Document OJkBb7Z0kNb5oBbaBBwjBg60K

PRODUCED' TO .EAGLE-PICHER JN LITIGATION DEC 1984 The Saranac Laboratory Of THE EDWaIO t. TRUOEaU POUNOaTiON Saranac LaKE. N.Y. Aithui J. Vomwal.0. M O CT " October 50^, 1947 ' O^BJfS ILLINOIS GLASS COMPANY " TOLEDO, OHIO r i i [ a. Interim Report on Animal Inhalatl Experiments with EA2YL SUMMARY: The first inhalation nt with tAs dust was begun in February, 11 to a high Incidence ol fatal pneumonia, another a Lai _ ment was started in August, 1945* Both of thes^/^&perix a were designed to test the effect of the dust alo tion in guinea pigs In August, 1945, also, an exp begun to test the effect of inhaled dust on tubereulous tion in guinea pigs* In October, 1945, a group of white as placed in the o^st room in order to study the^off^oV of Kx alation in this species* It say be sai ere is no evidence to suggest that the dust.alon into the lung produces pulmonary fibrosis in rats i period, or in guinea pigs in a SO month period* the experiment in which dust . exposure and tubere combined are inconclusive and further work^Shpuld fore a definite statement can be made as to/vrt'poasible of the dust on tubereulous in faction* (k11 these experiments are virtually completed, although several aMmals In thft original groups are betng kept alive for more proll INVESTIGATION! Petrographic and x-ray diffraction analyses of the dust supplied and used* Analyses were carried out initially and periodically during the course of the experiment. The eoaposlte of such analyses "`reveals the following: Samples from the supply carton Quartz <ljC (approximately) Chrysotile and Serpentine 15. - 20; largely fibrous* The fibers and bundles present are usually aeveral millimeters long and seldom are less/'OOlri'DS than 500 p . < . tC* . O'--* Fag 2 Over. Illinois Glass Company October 30, 1947 Interim Report - &AYL3 Balance consisted of clay-like masses which gave a definite x-ray diffrac tion pattern not unlike a hydrous PRODUCED TO - calcium silicate. . EAGLE-PICHER . IN UTlGATlON Samples collected from the atmosphere ~ of the chamber. ~ - DEC 1984 _ -- Quarts 1$ _Chry80tiie *gd~Syrp~c~ntlne---0 -- 15;-- ''"'let's'fibrous-'material--tha--ia-- _ sample above; many fibers are shorter than SO Calelte 10 Balance consisted of elsy-like masses whleh gave a definite x-ray diffrac tion pattern not unlike a hydrous calcium silicate. The analyses reveal several points of interest; The low quarts eontent (lf) indicates that the amount of crystalline free silica in the dust la practically negligible as a possible cause of silicosis* . Although diatosaeeous earth la one of the material* from which KAYLO Is made, only a trace, probably such less than l, is unaltered and appears in the final, product la Its original fora. early all' has been changed by the manufacturing process Into another chemical compound, calcium sili cate* Kost of the material consisted of cltyllke Basset which gave a definite x-ray pattern similar to that of a hydrous calcium silleate* About 10 to 15 of the atmospheric dust in the experimental dust rooa was chrysotile or serpentine* Both of these minerals produce the same x-ray pattern* Proa petrographic ex , animation of the original material, it appears that ehrysotlle, which Is a fibrous mineral, is present In such greater amounts than*the non- flbrous serpentine* In the atmospheric dust also, the fibers still predominate over the non- flbrous serpentine* " Page 5 Q-jens-Illlsols Glass Conoanv October 30, 1947 Interim fieoort - 2UVL3 - Ia the original material, Lne fibers fere pr esent la bundles often x everal allli- - aetara long; not many vert shorter than 300 ji The fibrous portion of the atmospheric sample^ however, consisted mostly of shorter bundles and of fibers, relatively few la number, whieb veP6 generally about 10 to 40 \u In length, produced to ^gle-P{cher B* Techniques of Investigation* Two species of animals were introduced Into the dust chamber assigned to this In vestigation* They Included guinea pigs and white rats, animals which the Saranac Labora tory has used routinely in its studies apropos pneumoconiosis* Appropriate quantities of the original material supplied by the company's ere introduced into the dust, generator of the cham ber and pilot runs were made to ascertain the character and concentration of tne air-borne dust produced* Having established the desire, concentration, the animals were then introduced and throughout the course of the Investigation dust counts ware taken periodically* The average concentration of the air-borne material was 122 million partielea per eubic foot of air* This figure varied somewhat from month to month, ranging from a lover limit of . 99 million to an upper of 122 million particles* This rang* is not unique for this material but - has been observed also with most agents which we have employed* Zt results no doubt from day to day variations in atmospheric conditions which eannot be precisely controlled with our present methods* Nevertheless, we have elected to continue with such methods, believing that to do otherwise would Introduce factors uncommon to actual Industrial exposure* The results of the experiments will be presented under the following headings: I tXXLO inhalation alone in guinea pigs* zz EAYLO inhalation alone In white rats* XII XAYLO inhalation and simultaneous tuberculosis infection in guinea ( pigs* 17 Conclusions* .~ O \ V ^ '' J ' EAGLi-PlCHSR ]`x LJTiGATiON DC iSS4 Tags 4* . Orans-Illlnois Pitas CoaoariT October 30, 1347 Interim Haport - XAYLO I. KATLO inhalation alone in pulnea pica: . m\ -'.The eourse of this experiment through the first fourteen months tu discussed in a summary written by Doctor Gardner In Kay;,1946* In this report he described the difficulty encountered pru^naonia In the guinea piss and concluded that It was an-aceiden.t.al _epidemic-^ hu enneniiiAn up<juwert by the ftrt that subsequently only f^Mr~*TiTff1TT~ff~~w^TTt -rtfed *>f ppwninnl.e-- TM.f_ occurred, however, only after all apparently sick guinea pigs were disposed of, the apparently well pigs Isolated, the dust rooa thoroughly cleaned, aired and disinfected, and all the cages sterilised. A third and new series of guinea pigs were intro duced after control of the epidemic described* Throughout the course of the study, all animals were exposed continuously to the ehaaber 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 course of the pulmonary reaction resulting froa the deposition of inhaled dust In the lung* The period elected for study ranged froc two to thirty aonths* Over this period no definite gross changes occurred in the lungs* The tracheobronchial lymph nodes hare enlarged slightly, but re sained soft* Microscopic study of the lungs reveal gradually Increasing numbers of dust-contalnlng macrophages* These inactive ''dust cells* are found charseterlstleslly 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 tlssua histiocytes and connective tissue* Fibrosis is not present*. Occasional multinucleated giant cells are seen* In addition, there are a very few elongated segmented brown structure which take an iron stain and are clearly asbestosls bodies* Microscopic study of the tracheobronchial lyaph nodes reveals clusters of inactive *dust cells* similar to those demonstrated by the lung* There is no apparent over development of connective tissue, and again fi brosis is conspicuous by its absence* * Results of the ehemleal analyses carried out on the lungs of the exposed pigs sacrificed at the various periods are re corded in the following table: TABLE I. 5 eagle-picher JN LITIGATION DC ^ . Par 5 _ { frwens-Illlncis Claae Company October 50, 194? Interin Report - P.AYL0 l Anisral Nucber CPIKICAL ANALYSIS 0? GUI 1*--A PIG LB>T,S Time Exp _____________ Err Matter___________ Asn sibo go CaO ------- ---------- -- --------- ________ Ash sloi * E?rO CaO < 2 6 10 XI 16 16 23 54 45 56 ' 45 45 67 63 105 106 49 50 2 CO 2 CO 4 CO 4 CO- 6 CO 6 CO 3 cc e CO 10 EQ 10 CO 12 CO 12 r.c IS r.o 13 r.o * 21 CO 21 DO 24 CD 24 CO 4-42 4,94 4,47 4,42 4.15 5.74 4.59 4.54 4,53 4,61 4.86 4.93 4.33 4 .90 5 *12 5.23 5.02 4.71 0.24 .20 0.07 06 0.06 . .00 .22 .00 .0= .19 .03 .06 .25 .OS .05 .20 .03 .10 .51 .06 .1? .26 .11 .11 4C .21 .14 41 .14 .11 .55 .22 .14 .51 0o . *1C .52 .26 ' .05 66 52 .10 .64 .54 .11 .03 .1? .15 66 06 15 60 .08 .14 5 .57 4.06 4.87 4.42 s.eo 5.36 6.84 5 .56 10.12 6.14 10 .SO 10.25 12.27 17 .65 12.54 10.30 12.70 12 .52 1.59 1.62 1.99 1.89 2.00 2.IS 1.77 if 4 .50 3.02 4.65 1.60 5.31 C .56 2.03 1.45 1.20 1.72 1.85 1.23 1.99 1 .65 2.00 2.66 2.75 2 .43 3.04 2.20 2.30 2.17 1.35 2 .06 5.68 2.90 2.54 2 .92 ( (a TAB12 I Page 6. * Qwacs-Illlnolg Class eagle-pjcher IN LITIG DEC Company October 30, 19-47 Interim Report - *AYLO In rle# of the observations recorded above, certain pertinent consents appear to be warranted# The microscopic studies of the lungs and tracheobronchial lysph nodes make it obTious^that dust wad suspended In the atmosphere of the chamber and that portions of\hat dust wars Inhaled and gained access. Into the lungs of the aixloala exposed* This Is further substantiated by the chemical analyses of the lungs which show a gradually Increasing amount of --t*if ^Kponents of the dust used In direct propor- --the--axpaaure_.tin.es._ _Xr__the_case of "SiOg, example, the------- pigs sacrificed after ten months exposure reTeTttvd* "* two*to tnree-- - - fold increase# The HgO and CaO content also increased but the increments are inconsistent and less manifest# The silica component of the mineral dust is of such low concentration that a significant pulmonary reaction would hardly be expeeted in consequence of its presence in the lung# This proved to be the ease even in those animals exposed for a period of twenty-four months alnd revealing an average total of 12.61^ SlOg of the lung ash# ?he character of the tissue response at thirty months is comparable and it is anticipated that the SiOg 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, however, cannot be verified sinee serpentine is a relatively benign dust e4 falls to produce reaction other than mere dust laden macro phages# This interpretation Is in accord with Doctor Gardner'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 occasional giant cell without other cellular reaction and without fibrosis# . . The presence of a rare curious body (asbestosis body) Indi cates that some of the air-borne fibrous component of the ehryeo- tile gained access to the lung# However, the~ab*enee~of cellular reaction or fibrosis-about the terminal bronchioles.would indicate that^the inhAled,,,quantity of such fibers was-of extremely low Intensity and insufficient to produce permanent damage# This view is'aupported by the petrographic and x-ray diffraction studies which disclosed that a-major amount of the ehrysotlle component of the dust consisted of structures too large for dispersion ipto the breathing atmosphere and for easy- access Into the lung. In consequence of the experimental studies with guinea pigs to determine the biological activity of KAYLO, it may be tentatively eluded that XAY10 alone falls to produce significant-pulmonary carnage . when Inhaled into the lung# . 5 ?&8 7 Oftcna-UllPols Class EAGLE-PICHER IN LITIGATION' DEC 1384 Coraoahv October 30, 1947 Interim Report - KJkVLD II. KAYLO inhalation alone la -white rata: This apeeiea vas used because the rat is known to derelop - j fibrosis from quarts Inhalation more rapidly than other animals. Forty rats were used in this experiment and, as indicated by Doctor Gardner Jja his report in Kay, 1946, no difficulty with yearly derelopaent of pneumonia was experienced* In fact, during ~ the first thirteen months of the experiment no spontaneous deaths occurred* Froa thirteen to eighteen asnt&s, about 40 of the---- anlnals "STed "with" TeralnAl bronchiolitis- and- lobular 7pnumonia- - Tnis is of no significance, however, because it is well known that laboratory rats beyond the age of one year are subject to this type of lesion* ' The results of exposure to KAYLO were followed by killing four rats at each of the following periods: six, nine, twelve, fifteen and eighteen months* The duration of the experiment vas eighteen aonths and all animals have been killed^ The reaction to the dust .throughout the experiment has 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 change vas an increase in the number of such foci* Slallar dust filled maerophages vere seen In the tracheobronchial lymph . nodes* In no animal vas there any tissue r action of the sort seen in silicosis or aebestosls* Chemical analyses of the lungs of these rats show that the S102 reached the average level of 16*6^ of the lung ash within nine aonths and subsequently diminished. This is difficult to explain, since in the same period the percentage of SIO2 In guinea pigs* lungs exposed simultaneously (sea Table I) vas increasing* The table of these results follows* TABLE IX* L 5 ~SG Pag* s. frrer.s-Illinois Glass Company EAGLE-P^fitrt rQATm October 30, 1947 Interla fieoort - TJOl LO ~ EAYLO XKKALATIOK - 3$ .* SILICA PETERHIJJATIOXS OP LUSS TISSUE IX RATS ORLY K Animal Ho* .Ash in Lry Croup Tissue S" SiOp in Ory SiOo in iissua --------ksh--------- nf 5. 4-------........ --Exposure 10X2-202 1012-201 IV IV 4.8 4.6 0.663 0.735 14.0 16.1 6 months 6 months 1012-205 1012-206 IV IV 4.6 3.8 0.36 0.523 13.3 13.9 9 months 9 months 1012-209 1012-210 IV IV . 4.2 4.4 0.55 0.655 15.2 15.0 1Z months 12 months 1012-215 1012-223 IV IV 4.2 4.2 0.443 0.562 10.5 13.3 IS months 15 months 1012-225 1012-234 IV IV 3.1 3.9 0.43 0.44 13.7 11.3 13 months 18 months TABLE IX - "* K.j 3 r- *** /t- PRODUCED TO EAGLE-PICHER , IN LITIGATION Page 9. Owens-Illinois Glass Company October 30, 1947 Interln Keoort - VLO , In commenting upon ?art II of the Investigation using white rant as the experimental animals, reference is oade to*the consents recorded under Part I, which apply. ` -IH. gAYTQ isolation sad simultaneous tuberculous infection ' in guinea pigs. --- - ------------ Tnis experiment was earrled out by the use of only one of the three methods which have been found valuable in determining the effect of dust on the course of tuberculosis. In this ease the animals were Infected with tubercle bacilli by Inhalation and im mediately placed 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 dust or the infection alone for several months, subsequently exposing then to the other agent. r?e nay then study the effect of a ceumulated dust on t he eaxly 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 which has been a problem throughout all the experiments with this dust. Pour others were killedbeeause they were In poor condition, and it was hoped to cut down the incidence of pneumonia by removing them from the room. These four also showed some evidence of pneumon Of the remaining sixteen animals, eleven were killed in order to follow the course of the Infection. One was killed at one month and two were killed at slz, nine, twelve, fifteen and eighteen months. Two died at fourteen months, apparently from progressive tuberculosis; these will be discussed further below. Two animals died, one st thirteen and one at seventeen months, of undetermined causes, but not from tuberculosis* Of the animals killed to follow the course of the Infection, 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, but were associated with the production of fibrosis, indicating a tendency to heal. ^ The interpretation of the reaults of tIs experiment is com plicated by the accessory factor that some of the animals were used for breeding during the experiment. This was done in an attempt to develop a strain of animals resistant to the Infection which was producing the frequent pneumonias* X review of the records shows that each of the two animals which died of progressing tuberculous infection had been used as breeders. In addition, the animal which was killed at fifteen months and showed widespread lotions bad also been used for breeding -n r- ' an