (Table 5). One of these 11 persons who is still alive, aged 43, was occupied in the PVC manufacturing area for 13 years. The rate of VC expo sure to which he personally was exposed during the last years was between 35 to 150 ppm. He was occupied for 20 to 30 hours per month manually cleaning the autoclaves. In addition, he some times took samples from the decontamination plant to the labora tories for testing. An analysis of 200 metaphases produced a rate of aberrant cells of 7.3% excluding gaps and isogaps, a significant increase com pared with a control group which had a rate of 2.1%. The rate of aberrant metaphases including gaps and isogaps was 16.6% com pared to the control group, with a rate of 5 5%. 10- rn a. oX TJ Cts 73 .5 5- 41 <o Ja<Z. oj E 11.2 5.5 Control ind. gaps Control exd. gaps Exposed ind. gaps Exposed exd. gaps <u X) Fig 5. -- Chfomojomt aberration rat* ol cipoMd worktn ihowing symptoms of VC Illness. Ao* CxpOSur# Symptoms Nr, inym \MwmM macaph. EKM*g me0*pi id. gaps *0 5* 6 Ltwr (ItWDM. OHOOtueu wtM, ttirameocyMewn, aptraimpaty 10 6 2 l 54 11 Uver fibrosis, portal hyper tension, tftromeocytoptftu, otenomegaiy. oesophagus vahoes 11 6 1 119 33 7 Uvar parenchyma damage, thrombocytopenia, splenomegaly r '2 - 1S8 37 5 Sderodermta, liver fibrosis, Rayheeds syndrome. eeroosleotyw*. Splenomegaly, thrombocytopema 7 41 160 32 w 42 1*2 60 Uvar parenchyma damage, thrombocytopenia 15 Thrombocytopenia, fiver parenchyma damage 30 Uvwr pwancftynw Oanugw, R*yn*ud* tynUinm.. Mnwnbocytoswiia. MwxDugu* wm 13 10 11 71 51 5- 164 53 I6S 64 19 Uwr flam*. mnxnbocylopwK* 22 Uv*r Dbtsita. ttwwnbocytoCMniw, * 11 41 42 167 52 29 liver parenchyma damage* thrombocytopenia 20 0 t 170 34 171 44 4 liver parenchyma damage, thrombocytopenia, splenomegaly 9 9 liver fibrosis. Rsynaude Syndrome, scrooiteotyis 11 31 51 172 52 5 Uwr curandiywn 4*m*g*, R*/iuikU lyndrema, nuombocsMbWM 8 3t 173 41 6 liver parenchyma damage. Splenomegaly * 3t 174 62 10 Liver parenchyma damage* ftaynauda syndrome 22 10 2 176 41 16 Uver fibrosis, thrombocytopenia, oesophagus varices 14 61 177 39 1*3 49 193 52 203 42 11 Tbiwnbocysopwii*.h*pMgm*gi1y 10 l*"tr parenchyma damage, splenomegaly 12 Uver parenchyma damage 8 Uvar fibrosis, splenomegaly 1* 7 6 11 2 5- 51 52 Fig 4. -- VC symptoms and rata of chromosoma abanation In tha eiposcd probands. Apart from our investigations on human beings, tests were also performed using bone marrow of Chinese hamsters. Two types of tests were undertaken: (1) inhalation tests with dosages of 2.500 ppm and 5,000 ppm, respectively, for four hours a day, 24 hours apart over a five-day period, and (2) intraperitoneal injections of 300 mg/kg and 600 mg/kg, respectively, given in five injections. 24 hours apart, over a five-day period. Table 6 shows a survey of all important data concerning the arrangement of our animal in vestigations. Both tests (Figs 6 and 7) showed that the frequency of structural anomalies, inclusive and exclusive of gaps in the bone marrow cells, was significantly higher compared to the control group.1 Discussion The mutagenic effect of vinyl chloride mentioned in various sources of literature, bacterially tested, investigated on mammals, sub-mammals and human beings, has been confirmed by our in vestigations on mammals (2.500 ppm and 300 mg/kg) and man (5000 ppm and 600 mg/kg). The most important point to mention is that an increased rate of chromosome aberration could be found only in those persons showing symptoms of VC illness, as well as in an angiosarcoma case. One very important fact must, however, be taken into ac count. Nine months before blood samples were taken (February 1. 1977), the patient with the angiosarcoma was given polychemotherapy which was comprised of applications of 150 mg Adriblastin, 120 mg Vincristirs, 3,000 mg Endoxan, and 6.000 107201 Journal of Occupational Medicine/Vol. 20, No. 8/August 1978 559 Table 4. -- Chrorooiom# Aberration fat* of Eiposed Persons Shenint Symptom of VC Illness. Quntien t Eieowrt Anatynd % Ab*rrnt MhjpKmh fe. 44* (Tun) WMaykaM taeL G^t (id C*p 90 S6 SI 54 IIS 33 tsa 37 160 32 161 42 162 60 16* 53 165 64 167 52 170 34 171 4* 172 52 17] 41 174 62 176 41 177 38 181 43 ISS 52 203 42 6 11 7 5 6 15 30 IS 22 26 4 s 5 5 21 IS 11 10 12 t too 10 100 It 100 1 too 1 too 13 100 10 100 II too s too 11 too 20 too s too 11 100 1 100 1 too 22 too 14 too IS 100 7 100 a too It 6 6 2 4 7 5 5 4 4 S 3 5 3 I 10 6 8 5 5 5 mg OTIC (Dacarbazin). It is, therefore, not possible to say whether or not the results we have obtained are solely due to the patient's exposure to VC or whether his chemotherapeutical treatment, together with the exposure rate of VC. affected our results.1 Summary Chromosome analysis was undertaken on lymphocyte cultures taken from (1) six workers with estimated exposure to VC and four workers with monitored exposure to VC (employed in the BASF); (2) 20 workers showing symptoms of VC illness with unknown exposure and one angiosarcoma case, due to VC ex posure (not employed in the BASF); and, (3) on bone marrow cells of Chinese hamsters after inhalation of 2.500 ppm or 5,000 ppm. Table 6. -- Tost Procedure. Anna! n*ct*s AamwTs ag AimusTi wtitbt Substance Tot l Tim el adimuttratiM Mtrvjl d adfiwwitratton 0o|f Tim imI4 uoilratf titer Int adiniohlratao Tot 2 Tm ol adminalnlkn Solvent Interval ol adimnnUation Quantity adtwMlerod Dosafi Tim uad saodnd alur last administration Q*me fuinttert 1014 mm 2630 ( VM cWoridt WnUtion 24 hours 2300 ppm/5 dqrf/4 tan SjOOO ppm/5 diyt/4 horn 6 bow i.f. nvKtten 01m orf 24 Inn 5 I 05 ml 300 mlAy body (00 miA* body Mift* ( ton or after intraperitoneal injections of 300 mg/kg or 600 mg/kg. The proband groups showing symptoms of VC illness, the only living angiosarcoma case, and the animal test show a significant increase in the rate of aberrations in comparison to the control group. References 1. Bartsch H, Malaveille. and Montesano R: Human, rat and mouse liver mediated mutagenicity of vinyl chloride in S. typhimurium strains. Int I Cancer 15:429-437, 197S. 2. Bauchinger M and Schmid E: Cytogenetische Verahderungen in weiben Blutzellen nach Cyclaphosphamidtherapie. Z Krebslorich 72:77-87, 1969. 3. Oucatman A, Hirschhorn K, and Selikoff l|: Vinyl chloride exposure and human chromosome aberrations. MuUl Res 31:163-168. 1975. 4. Fleig I: Zur Mutagenitat von VC -- Untersochungen an Sa'uger und Mensch. (Thesis). Heidelburg University, Federal Republic of Cermany, 1977. 5. Fleig I and Thiess AM: Chromosornen-Untersuchungen bei Vinylchlorid-Exposition. ASP 9(121:280-283. 1974. R&S 107202 Tabla S. -- Survey el Ani'tourcemes el the Liver Due te VC Exposure in the Federal Republic of Germany. Rif. No. 1 2 3 4 5 6 7 8 9 10 11 Plant 4 4 12 4 11 7 1 4 4 2 2 Octvpatnn PVC produdKn auteWm PVC product** artodtw VC IppItOtKA spny fill* PVC {roducfon worker PVC tvjtvtft dept. Ml uvitHt PVC product** itoft loremin T*dimcl *ppl tutocU* cleaner PVC production PVC production/ proettwf PVC production polymerisation VC proccssmf Oat* *1 OirtN 06/04/30 07/26/31 07/16/30 09/04/30 05/09/36 01/01/32 09/29/26 01/19/17 08/31/07 12/13/34 12/29/36 OacnnMd 01/25/69 12/14/71 10/10/73 U/2S/74 12/16/74 01/10/75 l1/13/75 12/25/75 10/20/76 Alive 03/77 A<e at Dia(nuiU 38 M 43 44 38 43 49 58 -- 41 40 Ported f Cxpomr* (Y**o) 12 Latency Petted (Tun) 1/2 11 2 15 1/2 17 59 12 1/2 211/2 201/3 92/3 1-1/2 5-1/3 141/4 Alwt 10 Hid 1971 5-1/2 560 Mutagenicity of Vinyl Chloride/Fleig and Thiess R&S 107203 l 10- Oasi UK 7.6 ii-W- gH Ita Control ind, gaps ,0<JJ*lxj,i Fl( 6. -- Chroimnoma aberration rate of Chlnevs hamstsri after Inhalation of 2500 and 5000 ppm VC, t, respectively. E s - 2500 ppm- -5000 ppm - 10- Fic 7. -- Chromosome aberration rate of Chinese hamitera after being Intraperitonoally injected with 300 mgAg and 600 mgAg VC. 80 40 Control kid. papa Control exd. caps 10.3 Exposed ind. gaps Exposed exd. gaps 6. Funes-Cravioto F. Lambert B, lindsten |, et al' Chromosome aberrations ir workers exposed to vinyl chloride Lancet 1:459, 1975. 7. Hansteen IL. Hillestad L, and Thiis-Evensen E: Chromosome studies in workers exposed to vinyl chloride. Fifth Annual Meeting, EM5, Florence. 1975. 8. Kilian Dl and Picciano 01: Cytogenetic monitoring of vinyl chloride workmen. Fourteenth Annual Som Cell Genetics Conference, Houston. Texas, 1975. 9. Lange CE. Schwinger E, and Veltman G: Zur Frage der Chromosomenveranderungen bei Patienten mit Vinylchlond-ICrankheit. Dtsch. Ges. f. Arbeitsmed.. Munchen 1975. 10. Leonard A. Decat C. Leonard ED. et al: Cytogenetic investigations on lymphocytes from workers exposed to vinyl chloride / Toxicol Environ Health 2 1135-1141. 1977. 300 mg/kg --------600 mg/Kg-------- 1 11. Loprieno N, Abbondandolo A, Barale R. et al: Evaluation of the genetic effects by vinyl chloride monomer (VCMI under mammmalian metabolic activation: Studies in vitro and in vivo. Mutat Res 40:85-96. 1976. 12. Magnusson I and Ramel C: Mutagenic effects of vinyl chloride in drosophila melanogaster. Mutat Res 38:115. 1976. 13. Purchase IFH. Richardson CR. and Anderson D: Chromosomal and dominant lethal effects of vinyl chloride. Lancet 2:410-411, 1975. 14. Rannug U. lohannson A. Ramei C. and Wachtmeister CA: The mutagenicity of vinyl chloride after metabolic activation. Ambio 3:194-197, 1974, 15. Thiess AM and Frentiel-Beyme R: Retrospective survey of the alleged diseases associated with vinyl chloride in the Federal Republic of Cermany. I Occup Med 17:430-432. 1975 Journal of Occupational Medicme/Vol. 20, No. 8/August 1978 561