Document xzV81QzqQy48mnNVYRQpReyDm

f. 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. Review of the Literature According to our knowledge, nine studies' 10 " relating to vinyl chloride (VC) 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 Department for Oct upational Medic me and Health Protection BASF Aktiengeseilst haft 6700 Ludwigshafen W Germany Corporate Medical Director Professor Dr Med A M Thiessi Paper presented at the Second international Conference on Environmental Mutagens lulv n-16 m Edinburgh medical department5 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 Germany 15 These values are 1945-1950 -- 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, m order to discard those subjects who have multiple ex posures We matched these sub|ects with healthy controls of the same sex (in this case male) and approximately the same age who Authon Fleig Thtess (1974) OuCdlfTian Htrscbbofn 3ehkoff U975i FuoesCravioto ei ai U975) Hansteen Hillestad Thus Evenson (1975) Kilian Picciano (1975) Lange Schwinger Veitman 119751 Purchase Richardson Anderson 11975) Fieig fl977) Leonard e1 fi 119771 Tabl* 1. -- A Survey of Roiults from VC ExpowO Persona. No of Probands 10 workers 11 workers 10 controls 7 workers 3 controls 39 workers 16 controls 203 workers 108 controls 12 workers length of Exposure (Veers) 4 26 4 28 9-29 -- Uo '0 30 3 25 21 56 workers 24 `onfrols 39 workers 20 onrrols 4 30 13 WOfkprs '0 "0n*rQiS 2 '7 % Chromosome Aberrations VC Exposed Controls (Average) 75 55 9 32 64 9 52 94 Mutagenic Effect Negative Positive Positive Pathological fteeuits (VC Syndrome) No pathological results --* 1 thrombocytopenia 341 39 3 l] 11 2 1 9 'PSO 23 79 Positive 5 Negative -- -- Negative Thrombocytopenia splenomegaly acroosteolysis Raynaud s syndrome sclerodermu 13 Positive -- 65 Positive Thrombocytopenia splenomegalv sderodermia Ravnaud s syndrome liver fibrosis esoohagus and stomach varices of the fundus 6 More severe aeroosteolyS'5 chromosome anomalies -- due to radiograph' Reprinted from Journal of Occupational Medicine August. 1976. Volume 20. No. 6 pp 367-561 :JOM 1976 SL 101717 ? 4-t Autnors Sansch MaMv*illa MontasafiO it975) RiprSijQ .J0hnnson Ram# WecMmeistar H974> k.opnano at il (1976) MapnuMpn Ramat (1975) Rurcflaaa Richardson Anoareon H975I rest - Mfltrioq ai TA 1530 TA 1535 <5 46 01 AMES-Test TA 1536 TA 1537 TA 1538 tl AMES-Tnt TA 1535 Of AMES-TmI TA 1536, TA 1537 TA 1538 i) AMES-Tst Sehiiosacctiaromyces pomde 0) AME5-Tt Saccharpmyces cerevtstaa Ci host-mechat?o-as$ay (Moutti MULLER'5-Mmod {Dfoaoofiiiai DOMINANT -lTHAL-T* iMouaai t.DQ4ura 2,300 20 000 nd 200 QOO oom 6 n and 48 h 200,000 ppm 30 60 90 Mina ai 3,000 ppm bl 3,QOO ppm Cl 700 mQ/kp oral net known 3 000, 10,000 and 30,QOO ppm MutapAbtC EffaCt ai positive b> nagativa ai positive bi fiagate** It positive 01 positive C) poaitive ooaittva nagatrve Aberration Type ________________________ Point mutations __________ Point mutations 1 Point mutations necaaaiv* wtfiai mutation# No dominant lethal mutation* not exposed to VC or -n inv otner known c hromosome damaging agent hut worked in the xame rat torv m dhterent departments ,eg, clerical 'tart trom the sales or technical department' The -ame questionnaire used tor obtaining data on experimental >ub|ects was also n-ed tor controls and the same criteria tor exi lusion from the studv was applied l.vmphocvte cultures were prepared from both groups and fmallv processed using the so-called micro-method ' One hundred metaphases per person were in vestigated making a total of 3 000 whu h were analyzed tor structural chromosome anomalies The frequency of aberrant Retg (1977) &ONE MARROW (H*msterl 2,500 5,000 pom positive Fig 1. -- A survey of the results using VC in various tests. Chromosome aOarTatton* metaphases totaled 3 1% excluding gaps and 7 5% including gaps The correspond ing values in the control group were 2,1% and 5 5%, respectively The individual datas Recording Point Fluid Mixer Fluid Mixer Exit Mixing-mill Mixing-mill directly above the nip Colander feeding point Colander take-off VC content m ppm Extruder jet Extruder feed-hopper 1-4 1-28 Remarks At headlevel in the position occupied by the operator At headlevei in the position occupied by the operator (after the mixing process At headlevel in the position occupied by the operator Position not occupied by personnel At headlevel in the position occupied by the operator At headlevel in the position occupied by the operator Only temporarily occupied by the operator Only temporarily occupied by the operator 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 ot ex posure. Within the framework of a re search program from the insurance associa tion ot the 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 than one symptom of VC illness The statis tical evaluation revealed a significant differ ence between the rate ot chromosome Fig 2 -- Recordings of VC concentrations in the atmosphere in the region of the KAE Technical Department 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%, Table 2. -- Chromosome Aberration Rite of Controls. respectively, in the control group (Fig 51, The control group was Amtyzad % Aberrant M*tapfia No. Ace Mataphaat* Ind. Gaps Exci Gap* 18 42 22 50 26 33 29 39 132 37 149 56 153 46 206 54 207 44 208 64 209 37 210 62 211 38 212 35 213 57 214 42 216 50 217 33 Z1S 52 219 50 100 100 100 100 100 100 100 100 100 100 100 100 too 100 100 100 100 100 100 100 6 3 5 3 3 -- 2 -- 2 -- 1 i 7 3 4 2 i 2 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(oma due to VC exposure According to information received Table 3. -- Chromosome Aberration Rate of Exnosed Reruns Showinc No Symptoms of VC lllnote. Duration of Exposure Andyred % Aban-ant Mataphaaaa No At* (Ypare) Mptapfwm Inct Gap* Exel, Gaps 5 51 11 100 1 6 19 7 100 8 14 57 20 100 10 T6 52 25 100 9 82 43 15 100 7 33 52 20 100 8 34 64 20 100 6 36 46 4 100 8 37 55 26 100 8 101 34 6 100 4 1 2 2 1 3 5 4 4 5 2 SL 101718 4 .. '0 - '6 40 Control nci gaps Control exc1 gaps Exposed met gaps 93 Exposed exci gaps Fig 6 -- Chromosome aberration rate of Chinese hamsters after inhalation of 2500 and 5000 ppm VC. i respectively 5000 ppm 10 -! Fig 7, -- Chromosome aberration rate of Chinese hamsters after being intraperitoneally injected with 300 mg/kg and 600 mgAg VC. SO Control met gaps Control exci gaps 10 3 Exposed met gaps Exposed exci gaps 46 6 Funes-Cravioto F IambertB Lmdsten I, et al Chromosome aberrations in worker1' exposed to vmvl chloride Lancer 1 459 1975 Nankeen IL Hillestad L and Thus-Evensen E Chromosome studies in workers exposed to vmvl chloride Fifth Annual Meeting, EEMS, Florence 19'5 8 Kilian Dl and Picciann Dl Cvtogenetic monitoring of vinyl chloride workmen Fourteenth Annual Som Cell Genetics Conference Houston Texas !9o 9 Lange CE Schwinger E and Veltman G Zur Fraqe der Chromo-'omenveranderungen bei Patienten mit Vmvlchlond-Krankheit Dtsch Ges 1 Ar beifsmed Muhchen 1975 10 Leonard A, Decat C Leonard ED pt al Cvtogenetm investigations on omohotvtes rrom workers exposed to vmvl i hlonde / 'oo >i fnv.roo HeJ'rh 2 1 1 35-1141 19?" iOO mg/kg--------- --------- 600 mg/kg -- -- 11 Lopneno N. Abbondandolo A, Baraie R. et al Evaluation of the genetic erfects bv vinyl chloride monomer (VCM) under mammmalian metabolic activation Studies in vitro and m vivo \4utat Res 40,85-96, 1976 U Magnusson | and Ramel C Mutagenic effects ot vtnyl chloride m drosophila melanogaster Vfutat Res 38 115 1976, 13 Purchase IFH, Richardson CR, and Anderson D Chromosomal and dominant lethal effects of vmvl chloride Lancet 2 410-411 1975 14 Rannug U lohannson A Ramel C and Wachtmeister CA The mutagemcitv of vmvl c hloride after metabolic activation Ambio 3:194-197 19-4 15 Thiess AM and Frentzel-Bevme R Retrospective survey of the alleged diseases assoc i.ited with vmvl chloride m the Federal Republic of Germany I Otujp Vferr 17 430-433, 1975 SL 10171Q Table 4 -- Chromosome Aberration Rate of Exposed Persons Showing Symptoms of VC Illness, Du retie' of Expotu M0 (Yearn 90 56 9! S4 119 33 153 37 160 32 '61 42 162 60 164 53 165 64 167 52 WO 34 171 44 172 52 ;,n 41 174 62 ' 76 41 'W 38 i S3 49 199 62 203 42 6 '1 5 6 15 30 19 22 26 4 9 5 5 71 16 11 10 12 8 Analyzed Metaphaaas 100 lOO 100 190 100 130 100 100 100 100 100 100 100 1G0 LQ0 100 100 100 100 100 % Aberrant Irtcl Gaos xci Gaps 10 6 .1 6 7 74 13 i0 5 11 6 94 11 4 20 9 93 11 5 83 83 22 10 14 5 18 8 5 85 11 5 mg DTIC (Dacarbazm), It is, therefore, not possible to say whether or not the results we have obtained are solelv due to the patient's exposure to VC or whether his chemotherapeutical treatment, together with the exposure rate ot VC affected our results1 Summary Chromosome analysis was undertaken on lymphocyte cultures taken from (1i six workers with estimated exposure to VC and tour workers with monitored exposure to VC (employed in the BASF;, '2! 20 workers showing symptoms ot VC illness with unknown exposure and one angiosarcoma case, due to VC ex posure mot employed in the BASFi, and 131 on bone marrow cells of Chinese hamsters after inhalation ot 2A00 ppm or 5,000 ppm. Table 6. -- Test Procedure Animal sperys An.mat s age Animal r Substance Test l Tpe ot aaTnnis!r3t.on intern M sommistranon Dosage Time until sacrificed after last administration Tatt 2 Type of administration Solvent interval of administration Quantity administered Dosage Time until sacrificed after last administration Cbmpse 'ams'ers 10 14 WPPHS 25 V : V -1. "f full 'nti3ia"on 74 "(Xrs 2 500 oom, 5 lavs 4 lours 5 000 ppm, 5 tdVS'4 hours 6 hours i o miection Olive oil 24 hours 5 x 0 5 ml 300 mg/kg nodv weight 600 mg/kg body weight 6 hours or after intraperitoneal mictions 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 m 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 1975 2. Bauchmger M and Schmid E1 Cytogenetische Veranderungen in weiben BluUellen nach Cyclophosphamldtherapie Z Krebsforsch 72:77-87, 1969 3 Ducatman A, Hirschhorn K and'"ehkoff l| Vinyl chloride exposure and human chromosome aberrations -fes 31,163-168. 1975 4 Fleig I Zur Mutagenifat von - Untersuchungen an Sauger und Mensch (Thesis), Heidelburg University. Federal Republic of Cermany. 1977 5 Fleig I and Thiess AM Chromosomen-Untersuchungen bei Vmylchlond-Exposition 4SP 9(12) 280-283, 1974 Rtf No 1 2 3 4 5 6 3 9 10 Tibia 5 -- Survay of Angiosarcoma! of tha Livar Dus to VC Exposure in tha Fadarel Rapubllc of Garmany Riant Occupation 4 PVC production autoclave? 4 PVC production autoclaver 12 VC application spray tiller 8 PVC production chemical worker 11 PVC 'esearch deot lab assistant 7 PVC production Shift foreman 9 Technical aopl autoclave leaner 4 PVC production 4 PVC production' processing 2 DVC production Dolvmewat'on 2 VC processing Data of Birth 06/04/30 07/26/31 07'l6/30 09/04/30 05/09/36 01/01/32 09/29/26 01/19/17 08/31/07 12/13/34 12 79/36 fTir--ri 01/25/69 12/14/71 10/10/73 11/25/74 12/16/74 01/10/75 11/13/75 12/25/75 10/20/78 Alive 03-77 Ace at Dlatneeb 38 39 43 44 38 43 49 58 -- 41 40 Parted of Eg--ire (Van) 12 Latancy Parted (Y*) 1/2 11 2 15 1/2 17 -- 59 12 1/2 21 1/2 -- 2Q-1/3 9-2/3 1 1/2 5 1/3 14 1/4 Alive 10 until 1971 5 1/2 st 11720 V 10 aia a> d c a 5- 75 5,5 Control incl gaps Control excl gaps Exposed mcl gaps Exposed excl gaps Fig 3. -- Chromosome aberration rate of exposed workers showing no symptoms of VC illness. from the Berufsgenossensc haft, 11 t ases from a total of 17 t ases or hemangioendothelial sarcoma, attributable to VC PVC exposure, were discovered in the Federal Republic or Germany Table 5' One of these 11 persons who is still alive, aged 43, was occupied in the PVC manufacturing area for 1 3 vears. The rate ot VC expo sure to which he personally was exposed during the last vears was between 31 to 110 ppm. He was occupied tor 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 tor testing An analysis ot 200 metaphases produced a rate ot 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 ot aberrant metaphases including gaps and isogaps was 16 f>% com pared to the control group, with a rate ot 1 1% 10 - CacC/a5T O x 5- 112 55 Control incl. gaps Control excl gaps Exposed incl gaps Exposed excl. gaps Fig 5, -- Chromosome aberration rate of exposed workers showing symptoms of VC illness. Rf Age EjpOSure Symptoms Nr in years 90 56 6 L<ver fibrosis oesophagus vances thrombocytopenia splenomegaly 9' 54 i i Liver fibrosis portal hyper tension thrombocytopenia splenomegaly oesophagus varices \ aberrant Tietapn Exchanges met gaps eici gaps 10 6 2 11 6 1 H9 33 7 Liver parenchyma damage thrombocytopenia splenomegaly 2- 15S 37 5 Sclerodarmia, liver fibrosis Rayrtauas syndrome acroosteoiysis 7 4 ' splenomegaly, thrombocytopenia 160 32 161 42 6 Liver parenchyma damage thrombocytopenia 15 Thrombocytopenia uver parenchyma damage 113 7 110 5 162 60 30 Liver parenchyma damage Raynauds syndrome thrombo cytopenia. oesophagus varices 11 5' 164 53 19 Liver fibrosis, thrombocytopenia 9 4 1 165 64 22 Uver fibrosis, thrombocytopenia, 11 splenomegaly 42 167 52 26 Liver parenchyma damage, thrombocytopenia 20 9 1 170 34 4 Liver parenchyma damage, 9 31 thrombocytopenia, splenomegaly 171 44 9 Liver fibroma. Raynauds syndrome, acrooateotysia 11 5 1 172 52 5 Liver parenchyma damage. Raynauds syndrome, thrombocytopenia 8 31 173 41 5 Liver parenchyma damage, splenomegaly 8 31 174 62 10 L/ver parenchyma damage, Raynauds syndrome 22 10 2 176 41 16 Uver fibrosis thrombocytopenia, 14 6 l oesophagus vances 177 36 11 Thrombocytopenia, hepatomegaly 18 82 163 49 10 Uver parenchyma damage, splenomegaly 7 5- 199 52 12 Liver parenchyma damage 8 51 203 42 6 Uver fibrosis, splenomegaly tl S2 Fig 4. -- VC symptoms and rate of chromosome aberration in the exposed probands Apart from our investigations on human beings, tests were also performed using bone marrow of Chinese hamsters. Two types of tests were undertaken1 (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 inactions of 300 rng/kg and 600 mg/kg, respectively, given in five infections, 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 4 Discussion The mutagenic effect of vinyl chloride mentioned in various sources ot literature, bactenallv 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 ot chromosome aberration could be found only in those persons showing symptoms of VC illness, as well as in an angiosarcoma case One verv 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 polvchemotherapv which was comprised of applications ot 150 mg Adriblastin, 120 mg Vincristin. 3,000 mg Endoxan. and 6.000 Si lOi ?3l !