Document 3NNkRGQYL1X9nLrQQZNg6zXG6

daioh Ruinos israiAL foi;d for cancer research PROGRESS REPORT SUPPORT EEIOKD SECOIID TEAR Titlo of Project: EIR'EROIS-SRITAL CARDER PRIRj 76 C The caaacitv of Irhslcd industrial dost to orcduco Amount to,7^6.27 pulmonary cane or in exporincntal anlmls. F7-0H Dccc-r.'ccr 1 Nome of Institution The Saranac laboratory 1952, to November '0 195 7 Church Street, Saranac Infra, Ftr.r Tork_________ Dates covered 'by report Rome of Project Director 0. W. H. Scherers, M.D.,D.Sc. 12/23/53 to 9/15/54 Title of Project `Jiractor Director, Saranac lob. Tears of support for this Frojcot , October 1. 1950 Ecsves ar.d Titles of Technical and Professional to Kcvg;~.bcr ~0.195^ participants in projects: P. C. Pratt, K.D. Pathologist. Total cr.ount of support E J. Urban, 1T*S*E Engineer from DRIC? $73.293 ` H. Roberts, H.E. Engineer(replacing Urban) D. Bailey A.3. Dslahant Petrographer Ressarch Associate Russel Clark Amount requested for next V.bJlace Gay year_____ $26,510 Edvard Gockeler Robert Liddy Technicians lilies Dcody (replacing J. Kaxconi) Jaccb Jacobson Oliver Boula SUKMART STATE'ZixT OF PROGRESS The study is still in progress. A number of rats remain to be sacri ficed in accordance with. ths orie-n&l .acd the cotailed histological and biocbcnical analyses of a cumber.of tba cninalB already sacrificed era yet to bo completed. Preliminary observations include the following: (a) Various typco of neoplsEmta have been provoked in rats by tho inhalation of beryllium-ovlpuato oust for continuous periods of 6 norvths. (b) Theso neoplcsmata include papillary adenocr.rcinena, raucous forming edeaoesreiuoxa and squamous coll carcincna. (c) Ab cany cancers resulted from the current exposure schedule lasting 6 ecutba only,as were provoked after continuous * oxposura for 14 months in a previous comparable experiment on rats. ** -1- DEFENDANT'S EXHIBIT JLao C'12751J Donon Runyon Memorial Fund Septeaber 25, 1954 lint of the Project: The statistical study, of the growth characteristics of transplanted lung cancers. A. systematic exploration, of the Eatabollic activity of the tunors end the reaction of the host theroto. A definitive and progressive investigation of cc- and anti-carcino genic agents. ' Freon si on of Facilities? The beryllium dusting phase of the investigation vill he ccnducted bry neons of another research grant, but the dust chambers built `by neons of earlier Damon Runyon grants will continue to bs used with aone raodiflcations. Specific instrumental acquisitions vill involve the purchase of electrophoretic apparatus and various ricroscopo accessories. Changes in personnel couprise the following: (a) Addition of two pathologist: to handle the clinical pathology phara (Olga DotrogorsUi, H.D. and J. Schmidt). (b) One biochemist (L. Scheel, Fi.D.), and one additional technician (F. Gordon). (c) One statistician. (S. Fascer, M.2.,.F.) in view of the increased significance of this phase. ilr. Kaszer has already been partici pating in tho preceding study. (d) One senior histologist (F. CreedonX with tsclmical assistant (E.Fear). (e) One. additional ehiruvl caretaker (R. Vhitelav). (f) One part-fino. typist (G-. Nesbitt). SUKIARY OF PROPOSED 5JDGF? FOR NrTT YEAR Personnol- u Percanent equipment Consucablc eupplies Other expenses Institutional overhead Total budget for year 12,500 1,300 2,500 8,236 1,924 26,510 Ftercnrks: A small suroi ia accuruie.ti in the existing budget tho status of which wi be reported on Nov.30, Signature of Project Director Darien P.unyon Kumcrial Fuad Septonber 25} 1954 . Progress Report on PROJECT DRIP.-75C SKVIR0:2-ii3TAL CANCER: THE CAPACITY OF IDHALED IHDU5TPJAL DOST TO PRODUCE PULMOHART CANCER III EIPERIKaiTAL >iIXMAL5 I. INT. RODUCTION The above current experiment at Tne Screnac Laboratory vas designedto evaluate various methods for the experimental induction of pulmonary neoplasm through exposure to .atmospheric dusts. The possibility t at industrial dusts of various compositions taa7 cause puixenary neoplasm in . industrial vorkers baa for cany years received considerable attention in medical and legal circles. Tissue specimens of numerous industrial vorkers vho developed pulmonary carcinoma are constantly submitted to The Saranac Laboratory for reviev, and consultation about such cr.sea is often eought in respact to claims for compensation based on the alleged carcino genic influence of various dusts. This etate of affairs creates a great-and need for definitive evidence concerning carcinogenesis in industry. This interim report summarizes the experimental 6tudy of animals vcich. has nov been in progress' for 16 tenths. Tnia study vas undertaken after completion of a previous study in vhich it vae found that quartz dust and iron-oride dust did not influence the incidence of pulmonary tumor in mice. That earlier study involved nice of two strains, strain C aac strain C57 black, having a high and lov eponteneoua pulmonary tumor incidence,reopactively, and on completion of tho experiment it vas noesiole to state to the Damon Runyon Memorial Fund,in a report dated October 20, 1952, .that "Meitbsr of these dusts is capable of eliciting tumors in nice of a strain highly susceptible to tumor dsvelopmsnt or. of initiating tuaor formation in nice of a atfsia of lov spontaneous tumor lnciuenco". In the proposal to tho Damon Runyon Fbnd Committee dated October 29, 1952, in vhich the present experiment was outlined, it vas stated that if . either Rucrtx dust or iron-oxide dust dotslnfluenca the incidence of pul monary carcinoma in man, as eoce observers suspect, tho souse is valueless for investigation of the relative potency, aeche-niem of action, or pro phylaxis of that effect. It vas reasoned further that efforts to identify an experimental technic -for tho investigation of these aspects of pulncuary carcinogenesis should bo continued. Discussion was introduced concerning various pocsiblc approaches to the -problem and it vas concluded that the tent avenue of approach vould be cno involving "preliminary preparation:, of cairaals -with a standard cnrcincgenic product fclloved b'K- their expoouro to the substance to be tested". - 4- 012754 Damon Runyon Utxorial fund September .25, 1954. Project DRIR-7'C Tie Btudy nov undervny involves tills t7pe of approach to the experi mental study of pulmonary carcinogenesis. It has been found that rats exposed to beryllium. compounds for 10 to 15 months develop true pulmonary carcinomas of the alveolar cell adenocarcinoma type. Since theso lesions are true anaplastic malignant tumors of a type sometimes seen in human lungs, they seen mors promising: for the study of pulmonary carcinogenesis than the peculiar nom-ancplastic, usually banign tumors occurring spontane ously in certain strains of mice. The use of animals prepared by exposure to beryllium for ths study of other agents as carcinogens is analogous vith the fact that in using tunar-susceptible strains of nice, a "neoplasia-genie0 factor is present, vfaether genetic or infectious in nature, so that any experimental agent, even in the mouse, is' being studied as a co-carcinogen. This is in further conformity vith the belief of cost observers that multiple factors probably combine in carcinogenesis in man so thet any single iraplicr.tsd agent should probably bo regarded as acting as a cc-carcinogea vith one or mors other agents. Beryllium has the added advantage that it undergoes no metabo lie change in animal tissue, so that as a co-carcincgen its effects are precisely definable. II. HTTHOD A poriod of 6 months exposure to beryllium compounds has been found to be a sufficient stimulus for the subsequent development of pulmonary carcinoma, during a period from about 5 to 10 months after the cessation of the'exposure. The current experiment is designed to reveal vhethor or not quarts or iron-oxide dusts introduced subseouently to the phase of beryllium exposure vill alter the latent period or the incidence of such tumors. The experiment vas started vith a total of 200 albino rats vhich vers exposed for 6 months in too special dust chambers to an aerosol suspension of berylliun sulphate. An atmospheric concentration of about 12 nicrogroas of BoSO, (equivalent to 1 micrcgram Us) par cu. ft. of air vs maintained and the dust was disseminated for 8 hours per dey, 5} days'par vaek- It vas expected that during this period of- exposure to berylliun sulnhato seme rata yould die in addition to those vbich vere to ba sacrificed. The total number of deaths from all- causes-vas 65, somevhat more than bad been anticipated. These deaths occurred in tvo episodes; the first shortly after introduction to the dust * chambers, and the second lovards the end of the dusting pheso. These remain ing 135 animals" vere divided into three groups of 45 anihals each. Tvo of these groups vere exposed in separate dust chcmbars to quart! dust and to iron-oxide dust in a concentration of approximately 300 nillion particles par cubic foot of air. The third group vao reserved for residence in normal air as a control. Animals from cr.cli of the above groups bsve_ been end are being sacrificed in samples of four at intervals of three raonthe freu the end of oxposure to beryllium. It vas originally planned that if it oppearsd at any sacrificing -5- . N C12755 Demon ftunyon Memorial Fuad September 25, 1954 Project DRIR-75C pei'iod that therevere differences between the incidence of carcinoma among the groups, a larger number of animals vould be sacrificed. Studies of the tissues of the tnijaals included gross end microscopic examination of the lungs, lymph nodes, spleon, liver, and kidneys. It vras planned that if at any tins there aro differences in the inci dence of pulmonary tumors, the lungs of the experimental animals and of ths controls vould be analyzed for beryllium.. This procedure had to he carried out bacauee of the possibility thet the inhalation of quartz or of ironoxide dust night interfere vith the excretion of previously accumulated herylliua. If excessive retention chould thus occur, it vould interfere vith the interpretation of the experiment and vould necessitate a revision of the technic and repitition of the overall study to ensure that the tissue content of beryllium vill bo comparable in ec.ch group. One important pre caution has been taken against this possibility. The animals vera kept in normal air for one week between the end of beryllium sulphate exposure and the beginning of the quartz dust and iron-oxide dust exposure, for the reason that during a period of bit one veek the greeter portion of therecently deposited beryllium, vould be eliminated vhile in subsequent months but little more beryllium vould be excreted. is no indication hod arisen at the tine of the 9 months sacrificing subsequently to the cessation of beryllium dusting that either tan quarts or the iron-oxide exposure vas exerting any material influence-upon the induction period of carcinogenesis or the numbers of pulmonary tumors being produced, and as the aaimalo had already been under -experiment for a total of 15 months and could not be continued cn this status ir.dafinitely, it vas decided to sacrifice half of the remainder in each group. This vaa the only material departure from the course of ths experiment as originally projected. The .observations cado at the various sacrificial periods are assembled in Tables' I to IV and in Figure Ii-' It trust be recorded that this interim report is incomplete insofar that it has not as yet been possible to process all microscopic preparations or biochemical analyses on the tissues of the recent large group of sacrificed animals. Therefore, the results presented, especially* as they pertain to these latter animals, are based on gvosa observation only end mist 'ha considered to be tentative. Toe inferences vhich nay bs drawn froa the tables ere that neither quarts nor iron-oxide, inhalation lias effected the latent period or-overall incidence of pulmonary carcinomata. Comprehensive microscopic study of oil the tissues vill allow further evaluation of tha result- in tenon of certain, additional criteria, such os 6ize, type, and location of tumors. It has already, however, been noted that various types of tucor havo occurred, including papillary adenocarcinoma, mucous-forming adenocarcinoma, and Bquenous cell corcinoaa. Tho ratios in vhich theca tumor types occur in the varioue animal sub-groups of this experiment yet remain to bo determined. -6- DcjaoQ Runyon ".enorie-L t uau. 9 September 25, 1954 Project DR1R-75C Table I ehows the macroscopic observations in each of the groups. The vertical "Duration of Exposure" column at the left is self-explanatory. The colum narked "Humber of Animals T>~ mined" records the number of animals examined at appropriate time intervals from each of the types of dust exposure, namely, VeryIlium sulphate followed by quartz, beryllium sulphate followed by iron oxide, ond beryllium sulphate followed by normal air. It shows also the totals of animals from ell groups examined at each interval. The column narked "Humber of Tumors Seen." presents figures for the number of tumors observed in each animal examined. Thus, in the second line (femanths BeSO^ and three months time lapse) there were 6, 4, and 3 animals obaorved in the quartz, iron, and normal air groups, respectively; and under "Kumbor of Tumors" there are groups of 6, 4, and 3 zeros to show that no tumors were observed in these animals at that time. Table II summarizes the pooled data, from Table I and reveals a remarkably sharp Increase in the rate of incidence of tumors between nine and tea months. Presumably this is an indication that following a six-month exposure to beryllium sulphate dust the induction period before the devel opment of tumors is comparatively consistent and is approximately nine months in duration. The peak onset of tumor production, therefore, commences approximately 15 months from the date of onset of beryllium sulphate duet exposure. It is of interest to note that in an earlier experiment in which exposure to beryllium was continuous the peak, rate of appearance of pulmonary carcinomata also occurred at about 14 months from the date of commencement of the dusting experiment. This phenomenon is graphically emphasized in Figuro I. Table III p'resents the comparative data by type of exposure for the occurrence of tumors in the groups of animals killed after 10 months* exposure to quartz, to iron-oxide, and to normal air , following the prep aratory exposure to beryllium sulphate dust. The data axe broken down aa indicated in the left hand column for the purpose of other comparisons which are presented in Table 17. In Table III the important figures to note are the totals. In the column beaded "Kuaber of Tumors per Animal" it is apparent that exposure of tha_rats to quartz or iron-oxide dust after the preparatory exposure to beryllium sulphate dust has not altered the incidence of tumors to a material degree. 7 Damon Runymn J*anorial Fuad Septenbr 25, 1554 Project DRIR-75C Table 17 presents the results of combining the data garnered thus far In this experiment in various vnys other than by type of exposure. Thus, two strains of animals were used and the first set of comparative data in Table 17 shows that there is a slight, .but lnmaterial difference in tumor incidence between the two strains. The original 200 rata were exposed to the beryllium dust in two separate dusting chambers, and the beryllium dust in two separate dusting chambers, end the second set of data compares the incidence of the resultant tumors. Again there is no significant difference. The third set of data compares the incidence of tumors in male and female rata and ones more no significant difference is' apparent. In & previous comparable otudy on mica conducted by means of a Damon Runyon grant, neither quartz nor iron-oxide dust appeared to influenca the incidence of spontaneous pulmonary tumors. Evidence now exists that at least for rodents of these two genera, neither quartz nor lron-roxide dusts act as co-carcinogens with beryllium sulphate or any other primary carcinogen. In view of the large numbers of human beings who are constantly being exposed to either quartz or iron dusts of various kinds, this oboervstion nay have a certain measure of significance. III. TRANSPLANTATION OF LUNG TUMORS A major result of this series of studies on environmental cancer has been to demonstrate that beryllium dust is a- reliable experimental tool by means of which lung cancer can be provoked'in rsts. It had not yet been established whether the presence of the inhaled beryllium was necessary for the survival of the neoplaam, i.e., whether the cancer cells used the . beryllium during some or other phase of abnormal metabolism or whether the new growth could continue in the absence of the beryllium, i.e., ' whether eome genetic change had been effected,chcnging the cancerous cell to a parasite rather than merely maintaining it ae a diseased cell. This point could not be proved readily on the tumors in the animal lungs because the progressive expansion of the tumor and the occurrence of multiple tumors rapidly killed the rats. The extension of the biochemical study of the tumor could only be pursued therefore if the tumor could be induced to grov more slowly or in another accessible tissue. Nor would any attempt at thcrapeusio prove meaningful until the experi mental cancers could be proved by histological methods prior to the institu tion of any proposed method of treatment. This could not be attempted in animals such as the rat unless the tumors could be transplanted. Fortunately, it was discovered that the beryllium-induced ncoplssmata possecs such biological vigor that they readily survive on subcutaneous % -H- C 127 Jxsjleatar'' ^5, '1954 Project DRIP.-75C YISIRZ I INCIDENCE OF TUMORS AT VARIOUS INTERVALS AFTER' EXPOSURE FOR 6 MONTHS TO EeS04 (froa TABLE II) X - DENOTES 6 MONTHS OF EeSO^ r\ \y >12730 'O c Damon Runyon M e m o ria l Fund September 25# 1954 P ro je c t DIUIU75C I -< o Qa n t Oa ~r-i |U 3 93 Uo 50=*2 2tm r\0 uo Rg t & u J8 II O O v-v O' ro\ 3a ? 88 1* 'O 2H 'HS iO-H' >"*> o. 3 sO ffl c^c. O3 (* & *0 c"i *0 *o V\ iH * ` *< * t- H *4 taso* f 'fl t* I! *".8 2U S x) C\ 3 a fci t. Q, b> o 3 H 4 O e O 0:0 sos s'* 5 u a i* 0 bO* A >fl O' O . r"` ao 3 2 o 55 3 E- 03 a 3a vO Q a ::"a3l. u A otv' pc f* X H CO z> Cl v-t v_> \ Daman Runyon F.cmnai. t una Septenbor 25, 1954 o Froject DRIIU75C TIZLE II SM1ARI OF DAT! OF TAELS I (All observations pooled at each tine interval) Duration of Exposure To BeS04 months 6 6 16 6 6 To * or to Quarts months 0 3 6 9 10 . . Kunber of Aninals examined Ifumber of Tumors (total) t 6 13 16 19 -43 - e ,/ 0 0 .5 9 130 Hunber of. Tumors por 6nip.nl 0 0 . 0.31 0.47 2 3.02 # O C127 N Damon Runyon lleionoi kuna September 25, 1954 Project DRIR-75C TABLE III COMPARISON OF TUMOR INCIDEICE AT 10 MONTHS ONLT IN RATS OF EACH OF THE GROUPS EXPOSED TO BgSO,, THE! TO CJJARTZ BeS07, THEN TO Fe20Be904, THEf TO HOHMAL^AIR. Data of iroomre Strain of sen Chamber eninal used Humber of animals examined Humber of animals vith tumors Runbor of tumors _ Kunber of tumors Dar eniral Experiment #1236 BoS04 for 6 months fo Lloved by quartz for 10 months Shaman F #10 Shaman F 11 32. 39 2 1Q She man H 11 1 0 0 Shaman H 10 3 3 7 Sar. Lab. a F 10-11 2 2 8 - TotalJ 11 10 3L 3.08 .Experiment 1237 BaSO^ for 6 months followed by Fe203 for 10 months She man F #10 4 4 16 She man F 11 2 2. 3 Sheman H '10 6 26 S&r* F 10-11 .5 3 6 Total1 17 15 fi 3.00 Experiment.L238 BoS04 for 6 months f<olloved by normal cir for 10 month Sheman T #10 Sheman F 11- 4 3 3 .3 ` 9 ' 10 Shaman H S&r. Lab. F 10 5 10-11 3 TotalJ_______ ______ 5. 3 U 19 5 43 2.37 O 03,276; {s ) L og*cros o ts v\ CQ I** d| o S S'8 Q CO & t U * u c e 0 Arj Wa| | B a S 1 m 0 *"1 H 00 u 5 0 cl M a1 Ee p1 fc *: H1 s; * QW SB wj M Q PQ S a. " 8 8 | 8* 3- s* 4 V* O ^ Iq "O o O* 0 4* S e3 0 H 1st JO ^ 0.. *1 Si 12 . t & M' -P O. Or M 3 a 0 q0 "s K ^* EM 8 r*f ta ** E ca no. O3 Cl 2 2 0 u. w u JS 1 a 18 o' fae a . B 8 0c 5 tV04| 'U $ 2? ft $^ *A >0 'r vO o CM 0 C*> ft sr s' ^ .0* cm A. 4U d s -i Ou I0 0 ca 0 * M r\ *> r- r% cm cv CM CO CM 'O *c\ r- ss 2? Sf vO w\ CM CM O so r-1 H * 55 CO vr\ CM un to ft CM <n ff *a CM O H *a C\ vO --s e*- a a rH-l a Oft . U* k 1a 1a n re-f HE ^ . > ac4 31 X fx T o ta l nunbar o f anim als examlnod 43 T o ta l number o f tumors 123 Tumor per anim al (o v e ra ll avarago) 2.93 cc t cv <_ N September 1, 195A HElORIKDUHl Does Silicosis Protect Against Carcinoma of the Lung? Referencei Zcntr. Allchea. Pethol. u; Pathol. Anat., Yol. 89, p 187, 195*. S. Soorleia At the University of Vuroburg, 353 cases of silicosis vere determined frc-a 14,571 necropsies performed during tne period 1930 to 1947. Thera was evidence of bronchial cr.reinoxa in only 7 of these cases. This gives an incidence of carcinoma a fifth tc a si.vth of that which could have teen calculated had silicosis been v'.tuout effect on the lung in respect of carci noma. Moreover, the i;reotest Incidence cf carcinoma vss a,aongrt tne cases vlth. tne least amount of silicosis. Thus, thore were 5 cases of bronchial carcinoma amongst 9 subjects with eligat silicosis, and 2 acongt loj with moderate silicosis, and none snongst 137 with severe silicosis. The inference is c.\de thut silica or eilitic acid, instead of prs-disposing to tho dcvelopssnt of bronchial carciacna, decidedly protects against it. ec: fa non IUmyoa fund exa^rinfent CVHSsgn iMnocvn >cmoor iVHYVIVC rYSOUYlOUA % o* ot 0i<s o% o`cw OCWVViHCWl Ok kfOfOfOC OCkrvXHfHA Ok kHA^icro OCL'FtfiHCWl Ok !OCHCh WOH XHC CriMlCYr YMO EXbEHtWGMXYr ?mOA Ok. tnrwoUVMA D12EV2C 1HE AtSnDEVn hOnUDVllOH . A < 0*7 x '/*1