Document dYeJ3dyEXgg4gMmM2kX3pN95Q

SKIS 33 iliUyJ lUvcj JpSSfiM STHfPUSIA [ S/ARC SCIENTIFIC PUBLICATIONS K IB Environmental Pollution and Carcinoqenn. Risks pollution de I'eimronnement at nsques canc^rogenes INSERM, 1976, Vol 52, pp 127-150 OCCUPATIONAL CHEMICAL CARCINOGENESIS : NEW FACTS, PRIORITIES AND PERSPECTIVES Cesare MALTONI Institute of Oncology and Tumour Centre, Vtale Ercolani 412, 40138 8ologna, Italy The first historical observation of an occupational cancer dates back to the sixteenth century: it was the so-called "mountain disease" m miners of Joachimsthal, which, only three hundred years later was known as pulmonary carcinoma, and which, only m the thirties, was recognized as due to radioactive pollution present in those mines. Since then, up to a few years ago, the policy has been often the sane: that is "let us '/ait and see" if an agent present in the work place is carcinogenic or not. Which means, if it produces career on man, on. the basis of an epidemiological evidence, of course validated by a statistical analysis. Tins is not any more the time of the policy of "let us wait and see". It must be rather the starting of a new, more correct, social and scientific approach. The reasons why occupational oncogenesis, which nowadays is mainly industrial oncogenesis, need a new approach which cannot anymore be delayed, are the following. First. It has been estimated that from 80 to of the tumours an human beings depends on causes present in the occupational and general environment, and that, therefore, cancer must be largely considered an ecological disease. Second. The oncogenic risk has been progressively increasing m the last century, in relation to situations depending on industrial trends (table 1). Nowadays the major problem is represented by the multitude of the products of synthetic industry, which are un known to animal and human protoplasm, and on whose effects we are therefore fully ignorant. Third, Factories, that is the work-places, represent m the present situation the crucial area: in fact, the most exposed population lives there, and from there the new chemical compounds are spread m the general environment as consumers goods and pollution. 127 0005749 BSB RSV0033707 Fourth. There are at present clear-cut facts ind knowledge on general oncogenesis vrtiose significance is self-explanatory (table 2). On the basis of these facts, it emerges that the only real measure to prevent industrial oncogenesis is to identify the potentially once gome agents, before workers and the whole mankind are exposed to ~ them. In other words the time has come to predict the oncogenic risk, so preventing the exposure of human beings to it. Are there nowadays available tools for this purpose' We believe sd. It is a fact that the four most important cases of environmental and occupational tumours, discovered m the last years, have been in directly or directly predicted m some way, experimentally. The first is the case of the clear-cell-adenocarcinoma of the vagi na in adolescence, found in girls born from mothers treated during pregnancy with synthetic nonsteroid estrogen therapy. If we go back, in 1938, Lacassagne (1) already reported that he had produced mamaary carcinomas in mice treated with stilbestrol and m the following years it was shown, by several scientists, that the same hormone was produ cing a variety of tumours m hormone dependent and non-dependent tis sues, among different experimental animal species (2). The second is the case of pulmonary carcinomas among workers expo sed to bis(chloromothyl)ether. The discovery of this new type of occu pational tumour came together with experimental evidence produced by Van Duuren, m 1968 (3), and by Laskm, in 1971 (4), showing respect ively that, when injected subcutaneously into rats, or applied to the skin of mice, the compound was producing subcutaneous fibrosarcomas and skin carcinomas, and when inhaled by rats it induced squamous cell carcinomas of the lung. The third and most evident example is the history of vinyl chlori de carcinogenicity (5, 6, 7, 8, 9, 10). Experimental bio-assays on the carcinogenic potential of this com pound have provided information on: - its oncogenic effects, - the target organs and the type of tumours, - the still effective dose levels. Following our early observation on the dysplastic changes produced by vinyl chloride on the cells of the respiratory tract m exposed workers, and the early results of Viola (11, 12) that rats exposed to 128 0005750 BSB RSV0033708 ts and knowledge on general sxplanatory (table 2). that the only real measure ntify the potentially onco mankind are exposed to ~~ predict the oncogenic risk to it. ' .his purpose? We believe so. t cases of environmental last years, have been mexp erim ental1y. adenocarcinoma of the vagimothers treated during n therapy, if we go bacJc, at he had produced mammary and in the following years he same hormone was produent and non-depandent tispecies (2), mo.--.s among workers expo s' of this new type of oceu ntal evidence produced by 1971 (4), showing respectto rats, or applied to the ocutaneous fibrosarcomas ts it induced squamous 2 history' of vinyl chlori ne potential of this com ers, splastie changes produced atory tract m exposed 123 that rats exposed to 30,000 ppra of vinyl chloride developed carcinomas of the Zjmbal seba ceous gland of the external ear duct, we started m 1971 a project of integrated experiments to study the biological effects of vinyl chlori. le, m order to predict its cancerogenic risk. A series of 17 experiments have been started m sequence. The data of the crucial experiments are now being presented m the tables 3-5. When administered by inhalation, vinyl chloride produced on rats Zynbal gland carcinomas, nephroblastomas, angiosarcomas of liver and of other sites, subcutaneous angiomas, skin carcinomas, hepatomas, brain neuroblastomas and mammary carcinomas. The compound shows car cinogenic effects down to the dose level of 50 ppm. In mice vinyl chloride produces pulmonary tumours, mammary carcino mas, liver angiosarcomas, other vascular tumours of different sites and types, and epithelial tumours of the skin, being effective in the species also at the level of 50 ppm. In hamsters the monomer induces liver angiosarcomas, melanomas, forestomach papillomas, acanthomas and trichoepitheliomas, and it seems to anticipate the onset of lymphomas,being the latency time 48 weeks m the treated animals and 82 weeks m controls. Vinyl chloride is active also when given by oral administration, producing in rats some of the tumours observed following inhalatory exposure (table 6). Koreover tumours correlated to vinyl chloride exposure have been observed on offsprings bom from mothers exposed during pregnancy to high dose levels of the monomer m the air (table 7). Before the end of 1972, a short time after it was known that vinyl chloride was inducing in rats not only zymbal gland tumours, but also nephroblastomas and liver angiosarcomas, these results were also made known to several interested industries, and then were communicated at an international meeting in April 1973. This information promoted the clinical observation which in December 1973, for the first time, identified a liver angiosarcoma m a worker of a U.S.A. factory pro ducing VC-PVC as occupational jn origin. Since then epidemiological investigations have led to the discovery of nearly 45 liver angiosar comas among workers of VC-PVC industries in the U.S.A. and several European countries. Host of these tumours arose before 1973 (the first known case dates from 1961), but m absence of experimental data, they were not linked to the occupational exposure, and as a natter of fact, they were often not properly diagnosed. 129 0005751 BSB 9 RSV0033709 i Vinyl chloride has caused m one or more of the three animal spe cies usel m our expenner.ts, all types of tumours, which at preserv^ OH the 03.S1E of avsilcole epidemiological evidence, are correlates to occupational exposure, i.e, liver angiosarcomas, brain end lurg tj. mours, lymphomas, leukemias and hepatomas (table 8). There are now going on, in our laboratories, several experiments * better assess the degree of ns!t from e:-posure to vinyl chloride m relation to doses ar.d length of treatment. '/e are studying the effect of a short exposure (100 hours variou-z distributed m time) to high doses of VC on rats (table 9). The potential oncogenic effects of exposure to 50 ppm of the mono mer by inhalation is now furtherly investigated (table 10), A further experiment studies the effects of vinyl chloride by inha lation at doses below 50 ppm, that is 25, 10, 5 and 1 ppm (tables 1l~ and 12), Finally, a fev. months ago, an experiment started to determine the effects of low doses of vinyl chloride by oral administration (table 13). The fourth example of the predictive value of experimental bioassays is represented by the carcinogenicity of chromium pigments. In 1969 we started a program of investigation on the cancerogemc nsk from exposure to several inorganic compounds and pignents. A senes of several of the compounds under studies, with the available results, is given in the following table (table 14). The results dealing with lead-chromate pigments and 1cad-molybdcnum chromate pigments were made known in April 1973 (5-13). i Data published a few months ago (14) and on going epidemiological investigations indicate and excessive incidence of bronchial carcino mas among workers producing chromium pigments. On the basis of the present evidence it may be concluded that car cinogenicity bio-assays, if properly done, have a high predictive value. 1 Therefore, they cannot be any more delayed. Up to the recent past too little importance has been given to pre I dictive bio-assays, for many reasons, among which there were: 130 0005752 BSB RSV0033710 c of the three animal spetumours, which at present evidence, are correlated ' sarcomas, brain and lur.g t.(table 6). nes, several experiments sure to vinyl chloride m t l * f 5 : J xpoDore (100 hours various! ! n rats (table 9). sure to 50 ppm of the mono gated (table 10). s of vinyl chloride by inha { 10, 5 and 1 ppm (tables 11 ,) scepticism on their validity, based upon past results; -j) fatalistic acceptance of the situation because of the huge number of newly produced compounds; .) complication of elaborate testings. As regards scepticism, we think that time has come to critically review methods and results of past experiments which, nowadays, may oe thought to have been inadequate. As regards the large number of compounds, it may be true that there ore already millions of newly produced chemicals, but the ones which urgently need to be tested, mainly because of their widespread diffu sion, actually number m the hundreds. Concerning complications in performing elaborate testings, it may be true that the more experimental tests on animals can reproduce the conditions of human exposure, the more relevant they are m man. It should be pointed out, however, that agents to be examined should follow a pattern of te3ts which progress in degree of precision, so as to filter out and expose the most dangerous. t started to determine the oral administration (table of experimental bioty of chromium pigments. Ration on the cancerogenic .ipounds and pignents. A tudies, with the available table 14). >igmeats and 1 ead-molybde'nl 1S73 (5-13). I on going epidemiological enee of bronchial carcino ts. j j ^ I i may be concluded that car have a high predictive ed, -nee has been given to pre, which there were! * ; ! i i In our opinion the future policy for the prevention of the occupa tional as 7/ell of the environmental oncogenesis, are the following! t) priority to experimental bio-assays for the scrutiny of compounds already produced and widespread and for the compounds to be still produced on a large scale; 2) basic studies to investigate the potentiality of experimental mo dels in determining the oncogenic risk with particular emphasis on easy and rapid tests which, however, should be carefully and criti cally evaluated; and 3) the study with proper long term bio-acsays already validated, of the most important and widespread compounds, whose effects arc still unknown, following a list of priorities determined by all the interested social parties. On this line m our Institute we are now studying severs' eonpounds, among which styrene, acrylonitrile and vmylidone chloride. The plans of these experiments are given m the tables 15, 16, 17. Ongoing experiments performed m our Institute on the caninogcmc potential of drugs have shown in last weeks that a drug widely used ir. tumour chemotherapy and also in the prophylaxis of neoplastic recurren cy, namely adriamicme, is a potent carcinogen (table 18). Concluding, occupational oncogenesis should not any more be consi dered a limited medical problem,but a wider scientific and sociale one, for whose control we look forward to the collaboration of all the into rested parties of society! scientific community, unions, industries and governments. 0005753 BSB 131 RSV0033711 SUtWARY There is need for a new active approach to the problem of occupational carcino genesis. It is no longer acceptable to adopt a "wait and see" attitude since- 1. evidence suggests that cancer is an "ecological" disease: 2. the carcinogenic risk has been increasing as industrialisation increased: 3. factory workers are the most exposed population and hazards spread from tne work-place to the general population as consuner goods and pollution. It is necessary, and possible, to identify potential carcinogens before workers and the general popu lation are exposed. Wiat predictive experiments could be done, is exemplified by the history of stilboestrol, bis(chloranethyl)ether, vinyl chloride and chromium pigments, all of which were reported carcinogenic in animal tests before any epidemiological results were available. Unfortunately, these results were received with sceptician because the test methods were thought to be inadequate. It is necessary to develop the future policy of prevention by: 1. establishing a priority for testing compounds already produced and widesoread in occurence and those about to be produced on a large scale: 2. establishing experimental models for determining carcinogenic risk, especially rapid screening tests; 3. study of the more important compounds by long-term bioassays. Pull details are given of the testing of vinyl chloride morarer and preliminary results are tabulated for tests on styrene, acrylonitrile and vinylidene chloride. RESUME Une nouvelle approche active du problems de la canedrogenftse professionne1 le est noessaire. Adopter une attitude d'expectative n'est plus acceptable pour les raisons suivantes: 1. des risques conduisent a penser que le cancer est une maladie "doologique"; 2. le risque cancfirogSne n'a cessS de croltre avec 1'industrialisation; 3. les travailleurs des usires sont la population la plus exposce et les risques se propagent du lieu de travail 5 la population gendrale conns les biens de consermation et la pollution. II est nScessaire, et possible, d1 identifier les cancdrogenes potentiels avant que les travailleurs ot la population en gfinfiral ne soient exposIs. La possibility d'effectuer des experiences i valeur predictive est mantrde par les examples du stilboestrol, du bis (chlorcmSthyl)ther, du chlarure de vinvlo et des sels de chrome, substances que les tests effectuSs sur les animaux ont signalers came cancrog6nes avant toute Stude SpidSmiologigue. Malheureusement, ces rfisultats ont Std accueillis avec scepticism pares qu'an estimait les radthodes d'exnGrimentatiC' insuffisantes. II uiporte, pour la future politique de prevention, de prendre les mesures sui"ar.' tes: 1. dormer la priority & 1'experimentation des cenposfis df-gS produits et largemcr.t rSpandus et de oeux qui sent sur le point d'etre produits a grande dobelle; 2. Slaborer des noddies experimantaux permettant de dyterminer le risque cancerog&ne, ypreuves de detection rapide notarrment ; 132 0005754 BSfi RSV0033712 } a problan of occupational carC,.r_ nt arc! see" attitude since; jical" disease; is industrialisation increased; ition and hazards spread fran t'< ds and pollution. It is neep. `ore workers and the general pop . 3. dtudier les ccmposSs les plus imeortants au moyen d'Spreuves biologiques longue durdo. 11 est donnS des resultats sur 1'experimentation du chlorure de vmyle moncnere les rC-sultats prSlminaires sent mis en tableaux pour des tests sur le styr&ne, urylonitrile et le chlorure de vinylidbne. exemplified by the history of ie and chromium pigments, all of fore any epidemiological results received with scepticisn because f. prevention by: 3s already produced and widesorej < rge scale; imng carcinogenic risk, especial. tg-term bloassays. chloride nencmer and preliminarv .trile and vinylidene chloride. References ';) Lacassagne, A.: Apparition d'adenocarcinomes mammaires uhez des souns males traitees par une substance oestrogene synthdtique. C. R, 5oc. Biol., 1938, 129 641. (2) Lacassagne, A.: Les cancers produit par des substances chnaiques endogenes, 1S50. Herman, Pans. (3) Van Duuren ,B.L., Sivak, A., Goldschmidt, B.M., Katz, C. and tlel chionne, S.: Carcinogenicity of halo-ethers. J. Nat. Cancer Inst., 1969, 43., 481-486. can. ,,.ogen8se professionnelle est t plus acceptable pour les raisert (4) Laskm, S., Kuschner, K., Drew, R.T., Cappiello, V.P. and Nelson, H.: Tumors of the respiratory tract induced by inhalation of bis(chloromethyl)ether. Arch. Environ. Health, 1971, 23, 135136. r est une maladle "ecologique": vec 1'industrialisation; an la plus exposes et les risques rale come les biens de oonscnru3, d' identifier les canc&ogeres m. en gdnfiral ne soient exposes. ! i , ! ! j Leur prGdietive est montrGe par :her, du chlorure de vinyle et is sur les animaux ont signalfes Malheureusanent, ces resultat* ait les methodes d'exnftrimentatic J j j | j on, de prendre les mesures sui^ar* raposfis d^3i produits et largmsnt uits b grande Schelle; do determiner le risque cancer- (5) I'altoni, C.: Occupational carcinogenesis. "II International Sym posium on Cancer Detection and Prevention, Bologna, 1973" in Advances m Tumour Prevention, Detection and Characterization, Ualtoni, 1974, vol.II, pp.19-26, Excerpta Uedica. (6) Kaltom, C. end Lefemme, G.: Carcinogenicity bio-assays of vi nyl chloride. I Research plan and early results. Environment al Res., 1974, 7, 387-405. (7) lialtom, C. and Lefemme, G.: Le potenzialita del saggi sperimentali nella predizione dei nsehi oncogeni ambientali. Un esempio: il cloruro di vimle, Accademia Nazionale dei Lmcei, Ren diconti della Classe di Scienze Fisiche, Katematiehe e Katurali, Vol. LVI, Serie VIII, Fasc. 3, March 1974. (8) Maltoni, C. and Lefemme, G.: Carcinogenicity bio-assays of vinyl chloride: current results, in "Toxicity of vinyl chloride-poly vinyl chloride", Selikoff I.J. and Hammond E.C. eds., 1975, pp. 195-218. Annals of the Her/ York Academy of Sciences. 133 j 0005755 BSB I RSV0033713 (9) Kaltoni, C., Lefenine, G., Chieeo, P. and Carretti, D.s La canco rogcnesi arabientale e professionals nuove prospettive alia luce della cancerogenesi da cloruro di vinile, Gli Ospodali della Vita, 1974, 1, 5-6, 4-66. (10) Ealtom, C., Ciliberti, A., Gianni, L, and Chieeo, P.: Insorgcmo di angiosarcomi m ratti, m aeguito a somramistrazione per , , - .via orale di cloruro di vinile, Gli Ospedali della Vita, 197!,, 2 1 65 66 (11) Viola, P.L.; Cancerogenic effect of vinyl chloride, in "X Inter national Cancer Congress, Houston, 1970, Abatr. vol. 29. (12) Viola, P.L., Bigotti, A. and Caputo, A.s oncogenic response of rat skin, lungs and bones to vinyl chloride. Cancer Res,, 1971, 31, 516-519. (13) Haltoni, C., Sinibaldi, C. and Chieeo, P.: "Subcutaneous sarco mas m rats following local injection of chromium orange and molybdenum orange". V International Symposium on the Biologic al Characterization of Human Tumours, Bologna, 1973, m Ad vances in Tumour Prevention, Detection una Characterization, Baltoni, 1974, vol. I, pp. 133-134, Excerpta Medica. (14) Langard, S. and Horseth, T.: A cohort study of bronchial carcino mas m workers producing chromate pigments. Brit. J. Ind. Med., 1975, 32, 62-65. 0005756 BSB 134 RSV0033714 OCCUPATIONAL CHEMICAL CARCINOGENESIS C. MALTONI Tables 1 to 18 0005757 BSB RSV0033715 Table 1 CAUSES OF INCREASED ONCOGENIC RISK IN HUMAN ENVIRONMENT I - Concentration of oncogenic agents already present in the surface of the Earth. II - Surfacing of oncogenic agents from the depth of the Earth. HI - Production of new potentially oncogenic compounds by chemical and petrochemical industry. Table 2 KNOWLEDGE AND FACTS WHICH MAKE URGENT A NEW APPROACH TO THE PROBLEM OF INDUSTRIAL ONCOGENESIS I - The potentially oncogenic agents in human environment are progressively increasing. II - Different oncogenic agents may have additive effects, III - Changes produced by oncogenic agents are largely irreversible, IV - Oncogenic agents may exert their effects on different organs and tissues and widely affect the target organs. V -A large part of the natural history of tumours take place without any clinically and sometimes otherwise detectable pathological changes. VI - Cancer la not a reversible disease. 137 0005758 BSB RSV0033716 \ Table 3 bll and M6 i Sxaaaura by Inhalation to VC In Ur. at 30.000, 10,000, 6,000. <,990, 900, <50, 90 ppn.. * bra. daily, 5 daye seehly, tor 5* ***. laaiilta aftar 1J5 "take (add af the <nirlitt). irauu mtiials m n TUbouM 6MUP3 J9D (amcoB<MLer ATS) Corrac tad 2/Mal gland ts> Anglo arcnsec 1 41 neeas n gienaa " TBSATMEPT Total nuabar <a> Average A/erase Ha. i latency tana 6o. 6 latency tint Pe. {seeks) (stake) AveragL 5 latency Use . >e. (seeks) K. "1 VC 30.000 ppe. 60 60 35 98 43 _. - . 18 30 53 1 <e * '"iff0 9 61 16 26 50 5 8 99 9 19 64 3 (f 4 VC 6.000 BP*. 72 60 7 12 62 47 5 13 22 70 3 <f 3 Repateaaa Praan ney rblaato= Vanaaml other careins type Total aw (t) He. bo. _. 3 ,1 Re. fa. be. . _ 91 (a) 7 1 4 (n) 3 _ 8 U) *, 51 38 31 VC 2,500 ppa. 74 59 2 3 _3> ... 6 to 74 13 22 78 3 (b 3 1 VC 500 vtn. 67 99 1 79 4 7 ej 7 12 61 * fi , 1 _ _ _VnVC <90 PH. . _ _ ,,VC 'VI5I0I ppa. *7 64 59 59 6 TO 80 7 79 7 U _ 4 _ 12 135 2T 135 1 (a T 1 Ve treatRcnt 68 98 total 541 475 64 - - 26 - - 65 - - 15 12 5 1 3 (r) 3S _ , 3 (a) 22 . t (t) 16 7 (v) 10 10 M 6 '5 8 64 206 a) Alive aniaele iftr 24 seeks, han the first tkuour (a iynbel {load cikidohJ iu observed, the percentages art referred to the corrected nunbar. b) iataetaaee to Lung. e) Metestssts to liver, lung, spleen and train. 4) Mdtaetseae to lung, e) 1 lung aagiesarao**. f) i utrabdoninal aagloeercoaa (next to liver); i eaglomreoaa of the lipe; 1 a^iosareoaa of the mm. g) 1 aaglooarccM in eubeutaaeoue fibrosing aagloe*, 1 ossifying pareMrteulsr angiaaereoaaj t iatrabdoaiaal aagioaereone (next to liver). h) 1 deifying &ngleeereaa of neckj 2 Intrvbdealaal aagleaaroQaas (1 neat to spleaa and 1 next to ovary). 1) 1 nncuaarcota of uterus; 1 lung angloaarccn*. 1) 1 lntrctMeainU eitflooarcaM (sent to spleen); 1 intrathoraele ossifying engieaercane, ) 1 IntmMor.jnel diffused angloenreeae. n) Several eases of breast fibresOenoaaa. adrenal end pituitary wisoure (generally adenoaat) have not bean confidered, alnea their distribution la tfea different groups doee net vary. e) 7 SjtftcU gland adenoaas) 1 ekn scuenoua eeretaanaj 1 suboutasaoua flbrosaglonai 2 aaoaaey eareiaeaaa< 1 hepatle angloaai 1 hapatoaaj 1 ovarian aagleaa; 11 fareetorach pspillonatj i brain seurobleatoaa; 1 Harderlaa gland carclneaa. f) 2 tphfeal glnnd ndanaaat) 1 evariaa ciatoadanooareinoaai 1 neurllcaaaM. e) tyiebol dlaad adenoma, 2 hepatle and i peritoneal angioam > ' eallvary gland adenooaralnoaa. r) 1 2}abal glana adenesa; 2 epandynoue. e) 1 Tulaenory fibm-areof-o, ? lyopheaac. t) 1 Zyabal rland ado-em, 1 lyephsae, u) ) Sy**al la*4 j0cTOe-., 1 subcutaneous angloperacltoan, ) uterine artenaeareut'tae {i vltfc. seruoaetau* taapment), t) Several aiuaaJa eith 2 or acre tuaerje. ?lCTr! 2 TMr'*' ->* RSV0033717 .stsooo . -**... ,, , of the llpa; 1 aagloaaraoat of the noae. f() l! *,nwe*>oa*<r*cr*'o"- i* -rt9bccuutauneiuou.iel flbDrroeenmog, --aapoaa--ai 1 oie.eifiyfyiinng, pai-aurlcular aapeearvonef 1l uiattrnabudenaulBa wlo-re-. bat to Hvorl 1 oeeifying angioelaeroowe of snt*eek|i 2 lntrranbbddemaiiMl eanpoeerooaaa ((1l ne*xxt to apleen and 1 anenxtt to 0o0v0a.1rr7)). 1) 1 anelonreo-io of uteruej 1 lung aagioenrcae*. 1 1) 1 latnbdealnal arpoaareana (next to apleen)t t utrrthereeio oaelfy&ag iMU ) 1 intrebdemnal difffUbooee4 uanagletaeanrenoiia.. '" 11 grJ?f ioa not wiy, "`TM*1 "d pi","iT " <.TMuy -- ...- --n--. .i... *.1, .,,.,, ,, tt. O)) 7 ilyTatbteal eland fatddBonfMoaOolt., 11 aelkriina immiMawU.i carelnMo.a.a>f i1 .e.kubc..a_t&em.o>u.B flbroo__n_glon ferattewech papillo*aj 1 brain nouroklaataao} 1 Herdenaa gland caroiaene. .. . ____ * bmb>7 eereinoaaat l hepatle i^isu, 1 kepatoi ; 1 ovarian aapoaa; *? \ %*** *"* edeaoeaaj 1 ovarian oyetoedanocarelnma; 1 ntunlrawM. q) 4 trrbai eland adenoma, 2 hepatic and 1 porttooeol enpemai 1 ealivary Pert rtaDoeercinsM. r) 1 ipdiu eland adanooto, 2 eptnlynoeae. a> 1 pulaooory fibrooareaaa, 2 lymphoma. t) 1 lyobnl (land ed*rcea( 1 lynphom. u) 3 lyabal eland adonoaeai 1 aubouttnaooa aaaxepertcltoaa, 3 utonna rteneearolnoma (1 alth_______ __________________ naasr** n .TM. Zable 4 Experlaeat Bt4 j ftrw by inhalation to VC u air at iQ,00Q, 6,000, 2,300 , 300, *30, 30 yj., for 4 hro. dally, 3 daya teVly, for 30 *oka. Roaulto after 81 wooke (and of tho experiment). ______ __________________ ______________________ osovs Annuls (SWISS BICE) fatal Corrected ttuaber (a) TKZATMBTT ?0* 9 f? f i -V--C----1--0- T.00T0-p*--pn. ID 10 $0 22 28 50 >0 so 26 28 V 30 60 2t 30 53 Ung brteura (V) Average no. * latency tin, (veoke) AXIBUS PITH fUBOUJtS ran* 7 MKlneoa, Co} Liver an-- Vaaeulep tuaeure Epithelial gtaaarca-- of other UP* tuaeure ef and/cr alte the akin Average He. * uateney No. ho. he. aad/or alte he. 35 70 )6 i) 47 V 79 * a 28 10 57 9 ? )1 8 a , -------- _______ 35 T1 9 (A) 9 (e) 12 (f) 3 to 6 (n) 3 (0) 2 <r> 4 <B) 2 (t> Total (*) ho. 36 39 - 31 . ja--ya,m*, 10 60 29 29 5B 16 so10 10 29 29 38 3? VI 10 30 60 27 30 37 ------------vjl ri~ ' 67 1 T 2 a fetal 260 230 310 230 241 471 164 66 57 3.5 6 - 4V 45 51 44 - 7 24 11 P 12 33 . so - 37 M 43 , 11 ____ ______________ 1 -to) - O 11 22 (h) 2 f> J (v> 40 1 13 (i) . . _____ L ______ 47 84 j 15 1 () 7 1*3 20 18 _U___ 220 a) Alive animala after 16 weeke, whan the first tumour (a aomnry carclrxea) oat observed. the percentages ar* referred to the corrected nmbar, b) adanearns. oemo of which undergoing aollgn^nt tmeferwetib, e) Sr fevalea. Adanacarcinoma frequently with araaa ef aquoneua aetaplaaia. 4) 3 aubcutaneeuc arponoa. 4 liver nbroaacleaas; 1 heart flbroaacloaai 1 aaoifylnc Uitaracapolar anpeaa. e) 1 aubeutaaeowa anpoaareeM; 2 liver fl*>roaAg*ca, 4 llvor nnffiaaaa,' > roaal fibwneioao, \ tlpnwa on^lom. f) 3 aubewtaneouo ar encore ou( 1 eubeutaneoua anpoaat 3 mr anfioaae; 2 latrabdeainal ancioaereabaei 2 renal ancleoareoaear 1 lung ancloaa. *) 2 aubeutaneoua aneiaiareenaat 1 ubcitnnaoLs flbrocnioaa, i subcutaneous ta(ie, 4 liver flbrcanglo^e: 1 liver annoaa: 3 intrabdoauml aopoaemtirniTi 1 lutraWoninal fibroandioaAs 1 ronal aaioaareMat 1 taatleular fibraeapona; l anpena of the eaoeua; 1 Iun fibroaapeaa, 1 lony anpoae. h) 1 aubeutaneoua anpom. 6 liver fibroa&cionae. 10 liver aaponaa; 2 latrabdoainal uipoaorconaa. 1 ovarian anponai 1 acrotal anflem, 1 luni anpoaa. 1) 1 aubeutaneoua enclooorcoaat 1 aubautiAcuuo fibroai^iooa, 2 aubeutaaoeua angiMaa, 2 liver fibroanpoaaa, 3 llvor anpoaaa, 1 lotrabdealnal ariena 1 ovtrtan upona; 1 IBtratboroesc fibraon^loaa, l anpoao of the asteracapular fat pad. 1) 1 anpaaarcaaa ef nteruo. a) 2 oqueneua oarelneaea*, 1 invaelve aoanthoaa. a) s aquenoua earel&oMet 1 aeantbena. 0) 1 aquMoua carcinonas 2 oeairtboaae. p) 1 aeanthoM. a) \ ana adonoearelno'ea. 1 baaollena. r) 1 zynbol part adenono; 1 forestoiaaeh paplUowa. a) 1 aubcutcaaeve leie*yo*rcona; 1 foraeteneieh papillom; 1 kardarian (lead adoaoaa; 1 lywphaaa. t) 1 foroetoaaeh pBoxllenai 1 peuretld part nixed tunour. u) 1 lyabal port edaronn. vj 1 lyrbal eland rta&ota; 1 leydig cell# tuaour; 1 lywpbona, w) 1 parotid port rtenoeare taana. y) \ aubeutanaoue ioieyoaarcw: 1 adenoae of epos: 1 ovarian cerelanaai 1 lolenyoaarooaa ef uteruoi 3 lyapbonae. a) Several caaoa with 2 er acre -paeuro. I 1 \ 0 0 0 5 7 6 0 BSB RSV0033718 table 5 Experiment BTB . Exposure by inhalation to VC in air at 10,000, 6,000, 2,500, 500, 250, 50 ppm., 4 hre. daily, 5 days weakly, for 30 weeks, ftpeultt after 105 weak#. AlfUtAlS PITH TWOURS GROUPS TREATUERT (GOLDEN HAMSTERS) total Survivors Skin trlLiver ehoepith* sjigiOB&r- lioaas and comas basaliomas (.) HO. Ho. Fc, Lymphomae forcrtomach epithelial tumours (b) Other type and/or sits Total lg) Ho. Ho. No. No. I VC 10,000 ppm. IX VC 6.000 ppm. Ill VC 2,500 ppm. iV VC 500 ppm. V VC 250 PPA. vi ?c 50 ppa. VII Ho treatment 35 32 33 33 32 33 70 1 -6 12 -1 23 -3 -6 -2 1 - 4 ^ (c) A 2 2 7 7W 10 - 1 10 3 (el 12 - 1 7 2 Ef) 12 -12 6 1 1 4 _ 10 2_ 5 Total 268 1 3 23 4 8 34 14 59 a) Several case* with aoantbosls and aoae undergoing malignant troBforuatioa. b) Papillomas, acanthomas, erne of which undergoing malignant traaformation. e) 1 mibcutaneous angioma; 1 gall-bladder adenocarcinoma. d) 2 hapatoassj 2 liver fibruang.pmas; 2 liver anglomae; 1 biliducts odenoeuelaoma. e) 1 hepatoma; 1 liver fibraanginn*; 1 liver angioma. f) 1 subcutaneous angioma; t bronchial carclnoo*. g) Savaral animals with ? >r more tumour*. Table 6 Experiment BTIt : Exposui e by ingestion {stomach tube) to VC in olive oil, at 50.00, 16.65, 3-31 agAf body weight, once daily, 4-5 days weekly, for 52 weeks. Results after 64 weeks. GROUPS AND TfiEAtMEHT ANIMALS (SPfUGUB-D/CVLFf RATS) Total Survivors 2 VC 50.00 mg/fcg SO 25 11 VC 16.65 mgAg So 33 111 VC 3.33 aA IV Cotjtr^ oljve BO Oo 39 37 Zymbal gland carcinomas Ho. ANIMALS WITH TUKOURS Nephrobla stomas Anio pLrcomae Liver Other sites No, Ho. Ho. . - e 1 (.) 1 15 _ __ - - -- Other type and/or site (b) Ho. Total He. 3M 11 Ed) 7 _ -- Total 320 134 1 1 13 t 3 18 a) 1 thymus angiosarcoma. b) Several case* of breast fibroadenomas, adrenal and pituitary tumours (generally adenomas) have oot been considered, since their distribution in the different groups does not vary. c) 1 majonajy carcinoma, 2 foreatooach papillomae, d) 1 animal with 2 tumours. 140 0005761 bsb RSV0033719 000, 6,000, 2,500, ekly, for 30 weeks. H TW-'OTOS yoreetoracn BAB epithelial tumours tv>) So. Other type and/or Bite Mo. To*aa (> ho. 4 7 TO 7 2 4 _ 54 Ml. 2 (*> 1 (1) 3 () 2 If) _ 14 t 10 1? 12 6 10 5 59 'arc toons. Table 7 laganacftt 6T5 t tocooeme by inhalation to VC in air t 10,000 and 6,000 ppm, of breederSi 4 hro. dally, for 1 week (froa 12th to 18 th day of pregnancy). 141 *Vb fand CROUPS AMD irat*ekt r ,c 10,000 pp". ----------ii YC 6,000 FPO. ahxkals (SPBACUE-DAffirf RATS) Total Survivors 2ymbal gland carcinonee Bo. 30 - 1 30 - - WOKALS Wlt TUMOURS MephroblaBtO'BftS Anglo sarcomas liver Other sitae Ho. Ho, Ho, - - 1 () - -- Ill vc io,000 ppm. 54 - 3 1 - 2 0>) - IV vc 6,000 ppm. 32 - 1 - - 2 () total 146 - 5 1 -5 Other type and/or 82 tc Ho. Total Ho, 2 Id) 3 - 2 () e 4m 6 8 17 *) b) 1 1 intrabdomlnal angiosarcoma. subcutaneous angiosarcoma; 1 angiosarcoma of the log* e) 1 subcutaneous angiosarcoma; 1 intrabdomlnal angiosarcoma* 1d) 1 e) 1 t) liver fibroangioma; 1 liver angioma, Zyvbal gi""* fibre eareoaa, 1 ovarian leiomyoearcoaa. Symbol gland adenoma.; 1 akin carcwsosa} \ subcutaneous flbrcanglgaa \ 1 maanaxy carcinoma. C in oliva oil, once daily, TUMOURS ircoma* Other altes Ho, Other type aid/or ite (b) Ho. Total Ho, 1 (a) _ 3 tc) * _ ii td) 7 - -- 1 3 18 (generally adenomas) have doea not vary. table 8 ITCOTES PncSBfTU- CGEBEUtED TO VC EXPOSCW: (W IHHALATIOn) t OB EXPERISOTAl fiODEKTS ABD KAB Specie* TUMOURS Angiosar comas of liver Tunoura of brain Tumoura of lung Lympho mas and leuke Angio^f nephro corns blasto and an- mas mias gionns of other Sebaceous Squamous laar- culuncous tumour* ry carcino of epi carol mas dermis nomas Heps Poreto otomach mes paoillg as and scan the IUt + + * * U) a + House + + + 4- Roaster + (*t [+> U) - U) * Ban <+> () <) M I 141 0005762 BSB i j RSV0033720 Table $ Exparia6t BT10 t Excopui by inhalation to VC for 100 hrs, , followuig different achedulee, Results after 59 week*. GROUTS and TREATMENT I VC 10,000 apr, hd/5dr./S ZJ VC 6,000 ppa* 4hd Ad*/5 III VC 10,000 ppn* 1hd/4dw^5w 2V VC 6,000 j.j, 1hd/4dtf/T5 V VC 10,000 pm. 4M/1dw/T5w VI ic 6,000 pp. 4bd/1dw/T5w VII No treataect ANlkALS (SMAGUE-DAIfLEY ____ _____ RATS] Total Survivors 120 120 120 120 120 170 249 64 85 89 85 96 97 160 Zynbal gland carcinomas No* 4 3 4 2 - ratal 969 716 a) 1 subcutaneous engicearcena, b) T lyaphoaa, e) 1 subcutaneous aagloperccitoaarcoBe* 1) 1 lyapfceaa.* 13 AF1EAL9 WITH TUMOURS Nephrobla atoaae Angiosarcomas Liver Other ites No. No. No, - -- - -- - -- - 1 (a) - -- - --_ _ .1 Other type and/or site No, 1 oo - - 1 (e) 1 (4) 3 Total No, 4 4 4 1 3 1 17 Table 10 Experlatent 8*9 t Exposure by inhalation to VC In air at 50 ppa,, 4 bra, dally, 5 days weekly, for 92 week** Results after 59 week*. OPOOTS 1XD TREATMENT ANIMALS fSPRAGUZ-DAVLEtt HATS) Total Survivors Zyiobel gland carcinoma* Bo. ANIMALS PITH TUrCUHS Hphrobl stow Angiosarcoma* Liver Other pi tea No. No, No, Other type and/or site No, 1 VC 50 PJ*. 300 216 1 --- - n lo treatment 100 72 - --- - Total No. 1 * Total 400 268 1 --- -1 v: VC VC I VC WO trc Total OPOLTS AtO ntjiYf'- J VC 1 t?=. 12 No treat" Total 142 0005763 BSB 1 RSV0033721 , following different schedules, A WITH TUM0U8S Angioaareoue Liver Other item No, No, Other type and/or aite HO. total Ho, -- - 4 -- 1 (b) 4 -- - 4 - 1 <) - 1 - ' (c) 3 --1 1 <d) _ 3 1 17 f Table 11 i t 4 Experiaont BI15 t Exrxxurt by inhalation to VC ix air, at 2J, 10, 5, 1 ppm,, 4 hr*, daily, 5 days weekly, far 52 wek*. Result* after if weeks. CROUPS AID IHZAI1IDIT X VC 25 ppa. n VC 10 ppm. 3X7 VC 5 PJ, TV TC t Ppo. V Ue treatment Total ahikais (SFRACUS'DAVL&Y RATS) Total Survivor* 120 110 120 111 120 113 120 112 120 97 $00 543 2ynbal gland careinoaaa Ho. AHDtAlS am TiatOCRS Jlephroblaatomas Ho, Angl0 aareamas Laver Other site* Ho, Ho. . _ _,, _ .. _ .. - - -- Other "type and/or Site Ho. _ Total Ho. _ _ _ _ -- t 50 ppm., 4 hrs. daily, TTTHTUroURS 'aoaareoaaa r Other eltsa Ho. and/or site Ho, -- Total Ho, 1 - -- -1 Table 12 Experiment BT17 i Brnosura by inhalation to VC in air at 1 ppa., 4 hra, dally, > dya aeekly, for 5? >cek*. Heeulta after 46 weeks. CROUPS AJID TRSATMEHT I TC 1 ppm. XI Ho trassent Total abimals (WISTAR RATS) Total Survivors Zyab&l gland careinoaaa HO, ahikals with iwcwrs Haphroblaatomaa Angiosarcomas Liver Other sites Ho. Ho, No. Other type and/or aite Ho, Total He. 120 96 - - - - - - 130 89 _ - - -- 250 185 - - -- - - I 143 0005764 6S6 RSV0033722 Table 13 Experiment BT27 : Exnogore by ingestion (stomach tube) to VC m olive oil, at 1, 0.3* 0.03 rag/Kg body weight, once daily, 4-5 days weekly, for 104 weeks. Results after 21 weeks. GROUPS AND TREATMENT ANIMA1S ( spraoue-dawley EATS) Total Survivors I VC 1 mg/fcg/bw 158 II VC 0.3 mg/Kg/bw 159 III VC 0.03 mg/Kg/bw 163 IV Control: olive oil 159 Total 639 147 145 147 148 587 Zytobal gland care inomas No. AHIKA1S WITH TtMOUES Nephrobla stomas Angiosarcomas Liver Other sites Other type and/or site Total No. No. No. No. No. - - -- -- - - -- -- - - -- -- - - -- -- - - -- -- 0 0 0 5 7 6 5 BSB table 14 RSV0033723 VC 0.03 mgAg/bw 163 147 - - __ XV Control* olive oil 159 148 - - .. Total 639 587 - - -- _ ----- Experiments oi and 8T2001 t table H 1 subcutaneous injection of 3D mg of -the compound in i cc of RgO* Groups Treatment I Chromite n NeoohrooiuB hi Chromium allumen IV Chromium yellow (load chromate) V Chromium orange (lead chromate) VI Molybdenum orange (lead chromate, sulphate and VII Cadmium yellow (Cadmium sulphide) VIII Iran yellow (Iron oxide) n Iron red (Iron oxide) X Control ji Titanium oxide XII Zinc chromate A&lAftlS (1) Sprmgue-Dawiey rata) tumours (sarcomas at the sits of injection) Total Survivors 17oof animals Average latency] with tumours time (weeks) I Histology lype 40 40 40 40 40 - 9 8 26 27 - Rhabdomyosarcomas 7& and fibrosarcomas Bhabdomy osarcomas 72 and fibrosarcomas Rhabdomyo saroomae 40 and fibrosarcomas Bhabdomyosarcomas 47 and fibrosarcomas 40 - 36 HhabdOBjroaarcomas 32 and fibrosarcomas 40 40 - 16 flfi Rhabdomyoseu-comas and fibrosarcomas Rhabdomyosarcomas 77 and. fibrosarcomas 40 140 - - - - 120 116 - - - 60 SO - - - Ended Experiments InAi) Ongoing experiments time from start (weeks) *33 149 137 '50 150 n7 143 125 '34 127 35 4 (1) 50i rales, 50jt fwslss 0 0 0 5 7 6 6 BSB RSV0033724 Table 15 PLAIT OF THE ECPEP-UdHTTS OR THE BIOLOGICAL EFFECTS OF STTRBTE Experiment* Species and age at start (weeks) BtiOl Rate (U) I II III IV No.of aninals at start Total *9 Route 60 30 30 Inhalation 60 30 30 60 30 30 m 60 30 30 - Trealatent Reeks Lose Stopped Ongoing Length start (weeks) 300 ppm + 44 44 200 ppm + 44 44 100 ppm 4- 44 44 50 ppm + 44 44 BT102 BT103 V 60 30 30 - 25 pp Untreated VI 120 60 60 (controls) - + 44 44 44 Total 420 210 210 Rats I II III 80 40 40 Ingestion 80 40 40 60 40 40 250 mg/Kg m olive oil 50 og/Kg in olive oil control: olive oil + 37 37 & 37 37 4- 37 37 Total 240 120 120 Rats I II 80 40 40 4 intrabdomlnal 50 mg in injections olive oil (x 4) + 80 40 40 control:olive oil + 38 38 Total 160 80 80 BT104 Rate (13) I II Total 80 80 160 40 40 1 subcutaneous 50 ng in olive injection oil 40 40 80 8o control: olive oil + * 38 38 TOTAL ........ 1___________ 960 490 490 ./ Table 16 0005767 BSB RSV0033725 BT103 Sets (1J) I 11 Total 80 80 160 1-1 40 40 4 wtrabdononal 50 Bg in injections Olive oil lie 4) 4- 40 40 - control: olive oil + 60 80 I 1 38 38 BT104 Rets (13) 1 n Total 60 80 160 40 40 1 subcutaneous 50 ag in olive injaction oil 40 40 * control: olive oil 80 60 + 4- 38 38 TOTAL 960 490 490 1, Tati*. 16 flak op tub mzsntBirrs on ns biological efpects op achylokiihile Sxpenante Species at start (weeks) Groups Vo, of amnia at start Total o' *1201 Sate (12) 1 11 62 32 30 6o 30 30 Route Inhalation - Trealatent Dose 40 ppm 20 ppm III 60 30 30 10 ppm IV 60 30 30 - 5 PI V 60 30 30 entreated (controls) - Stopped Ongoing Length (weeks) Weeks froB start 4 35 35 + 35 35 + 35 35 * 35 35 35 3T202 BT203 Total 302 152 150 Sata (12) 1 n 105 58 47 Ingestion, (l) 80 40 40 - 20 ag/Xg in olive oil control: olive oil + 6 35 6 35 Sate (10) Total I II 185 82 158 96 87 42 40 Ingestion (2) 62 76 5 og/Xg m olive oil control: ol^vc + 21 21 + 21 21 Total 240 124 116 TOTAL 727 374 I- (1) 3-5 tinea weekly (2) 3 tines weekly \ ' 0 0 0 5 7 6 8 BSB / RSV0033726 Tam* n plait o? thb msHnarrs on the biological zmcis or vnransns chloride Experiments BT401 Spociss end l([t at start (weeks') Bats (16) I XI in IV V VI Total So. of tainels at start stotal * i 120 60 60 60 30 30 60 30 30 60 30 30 60 30 30 200 100 100 560 280 280 Treetoent Bouts Doae Inhalation ISO ppn * 100 ppo - 50 ppn 25 ppn - 10 ppn Untreated (controls) - Stopped Ongoing + 4 4 4 Length (weeks) 21 21 21 21 21 BT402 BT403 Sice (16) M Bats (9) I II III IV V VI vn Total X II III IV 120 60 60 Inhalation 200 ppn 60 30 30 * 100 ppn + 60 30 30 - 50 ppn 4 60 30 30 - 25 ppn 240 120 120 * 25 ppn 60 30 30 * 10 ppn 200 too 100 Untreated (controls) - $00 400 400 100 50 50 Indestlon 100 50 50 120 ng/Kg in olive oil 10 *g/K in olive oil 100 50 50 200 100 100 * o5liv*ecAoitl in control;olive oil 2 days 2 days 4 days 4 21 4 19 4 21 4 32 4 32 4 32 4 32 BT404 Total K.t. (9) I 500 250 250 100 50 50 Ingestion 0.5 ag/Kg In olive oil 4 21 BI4C5 TOTAL Chinese Hanaters (28) I II Total \ 60 30 30 Inhalation 25 ppn 35 18 17 Untreated. (controls) - 95 48 *7 |o.o [1.0*1 1 .O"" r ^ 4- _. ) 13 teaks start 21 21 21 21 21 21 21 21 21 21 19 21 21 32 32 32 21 13 7 1 j 0005769 BSB RSV0033727 (9) 11 100 50 50 m 100 50 50 " IV 200 100 100 to ag/kcin olive oil oontrolsollve Oil + 32 3? + 32 32 32 32 total 500 250 250 7404 Hat. (9) I TOO 50 50 Zngeetien 0,g ag/Kg in olive oil * 21 21 3T405 Chinese (28) 1 II Iota! 60 30 30 Inhalation 25 ppn 35 18 17 Untreated (controls) - 95 48 47 + 13 ____________ 13 13 TOtJO. U.053 1 .028 1.027 i. 1 I r Table 18 EXperimant BT1001 t Treatment: 1 subcutaneauB injection of 2 mg of Adriamieine in 1 cc of olive oil. Results : after 38 weeks. GROUPS AND rEEATMEnr i Adriamieine 2 mg II Control: olive oil lotal ABDULS (SRRAGUE-DAWLEY RATS) Corrected Sex Number number Survivors ? 42 42 35 0" 40 40 30 9 40 40 o* 40 39 34 30 ? o' 162 161 129 TtMOURS (1) No. * 14 35 12 30 --- 26 WERAGE LATEHCY TIKE (weeks) First tumour 19 Average 31 25 32 -- -- (1) At the site of injection. 0005770 ir> CP RSV0033728