Document jBqR48JDjx670DgxVmG2mEKNy

CL.' OCL October 30, 1047 OXERS ILLINOIS CLASS COMPANY TOLEDO, OHIO ri Interim Jloport on Animal Inhalation Experiments with EAYLO SUMMARY; The first inhalation experiment with this dust rao begun in February, 1945* Due to a high incidence of fetal pneumonia, another similar experiment was started in August, 1945* Both of these experiments ears designed to test the effect of the dust alone by inhalation in guinea pigs* In August, 1945, also, an experiment vas begun to test the effect of inhaled dust on tuberculous infection in guinea pigs* In October, 1945, a group Of idiite rats was placed in the dust room in order to study the effoet of Xaylo inhalation in this species. It may b* said hero that there is no evidence to suggest that the duat alone when inhaled into the lung produces pulmonary fibrosis in rats in an 18 ^onth period, or in guinea pigs in a 30 month period* The results of the experiment in which dust exposure and tuberculosis were combined are inconclusive and further work should bo done before a definite statement can bs made us to the possible offset of the dust on tuberculous in* faction. All theso experiments are virtually eomploted, although several animals in ths original groups are being kept allvo for more prolonged exposure, INVESTIGATIONSi A, Petrographic and x-ray diffraction analyses of t ^ du3t supplied end used. Analyses vers carried out li. -.icily and periodicallyduring the course of the experiment. The composite of euch analyses reveals the following: Semples from the supply carton Quarts <l (approximately) Chrysotlle end Serpentine IS * 80 largely fibrous, Tho fibers end PLAINTIFF'S EXHIBIT bundles present are usually several uw-6olS?5 millimeters long end seldom are less than 300 u Calotte SJ* 08 006 1100 CLAIMED PRIVILEGED BY OCE1 Pago 2 Oetc .P 30, 1347 Owens Illinois Olaaa ComairrInterim Kooort - EAYL3 Baloaoe conslatod of elsy-llke masses whloh qeve a definite x-roy dlffraetlon pattern not unlike a hydrous oalelua silicate* Samples collected from the ataosohoro of too cnaaaar. Quarts: l Chrrootllo and Serpentine 10 - 15f!-; leas fibrous material than in sample) above; many fibers are shorter than CO |t. Calolto 10 " Balance consisted of olay-like aaoaes which gave a definite x-ray dlffraotlon pattorn not unlike a hydrous calcium silicate. The analyses reveal several points of intorests The low quarts eontont (1'-*) indlcatos that tho amount of crystalline free silica in tho dust is practically negligible & a possible cause of silicosis* Although diatomaoeous earth la one of tho materials from which KAYLO is made, only a trace, probably much less than 1a, is unaltered and appears in the final product in its original form* Nearly all has been changed by the manufacturing process into another chemical ooanound, calcium sili cate. Host of the material oonsioted of clayllko masses which gavo a definite x-ray pattern similar to that of a hydrous calcium silicato* About 10 to 15J* of tho atmospheric dust in tho experimental dust room was chrysotile or serpontino* Both of thoso minerals produce the sane x-ray pattern* From petrographic ex amination of tho original material, it appears that ohrysotllo, which is a fibrous mineral, is preaont in much greater amounts than tho nonflbrous serpentine* In the r. ..oosphorie dust also,, tho fibers still predominate ovor t.*:e nanfibrous aarpontins* 08 006 1101 I e c . Owana l e co p ie d , accordan v " In B ert OSOC SO o t to be xcept In n e t Oer ld .e,r tui toen ,d r C ourt le e . a In te ra c d is trib unde ib e m tia l d or duced t.i~ r n fid e n roduce P ro ca* Co rep Page S* Oaono-Ull ala S1*m Cogonn? claimed PRIVILEGE^ BY OCF October SO, 1047 Intorla Boport - YJ,71.3 Zn tho original material, the fibers woro pc* esent in bundles ofton o everol raillimotors long; not nary were shorter than 300 p* The fibrous portion of tho ataoophorle samplo, hovevor, consisted aostly of shorter bundles and of fibers ^relatively fosr in number, which were generally a bout 10 to 40 p in length* B* Techniques of Investigation. Too species of animals were introduced into the dost chamber assigned to this in vestigation* Tho? included guinea pigs end shite' rets, animals which tho Saranac Labora tory has used routinely in its studies apropos pneumoconiosis Appropriate quantities of the original material supplied by the company eore introduced into the dust generator of the cham ber and pilot runs wore cade to ascertain the character and concentration of tho air-borne 'dust prodnoed* Having established the desired concentration, tho animals were thon introducod and throughout the course of the investigation dust counts were taken periodically* Tho average concentration of the air-borao materiel was 122 million particles per cubic foot of air* This figure varied somovnat from month to month, ranging fron a lover limit of 9d million to on upper of 122 million particlos* This range is not uniquo for this material but has been observed also with most agonts which *o have employed* Zt results no doubt from day to day variations in atmospheric conditions which cannot be precisely oontrolled with our present methods* Nevertheless, we havo elected to continue with such methods, believing that to do otherwise would introduce factors uncommon to aotual industrial exposure* ' The results of the experiments will be presented under tho following headings t 1* II* III* IV* KATLO inhalation alone in guinea pigs* EAYLO inhalation alone in white rata* XAYLO inhalation and simultaneous tuberculosis infection in guinea pigs* Conclusions* 08 006 1102 co p ie d , accordance v . Q-- n * M l**: b* in not to *xc*pt In H urt OSOC SD r . tui toen ,d O rd* *1. t ba r*t rt rmi u o *, fios C q nd l r a I * r r tia l do d * un lfe n e n fid e roduc d e c u d Co rep o r P Page 4* ( Oworts-Illlnols Glass Company October 39# 1047 Interim Heaort - KAYLO X* KftYLQ Inhalation alono In rulnoa plrai claus'd Tho course of this experiment through the first fourteen PR1 ^GED months was discussed in a nummary written bj Dootor Gardner in BY 0CF Uajt 1946* Za this report ho described tho difficulty encountered with scute pnouasnla in the guinea pigs and ccnoluded that it was an accidental epidemic* Uis conoiusion la supported by the faot that subsequently only flour anionic havo died of pnouasnla* lhio occurred, however# only nftor nil apparently nick guinea pigs woro disposed of# the apparently sell pigs isolated# the dust roam thoroughly cleonod# airod and disinfected# and all the cages sterilised* A third and new series of guinea pigs vers intro* ducod after control of tho epldeaio described* Throughout the course of the study# all animals wore exposed continuously to the chamber cloud for oight hours a day# five and ono-holf days a week* Thus xer# a total of thirty-nine pigs have been killed in order to follow the oourso of the pulnonory reaction resulting froa the deposition of inhaled duet in the lung* The period elected for study ranged froa two to thirty months* Over this parlod no doflnlte gross changes oocurrod in the lungs* The trac'r.oobronchial lymph. nodos have enlarged slightly# but re mained soft* Microscopic study of the lungs rovonl gradually Insrsr.sin^ numbers of dust-containing aacrophegoa* Those inaotive "duat cells" are found ehoraotoristioelly in clumps within groups of adjacent air spaces* The intervening alveolar walls aro only slightly thickened by aoouaulatod lyaphooytos# but tnorc is no undue proliferation of tissue histiocytes end connective tissuo* Piorosie is not pro:<ont. Occasional suit!* nncloatod giant colls are soon* In addition# there ore a vory fow elongated segmented brown structure which take on iron stain and aro dearly asbestos!* bodies* Uicroaoopio study of ths tracheobronchial lymph nodes roveals clustors of inaotive "dust colls" similar to those demonstrated by the lung* There is no appuront over development of connective tissue# and again fi* broals is conspicuous by its absence* Results of the ohomioal onalysos carried out on tho lungs of the exposed pigs sacrificed st tho various periods aro re* corded In tho following tablet TABLE I. 08 006 1103- CLAIMED PRIVILEGED BY OCF Togo 5* P.jonQ-111' 'tola Olaoa Conpany Ootobor SO* 1047 Intoria noport - iy,vr/D CHLIilCAL AHALYSIS 07 001ISA PIG LOKOS Animal fluabor lao E39* 2 6 10 11 10 18 3 54 43 56 45 46 67 68 105 108 49 50 2 bo 2 co 4 no* 4 no* C BO* 6 BO* 8 BO* 8 BO* 10 BO* 10 DO* 12 BO* 12 BO* 13 BO* 18 BO* 21 BO* 21 BO* 24 bo* 24 no* Dry tiattep i\oh S SIOp "g CaO * 4*42 4*94 4*47 4*42 4*15 3*74 4*59 4*34 4*50 4*C1 4*06 4*89 4*03 4*90 5*12 5*23 5*02 4*71 0*24 20 22 19 25 20 31 28 *46 41 53 51 SO 86 C4 54 66 60 0*07 0*08 08 OC 09 09 00 08 G8 08 GO *10 03 13 11 11 21 14 14 .11 22 14 03 16 26 09 32 10 11 19 08 * 15 .06 13 06 14 Ash IdrO v> CaO 6*57 4*06 4*87 4*42 5*83 5*35 6*84 5*36 10*12 8*14 10*80 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*10 1.77 2*43 4*30 3*02 4*65 1.60 5*31 6.56 2*03 1*45 1*20 1*72 1 *25 1*23 1.00 1*C0 2.00 2*68 2*75 2*45 3*04 2*20 2*80 3.17 1 *83 C.OC 3.68 .90 2.54 2.92 TABIE Z 08 006 1104 Pago 6. Owens-Illinois Picas Company CLAIMED PRIVILEGED BY OCE October SO, 1947 Intorlm Booort -- KAYI/3 In view of the observations recordod above, certain portlnont oonnento appear to be warranted* The microscopic etudloo of tho lungs and traohoobronehlol lyspb nodes make it obvious that duet was suspended In tho etnosphero of tho ehcabor and that portions of that dust vers lnhalod and rained aoceas into tho lunge of tho animals exposed* this is further substantiated bp the chemical analyses of the lungs which show a gradually increasing amount of the various mineral components of the dust ueod in diroot propor tion to the exposure tines* In the case of 3102, example, tho pigs sacrificed after tan months exposure revealed a tvo to three fold increase* Tho HgD and CaO content also inoreaood hut the increments aro inconsistent and less manifest* -5 i i4 ITS k *? k :i|B fills Tho silica component of tho mineral dust is of such lev concentration that a significant pulmonary roaction would hardly be expootod in oonsoqusnoe of its presence in tho lung* This provod to be the oaso even In thoso animals exposed for a period of twenty-four months and revealing an average total of 126l Si02 of the lung ash* The ohoraoter of the tissue response at thirty months is oomparSblo and it is anticipated that the S102 content will not be appreciably different* Undoubtedly aoae of the dust eelle constituting the mild collulor reaction ob3orved may be in consequonoo of the aorpentins components of tho dust deposited in the lung* This, hovovor, cannot be verified since serpentine is a relatively benign dust and falls to produoa reaction other than mere dust laden macro phages* This interpretation is in accord vlth Doctor Gardner*s oosorvation that the sillcato serpentine is a biologically inert subatanco producing at Cost a mild rcaotlon consisting of email widely scattered local collections of dust laden phagocytes end on occasional giant cell without other cellular reaction and without fibrosis* The presence of a rare curious bedy (asbestools body) indi cates that some of the air-borne fibrous component of tho ehrysotile gained access to tho lung* Eovever, the absonoo of cellular reaction or fibrosis about tho terminal bronchioleo would Indicate that the Inhaled quantity of such fibers was of extremely low intensity and Insufficient to produoe permanent damage* This view la supported by tho petrographic end x-ray diffraction studios which disclosed that a major amount of the chrysotllo component of the dust eonslsted of atrueturos too largo for dispersion into the breathing atmosphere and for easy access into the lung* In oonsequonoo of the experimental studies vlth guinea pigs to d__e_t_e_rmmiTinKe* the biological activity of ZAYL9, it may be tentatively cox eluded that KAYL9 alone falls to produce significant pulmonary damage when inhaled into the lung* 08 006 1105 Pago 7. Oeem Illinois Gluse Corenanv CLAIMED October 30, 1947 Interim Roport - KAYL3 II. ; WLO Inhalation alone in tthlto ratal This apeolea vaa used bocausa toe rat la fcnotm to develop flbroala fhoa quarts inhalation noro rapidly toom other nnlnala* Forty rata aero uaod in this experiment and, aa indloatod by Doctor Cardnor in hla report in Uay, 1946, no dlffloulty with early development of pneunonla vaa experienced* In fact, during the first thirteen months of the experiment no spontaneous deaths occurred* From thlrtoun to eightoen months, about 40,1 of tho onlmla died with terminal bronchiolitis and lobular pneumonia. This la of no significance, hoeovor, because It la soil lawm that laboratory rata beyond the age of one year aro aubjoct to this typo of lesion* Tho .results of exposure to KAYLO voro followed by hilling four rata at oach of tho fallotting periodst six, nine, tsolve, fifteen and eighteen months* The duration of toe experiment was eighteen months and all on Iranis have been klllod* The reaction to t he dust throughout too experiment has oonalsted of focal collections in tho pulmanary alveoli of nacraphagaa containing particles and spicules of dust, is the duration of exposure lncreasod, the only observed chango was an increase In tho number of such fool* Similar dust filled macrophages verb soon in tho traehoobronohlal lymph nodes* Zn no animal vaa there any tissue r eaction of tho aort seen In ailioasis or cabastasls* Chemical analyses of tho lungs of theso rats shoe that the S102 roached the avorage lovol of 16*6 of too lung ash within nine manths and subsequently diminished. .This la difficult to explain, ainoe in the s&ao poriod tho percentage of SlOg in guinoa pigs* lungs exposed simultaneously (see Tablo I) vas increasing* The table of these results follows s TABLE II. OS 006 1106 reproduced o r d le trlb u te d except In eceordnnce C o n fid e n tla lin fo rn a tio n T n o t to bo coplod, vX I* *O:w* m In B urt 030C SO r . O rde *1. t rt o eC*o.u l luhnedrear t d duce B lM P ro Ca f P*0 O* Offcna-llllnola Clpm Coaasnr Oetabor 30, 1947 Intorla Popart - KAYLO KAYL9 ZBHAIATZ9B CLAIMED ^b^OC?0 SILICA DLT2RUU2A7IOKS OP LuIiG TISSUE HI RATS ORLY Ash la i<ry SlOo ir. iry SlOo in Animal E0 Group Tlasuo A'lsaua Ash -* ..-f>______ *- 1012*202 1012-201 17 IV 4.8 4.0 0*663 0*755 14*0 16*1 XXiratlon of IscnoBuro 6 sooths 6 Booths 1012-205 1012-206 IV IV 4.6 5.0 0*B6 0.523 18*5 13.9 9 Booths 9 Booths 1012-202 1012-210 IV IV 4*2 4.4 0*55 0*655 15.2 15*0 12 aonths 12 aonths 1012-215 1012-225 IV IV 4.2 4.2 0*445 0.502 10*5 15.5 15 sooths 15 Booths 1012-225 1012-254 IV IV 5.1 Se9 0*45 0.44 15.7 11*5 18 Booths 18 aonths TA3IE IZ 08 006 1107 CLAIMED PRIVILEGES BY OCF ( Pago 0* Owons-TllInals Glass Company Ootobor 30, 1047 Inter!~ flora ort - XAYLO In ecmaenting upon Part ZZ of the lnvootigr.tioa using elxlto rat* ae the experimental onluole, roforoneo Is node to tho 'consents rooordod under Part Z, which apply. III. KAYLO inhalation and alrsiltanooua tuborculoua infection in ralnoa p1,ts. This experiment va* carried out by the uao of only ono df tho throe methods which have been found valuable in determining tho affect el dust on the courso of tuberculosis. In this case tho were infected with tubercle bacilli by inhalation and im mediately placed in tho dust room. Tnuo, we are tooting the offoct of tha early aeouratlatlon of dust on the early course of tubercu losis. Tho othor two methods available consist in exposing tho animals to either the dust or the infoetlon alone for soverol months, srabsoqusntly exposing them to tho other agent. We may then study the effect of accumulated dust on l he early courso of tupcrculosls, or the offset of dust on healing tuberculous lesions. -S5 IdgL a 2 v VlfHk o o.. o i * -- - fs*1i-3e^as8* o3f 3e S2kS In tho experiment which has boon completed tvor.ty-flvo aninals were usod. Of thoso, five diod within ten months of tho pnounonia which has boon a problem throughout all tho experiments with this dust. Pour others ware klllod S ocauso they were in poor condition, and it was hoped to out down tho incidence of pneumonia by removing them from the room. Thaso four also showed soma* ovldonso of pnounor.is. Of tho remaining sixteen animals, eleven were killed in order to follow the course of tac infection. Jno xas killed at one month cad too were killed at six, nine, twolm, fifteen and eightocn months. Two died at fourteen months, apparently from pro^ossive tuberculosis; these will bo diseussed further oelow. Two in inula died, one at thirteen and one at aeventeon months,' of undetermined cauees, but not from tuberculosis. Of the animals killed to follow the oourse of tho infeotlon, the lesions were Isolated, encapsulated and regressing in all xoopt one of tho two killed at flftoen months. In this animal, lesions ware widespread throughout tne lung, but were associated with tha production of fibrosis. Indicating a tendency to heal. Tha interpretation of the results of this experiment is com pliceted by the aeoessory factor tnat aoao of the animals wore usod for brooding during the experiment. This was dona in an attempt to davalop a otrain of animal* resistant to tno Infeotlon which wen produolng the frequent pneumoniae. f A review of the reoords ahows that oaoh of the two animals which diod of progressing tutoroulous Infootion had boon uaod as brooders. In addition, the animal which was killed et fifteen months and showed widespread lesions had also boon usod far brooding. 08 006 1108 ' ? 10* Owens-Illinois 01 ass Comparer Oetobor 30, 1947 Interim nejart -- KAYLO Four otbor animals used for breeding during tho somo period did not show the extensive dleoeso oean in tho animals Just described* Thus* It aoeras hazardous to attempt to attribute the spread ing of tho tubcroUlous infection entirely either to the effect of the dust or to that of breeding* XV* Conclusions! 1* Xt Booms dear from the results presontod In parts Z and IX above that KAYL* acts as an inort dust uhon inhaled by guinea pigs and ratsf that la* it is phagoeytlzod by macropha^oo* but does not injure them or produce fibrosis' or anything suggestive of either ailioosie or asbestosls* This indicates that the dust rill probably be haraloso on inhalation in nodcrato amounts over prolonged periods by horaal human bolngs* 2. Tno results observed in the experiment concerning tho in fluence of tho dupt on tuberoulous infection are oqulvo cal. Although it is our feeling thot this dust exerts llttlo or no adverse influonco on tuberculosis* tho possibility that it played a pert in sproadlng the infection eennot be excluded* 3* In view of this possibility* further studies concerning this aspect of the problem should be carried out* Xt is imperative that tho original experiment with infection and simultaneous dust exposuro be repeatod in its entirety* One further study with a email series of guinea pigs is non in progress* Xt is designed to reveal tho Influence of the dust on partially healed tuberculosis* However* it is now felt that this experiment should be carried out on a larger scale by increasing tho number of animals* Should this expanded investigation be pursued* it nlct bo Identified cs follows! a* Repeat experiment to determine the biological activity of KATLO vhen lnholod into the lung harboring a tuborculous Infoetiea* vhleh otherwise would regress anu hoolf vix*s Does KAYLO stimulate tho disease? b* Repeat experiment to determine the biological activity of KAYLO when lnhalod into toe lung harboring a rograealng or quiooeont tuberoulous focus; viz*: Does KAYLO re activate a focus of infection? e* Introduce a now experiment to determine the biological activity of KAYLO inhaled into the lung upon lnfootlon; viz* i Does KAYLO predispose tho lung to tuboroulous infoetloa? 08 006 1109 claimed PRIVILEGED BY OCF Pace 11* Oaons-rillnala Glaaa Coaoany October SO, 1947 Intorln Kooort - KAYLO 4* In addition, It appear* Important to oxploro tho biological activity of hydrous cololua silicate oithur by inhalation or at loest pllot-wiso by injection methods should it be iaposslble to corporate a sumc loot cloud of etaasphorlo dust booause of tho charactor of tho aatorial* 5* These rococraondod investigations con be carried out sioultaneCusly and coiqploted in a period of elghtoon months to tool yoors* 7t is ontlolpatod that tueso studLos oouid roaove tho do.;bt ralaod by tha results of the original oxperinonts horoin reportod and could enable us to ooneludo finally that KAYLO is lnort not only in record to tho production of pulawnary injury, but aloo in relation to associated tuberculosis* AJVlIfl ce - \ Arthur J, Vorvald, M*D* Director of Research Cio Edoard L* Trudeau Foundation 08 006 1110