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ovu, J. \C iU'i1 prostate ulands. Js." The hiwj iiiate on IIrjul h \Jn m fmwi, \\M S'lninna. and Si a .^tceNMiry iimiiill (rsv." mi, C. < t l`>" 1 ' .11 \t by , of dirldllM *ii :r<- On* 2 1 ENviiiONMEsi a hkseahcii 7, 387-405 (H)74) 4 Carcinogenicity Bioassays of Vinyl Chloride I. Research Plan and Early Results1 Os,\M. Mai.tom anji Li-haiim- h.tituto di Oncolofiui `7'\ Mlthini" and C<nlm Tamaii, liulu^tta, Itahi }U (aired Man It Jl, 11)74 tin sc him stin.ilions .nr lonceilift! witli tlclci iiiininn the oncom-nic potentialities 14 of viinl chloinle (VI') m experimental animals. I lir elhit of this tnmpnimd is In-inn studied in M'lation In cartons experimental fin tins, Midi ns imiti' of ndiiiiiiistiJhon, dost- fusel, lenidh of licatment, species, strniii, nnd ujje of flic animals. Pleltmin.ny lesiifls Hie pirsellled. Wlleil ejseii hv inhalation, Vti indntr.s III rats, Zvinhal jjLiik.! e.ucommas, uepliiohlaslomas, and aiigiosaieoiiias of tlie liver ami of otlier aii.itoinie.il sites, ami in mice, piiliiioiiai s adenounis, mammary eareiooimis and liv'ei aimiosaieoiii.is. i 4 A dlteel ielidioiislup lias fieen ioond between dose and length of tiealment and i neoplastic lesponse. i Vinyl chloride (VC) (CU. -- (111(11) is a chemical t omp.uuul of im icttsiti^ ] i iudu.sUial importance. If is used in flic piothielion ol pnlyxinyl chlonde tc.sin, 4 as a copolymer in Saran ami other plastics, as a chemical infermcdiiitc, as a .solvent, and as a propellent, in the past, lor a short period, it was also used as anaesthetic. It is at present ptodueed at a rate ol near 12 million tons per xtwr. The followun' population mmips may he exposed to VC: (1) wmkux cm jinked in the pmductinn ol pulxxinx! elilmitle i PYC t .mil of V( ! and in tin- use ol VC lot ollui mtltt.slt ittl pmp.ises; (2; workers mamilacfm mu, l'VC, (di residents about factories producinu PY(I and \ (!, and Id) to a imic'i lesser t extent, people uutleiy.oiup eontael with tcxiiis uiatle with VC, with materials containing these resins or consumer piotliiels eont.i nin^ \ (as some piupell.mt t spt ax s. The picxcut leseateh ineliltles a senes ol touel.ited and iiileetaleil experiment . dealing with carcinn.ncmcity hiuassays ol this monomer. This leseateh billowed the prehmiii.uy tesults ol tin pumeeiimj, wotk ol lJ. H. V uila, presented at the X International Cancer C'iiil'icss (Houston, 1970) and publislied in 1971 ( \ tola et ui, 1971 ). Viola's results indicated that exposme to .>(),()()() ppm of VC, 4 hoots a clay, s o times a week ior 10 months, produced tumours in rats, and that these occurred , in skin, lunif.s anti hones. 1 -i Immediately altei the it pint in Houston, we communicated with Professor Viola, who kintllx pul at out disposal the m.muscnpl ol his exlensive tepoit i then in press, ami sexetal slides ol the tumours. \ On the basis of his results and mateiial, we teaehed (he following conclusions. i 'This project hits hrm siippnitccl In- the l.iimpcan (aiopciative Crimp (fX'.C) xvhicli i includes Montedison (Italy l, ICl ( C. k.), Sohay (Heljmnn), Rhone~Proyril (France). :1st faery rijfht (ft 1971 ttv Academic Press, Inc. All rights of reproduction m nox- form re vised. F-.r ( F | t I I I I t t 388 mu.ioni and i.i.ii.mim: (1) Under Professor Viola's experimental conditions, VC definitely showed a carcinogenic effect on rats; (2 ) 30.000 ppm of VC was an extremely high level of exposure; (3) The tumours dcsciibcd as cutaneous arose from Zymbal glands, which in rats aie responsive to a large' spectrum of carcinogens: c.g., 2-aeetylamiiio-fliioicne (Wilson ct til, 19-11), ben/idine (Spitz ct til. 1950), 3,2-dimethy]-4-aminol>iphvnyl (Walpole ct til, 1952), 4-aminostilbenes (Ifaddow ct til. 1948), 9,]()-diinothyl-1,2-1>on/anthraccno (Ceyer ct til, 1953), 3-mctIioxy-4-ammoazobcii/cnc (Miller and Miller, 1901), median (Tnimcillmimi Ct til, 1902), anil otheix. (4) The pulmonary tumours were most likely metastases from Zymbal gland carcinomas. These lesions were in fact morphologically similar to the Zymbal gland carcinomas, and all were observed in animals bearing Zymbal gland tumours. It appeared useful to further investigate the nature and extent of the carcino genic effect of VC, with particular emphasis on expeiimeutal factors related to exposure, such as route, concentration, duration, continuity or iutcrmittcucc, etc., which could give more direct information concerning the iixk of occupational and environmental exposure, and other factors related to the animals studied, such as species, strain, sex, and age. For these reasons, we were contacted bv Professor iiaitalini, Director of the Health .Services of Montedison who offered the support of his Companv for an experimental stndv of the biological effect of VC. Montedison was soon joined by SuIv.lv and Rhone-I'rogil. M.\ I l-.HI.U.S AM) Ml.TIIOPS The construction of an appai.ilns fm piogrammed and cuntinllcd exposure took several months. The eh,uubers uric designed to deliver coneeutraliom varying from 30.(KM) ppm to 50 ppm and to simultaneously treat nearly 1200 expei imeutal indents, Ihistcall}, it is built of si.unless sin I mu! gbiss. The control i > f VC couei III I .il ii ms IS b\ g.e. c III i on 11 oft .1 pli V VC lias been supplied bv Montedison txavh hatch has been anaiv/ed in the chemical research luboratorv nI Montedisoii before use to assess its degree of purity (99.99*). Tin: expeiimetils largely ulib/ed S])iagtic-Davv lev lats, Wistai iats, Swiss mice, and hamsters have also been used. All (lie animals, except the hamsters, have been Ined in mu Institute foi ve.us and, vvb.ilevei tlieii use, are all examined at death bv complete antopsv, giving us extensive information con cerning their current p.iflmlogv. The animals have been weaned and classified bv sex when 4-5 weeks old, at which time they have hern uumhered hy ear punch, and divided into groups !>y litter distribution hrom weaning, the animals have heon fed, titl libitum, ail adequate commercial diet. During the period of treatment, the animals are housed in groups ol 10 in stainless steel wiie cages with a solid bottom ol the same metal. After the period of experimental treatment, the animals are kept in groups ol > in inakrolon cages, with tops made ol stainless steel wire. A shallow kept in . Fourti at the o' results. I.xjicrinu Kxperi 10,000. f for 12 u as with at 25(H) luavimm /'. \ pi i im Kxpei i till- ellri I lie s.iiiii still s!:ov t'.xpcmn This V' const,mt /-. xpcnnii K\pe: in .ninth luglii : u I, xpiriiit, I las \ al im isjili I. xprinn Fvpei i /. ipi run, Tin's r as in Fv /. xprnuh Fxperi species, Expcrinu This vv effect at UCC 107042 I visvi. c.iuniimr < aik inooi m sis :W) v showed (Is. which , 2-acctylti, 1950), ostilbencs ver et ul., 1901), ii Zymbal lly siinilar n animals ic carcino* related to tcnee, etc., eupational ts studied, ntacted by ho oflered peal effect d exposure icentratiuns learJv 1200 i'lie coiiti'ol .zed m the degree of rats, Swiss ,e hamsters, ise, are all ination com urC'ls old, into groups libitum, an animals are atom of the als are kept eel wire, A shallow layer of white wood shavings served as bedding. The animals were * kept in a teinpeiuture eontiolled laboratory at 19-20C. ci Fourteen experiments have been started in sequence. Some, were programmed * 1 at the onset. 'I'lie need lor others appeared on the basis of initial experimental * results. The plan ol tin- espeiintents is shown in table J, 5 Experiment 1 Fxpciimenf I was to investigate the elleet of the atmospheric exposure to 10,000, 6000, 2300, 5(X>, 250, and 50 ppm of VC, 4 hours daily, 5 days weekly, for 12 months. 1 mi gioups of animals were added as controls: one untreated, as with all the following experiments, and one treated with vinyl acetate (VA) at 25(X) ppm, under the same- conditions, This dose of VA appeared to he the maximum possible dose1 lor a chronic exposure. Experiment 2 F.xpeiimeut 2 was pcilormed to obtain data in a larger number of animals on the elleet of doses between 250 and 50 ppm, i.e., 200, 150, and 100 ppm, under the same conditions as in K.xpt 1. It was begun after it became clear that 250 ppm still showed oncogenic effects. Experiment 'i This was planned to study the efFccts of a shorte r period of exposure, keeping const,ml the other conditions of I'lspt 1. s Experiment 4 Fspeiimcnt 1 studies the effect ol the same type of exposure as in Fxpt I, in another spei ics, ie, uuee. I he tleuluiiiil lasted 7 months in Clew ol the lughei molt,iht\. and the shorter life spun ol the mice of our hue. Fxprmnrnl 5 Tins was nuclei taken as a pilot investigation on (lie possible' efleets ol the * V atmospheiic e xposure during pregnancy, on offspring. # E\i>erimenl t> ** p.xpci ime nt 0 repioduced the conditions ol Viola's experiments. \ $i Experiment 7 i This experiment was designed to study the c'lleels ol I he- same type ol oxposme as in Fxpt l in another strain of rats, i.e., Wistar. I 1 Experiment H Fxperimeut 8 studies the effects o| the same .............. as in Fxpt I m a third I species, i.e., hamsters, for the same pci md as in Kxpt l. i4 Experiment 9 i 'Phis was planned to blither assess whether or not inhalation of VC lias any elleet at a dose' level of 50 ppm m experimental animals. i j 390 M A I . I O M A N 1> L E K M M IN E UCC 107044 TAHLK 1 Pl-TS OK TUI K XI'Mi l UK NTs TO MaHCII 11, 1674 No. of exjK.jri- ------------- mem limit? Treatment 1 K IM'.' ilf \ Ixnigl h HT1 HTd HTi BTti Inhala tion [nhalaI 1L1T1 Inhala tion InhaLiI ion Trait^pla cet! t;il Inh&Uttoll Inhala tion 10.0110, f> *htt 25tKI. 5i*> 250, oJi ppm I'ntrealed controlTreateil tunings: VA 2500 ppm 200, 1 10. 100 ppm l "iilreate i cinlrul10,000, ft "til. J-|i M). VlM. 2 il), .11 ppm Tntrcatcil cmitrmHI.OOll, ft *KI, 2KK) ,5>M 2,111, .It* ppin I'lomUed cimirtiU ltl,01)0, ft X) ppm 30.000 ppm 10.000, ftnOO, 2100. .ic1. 210, .10 ppm k'nt reined (mihp'Is 4 h utrs dalv, 5 dayweekly, 52 week** 4 1. urs di.ly, 5 da\" *eekl\ t Vi wi*ek^ 4 h mis tliii ]y% 5 day*. * ecklv r 17 weeks 4 h >urs dad\, 5 days weckh, 50 weeks 4 h mr' d':Y, 7 iyv, yn-4' t \ 4 h mrs dally, 5 da\s weekly, 52 weeks 4 h >urs daily, V days weekly, Vi weeks Ai. fijX'l'il'S St r :iin A piiVlVt.'. 1 Km Sprague- 1 -1 1 );iwk'V Kenpth No. nf ex- JKTITo- Ker monk t.-il pimip wee]. 2(> >011 577 01 00 l.il Kat Sprague 11 Hawley Kat Sprague- 21 Hawley Mouse Sis iss n 2MJ 265 545 120 HI 31 202 2 vs 110 <>U KNt 60 2.10 200 all) 00 -11(1 55 11,Hi Sprague- 10 Bteeiers 110 lift 140 30-14 00 Dnwley 12 days Kinbryu^ Kat Sprague - ' " lid :io (ill 60 55 1) a tv le v Kat Wiatar n -- 220 220 30- 40 31 BT* InhaLitl )|! Bni BTill Inlmhtimi Inh.dntimi 10,006, 60Xh 2500. 500 250, 511 rprn \ ntreated ooitrul* >o ppm U { ntre Oi*d rmilruK !r1 ! Oy h i, 0u< *o ppm 1 novated iinun'b 4 heirs daili. 1 days weekly, ,'io aivks 4 h( 01-= (lull;. , 1 days weekly, 12 weeks 1 Ik 1 or - ilit 11 \, 1 davs weekly, 1 week-: 4 hi'tir- daily. 1 day weekly, 21 weeks; 1 lio'ir dailt. 4 d.u s Ham ster (loMen 11 Kat Kat Sprain:' 1 hm lev Sj mpite Hawley H 1i 20's 208 32-71) 22 200 21X) 400 100 (cl noil if > 420 421) M0 120 i 7 liTT 4 II HI Inhnlatkm lit,000, 0000, 2500, .-,<10, -oil, 50 ppm Untreated controls t-i.-K._t , <lw M l K4 hours daily, 5 days weekly, 52 weeks Hat W i-tar ]I 220 220 50 10 35 . * E.M. i 'S1 .`--W 411 t- * : ...Ti- I " * HP. Illlltdu1 ll III HP.) HTlt) Inhala tion In halntinii 10,000, (iOOil. 25IHI. 2,011, 250, 2,0 ppm l litre.Med omtrid50 ppm (/) Uni rented emitroL- irt bUKhi. 6(MKl ppm I'mie.itod com rob UTI1 BT12 BTi i HT14 1nt;e>- Hub mg; 3,32 nig, Ml 1 ion mg kg lately weight in olive ml ( ikilln*[' olive oil f ,ndi- 4.25 n.c m 1,0 re olive oil pei ili>- 'Ej 111 1- 1,11 c r tdive oil ia-al iitje*- t5i HI > ilinitaneiiii' 4,2'j mg in 1,11 rv olive oil (Ymio!- 1 rt> (r olive oil inje. - turn InhMat in Hl.'tOO, 00!M) ppm 4 hours daih, 2i davs weekly, 30 weeks 4 hours daily, 5 dayweekly, 52 weeks 4 hours daily, 5 ilavweek!., 5 weeks: 1 hours daily, 1 dav weekly, 25 weeks. 1 hour daily, 4 davs v. eekh , 22> weeks 5 times weekly, 52 weeks 4, 3t 2. limes by two month', ami onee 1 injection 4 honn dailv, 5 davs weeklv. 5 weeks Ham i hildeii ster 11 lint Spi ague- n D-.wley lint Sprague- 11 1 lawle' liat Sprague 13 1 >awlt*v Hat Sprague- 13 I *awiev Ji a E Sprague- 21 Daw lev Ja t Sprague- 1 day Dawlev - .4... . t- **``1uattk 4i ''*. !i'tfci 2CS 32-70 200 200 400 l'O tc) 3j*1 It i 420 420 MO 120 ] (Vi \m ,T>i> >n 150 ]5fl 300 CO SO 70 150 7' 45 44 S2 43-40 ' ,,>> yi**. r n ro >n Z X T z cr <7. Ccco ^^^ "i ' ** <4 . -'j- II 392 MAI.TOM AM) I.KH'.MINT. Experiment 10 Expeliiiiciil 10 studies (lie effect ol a Ini'll, but short iiiid/intennittent atinospuciic exposme. Experiment 11 This was performed to evaluate possible effects of VC by ingestion, Doses have been chosen on the basis of early data on the release of VC from food containers made of PVC, and following suggestions of EEC. Experiment 12 This experiment studies a third geneial route of exposure namely the peritoneal cavity. Experiment 13 Experiment 13 was started to assess whether the VC acts on tissues directly or through metabolites. Experiment 14 This experiment has been included to study the responsiveness of newborn animals. The animals are examined weekly, and weighed every 2 weeks during the period of treatment, and monthly after the treatment is completed. All de tectable gross pathological changes are recorded during examination. All ani mals aic kept under observation until spontaneous death. Moribund animals are isolated, in order to avoid cannibalism. A complete autopsy is made of esery animal. Histological specimens include Zynibal glands, interseapnlar brown tat. salivary glands, tongue, lungs, liver, kiduess. spleen, stomach, different segments of the intestine, bladder, brain, bones of the legs and feet, and any other organ with pathological lesions. The tissues are fixed in alcohol, processed, sectioned, and routinely stained by hema toxylin and eoxin and with Van Ciesou's method, in selected cases, other stain ing techniques have also been used. All animals exposed to the highest doses (30,(XK) and 10,000 ppm) with or without tumours, are examined radiologieally, during treatment and/or at death. Moreover, radiological examinations are made of all animals bearing tumours, even il exposed to the lower doses. HKSULTS Preliminary results and other current data nt Expts 11T1, BT3, and BT4 are gisen in Tables 2--1. Special'aspects of Expt BT5 and BT6 are discussed in the text. Insofar as the other experiments are concerned, results aie awaited. In 1 able 2 are listed the observations on the nncngcuic effect of graded concen trations of VC within a range of 50--10,000 ppm. ft is evident that tfic overall occurrence of tumours decreases with decreasing concentrations from 39' tumour vi 1\\warn Animals l>uwiev rul*) (troop* iiiid tt'eallnehl Survi Total vor* T v'a `..',700 pjHD n VC 10,01(0 ppm 111 VC 0000 ppm IV VC 'Ja(M) ppm \' V( * r>00 ppm VI VC 250 ppm VII VC all ppm VIII No (rentmenl on 00 *>) 74 to 07 04 tl.S __ | .1 1 Total .`<77 a d Mola>litses lo lung, h Melius time* to liver and, or U Metastasis lo ,J Angiosarcoma in Mitfcuianeoi f IVo intrn-nhd ommai annioMi val miiKi of ihm I., Out* j M f 1111 < 11111IV lUjJ^ifivfll 'i i|n; J I )l H' Hill H-fll H Jof HIH.U * J VVo gland adenoma jnlfmh<,tf11<jmiit of ovurv. 1 Sfhurwms gland eamnomu o' * Zymtial gland adenoma., 1 Mmiiiml deviation hepatoma 1 Out* Zvudml gland adenoma, Join! No. o| iuiiktHir*, % incidence at K),(X)() ppm. 29 %1 ppm, i c.spectiu lv. Fifty (5 I liimted number of animals s controls and in rats exposed \ blastemas and angiosareom tumours. 'I he development * on the concentration of the of exposure. If exposure tim 1 ( Fable 3) the number of ti 4 mittcnt atmo- .estion. Doses 'C from food 4 1 Clrmip> niiti titmitJK'Mi VINYL CHLOIUDK C.AHCINOCKNKS1S 393 'I'AHI.K 2 I'Xi'i iiimi. nt UTI, I{I'.mci.ts M-n.n i::i Wi'.i i,h AhiimmU (Sprague l MiwU-v 1 tlln ) 1 ul ill Stu vtVul s Animals will) 1 uiiHHirs Zymlml k1;uh1h Nephrublu*- cur run iinn.V` Ionia*'' No. No, Aiij'iohjirnifiiji* Ollier Ivpo -- --------- --------unti/or Tu- biyer* OUu*r*iie* Min ini Nu, N11. No. No,"1 namely the ics directly or s of newborn as during the eted, All deition. All anibund animals imens include , lungs, liver, ladder, brain, 1 lesions. The ned by herna* s, other stain* ipm) with or d/or at death, .ring tumours, and BT4 are Insofar as the graded concen* lat the overall no 39* tumour VA J.iOO ppm ft VC 10,000 ppm HI VC 01 MSI ppm IV VC 2">00 ppm V Vt ` .r>00 ppm VI VC goO ppm VII VC fit) pprn VIII No treatment UG G'l 4 '1 74 t'u 1.7 04 i`>S Toiul >77 1 .`I 1 .7 ip ,r> a g:t - G-- 5* 27 a 11 \d ] 21 n * 0 21 :! 7 U/ / 2 21. -- -- 1* 10 Ol 11 -- -- :w> s -10 00 ' iMclnAlir-r.* In lun^ b to Jivt-r tilt 11, of to |tuK juul lo ItlUtf. J AnniDMvrnwjm in Mibruinunoiis fibrosing nntfhMi.T '1 Wu iut ru'iibilnmiiiMl /mj/io-'Mi tuniiH { I mnI i^umi-> 11 spItM-n itml 1 I u uvut y ), 1 ukmI'Vtu^ MiKio* Mu toma of neck. ) Out* ptilmonary ati^itHurcoina; 1 uiiKioMimimu of iilcms. " (>m.` in t la-alaloiniim! an^iai rmnu (near U. -pliMMi); I in 1 rat burarir o>~if vintf illicit ism mum J A Two Zyiiibul iciiitk< 1 uflriitttioiM, 1 ittkiirilriittnoinit of I)h* <*nr; ) iiiiiiiiiiitirv niiTinomn; 1 rvHln- (ulfiHM iii nimma <*i uviirv, 1 Srhucooiin ulaioi nirnmmm of -kin. ' Zunbnl yinm! mitninmn. L Minimal ilrviafion hrpnluiiitt * t )rif Z wnbn! jflimd a> ion* on a I sail vui y ^iam I i at <m< non. " Tula! No, of tuniuui>. incidence at 10,(K)0 ppm. 2OT at 6(XX) and 2300 ppm to 2.\% and 16" at 500 and 250 ppm, respectively. Fifty (50) ppm did not cause tumour development in the limited number of animals so exposed, and no neoplasms weie found in untreated controls and in rats exposed to VA 2500 ppm. Zvmbal gland carcinomas, nephro blastomas and angiosarcomas of the liver anti oilier sites were the pievailiug tumours. The development of the hitter 2 neoplasms was not always dependent 1 on the concentration of the toxic agent, but to a higher degree on the duration * of exposure. If exposure time was reduced from 52 weeks (Table 2) to 17 weeks '"S (Table 3) the number of tumours after tit) weeks is markedly smaller: their per- '' 4 A 4 VI *s -i I'AI'I 1; 1 V An ! 1' T l ei l oil1 (croups fuel treittineni T..- V i-it i VC 10,0011 ppm n Vi; t'lUOIl ppm III V(` ~-"00 ppm IV V( * "WM> ppni V VC LTill P1>[I1 M V( ' oO ppm VII No 11 enl meti f ;;o ;to GO :;p GO :>' GO ;j' t'l 1 :n GO ;:o ;iu GO SP 70 l.`>p Tui :il g50 510 b Ti.lal No. of 1 ll[lloiUN t*.-i it it J4L is 5V at VC 10,000 ppm am! 1.7'* at 6000 ppm; at this time no tumours wore found alter administration of the lower eoneentiation.s. It is also worthy of note that all malignancies in this series were Zymbal ''land carcinomas. After .35 weeks of exposure to 30,000 ppm (Expt BTO) 2 Zymbal gland carcinomas have been observed among 60 i.its (53 survivors). In Expt UT5, two groups of 30 mature breeder rats were exposed to VC 10,000 ppm and 6000 ppm for 7 da\s during pregnane)'. No tumours were obseived uttei Oil weeks among the 23 survivors ot each group. Angiosarcomas of the .subcutaneous tissue oc curred in theii offspring; one tumour m one ol (lie 54 rats ol tin- 10,000 ppm group at tin- age of 21 weeks, the other tumoi in one of the .36 animals of the 6000 ppm group was found at 22 weeks of age. fifty ami 30 rats, respectively, of the litters are still alive and under observation Table 4 indicates that mice lespond to VC inhalation ol 10,000-50 ppm in a similar manner as rats. While 50 ppm is again ineffective alter .35 weeks 30V, 15V, and S.4V neoplasms were found after 10,000-6000, 2500-500, and 250 ppm, respectively- It is peihaps not surprising that in mice, pulmonary and mammary tumors were more frequent than other localizations, but the presence of angiosarcomas of the liver also oecimcd in this genus. At present, 5 distinct types of tumours induced by VC have been observed; i.e., angiosarcomas, particulaily of the liver (in rats and mice), nephroblastomas (m rats ), Zymbal gland i,niinii:ius (in i.ils ) and piilmnnai v adenomas, soiik; of '"i " III IfMIfllo u One "kin M|U:iinnn*llitlnr ruM'ir *" One >kin M|iiij.mufeiiti|;u etirett `'Total N<. of hmiuis which undergo malgnant ti Macroscopic and microsco in Figs. 1-25. The frequent Zymbal glam are somelimcs bilnbated am can show different patterns (Fig. 2) appears ulten in a present various different hi (fig. 4*), squamous, anapl metastases earn the eharact. blaslomas (f igs, fi-3) lead I but can also involve spleen the liver in rats (Figs. 11*, growth and licmorrliagic ; metastases ( Fig. 15 ). Occa; blastoma oeeuned in the sa. appeal genei.dh as mole i 1 Figures iiiurki-il l.v .isteosks VI\Y|, CIN.OHIDF ("'AHCIXOC M SIK m Itlier type jind/or si t c Total No. No* <Ironp.- and trontuimt TAl'1,1: j m'ti a w 11k - AnminU (Ms i. i T ) I iraic. >Hrvivnrs To Tn- tal i ill \mhum;> wi. )i 1 amours 1 *i i !fti i- Main- Livn IIMlA' Itr.i S m. Mni mu ninui'-s ii , i,,i1`ofna.** No. No, No l Itlier 1 \ pi' and oi silt* No. Total No.* (4) 4(17) -- 1 () -- (1) - (5) - (1) (3) -- -- (1) () 4 04) 111,0(1(1 ppin It (Jill 10 ppm III grain ppm 1\ til III ppm V .'an ppm \1 an ppm \ ll treatment ;tf i :m :t i ,i' :tn :;o so Total 2m (id .in r>o i r.M -it) u i -51 00 fiO 7v) l.V' ;>io i; IS 11 11 ) > 11 2'> Oo 10 in 21 111 til i:;t Is? 20 :is 12 12 i 12 1 : in I 2` ::' in 2` IS 0 U <r o till lie no tumours is also worthy diiormis. After ml carcinomas 5, two groups 6(KK) ppm for weeks among mis tissue oe- ic JO,OCX) ppm animals of the s le.speetivoly, 5-50 jjpin in n I' i 35 weeks, 2o00-o(X), and ce, pulmonary tions, lmt the s. K'rn observed; plnoblastomas imnas, some of " In ftMimlrn tm.mi Ipitiwt; l -ulinilmiri iT' hMimiiffi'Uim; ^ < ffir kin pliiituM'i'JIfihir rai < nmiiiii, 1 i liMUir Is, in}>ln>-viH`iana. d Toial No. of tumors. which undergo mal gnnut tumsturmatinn and mammary carcinomas (in mice). Macroscopic and micioscopic illustrations ol the tiiinotus induced are sliown in Figs. 1-25, J I lie frequent Zymba! gland caicinomas can reach considerable size (Fig. 1),,, are sometimes hilohated and show a tendency to hemorrliages. Their structure * can show diilerent patterns. The arrangement ol the normal Zymlial gland (Fig. 2) appears often in a disorganized form in the tumours. The carcinomas piesent various diflereut histological patterns, e.g,, glandular (Fig. 3), solid i Fig. 4). squamous, anaplastic, and poly moiplants. The lieipu nt puhnonary inetasta.se.s carry the chaiaetcristics of the primary lesion, (l'ig. 5). The nephro blastomas (Figs, (i--S) lead to numerous melastases in the liver (figs, d, 10") -hut can also involve spleen ami lungs. The Ileqm-ntly lound anginsuieninus ol the liver in rats (Figs. LI, 12, Id, ami 11) aie chaiacti rizetl hv the vascular growth and hemorrhagic appearance which is also present m pulmonary metastases (Kig. 15). Occasionally, both angms ucouu ol the livci and nephne blastema occurred in the same animal (Fg If.), \ug os,ire an is of othei sites appear gcneiallv as more or less heaio.ibag c masses developing in the ab- ' Figmvs marked by astciisks i*i appeal on the mini plates F J ^' K' Vk M ft if r* V ' 1 \ \ i t i i t i r r i f UCC 107049 3% \l \l T(l\ I AMI lll'I.MIS'l (loinin.il ca\ 11 v (Fig. 17;, in tin- Milieu!,moons tissue (Kin. 22" j in llie uterus (Fin. 1 -S), or in the limn (Fin- 19) Tlio ossifying angiosarcoma ( Fins. 20, 21*) mid llie s.ihv.ny y'land lint umni.i ( Figs -2d mid 2 I) nmiunl imely (see Table 2, group VI). Finui'e 275 shows a liver niigiosaicomn of a mouse. CONCLUSIONS *,M) DISCUSSION l'loin llie lesulls piesenteil in die tallies, limn llie pathological and histological observations, the lollowinn earls eonelusions may lie drawn. (1) VC is oncogenic under our experimental conditions. It induces, in rats, carcinomas of the Zymlial glands, nephroblastomas and angiosarcomas in liver mid other sites; in mice, livei angiosarcomas, pulmonary adenomas and mammary carcinomas. (2) A duect relationship exists between the dose and lennlh of lieatmcnt and the neoplastic response. (3) Zymbal gland carcinomas and nephroblastomas may be bilateral. (1) Liver angiosarcomas are often mnlticentric. ("i) lllood vessel eetasias and endothelial hyperplasia, associated or not with cellular atypia, are often observed in liver and in other organs and tissues in treated animals, with or without angiosarcomas. There fore, the effect of VC on blood vessels and endothelia should be consideied systemic, (f> 1 The onset of 2 ossifying augios.neomas suggests a new orientation in the pathogenetic interpretation ol aeroosteolvsis in workers exposed to VC. (7) To the pieseiit no acroosteolytic lesions have been observed in our ex posed Ullihl.lls. (N; No tumours have been observed in rats exposed to VA (27500 ppm I Zymlial gland carcinomas, iicpbioblasloinas and liver angiosarcomas have ueser been observed to occur spout.mcmisly in our breed of Spraguo-Dawley rats. Although Zymbal glands, in rats, are responsive to a large number of carcino gens, only 3 spontaneously occurring eases ol Zymbal gland carcinomas have been recorded in this species (Tanneubaum rt at., 1002), To oiu knowledge, no spontaneous nephroblastomas mid liver angiosarcomas of rats have been reported I. Hat with /ymlu! j^LhhI Liircinobi l Fn; I. Xyinhul idand carcinoma solid poltem, in rat. II-Is X 158. Fu; 0. Hut with nephroblastoma, melastases in liver Fio. 10. Liver metastasis from nrphiohlasfoma, in rat II F X 158. 1m(;. 11. Hat with liver angiosarcoma. Ku:. 12- Hat \ulli hver an^iosarcotu.i with lung metastasis. I'M- 13. Liver aiiKuiAdiioiiL, chaiacttii lie pitleiii; in rat * !-'' v' 158. Fn.. 15. Pnhnonaiy metastasis of liver ambosau oma; in lal. I) F x 158. Fio. 10. Livei aokpnsaiconia 4 nephroblastoma in lat. Fu:. 20. Lafeiorrrvn ,il ossifvlmr ant'ios.m oma, in rat. If F, x |58. I'm. 21. I'nlnmnary metastasis ol nssdvnn; uni'iosauonia, in rat I l-ls x -10. 1'io, 22. Hat with MihentaMeniis angiosarcoma. s !;. 7, Ncpliruljla,'. 1 l-'ii. (. pl.ml.l III 1 .(i l'H,. S. N( jiIooliListunu of ,i H-K x 470 UCC 107054 1 VINYI, (11101111)1 ( NIK INOCI M SIS 399 is'. F t. M-E x 46. 'rrt Err;. 7. Nephroblastoma of rat, clunucteristU pattern; I HO X 185. r fIc I \ i` r Fir.. 8. Nephroblastoma of a rat, m pliin^t-nu blastema rhlh u nlinlinj> in a glomerulus 11-F. X 470. VINYL Cl: Kit.. J J.iwt angiusuiconi.t, duruetenstk' pattern m a rat. H-K X 185. Im<;. J8. lT(rnis ; If>. An^iosar* % :.. 1 IrUi. IS. Uterus angiosarcoma; in rat. II-1. x 470. Kk;. 10. Aiu;i()s,ir< i)tiiii of lung, in r;il, II- I! y 1S5, 402 \i\i insi \VI) 1 M l Ml\i 'W. 'J-'l*-c&U'*' :t -p J-L, # i i 4 \ l\ Kll.. 2.1. H.ll will) ill \ (iI.uk! ailrlllKUieilltllll.l. If It,. in literature and us lar as \ lepoi lid as bemy espeiimei Hepatic airainsnicomas li, ;md Lee, 19(>S; lladjiolov, lfj 1973 i and in iiamsU'i's (Tot been induced b\ e-aminoa/i ami by p-dime(!i\ laminnben. Ill the labhll ambosarcom and Speiser, 19.7 1 I. In man, ani'iosarcnma.s a: Cases Wile inlleileil In 19.'/ II VC I', 1 iidiii I uni ni Ii v f i anysi >sai a I i \ i T anyiosa I enma billnwt Cast's nl liver am/iosarcoiua lliomliasl ( MacNialioii cl til Horta cl ill., 1 ?Jf>-">j and <-\pi I lie i< Mills nl uni invi \li meinbers nf tlie Kmnpe.m C a liall ii.Ho, it became delimit nephroblastomas and other to major \mei lean mamilaeli /1 Vi\\ i, ( in < mini ( \m imh,i ` isis -KW (V i'-i l "t 4> m >*, j Os 7 .ivi'r amjiiiMiu .im.i in ii miiusc. It 1-1 >' 155, in literature ami us hu ns ue know, kidney nephroblastomas have never been repoitell as bcnii' etpei'imenlalh induced. Hepatic auxins,ucomus have been etpenmeulaily induced m nils ( licubei ant) I .of. 1965, ! ludjiolnv, 1972 >. in mire ( lino anti -Salam.m, 1951- (laidesu vA <ll., 197-31 and in humstcis (Toth, 1972,). In mict . ostruheputio uneiusarcomas have been induced In' n-aminuu/utnluene (Andeivout ct til., 1917, \ndni vont, 19-70) .mil In /idiinellit I nnmnber/i m i urn 2 uuplitliulcne (Millay and Nason, 19-7.3). in * 11 o rahliil, um'insui comas lute Inf ] i piudiiced by 11 mi nl l aat ( Zeitlholei and Spoisor. 195-1). In man, unthosurcomus arc (|iut<` rare One liimdrednotonloon dnouinonlod oast s woio cnlloclod In 195.5 (I .aiitlolls, 197,5 ), ol which only 21 arose in the liver. Induction of livor un'juosaicnmu.s ha\o Iioon jepurtod in Innnans, In one case, a livor ump'osai'cnma (ollovvotl (iio iinoilion nl a radium nootllo ( Hoss, 19-'>2), Cases of liver aniziosaroonus hate Iioon lopoited foliowinu; administration ol thorotrast ( MaeMahon ct al., 1917 l.ndm, 19-7,3, liusorj'a cl til., 1900; Da Silva llorla cl dl,, 1905) and esposme lo nr mho (both, 1957). The ie.sulls of our iii\e-.lit;,ilinns li.it t horn ponotlit allv liansnutlod to the members of the Kuiopo.m ( -impei.ilive flump \\ lion, ap|>io\imaloly a yar and a hull aoo, it heeanic delim'fels oloar that \ 11 tv.is able to induce an'.dnsuicomas, iiophrolilaslomas and ntln i iiinlo'ii-iiu iot, mn o-Milts won- also oonimunioatod |o ma|< a Aiiici lean inaiml.u luieis nl V< 1 and l'\ ( 1, to orient tlmio,il obsei vat inns 4U4 MAI.IOM ANL> LI.H-.M1NL and epidemiological investigations, and to guide industrial hygiene measures needed for prevention of diseases which might be associated with VC exposure. SUM MA11Y 1'his project ul investigations is concerned with delei mining (lie oncogenic potentialities of vinyl chloride (VC). The effect of this compound is studied in relation to various experimental factors, such as route of administration, dose level, length of treatment, and species, strain and age of the animals. The preliminary results ate presented. When given hv inhalation VC induces, in rats, Zymbal gland carcinomas, nephroblastomas and angiosarcomas of the liver and ol other anatomical sites, and in mice, pulmonaiy adenomas, mammary carcinomas, and liver angiosarcomas. A three! relationship between (lose and length of treatment, anti neoplastic response has been found. REFERENCES Andewvont, H. B., Grady, H. G., a no EowAito, J, E. ( 1942). Induction of hepatic lesions, hepatomas, pulmonary tumor.',, mid liemangio-endotlieliomus in mice with o-aminotizo- tolucne, ). Natl. Cancer Inst. 4, 131-153. Andeiivont, 11. B. ( 1950). Induction of hcniangio-endothcliomas and sarcomas in mice with o-aminoazotoluene. ]. Natl. Cancer Inst. 10, 927-941. Ra.skmga, It,, Yoioo, If, ano Hknecaii, G G. (1900). Thorolrast-induced earner in man. Cancer 13, 1021-1031. Caiuiesa, A., Pour, P., Alhioff, J., ano Monti, U. (1973). Vascular tumors in female Swiss mice after intraperitoneal injection ot dimcth>lmtrosaiuinc. ]. Nall. Cancer Inst. 51, 201-208 Da Sii.va IIoiua, Y., Ahuat, J. D., Cayoixa Da Mocia, I.., ano Rohiz, M. L. (1905), Malignancies and other late effects following administration of thorotrast. Lancet 2, 201- 205. Geyeii, R. P,, Bryant, J. E., Bleim.u, V. R, Peirce, E. M., ano Si are, J1'. J. (1953). Ef fect of dose and hormones on tnmnr piodoelmo m rats given emulsified 9,10-dimethyl-1,2- benzanthracene intravenously Cancer Res. 13, 503--500. IIaoiaiw, A., Harris, R. J. Kos, G. A. R, ami Rim, E M. F. (1948). the growth- inliiljitory and carcinogenic piopcrties of l-ammostilhene am! drrivalivrs. Phil, Trans. Hay. Sue. I.amlon, Ser. A. 241, 147-195 Hadjhjlov, D. (1972). Ilciiiungiiiciidolhc'hul saitomas ol tin liver in iats induced liy diethvlmtrosamioe. Neaplusma 19, 1 I 1-1 14. Landelus, J. W. (1958). Malignant tumours of the In-art and hlood vessels. In (R. W. Raven, Ed ), "Cancer,'' Vol. 2, pp. 512-524, RuttiTWorth. London I ,o ol N, M, Jn (1953), Ilaema.ngio 1 udot In Uonialo'.v son l,i4,rr und Mil/ hel 9 hmoliast. speieln ning. Schweiz. Z, Allfinrn. Pathol 10, 987-99) Mai Maiius, II. E., Mtmt'iiY, A, S., AMI Haies, M 1 (1917) Endothelial-c ell sarcoma ul liver following thorotrast injections. Ami r. J. Pathol. 23, 58,5-595. Milutr, J. A., ano MiM-ek, E. G. ( 1901 ). The carcinogenicity of 3-inrthoiy-4-umii>o a/olien/ene and its N-nit`thyI derivatives for estrahepatie tissues of the rat. Cancer lies, 21 1008-1072. Miu.ay, A, S, ano Sax&n, E A ( [tl.i.li l.rsioir in'lin i il in nme hv painting with p-ilimcthvlamin(>azoben/'ono-l-a/o-2-naphlliulene. ]. Natl Cancer Inst. 13, 1259-1273, Reoui.r, M. IV, ano IjEE., G. W. (1908), h.lieel of age and si * mi hepatic lesions jn Buffalo strain rats ingesting diethyliutrosainine. ]. Natl Cancer Inst 41, 1133-1140. Roe, F. J. C., and Salaman, II. H. 11951). A i|nantit.ittve study of the power and 11 VINA persistence (if tumour-initiating ^ Cancer 8, M>(>-07fi. . Ross, J. M. ( 1932). A case ilh. Bacterial. 35, 899-912, Roth, F. (1957). Arsen-Lehe; 408-503. Spitz, S,, Maguigan, W. H., anii (line. Cancer 3, 789-804. , Tannenhaum, A., Vessecinoviti Multipotential carcinogenicity ot Tumi. I). (1972). Morphologic hydrazine dihvdrochloride in 5v Vioi.a, l`. I.. (1970). Cancer. Ccmgre.r.T, Houston, Ahstr. Vol, Vioea, P. I,., Uiuorri, A., ano C and boors to vinyl chloride- Ci Wai.ioei:, A, I.., Wiij.iamh, M. action of 4-aimuodiphenyl anr 25.5-20.1. c Wilson, U. 11., he Eds, F., and activity of 2-ac'etamino(luotene. ! ZKrn.linFP.il, J., ANO Sl'KISKll, P, experimentelW Thorotrast-veral A * UCC 107060 ik'w measures i VC exposure. the oncogenic ul is studied in uistration, dose als. on VC induces, ireouias oi the mas, mammary and neoplastic of hepatic lesions, with o-aminoazo- sarcomas in mice -d cancer in man. tumors in female Cancer Inst, 51, M. L. (1965). Lancet 2, 201- J. (1953/. Ef9.10-dirncthyf-1,2- H) The growthnci. I'hll Trims. rats induced by cssels. In (R, W. ilz hei Thorotrast- uelial-cell sarcoma )-methoiy-4-amino t. Cancer Res. 21, h> jv,tinting with 13, 1259-1273. hepatic lesions in t, 1133-1140. if the power and VINYL CHLOIUPK rAHUNOOKN'KSIS 405 persistence of tumour-initiating effect of ethyl carbamate (urethan) on mouse skin. Brit. }. Cancer 8, 006-676. Ross, J. M. (1932). A case illustrating thr effects of prolonged action of radium. /. Pqthol. Bacterial. 35, 899-912. Hom, E. (1957). Arsen-Lelier-Tiimnien { Hanuingincnduthclium). Z. Krehsfursch. 61, 468-503. Srirz, .8,, Maocucan, W. II , ani> IIoiiium-.u, K. ( 1950). 'Ihe carcinogenic action of benzi dine. Cancer .1, 789-804. Tanm'niiaum. A., Vinsii.inoviu'.ii, S. U,, Mai.ioni, ()., and Mitchell, D. S. (1962). Multipotential carcinogenicity of urethan in .Spraguc-Dawley rat. Cancer Bet. 22, 1302-1371. Toth, B. (1972). Morphological studies of angiosarcomas induced by 1,2-dimethyl- hydrazine dihydroehloride in Syrian golden hamsters. Cancer. Br.t. .32, 2818-2827. Viola, I', (1970) fliineenigeuie eifret of vinyl ehloiiilr, Inlernathmal Cancer Congress, Houston, Ah-.tr. Vnl. 29. Viola, P. L., Unarm, A., ash Caputo, A. (1971). Oncogenic response <i{ rat skin, lungs and bones to vinyl chloride. Cancer Bet. Ill, 510-519. Walpole, A. I.., Williams, M. M. O,, ani> Uoni'.nrs, I). ft. (1952), The caicinoginic action of 4-aminodiphenyl and 3-2'*diinethyl-4-aminodiphenyl. Brit. J. Indutt. Med. 9, 255-203. Wilson, R. H,, i>k Eiis, K., asn Cox, A. J., Jn. (1041). 71ic toxicity ami carcinogenic activity of 2-acetaininofhiorenr Cancer Bet. 1, 595-008. /EiTLiiorEK, J., ash Speiseii, I1. (1954). Ilaiiiangiiiciidothcliiuiiatose beiin Kaninchen nach cxperirnenteller Thorotrast-verabreichiing. Z. Krehsforsch, 60, 161-108,