Document ZnOK798ynoNXq6Z0zKr0gojmd

V/ / The Saranac Laboratory OF THE EOWARO l. TRUDEAU FOUNDATION Saranac Lake, N.Y. Arthur J Vorwald. M O oiitacTow . October 30, 1947 0UTEK3 ILLINOIS GLASS COMPANY TOLEDO, OHIO Interim Report on Animal Inhalatl Brperlmenta with IATYL SUMMARY: The first inhalation begun In February, 1 fatal pneumonia, another slmlj August, 1945. Both of tbes the effect of the dust alo: August, 1945, also, an exper of Inhaled oust on tuberculous October, 1945, a group of shite in order to study the^effvot of t with thsfs dust vas to a hlgn incidence of meat vas started in s were designed to test tion in guinea pigs. In begun to test the effect tion in guinea pigs. In placed in the dust room elation in this species. It may be sal re is no evidence to suggest that the dust alom Into the lung produces pulmonary fibrosis in rats 1 period, or in guinea pigs in a 30 month period, the experiment in wnlch dust exposure and tubero 'combined are inconclusive and further vork^&hould fore a definite statement can be made as to/hSpossible e; act of the dust on tuberculous in* faction, these experiments are virtually completed, although several original groups are being kept alive far more pro!' Petrographic and x-ray diffraction analyses of the dust supplied and used. Analyses vers carried out initially and periodically during the course of the experiment. The composite of such analyses reveals the following: Samples from the supply carton Quarts <l (approximately) Chrysotile and Serpentine 15 - 20#; largely fibrous. The fibers and bundles present are usually several millimeters long snd seldom are leoo than 300 p oalc.te 5 a Asbestos Litig. Saranac Lake 1989 Depos. PvViiHif n IF / Page 2 Owens Illinois Olaaa Company October 30, 1J47 Interim Hoport - KAYL ) Balance consisted of clay-like masses which gave a definite x-ray diffrac tion pattern not unlike a hydrous oalciua silicate* Saaplea collected from the atmosphere of the cheaper. Quartz 1% Chryaotile and Serpentine 10 - 15; less fibrous naterlal than in sample above; many fibers are shorter than 20 p. Calclte 10* Balance consisted of clay-like masses which gave a definite x-ray diffrac tion pattern not unlike a hydrous calcium silicate. The analyses reveal eeveral points of interest: The low quarts content (l*) indicates that the amount of crystalline free silloa in the dust la praotieally negligible as a possible cause of silicosis. Although diatomaoeous earth is one of the materials from which KA7L0 is made, only a trace, probably much less than 1*, is unaltered and appears in the final product in its original form, nearly all has been enanged by the manufacturing process into another chemical compound, calcium sili cate. Host of the material consisted of claylike masses which gave a definite x-ray pattern similar to that of a hydrous calcium silicate. About 10 to 15* at the atmospheric dust in the experimental dust room was cnrysotile or serpentine. Both of these minerals produce the same x-ray pattern. Proa petrographic ex amination of the original material, it appears that ohrysotile, which is a fibrous mineral, la present in much greater amounts tnan tne non- fibrous serpentine. In the ataospneric dust also, the fibers still predominate over t.ie non- flbrous serpentine. } VI'A 5 .3 i % i 4 i ** I i i * 000251 Asbestos Litig. Saranac Lake 1989 Depos. / T I % v. c 5. Jol^or 20, 104^ iO:;a-Illln3ia diass QjnoanyIn'-ari. : t - ..... L '. la tils original material, t..e fleers were present In bunules of ten 3 everal .lllli- aeters lon0; not many were shorter c.'.an 300 Tae fibrous portion of '-he atmospaeric sample, however, consisted mostly of snorter 0u.nulc3 and of fibers, relatively few in numoer, wuich were generally a oout 10 to 40 n in length* 3. Tecnnlques of Investigation. Two species of animals were introduced into the dust caaaber assigned to taia in vestigation. They included guinea pigs and white rats, animals wnich the Saranac Laoora- tory has used routinely in its studies apropos pneumoconiosis. Appropriate quantities of tae original material supplied oy tue company were introduced into the dust generator of tne cham ber and pilot runs were made to ascertain the character and concentration of tr.e air-borne dust produced* Havln- eataolisned the desired concentration, the animals were tuen introduced and throughout the course of the investigation dust counts were taken periodically. The average concentration of t.ie air-borne material was 122 million particles oor cubic foot of air. 'This figure variod somewhat from month to month, ranging from a lower limit of 99 million to an upper of 122 million particles. This range is not unique for this material but has been observed also with most agents which we have employed. It results no doubt from day to day variations in atmospheric conditions which cannot De precisely controlled with our present methods. Nevertheless, we have elected to continue vita such methods, believing that co do otnerwise would introduce factors uncommon to actual Industrial exposure. The results of the experiments will oe presented under the following headings: I. II. III. 17. AAYL0 inhalation alone In guinea pigs. KAYLO inhalation alons in white rats. KAYLO inhalation and simultaneous tuberculosis infection in guinea pigs. Conclusions. nOOi Asbestos Litig Saranac Lake 1989 Depos. Exhibit p.49 Pa^a 4. J'wena-IllimLa Jlasa Cp-nanv October 2J, 1./4" Intori a .looort - LJ I. XAYLQ Inhalation along In ulnoa j! s: ihe course of tills experiment througn tae first fourteen nontns was discussed In a su.-n.Mry written oy doctor Gardner in May, 1946* In this report he described tna difficulty enco-ntjr-eu witu acuta pneumonia in the guinea pigs and conduced tnat it -as an accidental epidemic* dis conclusion is supported by tn_ fact tnat subsequently only four ani.-aals nave died of pneumonia. Tnis occurred, aowever, only after all apparently sick guinea pigs were disposed of, the aoparently well pigs isolated, tae dust room thoroughly cleaned, aired and disinfected, and all tae cages sterilized. A third and new series of guinea pigs were intro duced after control of tue epidemic described. Throughout the course of the study, all animals were exposed continuously to the chamber cloud for eight hours a day, five and one-half days a week* Thus far, a total of thirty-nine pigs have been killed in order to follow the oourse of the pulmonary reaction resulting from the deposition of inhaled dust in the lung. Die period elected for study ranged from two to thirty montns* Over this period no definite gross ohanges occurred in the lungs* The traeheobronehial lymph nodes have enlarged slightly, out re mained soft. Microscopic study of toe lungs reveal gradually Increasing numbers of dust-containing macrophages* These inaotlve "dust eells" are found characteristically in clumps within groups of adjacent air spaces* The intervening alveolar walls are only slightly thickened by accumulated lymphocytes, but there is no undue proliferation of tissue histiocytes and connective tissue* Plbrosis is not present. Occasional multinucleated giant eells are seen* In addition, there are a very few elongated segmented brown structure which take an iron stain and are clearly asbeatosls bodies* Microscopic study of the tracheobronchial lymph nodes reveals clusters of inactive "dust eells" similar to those demonstrated by the lung. There is no apparent over development of connective tissue, and again fi brosis la conspicuous by its absence* Results of the oncmieal analyses oarried out on the lungs of the exposed pigs sacrificed at the various periods are re corded in the following table: TABLE I. 00025, 4I f 1o Fa-c 5. Oner, a- 11Lap La Glass Company October 50, 10-L7 Interim Ft por t - 11 vtc\l a::ai.y:>io v gut?r?j.A PIC- LIP-''C An i-nal Murder Time AStl % 9ry Hatter 5I0o % H0 CaO nc. 6 10 11 16 18 28 34 43 56 45 46 67 68 105 108 49 50 2 ao> 2 ao. 4 30. 4 r.o* r mo. 6 30* 8 no. ? 30 10 too. 10 no. 12 no. 12 so. 13 no. 18 RO. 21 mo. 21 mo. 24 mo. 24 mo. i 4.42 4.94 4.47 4.42 4.13 3.74 4.59 4.54 4*50 4.81 4.86 4.99 4.83 4.90 5.12 5.23 5.02 4.71 0.24 .20 .22 .10 .25 .20 .31 .28 .iC . .41 .55 .51 .59 .86 .64 66 60 0.07 .08 .09 Ww .08 .00 .08 .11 .21 .14 .22 .08 .26 .32 .11 .08 .06 .08 0.05 .06 .03 .00 .03 .10 .13 .11 .14 .11 .14 .1C .09 .10 .13 .15 .13 .14 SIO^ 4U Ash ' ~0 CaO *7 ` 4 .r-'- 1.22 4.37 . *r s *1 <5 WC *^0 5.36 i .:? X* - J 2 .00 10 6.34 o *26 1.77 C jL ^ 12.12 2.14 'i Iv'J 3.02 10.80 12.25 4.65 1.G0 12 .27 17 .65 5 .31 6.56 12.54 10.30 *02 1.45 12.70 12.52 1.20 1.72 1.35 1.23 1.99 1.39 2.00 2.68 2.75 2 .43 3.04 2.20 2.80 3.17 1.83 2.06 5.C8 2.90 2.54 2.92 TAflIZ I 00023 Asbestos Litig Saranac Lake 1989 Depos. Exhibit p.54 / 3age 6. )ctjbar ;-J, l'..)47 - er.s-Illlnsls Jlaaa Gomany____________________In^crir. ^pji t In view of the observations recordod aoove, certain pjrc^.-.ant comments appear to be warranted. The microscopic stuoi.c o:` one lan-jS onu trac .eobroncnial lymph noues make is oovions v.r. t :..ot was suspended in the atmosphere of t:ie c..a,ooor ana tnat port! no of tnat dust mere inhaled and gained access into bin: lun .3 of o..e animals exposed. This is further substantiated oy t ie c.-^iical analyses of the lungs waica snow a .gradually increasing ar.oo.no of tns various mineral components of the dust used in direct propor tion co cue exposure times. In tna case of 3102# f^r example, t e pigs sacrificed after ten months exposure revealed a two to three fold increase. The MgO and CaO content also lncr asau a-il t..-. increments are inconsistent and less manifest. The silica component of the ilneral dust is of sucu low concentration that a significant pulmonary reaction would haruly be expected In consequence of its presence in tae l_n._. Puis proved to be the oase even in those animals exposed for a period of twenty-four months and revoallng an average total of 12.61, 3102 of the lung ash. The cnaracter of the tissue response at thirty months Is comparable and it is anticipates, tnat tue 3U2 content will not be appreciably different. Undoubtedly some of the dust cells constituting the mild cellular reaction observed may be in consequence of the serpen tine components of the dust deposited in the lung. This. no./ever, cannot be verified since serpentine is a relatively benign dust and fails to produce reaction other than mere dust laden macro phages. This interpretation is in accord witn Doctor ua: oner's observation that the silicate serpentine is a biologically inert substance producing at best a mild reaction consisting of small widely scattered local collections of dust laden phagocytes and an oceasional giant cell without other eollular reaction and without fibrosis. The presence of a rare curious oody (ssoestosis oody) indi es tes that some of the air-borne fibrous component of tao chrysotile gained access to the lung, however, the essence of cellular reaction or fibrosis about the terminal oroccniolea would Indicate that the inhaled quantity of such fibers was of extremely low intensity and insufficient to produae permanent damage. This view is supported by the petrographic and x-ray diffraction studies which disclosed that a major amount of the chrysotile oomoonent of the dust consisted of structures too large for dispersion into the breathing atmosphere and for easy access into tne lung. In consequence of the experimental studies witn guinea alga to determine the biological activity of XAYLO, it may be tentatively eluded that KAYL0 alone falls to produce significant pulmonary uaaa..e whan Inhaled into the lung. Asbestos Litig. Saranac Lake 1989 Depos. Exhibit p.56 0002bC .',a_-o 7. Dwen3-Illlnol3 dlasa Comoan7 Dctooer 30, 1347 Interim ueoort - 'J%. 0 ) II. KAYL3 Inhalation alone in wnlte rat3: This species was used because tne rat is known to develop fibrosis from quarts inhalation more rapidly t .an ocner an.tala. Forty rats were used in this experiment and, as indlcatoa by Doctor Gardner in his report in day, 1346, no difficulty wltn early development of pneumonia was experienced. In fact, uurin.: the first thirteen months of tne experiment no spontaneous ueat.is occurred* From thirteen to eighteen aontns, about 40,. of tno animals died with terminal bronchiolitis and lobular pneu.Ionia. This is of no significance* however, Dscaaso it is -ell knnrn tnat laboratory rats beyond tne age of one year are sucjoct to this type of lesion. The results of exposure to XATLO were followed oy killing four rats at each of the following periods: six, nine, twelve, fifteen and eighteen months. The duration of tne expori :.ent was eighteen months and all animals have been killed. The reaction to the dust tnrougnout the experiment nas consisted of focal collections in the pulmonary alveoli of macrophages containing particles and spicules of dust. As the duration of exposure increased, the only observed c:.an_e was an increase in the number of such foci. Similar dust filled macrophages were seen in the tracnsobronealal lymph nodes. In no animal was taere any tissue r eaction of the sort seen in silicosis or escestosis. Chemical analyses of the lungs of these rats show tnat the 313s reached the average level of 16.5^ of tae lung ash aitain nine months and subsequently diminished. This is difficult to explain, since in the same period tne percentage of SiOg la guinea pigs' lungs exposed simultaneously (see Table I) was increasing. The table of tnese results follows: TABLX II. Asbestos Litig. Saranac Lake 1989 Depos. Exhibit p.58 4# / ?a - > 3. ois uiaiis uomoanT Octooer DO, 1347 KAYLO IHHALATIOH 3IL ICA DDT'IRJINATI3NS OF LUUG riSSUL; in RATS ONLY Animal iio. Ash in Lry SiOo in Dry Si02 in Qroup Tiasu Tiasu Ash ______S_____________ Li------------ * 1012-202 1012-201 17 rj 4.3 4.6 0.668 0.736 14.0 16.1 Duration of ^xnoaur 6 months 6 aonth3 1012-205 1012-206 IV IV 4.6 3.3 0.86 0.528 18.3 13.9 3 months 9 months 1012-209 I) 1012-210 IV IV 4.2 4.4 0.55 0.655 13.2 15.0 12 months 12 months 1012-215 1012-223 IV IV 4.2 4.2 0.443 0.562 10.5 13.3 15 months 15 months 1012-225 1012-234 IV IY 3.1 3.9 0.43 0.44 13.7 11.3 18 months 13 months TABLE II / OOG `7 * b , Asbestos Litig. Saranac Lake 1989 Depos. Exhibit p.59 ?a^Q 9. Owens-Illinois ula3a Company October 30, 1947 Interim Report - KAYLO In commenting upon Part II of the investigation using white rats as the experimental animals, reference is made to the comments recorded under Part I, which apply* III* KAYLO inhalation end simultaneous tuberculous infection in guinea pigs. This experiment was carried out by the use of only one of the three methods whleh have been found valuable in determining the effect of dost on the course of tuberculosis* In this case the animals were infected with tubercle bacilli by inhalation and im mediately plaoed in the dust room* Thus, we are testing the effect of the early accumulation of dust on the early course of tubercu losis* The other two methods available consist in exposing the animals to either the dost or the Infection alone for several months, subsequently exposing them to the other agent* *e may then study the effect of aceumulated dust on t he early course of tuberculosis, or the effect of dust on healing tuberculous lesions* In the experiment which has been completed twenty-five animals were used* Of these, five died within ten months of the pneumonia it which ioas been a problem throughout all the experiments with this dust* Four others were killed because they were in poor condition, and it was hoped to out down the Incidence of pneumonic by removing them from the room* These four also- showed some evidence of pneumonia i Of the remaining sixteen animals, eleven were killed in order to follow the eourse of the infection. One was killed at one month and two were killed at six, nine, twelve, fifteen and eighteen months* Two died at fourteen months, apparently from progressive tuberculosis; these will be discussed further below* Two acimals died, one at thirteen and one at seventeen months, of undetermined causes, but not from tuberculosis* Of the animals killed to follow the eourse of the lnfeotion, the lesions were isolated, encapsulated and regressing in all except one of the two killed at fifteen months* In this animal, lesions were widespread throughout the lung, put were associated with the production of fibrosis, indicating a tendenoy to heal* vh+ interpretation of the results of this experiment is com plicated by the accessory factor that some of the animals were used for breeding daring the experiment* This was done in an attempt to develop a strain of animals resistant to the infection whleh was producing the frequent pneumonias* A review of the records shows that each of the two animals wuieh died of progressing tuberculous infection had been used as breeders* In addition, the animal which was killed at fifteen months and showed widespread lesions had also been used for breeding* Asbestos Litig. Saranac Lake 1989 Depos. OH HOC* U1/1740.., Exhibit p.62 ?a ,9 10. Qjena-Illlnola Glass Company October 30, 1947 Interim Report - KAYLO Pour otiier animals used for breeding during the same period did not jho the extensive disease seen in the aniaals Just described. Biusf it seeos hazardous to attempt to attribute the spread ing of tne tuberculous infection entirely either to the effect of the dust or to that of breeding. IV Conclusions: 1. It seams dear fro* the results presented in parts I and II above that KAYLO acts as an inert dust when lnnaled by guinea pigs and rats; that is, it is phagooytlzed by macro phages, but does not injure them or produce fibrosis or anything suggestive of either silicosis or asbestosis. This indicates that the dust will probably be harmless on Inhalation in moderate amounts over prolonged periods by normal human beings. 2. The results observed in the experiment concerning the in fluence of the dust on tuberculous infection are equivo cal. Although it la our feeling that this dust exerts little or no adverse influenoe on tuberculosis, the possibility that it played a part in spreading the lnfeetlon cannot be excluded. 3. In view of this possibility, further studies concerning this aspect of the proDlea should be carried out. It is imperative that the original experiment with infeotlcn and simultaneous dust exposure be repeated in its entirety. One furtner study with a small series of guinea pigs is now in progress. It is designed to reveal the influenoe of the dust on partially healed tuberculosis. However, it is now felt that this experiment should be carried out on a larger scale by lnbreasing the number of animals. Should this expanded investigation be pursued, it might be Identified as followsi a. Hepeat. experiment to determine the biological activity of KAYLO when inhaled into the lung harboring a tuberculous infection, which otherwise would regress ani heal; vis.X Does KAYLO stimulite the disease? b. Repeat experiment to determine the biological activity of KAYLO when inhaled into the lung harboring a regressing or quiescent tuberculous focus; wls.t Does KAYLO re activate a focus of lnfeetlon? e. Introduce a new experiment to determine the biological activity of KAYLO Inhaled into the lung upon infootlon; vis.: Does KAYLO predispose the lung to tuberculous infec tion? Asbestos Litig. Saranac Lake 00036 1989 Depos. - Exhibit p.64 U. Owens-Illinois Olaas Company Oetober 30, 1947 Interim Report - KAYLO 4* In addition, it appears important to explore the blologioal activity of hydrous oalelua silicate elthar by inhalation or at least pilot-wise by lnjeetlon methods should it be impossible to generate a sufficient eloud of ataospherio dust because of the character of the material. 3. These recommended investigations can be carried eighteen to two years. It is anticipated that these studies would remove the doubt raised by the results of the original experiments herein reported and would enable us to conclude finally that EAXLO la inert not only in regard to the production of pulmonary injury, but also in relation to associated tuberculosis. - - ?I iJTtU * Arthur J. Vorwald, M.D. Director of fiesearch She Bdward L. Trudeau foundation Asbestos Litig. Saranac Lake 1989 Depos. Exhibit p.66 a 00026c