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study of ilomatous : 745-752 elien auf uolymere len 2083: G. Size>iological '79). :pendent mineral ARC Sci. lency for ply Jhi m W, p' Mt. Environmental Health Perspectives VoL 51, pp. 73-80,1983 ^logical in Vitro and in Vivo Responses Chrysotile Versus Amphiboles a' J. Bignon* and M. C. Jaurand m*- gaff. m1"'!hf` te I%' Although all commercial forms of asbestos have been demonstrated to be carcinogenic in animals, so far epidemiological data are controversial concerning what asbestos types are the most carcino genic and fibjjrpgenic in humans. In order to understand the early cellular events induced by fibrous particles, different in vitro studies (hemolysis, release of enzymes by macrophages,,assays on cell cul ture systems) have been carried out in several laboratories; most of these studies have shown that cell and subcellular in vitro responses were different depending on fiber types: chrysotile versus .amphi boles. This presentation compares the. results of different laboratories with our data obtained by using a model which modifies the chemistry of the fibers by acid treatment.The acid-leaChed chryso- tile and acid-treated amphibole fibers showed different biological responses in several in vitro systems used in comparison to unieached fibers. Thefte differences in the in vitro reactivity were related to the chemical state of the fibers and might explain the differences in their. effects in animals after intrapleural injection as assessed bjf thfepercentage of mesotheli6ma,.the latency period, the survival time and the degree of pleural fibrosis. The careinbj;ehiC'dffeit of the fibers i's discussed in relation oj(tJ|p^jbi.pffr(>fnC|nifi*fiahsr^i|ii cytotoxic respoifted.' : - motion , the fibro^ettic and ^tieistos dusts are &iiV^^y*aeq^a,' tijj&e ,Jj).Considerable debate regarding bi 'tiie oiie Bjmectanisms of fibrpgCriesiS and carcinogen- on the other, the gtadieht in pathogenicity f$fbnt types of fibers. Bp''regard to the mechanisms of asbestos-re||iip$i$es, over the past 10 years considerable has been placed on the e^jfjmiehts of vmet al. (1,2) and Pott.et ai. (J, t)tyhich demon- pisofc physicochemical' parameters' fer behind Ijw&yer, we will see later on that the rOie of and physical constituents and particularly available at the.surface of the fibers must also a tole, but have been insufficiently assessed. jjTwrioupe de Recherches et d'Rtudes sur les Affections Respiet l'Environnement (INSERM 0 189 et ERA CNRS i). Centre Hospitaller Intercommunal, 40, Ave de Ver194010 Oreteil Cedex, France. From human and animal dat/thOi?iO'i& -strohg'eVi- - dencethat the jtynes/of jasbestps* the serpentine imrysotue and ihfe' ampnibolOsi croci- doliteand amosite;ar0all responsible tor.lung and plOuralfibrosis andfor lung arid mCsothelial cancers (ffi. Hbwever, there is still controversy about the gra dient OfpathOgehicity of these three types of asbes tos. Several epidemiological studies on human popu lations which have.been exposed to one type of fiber have persuaded many people that crocidolite {7-9), and perhaps also amosite (10,11), is much more car cinogenic towards the pleura than chrysotile (12). Previously, a group of experts at the 1976 IARC mebtaUg(6) cottclUdedthatmfcCupational.exposure to chii^sOtUe was more likely t6 causehingfibrosis and luttgcaricer than exposure to'ampWholes. Occupa tional Ofcpostmeto crocidolitemridamosite, however, was^moreoften associated with pletifal and perito neal mesotheliomas thtai exposure to chrysotile. In a reteent editorial, however, Liddell (IS) gave another opinion, pointing out that amphiboles were not only the most carcinogenic fibers in the mesothelium, but were also more fibrogenic and carcinogenic in the lung. Many authors are not convinced by this asser tion, especially after the epidemiological-demonstra tion by Peto that the incidence of pleuralrriesothelio- ma was almost as high in a cohort of workers ex- 74 BIGNONAND JA URAND posed mostly to chrysotile as in cohorts exposed only or mostly to amphiboles (14,15). Experiments in animals have also shown discrep ancies in the fibrogenic and/or carcinogenic potential of asbestos according to the type of fiber, but in all studies other factors intervened, such as defees,'mode of dust introduction (inhalation, intratracheal instilla tion, intrapleural implantation of injection, intraperi- toneal injection), type of diseases induced; animal strain, age and survival time. However, the results showing a modification in the carcinogenic effects of chrysotile after acid treatment (16, IT) raised the question that other factors, besides shape and size (chemical composition, surface physicochemistry), may play a role in the induction of fibrosis and cancer. The controversial position of scientists concerning such an important 'point needed aicritical review- in an attempt to evaluate significant information from the comparison of the biol^gicql respdnseVof chtyso- tile and amphiboles in vitro^ w.ejl as,in, vivo. The ef fect of acid treatment of the .fibers will alsobe taken into account, since it can lead to a better understand ing of the mechanisms of fiber .carcifv'ohbiiesis. Our provisional conclusions will be ?9P<:pr- dant results obtained in our laboratory and in others during the last decade. RMCIWts^BiRea SlootJ Ce8 ^' ' The hemolytici assay provides; a rApid way for investigating.the interactionhetweendustsand biolog ical membranes. Using this system, !several authors (18-22)iouad different responses witfrchrysotUe and the amphiboles, the former being, more'hemolytic than the latter. Generally speaking, after acid treat ment chrysotile was less hemolytic (16, 21, 22), whereas acid-treated amphiboles were foundi to be more hemolytic (21,22). Thus, if hemolysis explores the interaction between .fibers and(Cell membranes, chrysotile appeared as the. mostreaCtive fiber(type in these experiments. This discrepancy wasvaiso found when studying the adsorption of(phospholipids on fibers, .and this was greater, with chnyswtile than. with, the* amphiboles: {Jaurand, et.ah, Unpublished data) (Fig. 1). Jaurand et al. (24), when studying the kinetics of hemolysis by chrysotile, have shown a self inhibition of the reaction due to adsorption of. the membranes. This correlates with the observation of a decreased hemolytic activity after incubating fibers with phospholipids (24) andprob&bly relates to a decrease in the zeta potential (26), Indeed, Light and Wei (21) have demonstrated that the hemolytic activity was related to the absolute value of the zeta oC, .Ch CD Pjp/SI!piXPS) Figure: 1. Adsorption isotherms of liposomes of dipalmitoyl phosphatidyl choline (DPPC) on chrysotile (Ch) and crocidolite (Cr) fibers. Variation in the amount of DPPC ad sorbed on the fibers with the equilibrium concentration CBq. In the squares are indicated'the values of the ratio PMP determined by photoelefctron spectrometry anal ysis (XPS) W). potential of fibers. With chrysotile it decreased dur ing leaching, whereas with crocidolite it increased during the same treatment. ` '-i> < ,ui ( Reactivity With Mtic^pha^es Studies <cartied out in different laboratories over the lastfive years, describing the release of lysm somal acid 'hydrolases frbni the,amphiboles.,, . .. Davies et al.' (27) and, nxore. Recently in our labora tory, Jaurand et,al. (22) have cfearly shown that chrysotile works as an inflammatoi'y stimulus, induc ing a selective release of lysosomal, acid hydrolases. Howeveb, there was no release of cytoplasmic en zymes such as lactatfe dehydrogenase (LDH), which, in the case of peritoneal macrophages, showed 1.-1______.. ..... 11-.. . - .* * i obtatheqwith pmpr parties such as zympsap which we Know. $8). ' . - Tips .type; of infiammafobyresponse,.-cpfil^ ,'l?e re lated of the;fibers,;wHicli has lost most'of ,its ^Sg, d!d hjit release; lysosomal en zymes $8).and even released Lf)H, indicating a cyto toxic effect'^2,29). In contrast, untreated amphiboles (crocidolite and amosite) seemed to be cytotoxic, releasing both lyso somal acid hydrolases and cytoplasmic LDH. Acid- treated amosite and crocidolite, however, enhanced the release of lysosomal hydrolases, but this was as sociated with the release of LDH (22) (Fig. 2). .(XPS) EE of dipalmitoyl tile (Ch) and nt of DPPC ad- concentration <?s of the ratio trometry anal- creased durit increased ttories over ise of lysoor alveolar vve clearly ysotile and our labora;hown that ulus, ipduchydrolases. )lasmic en)H), which, s, showed i enhanced have been )san which iuld be reiperties of which has >soma! ening a cyto- dolite and both lysoDH. Acidenhanced is was as2). m: fe. 40* Ch 20H 50 SO 20H CHRYSOTILE VS. AMPHIBOLES 75 Am 100 JQ 300 330 Cr 1001 100 Qz JSL 100 300 Hg/mi 1.3 .76 .06 .85 .5 1.7 .6 0_ 1.52 1.72 0 |i''Release of enzymes from rabbit alveolar macrophages cultured with chrysotile (Ch), amosite (Am), crocidolite (Cr) or z,I)Qi2 (Qz) either unleached () or oxalic acid-leached (). Percentage of LDH atpj /J-galactoSidase (f) Gal) released'in the j^|^|^(ium,if9llowing20hrofcontactwiththeparticles(conceptration5p,'lQ,Q,or.^09(fi|f/m.U, t;.; . !).*'* . - * jiajspjopco___i_n___e___c_ Ile.*ar thatll.>. i- m>a. crophag/uen?rl secrete . |jhf!iprpducts (enzymes, mediatorsl aftef;in- ""m-mbto with asbestos andthat several of Mppmes may be directly involvfed in chronic Jltory responses. Chrysotile has been shown ^(highly significant increase in macrophage ' activity and prostaglandin synthesis induce the secretion by alveolar macro- jjm}a ichemotactic factor which attracts poly^.vjf^ar leukocytes (31). Macrophages from .. * i'.'-fv.itiO,vi S n 1-A <k AM*All **4" I /W\ vWrt A xl .'wVl. wS* l** _ i the significance of'the in 'vitro .responses of macro phages in different biological pathways according to fiber types, in relation 'to tttfe type and intensity of fibrogenesis and' carcinogenesis 'in vivo: however the relationship between the in vitro macrophage response to fibers and pulmonary or pleural carcino genesis, however, is far from being clearly under stood. ' .... , sn cultured in vitro, whereas latex parti$|p$fc\yield such an increase ($2). All these in- indicate clearly that chrysotile. Works.j^ent.activating factor on alveolar macroIPpiyeyer, other studies did not confirm the ^pppfilatory effect of chrysotile. Thus;. Sp||i|Cuh:h (S3) found an increased secretion of ip^'-biy peritoneal macrophages under the acL^flcjffiry'sbtile and crocidolite, but the high doses Pp'tiiis experiment do not allow a comparison tllb: effect. Moreover, in a recent experiment the oxidant production by guinea pig fr macropKages in vitro (release of 02` and Opposite pattern was shown, e.g., the 'ijuife|e asbestos were more effective than -'^le to cause macrophage oxidase activation iier in vitro experiments are needed, taking Reactivity with Proliferative Cells Several.authors. have usedicfeUvlines in short-term studies of-asbestos cytotoxicity.' Most of these stud ies are summarized in Thblfe l'. Which indicates the cell lines used by. theuuthbrs.Mfchfe cytotoxicity as says, the doses of asbestos tested and`the gradient of toxicity according to the type (chrysotile versus amphiboles) of fiber (35-41). In most studies chryso tile was more "toxic" than the amphiboles. More over, the acid treatment decreased the cytotoxicity of chrysotile and increased the cytotoxicity of croci dolite and amosite (41). Up to now, few experiments have been conducted with normal tissue or cells in culture. Some of them were carried out with normal tracheal tissue ex plants cultured in vitro (42). Only amphiboles have :V fv: * _ * / ,t - ' ' , 76 BIGNON AND JA URAND Table 1. Asbestos cytotoxicity on ceil lines. Epithelial-like cell lines Cytotoxicity assays Asbestos doses, j^g/mL Gradient toxicity3 Reference Macrophagelike P 388 D1 cells Human lung fibroblasts W 38 cells Rat liver-derived K 22 cells Chinese hamster ovary CHO cells Chinese hamster lung V 79-4 cells Growth inhibition Morpho changes Growth inhibition Colony efficiency Trypan blue exclusion Colony efficiency Human alveolar lung A 549 cells Growth inhibition Chinese hamster lung-derived CHL 39 cells Colony efficiency Chinese hamster ovary CHO cells Growth inhibition Human intestine-derived I 407 cells Colony efficiency Adult rat liver-derived ARL 6 cells Mouse colon-derived MCE 1 cells I 10-100 100 10 10-50 (cells seeded with fibers) 10 10 250 Ch > Cr Ch > Am Ch Cr>,,,,, (35, 36) (37) Am > Cr > Ch (38) Ch > Cr> glass fibers > LCh Ch > Am Cr Ch > Cr, Am Ch 10 times > Am-Cr I 407 > ARL 6 LCr 4 cytotoxicity LAm & Cr t cytotoxicity Cr > Ch, Am (39) (40) (41) aCh = chrysotile; Cr = crocidolite; Am = amosite4LCh,LCr, LAm = leached chrysotile, crocidolite, amosite. i! Pi iid I 1 ^1. been tested in this model. In our laboratory, we have developed a model using cultures of normal rat mesothelial cells for testing the reactivity with different types of fiber (43). This test studied the morphology and the growth characteristics of meso thelial cells treated with chrysotile and crocidolite which were either oxalic-acid leached or unleached (44). When , the samples are compared weight to weight, the results'ftg^ee with-those obtained by others- whp used; epithelial-like;;cell lilies (57, 41). Thus, chnysotile seems.' to; be more reactive and cy totoxic with epithelial^fe cell lihes1than: crocidolite; leaching of chrysotile-fibers' decreased the-reactiv ity; conversely, leaching-the crocidolite increased the cytotoxic effects on the cells. In vitro studies have also been carried out with cultures of lung fibroblasts which were stimulated to produce fibrous collagen under the action of dif ferent types of asbestiform minerals (45). In these experiments, chrysotile was tfye. most reactive, fol lowed by anthophyllite and amosite/crocidolite. This effect was dose-dependent, but the-response was not constantly .the same. In contrast, the acid- leached- chrysotile;.particularly when 80% of the magnesium was depleted, was much less active on collagen synthesis. the HGPRT mutant phenotypic test (39), induced chromosomal damage (45) and slightly increased sis ter chromatid exchanges (38). However, no differ ence was noted between chrysotile and the amphiboles. Althougjv. it has been demonstrated that chryso tile asbestos was much more active than thp amphiboles in. bindihg lgG ^i?), no difference fWdfrAdt&i betweeit chrySotile and the amphibolesifdr the acti-1 vation of the clhssical and alternative) pathway^ of complement1 49). Apparently, -. cdmplembnW acti vation whs not related to reactive Sites at'the fiber surface, sinice there was no difference between chrysotile and the amphibdles or between chrysotile and leached chrysotile (49, 50). Nevertheless, surface properties seem important for the adsorption of macromolecules by asbestos fi bers, as suggested by the results obtained-in our laboratory with chrysotile and oxalic acid-leached chrysotile. The adsorption of albumin or dipalmitoyl phosphatidyl choline on Mg-depleted chrysotile fi bers was characterized by a bulk incorporation of the macromolecules into the fibers. However, these results are different from those of others (51, 52) who found that albtimin had a decreased affinity for Mg-depleted- chrysotile. Stibcellular Effects It is still controversial as to whether or not asbes tos can bind to DNA and induce damage and muta tions. No mutagenicity was demonstrated by means of the Ames tests on bacteria (46)-. -However, tests carried out on mammalian cells in culture have shown that asbestos fibers may interact with DNA, since they gave a weak mutagenic response with Animal Studies Differential Fibrogenesis The early animal experiments did not clearly de fine the relative importance of asbestos fiber types in the production of lung or pleural fibrosis (53, 54). However, since the work of Wagner et al. (55) and more recently of Davis et al. (55), it appears that CHRYSOTILE VS. AMPHIBOLES 77 sotile given by inhalation causes far more lung fls than crocidolite, which in turn is more fibro id than amosite. This fibrogenic gradient was jfSiind to be the same when the number of fi$is adjusted to an equivalent number in dust mips (56). This gradient may be related to fiber -,i discrepancies between asbestos types as sug- by many authors. Davis et al. (56), who used Intensive fiber-length distribution, showed that 'f'i'chrysotile clouds in the chamber had many ,, 6: fibers over 20 pm in length than either of the <'|Miibole clouds in their experiment. It seems that '`^-fibers, less than 5 pm in length, are phagocy- ,without causing fibrosis, while fibers longer f'Jj,pm in produce foreign body granuloma with ml-In a recent, well-controlled animal experiKpiirig inhalation, Lee et al. (57) found that amot|as''at least 10 times more fibrogenic than poJi'pi:' octatitanate (Fybex) fibers, although con- '||jons and lengths of these man-made organic 'were many times higher in the clouds than , ,,jMk,mosite. These findings suggest that physi3m$V properties of the surface of the fibers an important role in fibrogenesis. 1Mt'V ' t ^Carcinogenesis ''`llp^erim'ents, some of them large-scale, 5TJ?qk^ed bu't in different species in order A|Mf^ferential effect of fibers introduced J||||^ral or peritoneal cavities, either by in * "Vjl^'itoplantation. The intrapleural or iritra^ ip/ihoculation of dusts has the advantage J|fepiments can be conducted with small ^^Cmaterial which allow, the comparison of " jhbiples of specially prepared or modified jjyOver, the experiments using the inhala_|PMi through the airways are more realistic `Mt^lared with human exposure: the ideal is |mi^ation in a special chamber. animal inhalation studies did not find ^4?i',;resulls in the production of bronchial mesotheliomas virith different asbesf^S-60). Wagner et al. (55), in a series of ex^fepats using amosite, anthophyilite,.cro- two varieties of chrysotile, found that Sfc mean survival time after first exposure ^Ijlfyed With chrysotile, particularly the Canafollowed by crocidolite and amosite. In the .lay, the highest number of malignant tumors srved in animals treated with Rhodesian ii^ aqd the" lowest in those treated with amoThpphyllite, crocidolite and Canadian chrysoY$ about the same number of tumors. The Oogenic effects of chrysotile versus the were observed even though much less dust was retained in the lungs exposed to chryso tile. Davis et al. (56) found clear-cut results after in halation studies in rats comparing UICC chrysotile A, crocidolite and amosite. UICC chrysotile A was more fibrogenic and carcinogenic than UICC croci dolite and UICC amosite, since all the malignant lung tumors were found in animals that had inhaled chrysotile dust. Only two mesotheliomas were found in this study, one with crocidolite, and one with chrysotile. Regarding pleural carcinogenesis, it also appears that chrysotile is the most carcinogenic--or at least as carcinogenic as the amphiboles. As early as 1969, Wagner and Berry (61), in a large-scale experiment comparing the effect of chrysotile, crocidolite and amosite on specific pathogen-free (SPF) and stan dard rats, found clear-cut results, in that all types of asbestos produced mesotheliomas. Chrysotile and crocidolite produced about the same percentage of tumors, the percentage in SPF animals with mesotheliomas being 61% for chrysotile and 59% for crocidolite, while in standard animals the corresponding percentages were 69% and 68%. The fewest mesptheliomas wep. produced by amosite (4Q% of the B^F and 3l7o of the' jslphdard mesothehi f tali'ty .'due chrysotile, ................ . ti. ... . .... time's (f>9,8"days fpr SjP$ pn$,'6l^ <fays! fop standard rats). This, was significantly less than crocidolite (718 And $55, respectively).and piuch Jess than amo site (811 and 801 days, respectively). In contrast, the survival of SPF and standard rats without meso thelioma, after elimination of the effect of mortality due to mesothelioma, was not different. Wagner et al. (55), however, using intrapleural inoculation of various dusts in rats, found that among the UICC standard reference samples (experiment 3), UICC crocidolite wks the most carcinogenic, being three times as actiye as.UlCC cjp-y^pt$e. But, in this very paper, the resdltS of' e^periirient 1, where SFA chrysotile wasconipared.to crocidolite in a dose-ef fect relationsJiip, Were in contradiction with the above conclusion. There was a relationship between the number of mesotheliomas "and the dose (from 0.5 to 8 mg) for both SFA chrysotile and crocidolite, but if we total the number of ratS'with a mesothelioma, there were 21 out of 59 animals with mesotheliomas in the SFA chrysotile group while there were 11 out of 59 animals with mesotheliomas in the crocido lite group. In a recent study carried out in our laboratory, af ter intrapleural injection of different , dusts in the rat, chrysotile and crocidolite produced about the 78 BIGNON AND JA URAND same number of mesotheliomas; the most striking difference was a more marked initial inflammatory reaction of the pleura with chrysotile, with a greater percentage of animals dead. from, other causes than cancer. Moreover, the latency period was shorter in the chrysotile group than in the crocidolite group (17). Conclusion Obvious discrepancies exist between the biologi cal effects of chrysotile and the amphiboles either in vitro or in vivo. Chrysotile seems to be the most re active in vitro as well as in vivo. These findings question whether or not it is sciehtifically coirrect to apply the. Stanton theory generally to carcinogene sis induced by, fibers, sipce' it tafes iiifoacgount only the fiber size jia'raine'tbi's^' . In this respect, it is odd thWt'fWe jiap'ei5 In memory of Staton (62) takes into account only artiphiboles and amphibole-like fibers, excluding ChryBbtile fi bers, which according to our results anil to tliose of other laboratories, appear as the most patent in- flammatdry stimulus. The striking modification in the biological response of acid^treateilj ahtiestol'sug' J) * although fibers were short (55). This underlines the necessity of pursuing basic research on the,, mechanisms of the biological effect of fibers because it seems that the Stanton hypothesis does not. explain all situations. REFERENCES 1. Stanton, M. F., and Wrench, C. Mechanisms of meso thelioma induction with asbestos and fibrous glass. J. Natl. Cancer Inst. 48:797-821 (1972). 2. Stanton, M. F., Layard, M., Tigeris, A., Miller, E., May, M., and Kent, E. Carcinogenicity of fibrous glass: pleural response in the rat in relation to fiber dimension. J. Natl. Cancer Inst. 58: 587-603 (1977). 3. Pott, F., Huth, F., and Friedrichs, K. H. Tumorigenic ef-, feet of fibrous dusts in experimental animals. Environ. Health Perspect. 9:313-315 (1974). 4. Pott.'F., Friedrichs, K. H., and Huth, F. Results of animal experiments concerning the carcinogenic effects of fibrous dusts and their interpretation with regard to the carcino genesis in humans. Zbl. Bakt. Hyg. I. Abt. Orig. 162: 467-605 (1976).' 5. Harington, J. S. Fiber carcinogenesis: epidemiologic ob servations ,and the Stanton hypothesis. J. Natl. Cancer Inst. 67:977-987 (1981). , 6. IARC. Asbestos (Monographs on the Evaluation Of Car-, cinogenic Risk of Chemicals to Man, Vol. 14), International, chrysotilfe was S^- flammation and subsequently cancer,' wjtfj&reai^oci- dolite needed softie in vivo ipodificatwn tb bOfcbme inflammatory and carcihogOmc. ' This puzzling biological ptbblem tyuhes the inter pretation of human data difficult; FHrbt, fiupians have usually been exposed to niiked fitiOrs associ ated with different cofactors. Peto et ail. ana lyzing epidemilogical data, obServed,l that thO risk of developiftg mesothelioma was substantially IbWer in humans Whose exposure tb/Olra^jpe^TOi^^ced. or ceased than in th'OSe Wheh^exppSiiiTb 'rii&ifr tained; by contrast, Oven b^ifef ex^Si^e. to b^ldb-. life could produce a substantial ,incidefi^ nieso1 thelioma many years lhttsr {SjrPetb^'t all'^Irsdg- gests that this difference could be diie either to'the fact that chrysotile was largely eliminated from the lung whereas amphiboles remained alfriost in definitely (55) or to the fact that chrysotile fibers are leached in vivo - (54) and thus cease to be biologically active in the body, while crocidolite fibers remain active or even become more active. Recent experiments in animals have shown that other fibrous minerals such as erionite-ze'olite' Were also carcinogenic, even more than asbestos, - - lap 9.- McHoijftldi.J; C.'Mesdtheliqrna after crpcidb|ite,pxposfire during gap mask manufacture. Envi ron.'Res.' 17:340-846 (1978). ' 10. Achesort,' E. t)!, Gardner, M. J,, Bennett, C., and Winter, P. D1. Mesothelioma in a factory using amosite and chryso tile apbeptos. Lancet: 1403-1406 (1981). 11. McCullagh/.S. F. Amosite as a cause of lung cancer and mesothelioma in humans. J. Soc. Occup. Med. 30: 153-156 (1980).' ' 12. McDonald, J, C., Liddell, F. D. K., Gibbs, G. W,, Eyssen, G. E., and McDonald, A. D. Dust exposure and mortality in bhrysotile mining 1910-75. Brit. J, Ind. Med. 37: 11-24 (1980),' , 13. Liddell,..j>, .Asbestos and pubjip health. Thorax 36: 241-244(1981): " '` . tiohal, Agency' fOr Research on Cpnper, Lypn, 1980, pp. 703-711. ' "' 15. Peto, J,, Seidman, H., and Selikoff, I. J. Mesothelioma mortality in asbestos workers: implications for models of carcinogenesis and risk assessment. Brit. J. Cancer 45: 124-135(1982). 16. Morgan, A., Davies, P., Wagner, J. C,, Berry, G., and Holmes, A. The biological effects of magnesium-leached chrysotile asbestos. Brit. J. Exptl. Pathol. 68: 465-473 (1977). 17. Monchaux, G., Bignon, J., Jaurand, M. C., Lafuma, J.. Sebastien, P., Masse, R., Hirsch, A., and Goni, J. Mesotheliomas in rats following inoculation with acid- iderlines the eh on the >ers because is does not lisms of meso>rous glass. J. tiller, E., May, s glass: pleural elision. J. Natl. umorigenic efmals. Environ. suits of animal fects of fibrous to the carcino.bt. Orig. 162: demiologic ob . Natl. Cancer uation of Car '. International nd, P. Diffuse North West'-271 (1960). P. G. Factory A continuing thelioma after lacture. Envi- .. and Winter, te and chryso- ng cancer and d. 30: 153-156 W., Eyssen, and mortality led. 37: 11-24 Thorax 36: ma in chrysoal Effects of Ed.), Internaon, 1980, pp. Mesothelioma for models of f. Cancer 45: rry, G-, and sium-leached 58: 465-473 Lafuma, J., nd Goni. J. n with acid- . CHRYSOTILE VS. AMPHIBOLES 79 f>;- lpached chrysotile asbestos and other mineral fibres. Carcinogenesis 2:229-236 (1981). BSf*S<:iinItzerf R. 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Paper#! presented at Symposium on Asbestos, Montreal, May 24-Vl 27.1982. EFFECTS OF MINERAL DUSTS ;f: - i itSF Wealth ncc^center AT D* * AS NOV i 9 1984 NOT TO 3E TAKEN FROM LIBRARY HEALTH PERSPECTIVES ILS. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service-National Institutes of Health National Institute of Environmental Health Sciences VOLUME 51, SEPTEMBER 1983