Document wD6pYVmEN1Nzydz2amKw9aYd

FILE NAME: Oil Industry and American Petroleum Institute (API) DATE: 1954 July-Dee DOC#: API033 DOCUMENT DESCRIPTION: Letters, Memos, Reports, Financial Reports & Other Relevant Documents from July-Dee, 1954 BUDGET OP THE KETTERING LABORATORY FOR THE INVESTIGATION OF THE POTENTIAL CANCER HAZARD OF THE PETROLEUM INDUSTRY SPONSORED BY THE MEDICAL ADVISORY COMMITTEE . of the AMERICAN PETROLEUM INSTITUTE J u l y 1, 1953 - June 30 , 195U Salaries (Based on proportion of time actually spent on project) Direct Salaries $38,500.00 Indirect Salaries Histopathological Preparation $ 5000.00 Other Services $28,1+50.00 $ 71,950.00 Miscellaneous Expense Purchase of Animals $ 2,500.00 Special laboratory supplies $ 6,800.00 Travel $ 5,000.00 Overhead (Proportion of heat, gas, electricity, steam, telephone, general laboratory supplies, postage, annuities, pensions, maintenance, etc.) $ 1 7 ,823*00 Total $ 32, 123.00 $10i+,073*00 API 05873 BUDGET OP THE KETTERING LABORATORY FOR THE INVESTIGATION OF THE POTENTIAL CANCER HAZARD OF THE PETROLEUM INDUSTRY SPONSORED BY THE MEDICAL ADVISORY COMMITTEE of the AMERICAN PETROLEUM INSTITUTE July 1, 1953 - June 30, 1951; I. D e t e r m i n a t i o n of the l o c a t i o n and extent of the cancer hazard from refinery intermediate and product streams Direct Salaries $ Indirect Salaries Miscellaneous Expense Overhead 1;,000.00 3 ,$ 0 0 ,0 0 2,000.00 1,860.00 $ 11,360.00 II. Development of semi-quantitative biological testing methods for measuring the relative carcinogenic potencies of petroleum products Direct Salaries $ Indirect Salaries Miscellaneous Expense Overhead 3,500,00 .,200,00 2,300,00 1,630.00 $ 11,630.00 III. Determination of the relationships between the rapidity of induction of benign and malignant tumors in experimental animals a n d v a r i o u s e x p o s u r e fa c t o r s s u c h as frequency of application, dosage per application, area of exposure, number of applications, special irritant or toxic properties of the oil applied, etc. Direct Salaries $ 1;,$ 0 0 ,0 0 Indirect Salaries $ ,2 0 0 ,0 0 i Miscellaneous Expense 2,300.00 Overhead 2,090.00 $ 11;,090,00 Indirect Salaries include Histopathological Preparation and other services. Miscellaneous Expense includes purchase of animals, special I laboratory supplies, and travel. API 05874 - 2 - IV. D e v e l o p m e n t of r a p i d a n a l y t i c a l m e t h o d s f or estimating the relative carcinogenic potency of any given refinery intermediate or product A. I s o l a t i o n and i d e n t i f i c a t i o n of c o m p o u n d s responsible for observed potency of various types of oils and determination of physical and chemical properties of these compounds which could be used in the development of a widely applicable analytical tool (including associated animal testing) Direct salaries $12,000,00 Indirect Salaries 10,200,00 Miscellaneous Expense 3,050,00 Overhead 5#500,00 $ 30,750.00 B. Semi-empirical correlations with carcinogenic potencies based on the chemical or physical properties of known or suspected carcinogens in the oils (including associated animal testing) Direct Salaries $ Indirect Salaries Miscellaneous Expense Overhead 5,000.00 3,700,00 1,350.00 2,330.00 $ 12,380.00 V. E p i d e m i o l o g i c a l P r o g r a m A. Collection and correlation of data on cases of cancer among employees of the petroleum Industry Direct Salaries $ 5,000.00 Indirect Salaries 3,500.00 Miscellaneous Expense 2,350.00 Overhead 2,320.00 $ 13, 170.00 B. Special Investigations of cases of cancer In particular plants In the Industry or among customers of the Industry Direct Salaries $ 2,500,00 Indirect Salaries 1,750.00 Miscellaneous Expense 850.00 Overhead 1,163.00 $ 6, 263,00 C. Survey of pertinent literature on cases of cancer occurring among workers exposed to tar or oil in their occupation Direct salaries $ 2,000.00 Indirect Salaries 1 ,14.00,00 Miscellaneous Expense 100.00 Overhead 930.00 $ k,k$ooo API 05875 J A merican SO P W e ES t T r o l St h e um STRE E T I n s t i t u t e N E W Y O R K SO. N. Y. ,a rT M E N T o p t e c h n i c a l s e r v i c e DAVID V. TROO P. DlMCTOR July 2, 195* To Menders Medical Advisory Committee Gentlemen: Chairman George M. Saunders has noted Dr. Eckardt's letter of June 22, 195*, copies of which were sent to all members, and has requested me to set a dead-line for the receipt of comments or objections on the publication of the manuscript entitled "Carcinogenesis of the Skin, I". Accordingly, we shall assume you have no objection to publication unless your specific objection is received by Dr. George M. Saunders, with carbon copy to me, on or before JU-y 31, 195*. Very truly yours, DVS:Js API 05876 rr /w * h >" DEPARTMENT OF PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH fHg FETTERING LABORATORY ALLEGE OF MEDICINE-- EDEN AVENUE Cin c in n a t i i s . o h io Ju ly 6, 1954 CASLE ADDRESS. KETLAS. CINCINNATI TELEPHONE: CAPITOL 1414 M r. D. V. S tro o p , D ir e c to r Technical Services A m erican P etroleum Institute 50 W est 50th S tre e t New Y ork C ity 20 D ear M r. Stroop: I am retu rn in g to you herew ith four copies of the ag reem en t betw een A m erican P etro leu m Institute and U niversity of C incinnati, which have been executed by the B oard of D irecto rs of the U niversity. I should ap p reciate your returning two copies to us when they have been signed by re p re se n ta tiv e s of A m erican Petroleum Institute. Very truly yours, ef Enc. E. R. F o rtla g e , S etfretary to D r. Kehoe API 05877 A merican P etroleum Institute SO W E S T SOTH S T R E E T N E W Y O R K 2 0 . N. Y. June 23, 1954 Board, of Directors University of Cincinnati Cincinnati 19, Ohio Gentlemen: This communication is authority to continue vork on investigation of the toxicity and mode of action of certain petroleum products for the year ending June 30, 1955, according to the terms of the agreement dated January 8, 19k6 , and on the basis of a maximum budget for operating expenses of $72,250. Upon receipt of your approval of the continuation of the project vith this maximum budget.ve shall pay you the sum of $36,125. The balance of $36,125, less the unexpended balance in the fund on June 30, 195^, will be paid on January 1, 1955. It is understood that any balance remaining in the fund on June 30, 1955 will be returned to the American Petroleum Institute, and that no excess over the amount of the budget is to be obligated or spent without written authorisation. Very truly yours AURI C A ! PETROLEUM INSTITUTE Accepted and Approved: UNIYEBSIT OF d H C O T A T I Through its Board of Directors By /f. Renton K. B rodie, C hairm an Ralph C. B u rsiek , C lerk API 05878 A merican P etroleum Institute 5 0 W E S T SOth S T R E E T NEW YORK 20 . N. Y. June 23, 193k Board of Directors University of Cincinnati Cincinnati 19, Ohio Gentlemen: This communication is authority to continue work on investigation of the toxicity and mode of action of certain petroleum products for the year ending June 30, 1955, according to the terms of the agreement dated January 8, 19^6, and on the basis of a maximum budget for operating expenses of $72,230. Upon receipt of your approval of the continuation of the project with this maximum budget we shall pay you the sum of $36,125. The balance of $36,125, less the unexpended balance in the fund on June 30, 195^, will be paid on January 1, 1955- It is understood that any balance remaining in the fund on June 30, 1955 will be returned to the American Petroleum Institute, and that no excess over the amount of the budget is to be obligated or spent without written authorization. Very truly yours, AMERICAN PETROLEUM INSTITUTE Accepted and Approved: UNIVERSITY CP CINCINNATI Through its Board of Directors Renton K. B rodie, C hairm an By Ralph C. B ursiek, C lerk API 05879 file: Medical Advlsary Caaaltte - HIV. (V CUCHtHAJI Sxpenditures * n e 23, 195* U n lw n lty at Claeianetl Ttm X a tta rla c L ^ c ra to a j Collapa of M o ld a s - M an to C lacln a a tl 19, Cfele gaolnaa yo* 111 fia d fo a r copla* c f ha M d l w l l o ^ w i m rt o-arla ATI Martim i a-- rch p ro ja e t 1 fo r II fla a a lyya a r ada *mm JO, 1955, A* t iim k r y l a--y altar 111 -- cnk cf ma a U l Jh l? 6 -- d ttta * -- t a ha acatara** ip il aa tah alf of ha ai--aralty m --tura t e a o aa ao a ay aaad. yaa oaa ahaok f -->ha fUrat half <* a hodm * tm ha a-- fia a a l ja . a h a ll aaaA ya* ae capta* o f haagra* a lha r. f-- y tauly youm. BVhtJa (*> M. M. 0, API 05880 JUaa 23, 199k Beavi etf Blzvetars U&lvarslty at Ciacinnati Cincinnati 19 * O astlaam : Lovaatlgatiaa af ttaa tca la itjr and aadn cf actloa a t oartalm patralaeai producta fc r d a paar --ding A m JO, 1999* anaordiag to tha taraa at t h a jg i-- t i ^ d ^Jdawary 8, lfd6^aad^oa da Upoa ra e a lp t at paar agpraaal o f tha nnntlnnrM nn o f tka p ro ja e t w ith dia aaatlaaa ta d p t ob a b a ll pay pou ttaa aaa a f 0 6 ,U 9 . a dalanaa o f 06 *1 *9 * tta law^ panrtad dalaaaa in ttaa tw* oa Jkaa JO, 1996, U li da paJA oa fc iaiar y 1, 1999* 1 la M iin tia l hat aoy alaaaa taaalalng in da fand oa *am JO* 1999 v i l l da fatw a ad to ttM AM srlaaa pateojaua ^ to da d ltp ta d e r apaatt o ltb e u t w ltd a a o a ttn rlta tlo a fw y tra ly paura. -- pici F i w i n i n u r r a m Acoaptad u u iro a a ra a r c u c im a tx ita *y bt API 05881 S H E LL DEVELOPM ENT COMPANY EM ERYVILLE. C ALIFO R N IA July i , 1954 m MIL a. E. Ickardt, M.D. Esso Lahors ta r las Standard O il Drrslopnsnt Company P .0. Box 51, Linden, Nov Jersey Dwr Dr. Eahardt: haforanoa 1 nods to your lottor of J\ma 22 la hioh you request approval > f th ragart on the a ctiv itie s of tb #.5\*oc--Ittoo on Carelaotew lally uhiah 1 to he rahaltted to Dir, X Saunders. 8jn rg'dCdlm ^ noanranitun of the A p ril 27th aaotiaf of the Operating Saint-- Ittoo of ths Mrtl 1 Advisory Ce-- it tes, lt la Indleatad that the MMloal and Haelth Cean&ttee has M M that aoeb rapart on tho euheo--ltteo ooatain a stato--nt otaioh eould not only define tho aoopo of tha projoct and ita Ju stificatio n , hot also a prediaiioo of tha antieipeted benefits to tha o il Industry. Za lig h t of th is request, 1 hallovo that as shslraon yon nicht add a atatanawt okdoh oonll doserihe tha honaflta otdeh tho study oadtrU hsn ty tha IM m nalttaa on Caroinofoniaity la oapaotad to aocoapllah for tha o il ladnatry . Othor than for th is ouggaotloe, Z appa i:it s of tho report ahloh tha tMhii-- tttan on Cardaofon io lty hao pr ^ u i< under your diraotion. Z a i n ha happy to ana--a eheln--ship of y-- r subocaadttaa for its oart aaetlag, Friday, 3p lantar 10, at tha Conrad Hilton la to l in Chisago. Vary tru ly yonra, *. -. >*. Cffl/Jch bo: T.B. Albin H. Oarahlnowlts J.H . Morrison T.B. Randal Q .li, Saunders D.V. Stroop C. H. U na, M.D., Fh.D. Consultlag Torlaologlat API 05882 rH{ RETTERIN# LABORATORY .oL L 8 I OP MEDICINE-- COIN AVENUE , rCINNATI I I . OHIO UNIVERSITY OF CINCINNATI DEPARTMENT OP PREVENTIVI MEDICINE ANO INDUSTRIAL HEALTH Tiu l y 12 , I 95I4. CARLI ADDRUS: KITLAB. CINCINNA^ ^ TILER I Mr. D a v i d V. Stroop, D i r e c t o r Department of Technical Services American Petroleum Institute 50 West 50th S t reet New York 20, New York Dear Mr. Stroop: ,, I herewith acknowledge with thanks receipt of a continuation of agreement dated June 2J, 195^# between the American Petroleum Institute and the University of Cincinnati,to gether with their check in the amount of $36 ,125 covering expenditures to be made on their behalf. Very truly yours, ef/phd API 05883 I Copy From D. V. Stroop For Information of Members of Medical Advisory Committee ________________________________________________________________ July 12, 1951 ESSO LABORATORIES STANDARD OIL DEVELOPMENT CCMPANY P. 0. BOX 51 LINDEN, N. J. June 30, 1951 Mr. D. V. Stroop American Petroleum Institute 50 West 50th. Street New York 20, New York Dear Mr. Stroop: I am sending you herewith the minutes of the fifteenth meeting of the Committee on the Carcinogenic Action of Mineral Oils of the Medical Research Council of Great Britain (MRC.5I/I5I) forwarded from Colonel Auld. I thought you might want to have this duplicated and distributed to the members of the Medical Advisory Committee. Very truly yours, /s/ R. E. Eckardt R. E. ECKARDT, M. D. REE:ilk Enel. API 05884 c o P Y MEDICAL RESEARCH COUNCIL For Specified (Normal Committee) Circulation MRC.5h/k5h COMMITTEE OF THE CARCINOGENIC ACTION OF MINERAL OILS Minutes of the Fifteenth Meeting, held at 38 Old Queen Street, London, S.W.l, on Thursday, 29th April, 195^> at ^.00 p.m. Present; Professor T. Ferguson (Chairman), Colonel S. J. M. Auld, Professor J. W. Cook, Professor H. N. Green, Dr. I. Hieger, Dr. J. 0. Irwin, Professor F. Morton, Professor J. R. Squire, Dr. D. L. Woodhouse, and Mr. J. C. R. Hudson (Secretary). Attended: Dr. B. S. Lush. An apology for absence was received from Professor R. D. Passey. 127. Minutes The Minutes of the previous meeting were approved and signed. 128. Progress Report by Professor Cook (CAM0.5^/l) Professor Cook said that Dr. Carruthers had been concentrating on two particular sub-fractions and, using methods agreed with Professor Morton, had been making aromatic extracts. From these aromatic extracts he had isolated 2:7-dimethylanthracene and 2:3:6-trimethylanthracene and two other anthracene hydrocarbons which had not as yet been identified. It was not known whether any of these compounds were carcinogenic, but this was the first occasion on which the first two had been isolated from petroleum. The average molecular weight of these compounds appeared to be about 220. T M s was not inconsistent with Professor Morton's estimate that the average molecular weight of the fractions was about 300, since aromatic compounds tended to have higher boiling points than aliphatic. 129. Progress Report by Professor Morton Professor Morton had circulated copies of his preliminary report prior to the meeting. In explanation of the nomenclature he said that the samples designated by the letter "D" were from the K^S fraction. The suffix "R" indicated a raffinate, and the suffix "E" the aromatic extract. Professor Morton added that not too much reliance should be placed upon the figures given because the method had not yet been confirmed as applying accurately to the samples which they had been investigating. For example, the proportion of aromatics appeared to be unusually high, but this may be due to a high sulphur content. However, this would not be known until the sample had been subjected to a full analysis. He said he hoped that the continous extraction process would soon be in operation. API 05885 - 2 - He recalled that it was originally thought that the Lagunillas fraction, which showed the greatest carcinogenicity, was distilled at an overhead stream temperature of approximately 210C. At present this cannot be done on the Steadman still without exceeding 300C in the distillation pot. At. the moment he was attempting to modify the still so that this could be achieved, but failing that it might be necessary to distill the Lagunillas fraction on a laboratory scale. 130. Report by Dr. Irwin (CAMO.5W 2) Dr. Irwin said that the figures were not, in fact, new, since they had been submitted at the last meeting of the Committee, but he thought that it was desirable to record them formally. It was significant, he 6aid, that the response to the standard carcinogens by mice and rabbits was different, but their response to the various fractions was similar. It was generally agreed that it was preferable to use rabbits as the experimental animals as their response was more uniform and they are less liable to inter-current infections and other hazards. 131. Progress Report by Dr. Woodhouse and Professor Squire (CAMP,5^/3) Professor Squire emphasized that thi6 report covered thirty weeks only and was circulated in case it should be helpful to the Committee in deciding on their future plans. He said that the log time response curves were reasonably straight, but to be useful for purposes of bio-assay they should also be parallel. They were not yet parallel although they were more so than in previous experiments. Dr. Hieger asked why it was that the rabbits appeared to be more susceptible in the y e w 1953-5^ than in the year 1952-53* Professor Squire agreed that the response did seem to be greater in the later year, but as the standard carcinogens had been used on each occasion the results were not invalidated since the relative potencies remained the same. Professor Cook thought it waB worth while to draw attention to column VII of Table 3 in the report, which showed for the first time in this investigation that carcinogenicity was concentrated in the aromatic fraction. He asked whether the different responses by the mice and rabbits was due to a different response to a single carcinogen or whether there might be another factor - a co-carcinogen or an anti carcinogen - which affected one species only. Professor Squire said that this could be decided by adding 9 :10-dimethyl-1:2-benzanthracene to one of the Kuwait fractions and then applying it to both mice and rabbits. This would then demonstrate whether the carcinogen was different in its action from the dimethyl benzanthracene or whether there was an anti-carcinogen or a co-carcinogen present. Professor Cook said that he had 3 S* K j ^ b and 9 6* KjjSgb which contained substantially all the anthracenes. It was agreed that he should send 1.5 g* of the first and 5 g. of the other to Dr. Woodhouse to be used in the September series of tests. 132. Future Plans Professor Squire said that they were able to do twenty-one tests per year, of which three were standards, leaving eighteen for testing substances. It was agreed that Birmingham should concentrate on the Kuwait fractions and that tests on the Lagunillas fractions should be carried out at Leeds. API 05886  -3 - Professor Morton undertook to proceed with the distillation and production of extracts from Lagunillas samples which he would send to Leeds for them to apply in a series of animal experiments "beginning in January 1955 Prom the Kuwait fractions he would concentrate on the aromatic extracts KLS]A to K^SSA, from which Dr. Carruthers would prepare (a) anthracene fractions by maleic anhydride treatment, and (b) a picrate forming concentrate from the remainder. This would mean twenty-four samples, but as only fifteen tests were available (3 of the 18 available were to be used for testing the Glasgow Compounds - Minute 13 1) he might blend some of the lower aromatic fractions, for example K i ^ K^Si^ It was agreed to wait for the preliminary results of the animal experi ments on the anthracene sub-fractions before deciding whether it would be necessary to synthesize larger quantities of the anthracenes already isolated and identified by Dr. Carruthers. Colonel Auld said that he hoped the Lagunillas and Oklahoma oils would not be overlooked in concentrating on the Kuwait oil,since all three were originally selected with great care as representing particular types of oil from various fields. He was also rather disappointed at the apparently slow rate of progress and wondered whether it would be any help if some of the tests could be undertaken in America where it was now accepted that the fundamental approach by the Committee was the correct one. It was agreed that it would be unwise to ask the Americans to undertake any of the tests because of the difficulties of communication which were already sufficiently awkward between the few centres involved in this country. It was also agreed that it was important to bear in mind the need to investigate the Lagunillas and Oklahoma oils, but this might be easier and done more quickly once the Kuwait oil had been fully studied. 133- Date of next meeting It was agreed that the next meeting should be held at 11:15 a.m. on Friday, 23rd J u l y , 195^* I i I - JL^ ,r t .v ESSO L A B O R A T O R IE S STANDARD Ohi DEVELOPMENT COMPANY P. O. BOX 51, L I N D E N , N . J . MEDICAL r e s e a r c h d i v i s i o n ,-a g R T E ECKAROT PH D RCl M .D D IR E C T O R CE W G ER AR D E PH O . M O H HEAO TO X IC OLO G IST in d u s t r ia l h y g ie n e s e c t io n N A T H A N v. H E N D R IC K S . C H E HEAO OF SECTION F R A N K L IN W . C H U R C H . M .S . G E O R G E M . W IL K E N IN G M .S . IN D U S TR IA L H YG IENISTS 13, 19$k to t i ftom-- i t o l i m C r t H n -- a lly t e Uh t o t e 1 to Ito API. d a n f t o it o B im t t o f ip Ito w rt tolto tea Ito far tea* t o i) Ito -- ton-- a S to ito to i , 1 t e c to to tow t e r Ito -- p i-- to tori trimt e t t a , fbr Ite p a rt* M x 1* U * S -t e 3 , l* * . Sto to ll t e t e Ito -- to Ito t i t e la Ito t o t e to tfMOO to a u lit e * ito Ito to tte -- fc to Ito f t H r lu Ltoartoary to rta i ito p r ia i to y 1, 19f$ tola 30, , z rtop-- fa tly t o r i l Uria Sili ilk it ? R. S. BURST, K. B. k 4tef*fiartaa & A. Katoa 3. T. Straap ft.pl 05888 I THE REPORT OP THE SUBCOMMITTEE OB CARCIHOQEHTCITT TO THE MEDICAL AD7IS0RI COMMITTEE OF THE API The Subcommittee on Carcinogenicity of the API Medical Advisory Committee, one of the first subcommittees created, was established early in It vas formed after it became known that certain petroleum oils and fractions had been demonstrated to possess fluorescence similar to certain known carcinogens, and, when painted on mice, produced tumors and cancers. Negotiations were then begun between this Subcommittee and Dr. Kehoe of the Kettering Laboratory which culminated, early in 19U6, in an agreement between the API and the University of Cincinnati, under which the Kettering Laboratory of Applied Physiology would "investigate the toxicity and mode of action of certain petroleum products". A ^ The investigation began with the testing of 8 oils supplied by three participating companies, in addition to negative controls of water and white oil and positive controls of methyleholanthrene. of the 8 oils tested, only 2 produced skin tumors in mice. One of these was an unflashed thermal residue, the other a clarified oil. These investigations were carried out at a cost of $6,000, and were reported in detail to the Subcommittee by the Kettering Laboratory on November 5, 19U8. However, when the results were becoming apparent early in 19U8, the Medical Advisory Cosmittee, on the advioe of the Subcommittee on Carcinogenicity, forwarded to the members of the Safety Committee of the Board of Directors of the API a resolution dated April 5 19U8. This resolution called upon the Executive Committee of the Board of Directors "to establish a comprehensive research project for the development of fundamental facts, analytical tech niques and related information pertaining to cancer-causing compounds which may occur in petroleum or be produced therefrom". This resolution called for the solicitation of contributions to a special research fund in the amount of $110,000 for the first year, of which $90,000 would be a non-recurring capital expenditure for construction of extraordinary experimental facilities at the Kettering Laboratory. This resolution was presumably approved by the Executive Committee and on July 1, 19U8 a cheok in the amount of $65,000 was sent to the Kettering Laboratory, $0,000 ef this amount as a grant for animal quarters. With the expansion of this research project in 19U8, A. W. Horton, Ph. D. was added to the staff of the Kettering Laboratory to supervise the chemical and biological work on petroleum and petroleum fractions, and the services of J. J. Phalr, M. D., Professor of Preventive Medicine, University of Cincinnati, College of Medicine, were made available to the staff to develop and supervise an epidemiological survey of the incidence of cancer in the petroleum industry. From this point on, the research project can be divided into two phases, nsmsly (1 ) the chemical and biological work, and (2 ) the epidemiology. API 05889 1 The Chemical and Biological Work Under Dr. Horton's supervision, an expanded program of biological testing for carcinogenicity was begun on a variety of petroleum fractions selected and provided by the technical staffs of the participating petroleum companies. It soon became evident that in order to express the relative car cinogenicity of different petroleum fractions in a quantitative fashion, it would be necessary to develop standards of reference to which the petroleum fractions could be related. For this Dr. Horton began biological work with solutions of a recognized carcinogen, methylcholanthrene, in varying concentra tions in various solvents. This work clearly showed that a biological assay for carcinogenesis was a highly complicated test which was affected by a great number of variables, such as the general health of the animals used in the test, the genetic strain of the mouse used for the test, the toxicity of the material applied in the test, the presence or absence of lice or mites on the animals, fighting among the experimental animals, etc. Therefore, steps were taken to control these variables insofar as possible, although it must be recognized that even after It or 5 Fears of research it has not been possible to eliminate or completely control these variables but only to minimise them, ttiere these variables were incapable of control, this early research did permit the bio logical experimenter to recognize their effects so that he could throw out as unsatisfactory those biological tests in which these variables have played too great a role in the final results. This biological research on the develop ment of standards of reference is summarized in the report "Carcinogenesis of the Skin, I. Basic Methods Bqployed in Testing Complex Hydrocarbon-Type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies" by A. kfesley Horton and Dorothy T. Denham. This report has been proposed for publication in a cancer research journal. The seoond achievement of this biological research has been the demonstration that solvents influence results obtained with pure carcinogens. At present two types have been recognised. The first type, represented by benzene, has no effect on the activity of pure carcinogenic hydrocarbons. This type of solvent has been called a non-accelerating solvent. When the pure hydrocarbon, methylcholanthrene, is dissolved in this type of solvent, the carcinogenic activity in animals is directly related within limits to the con centration of the carcinogen in solution. The second type of solvent, repre sented by dodecyl benzene, has an additive or accelerating effect on the activity of pure carcinogenic hydrocarbons. This type of solvent has been designated an accelerating solvent. When the pure hydrocarbon, methylcholan threne, is dissolved in this type of solvent, the carcinogenic activity in ani mals is not directly related to the concentration of the carcinogen in solution but rather appears to be more dependent on the amount of accelerating solvent applied with each application. The research work leading to this discovery is soon to be summarized in a paper which the laboratory will propose for publica tion, also in a cancer research journal. API 05890 - 3- In addition to the work on reference standards and solvents outlined above, the laboratory has tested biologically between 1$0 and 200 petroleum materials and fractions of these for their carcinogenic activity in mice. This work alone required some ii,000 mice for test purposes. A summary of the results of these investigations will undoubtedly be prepared as a manuscript for publication. The laboratory has also investigated the effect of hygienic measures such as washing and protective creams on the carcinogenic activity to mice of several petroleum materials. It is anticipated that these results will be used in the framing of practical hygienic control measures. The chemical work has encompassed the separation by distillation, chromatography, and chemical methods, of petroleum materials into various fractions in an attempt to define precisely the agents in petroleum responsible for this carcinogenic activity in mice. This work has already led to the sub mission of one manuscript entitled "The Use of Catalytic Iodination on Activated Alumina for the Determination of Benso (a) Pyrene in Complex Mix tures". This manuscript was 'authored by R. Tye, M7 J. Graf and A. V. Horton. It has been reviewed by the Subcommittee, the Medical Advisory Committee, and the Medical and Health Committee of the Board. No objections to publication were voiced and it has been submitted to the editors of the Journal of Analytical Chemistry for publication. In addition, the following general information has come out of the combined chemical-biological studies! a. Most petroleum materials boiling in the range 500-650*F probably contain accelerators. This includes paraffins from normal decane to hexa- or hepta-decane and naphthalenes which have at least a butyl chain attached. b. Carcinogenic petroleum materials boiling between 700-725*F probably contain phenanthrene-like carcinogens similar to benso (c) phenanthrene. c. Carcinogenic petroleum materials boiling between 725-800* F probably contain anthracene-like carcinogens, possibly cyelopentemo bensanthracene. d. Carcinogenic petroleum materials boiling above 800*F probably contain benso (a) pyrene. API 0589 2. Epidemiology - u- It was early recognised by the Subcommittee on Carcinogenicity that the chemical and biological work outlined above would give only a partial answer. It was expected that the epidemiological work would develop informa tion on the following two questions. a. Does work in the petroleum industry or at any specific job within the petroleum industry subject the worker to an increased risk of developing cancer? b. Have newer refining processes, such as catalytic cracking, or increased severity of thermal cracking introduced any new risks of developing cancer to any segment of the petroleum workers? It is in general these newer processes which have been shown to produce materials which have a high degree of ^ carcinogenicity, to mice. A central registry far the reporting of cancer cases occurring in the petroleum industry represented by the participating companies of the API was established at the Kettering Laboratory under the direction of Dr. Phair. In addition to reporting such cases, it was deemed essential that a complete occupational history for each case also be reported so that correlations with occupation could be made. In the course of setting up this registry, it soon became apparent to Or. Phair that the medical departments and medical records of some petroleum companies were inadequate to permit accurate reporting of cancer cases. Ini tially, therefore, Dr. Phair a major effort was in assisting medical directors, or refinery managers where medical directors were unavailable, in organising their departments and/or records so that good case reports and occupational histories would be received in his office. Therefore, it has only been in the past year or two that an approach to effective reporting of all types of cancer cases and occupational histories has been achieved. Despite incomplete uni formity of reporting and participation, over 1,300 cancer cases hove been reported to Or. Phair 15 companies, and are in the process of statistical analysis. It is believed that worthwhile statistical analysis of these cases will require maintenance of this registry for a minimum of 6 years. In the interim, Dr. Phair continues to act In an advisory capacity to the petroleum contantes and their medical directors for the collection and reporting of these cases. API 05892 T - 5 - Expenditures of the Subcommittee on Carcinogenicity 191*6 191*7 191*8 191*9 1950 1951 to June 30, 1952 July 1, 1952 to June 30, 1953 July 1, 1953 to June 30, 1951* * 6,000.00 0 65,000.00 (includes |1*0,000 grant for animal quarters) 1*9,1*13.00 89,963.39 59,065.12 31,692.1*2 10l*,760.00 79,000.00 *1*81*, 893.93 Budgeted July 1, 1951* to June 30, 1955 I 72,250.00 ' Future Work As indicated in the shove outline of expenditures, a peak of activity was reached in the period July 1, 1952 to June 30, 1953* Since that time there has been a gradual tapering off of expenditures for this research effort. The Subcommittee recommends that future work in the chemical-biological phases should be directed towards 1. Work on physico-chemical methods for assaying the carcinogenesis of petroleum materials. These methods will be validated by a. Determining the relative inportance of carcinogens and mixtures of carcinogens and accelerators in the process of carcinogenesis. b. Determining the relative carcinogenicity of additional petroleum products and materials, not adequately covered to date. c. Isolating and identifying additional carcinogens and/or accelerators in petroleum materials. 05893 T - 6- d. Continuing work on the improvement of standard bio-assays for carcinogenesis. 2. Effective hygienic control measures for use by the petroleum industry. 3. Work on possible carcinogenic inhibitors. In the Subcommittee*s opinion, this type of research should continue until at least 1959. However, expenditures will continue to taper off during this interval and should stabilize at levels not in excess of $25,000 after June 30, 1956. The epidemiology should be continued until at least 1959. The budget for this phase of the work will probably run between $15,000 and $20,000 per ^ i year. In the opinion of the Subcommittee, if full cooperation with Or. Phair v in the epidemiological work has not been achieved by June 30, 1955, the Sub committee would recommend that epidemiology work be discontinued since it would be likely that full cooperation would not be expected to be achieved after this date. If cooperation has befai achieved by this time, expenditures should stabilize at a somewhat lower level, since the work will consist principally of collecting and analysing statistically those cases reported. It is thus estimated that the anticipated annual expenditures for 1956-1959 will be in the vicinity of $l0,000-$ii5,000. Estimated Budget July 1, 1955 to June 30, 1956 The Subcoanittee recommends that the Medical Advisory Committee request funds in the amount of $65,000 for the continuation of this work during j the interval July 1, 1955-June 30, 1956. A brief breakdown of expenditures is approximately as follows t ^Biological Work $20,000 Technical Wbrk 32,250 Epidemiological Work 12,750 $65,000 Another breakdown of these figures can be given as follows: Salaries Direct Indirect Histqpathology Others $22,000 2,000 20,000 $,000 API 05894 - 7 - Salaries Miscellaneous Exp Animals Supplies Travel Overhead $ 1,250 5,000 U,ooo i o , 75o Composition of Subcommittee R. E. Eckardt, M. D., Chairman Standard Oil Development Company E. V. Adams Standard Oil Company (Indiana) L. C. Beard, Jr. Socony-Vacunm Oil Company, Inc. Allan E. Dooley The Texas Company F. D. Qassamy, M. D. Qulf Oil Corporation L. M. Henderson The Pure Oil Company ( C. H. Hina, H. D. Shell Development Company E. K. Linder, M. D. The Atlantic Refining Company R. C. Mithoff California Research Corporation M. H. Neequist, M. D. The Texas Company $W,000 21,000 $65,000 API 05895 T 8 - James W. Osborn, M. D. Standard Oil Company (Ohio) F. J. Sanders Standard Oil Company (Ohio) J REEsilk June 21, l?5ii Jl API 05896 Uyl . FW ' ; l* J # * P * * m . irt lu 1 g * - * U 13. Br. Bobrt A. U km V K i m v l t j or e i M i w t i *jr9, 195* U t& W API 05897 rii*: U nir, of C incinnati Ixpanditur* / V 3 * 1 7 9, 195* Or. Bobert A. tteiveralty a t ClM law U tttttr la c r A k n lr y of CIm I m hU 19 , Qi t e M * m . n , 191* a IM tea* * la a * M f c la tte m m * tf * 9 f U 5 99 * (9) s* p. a. a , a. . aitar'a A fila Valeerelty te M i a i fU c rj wrs API 05898 ESSO L A B O R A T O R IE S STAND ARD OIL DEVELOPMENT COMPANY P .O. BOX Ji, LINDEN, N.J. MEDICAL RESEARCH DIVISION oOSERT ECKARDT . PH O MO D IR E C T O R h 0 r ACE W G E R A R D E . PH O . M O HEAD TO X IC OLO G IST Jtoy 16, 1951* in dustrial hyg iene section N A T H A N V. H E N D R IC K S . C H E HEAD OF SECTION F R A N K L IN W . C H U R C H . M S. G E O R G E M W IL K E N IN G M .S . IN D U S TR IA L HYGIENISTS 3 *. Lao d a d a ttm Standard O il 0-- n j1 15 toat 5la t Stroot Baa lark 19, lav lark Our St. wadoi Yarn ooaamta eenaamiac tha firto paragHph of or. Bortaa'a a d Mias O nw H i por m Carpino-- la of tha Skia I.* 1la* m x U d a oonaldarahla a m t of aaaaaat amk m mtoTBfnr Tharo la ^yarontly a la aoa aonoorn toast h a jroar nbm bU a a ba roaolvad a t t i tha la tta ri* ! laharatary. am accatti on la that tha laat aaataaaa of thia p a r -toh ham tha phraa *ta aiaa* addad a tha and af It. If th ia atiaaia am id m n a tto ahjaatlana ridak yw ham ta thia paragraph, I m ara that tha la tta rla c Ltoartoary attori! ha -- nabla ta th ia a i-- ah m . If tha aham ahaac toaa aat aatlafy tha ah>atlana to d * m ham to thia para- cnyfe, i t avoid ha w aacfaalla* that ja a rmarlto tha flra t aa ym said alto ta ma it aardod, md that yva atotot thia v n r lt t a i ta Br. Saaadara with a aorta* aapy ta Br. Bahaa (hr. lartaa) ta aaa If tha l ottar e latoratw y m aid ha ornatola ta rw ritia c tha flra t p r a r t o aa ym mtod to * it r--rittoa. ZB tha ornato that tha la tta rla c Laharatary aaald aat a n t to ym r raaarlto, it m aid than app-- ta m that rito* th ia p-- r la f ar -- dad ta tha Camdttoo af tha Board prlar ta pah! H a th a , aa aaald tor raaaaa-- d, aa tha M alati Bdtoaary Ca--itta a , that y-- r-- i ta mtod ha prafarahla to tha am that tha lattarlac Ltowat ary prapim a. A t Baard Qaatottaa m aid thaa ha la a p a ritiaa to a-- tal oltbor that tha Eattaaiac laharatary m attai -- that y-- aaatoai m aid ha -- o profarablo to thaa. SaharaUy* tha fattarla fatar atary aaald ha la a paaitlaa aat ta aaa-- toth tha raaaaamadaklaaa af tha Baard Qaatottaa, hto hmtoac Kabo md athan to tha la tta rla c laharatary, Z bollava that thay mtod elm aoaaldarahla thaaght ta th ia I f tloy datodad to p tollto th alr aa -- tla c lnataad af that jtrofar r od ty tha Baard 0-- dttaa o d m aid mdatotadQy ham m ry atroce roaaoaa far otoor1n i ta th alr aw -- dla. I ritm a tili to dlaaaaa thia toth ym w tha talaphana today, hto alaaa yw vara ovt of yw r afflao Z thaacfct it aaald ha hwdlad hy a o ll, toaaa Z w f r is i oa t , v o ta ti ita rtla c w tha ITth wd t o ll ha to af tha afflaa far tvo moka, I | m a li ba happy ta dlaaaaa th ia fa rti-- lf yw fool l i aaaaaaary aftar I ra ta I irta w m aatloa and l f i t has aat almady barn aattaod la tha m anti. S ln a a rto y y-- a , RZEiilk J r . Q . M. S c a n d a R . L . BCKARJT, M. D API 05899 July 6 . 19Jb Ur. Ooorge StttMh, Chairaeo Hodleal k&r&krr Ooaadttoo Aaorleap^Potreloua Znatltuto JO teijCtk H m t i# 'T o rk , Mw Tart .* Dm t Doctor l io r iif i Or. b k u t t Mot aa a n r t of hl> le tte r to Mr. Streep aanoora lac the propoood pdhlloatiea hjr Mr. Rortoa u l IU o Doataua rogardii* oaralnofamaia of the Ida. MMt of tte h c im U im m hero aedo hare been properly rooocalaed. Vo aro t ill of tho oplaloa H ut the openly paragraph la 1 loading aa* aot la otrlo t heaping with tho fkota of the itaatioe. U ntil nak tlao ao a definite d la lo a l eerreletloa baa booo ootabUabod between f i ir i to h i# boil tag areaatle fraotleaa and tho oooorronoo of akla oaaaor. Z bailee theoo aaiaal data aro enly H iiM tlT t. It la ay te r ito a tln that M . Barbae itje rta to aay rawieian of thla paregrmyh. I f oaa aeeld ho aoro that tho aaaaeerlpt il l atay la a trla tly profhooioaal M a a o li, hia ahjeetlaae aro a o ll takaa. Fran paat oaporlaaoo, heooeor, Z A at m a a iM y aoro that thla M aa^ptlia aaaaot ho relied pea to prataatdp ta n a f a e t lf la i aapl^roo rolatlaaa prohlaaa. Tory truly yoara, EWjJak 01 Dr. X. I . Bekardt Mr. WllUaa Radon Mr. D. V. Stroop ZJO WADS, M. D. API 05900 V SH E LL DEVELOPM ENT COMPANY EM ERYVILLE. C A LIF O R N IA July 21, 1954 m VM L R. E. Eckardt, M,D, Esso Laboratories Standard Oil Development Company P. 0. Box 51, Linden, Rev Jersey Dear Dr, Eckardtt 1 know that you, as o h a i m a n of the Subcommittee on Carcinogenicity, will be Interested In this article on'the effects of benzpyrene when applied to human skin. While the reproduction of the histologic seotlons leaves something to be desired, I think that you may wish to have It distributed to members of the Usdical Advisory Committee because of its general interest. Very truly yours, CHH/Jck Enclosure be: D.V. Str`oop w/enccll. v ^ Kettering Lab R.A, Pratt C. H. Eine, M.D., Ph.D. Consulting Toxicologist API 05901 / THE EFFECTS OF 3:4-BEXZPYRENE OX HUMAN SKIN PROF. GIAN' BATTISTA COTTINT and DR GIAN BATTISTA MAZZONR (From the Division of Dermatology and Syphilrlony of tkr University o f Catania, ital\ Prof. F. Flnrrr, Director) The carcinogenic potency of 3:4-benzpyrene and other hydrocarbons ha- been studied in the common laboratory animals, as the mouse, rat, rabbit chicken, pigeon, and guinea-pig (I I. but no attempts to ascertain the reactive capacity of human skin to the carcinogenic agents have been recorded. | t was believed, therefore, that a study of this aspect of the problem would be of interest. Three considerations are of prime importance in this type of ex. HTiment: the extreme cahlion which must be exercised, the correlation with established clinical concepts, and the evaluation of the data obtained. Regarding the safety of the treated individuals, possible complications may be forestalled by daily observation. As circumscribed and readily ac cessible areas of the body are utilized, the potential danger is minimized bv the ease with which the area can subsequently be excised if necessary. ' The approach to the experiment as a problem in clinical dermatology is different from that of the experimental investigator. T ar is^n e of the most commonly employed trniedies in such skin conditions as psoriasis, lichenoid eruptions, kraurosis, and prurigii. and isTTfequently applied for long jteriiMls and not rarely in a concentrated form Xcuplasia as a result of such applica tions has never been observed. Related to the purely experimental aspect of the problem are the investiga tions of Ifaddow and Robin-on 11 ) and of Morelli and (ma.-talla t .i ). I h,-H. authors described an inhibitory effect if benzpyrene on the growth of trans planted tumors. Injection- of the hydrocarbon caused diminution of tin rati of tumor growth or regression of v-tabli-hcd tumor-. Regri--ive phenomena also occurred after local application- to pmliterating external neoplasms. Such observations suggest the pos-ibility ot therapeutic application of tht hydrocarbons in malignant di-ea-e. Although in view o f the m n - i d e r a t i o n - e n u m e r a t e d a b o v e the treat nun; appeared relatively harmh--. it wa- thought advi - abl e for the early c\p, ri- ments to select adult s or tho- e of a d v a n c e d age alter ted with -t: h -eriun- nr fatal disorders a- pe mphi gu- m n - j o s i - luugoide-. or pa ti e nt - with advanced neoplastic di-ea-e. as -cjuamou- . t i l m b.i-.rl cell e a r n e r , m e l a n o ma , or xero derma p ig m e n t o s um, or with e r upt ion- km a n a - br i ng a m e n a b l e to t ar Ihr rape, as psoriasis or porokerato-i- W in n tin- action of In nzpy rerre in thi- group was ascertained, the ob - e r va t i on - w i n ixt et r r h d 'o y o u n ge r imIixirlual- with less serious conditions. Tile rc-nlting d a t a are n i c e --arily c oi n me n - u r a t i with the limited scope of tile e x peri ment , which -ought to d i - d o - e the following: 1. The gross change-produced by benzpyrene in normal human -kin. 2. The possible relation ol tile change- to the type of skin or the age of the subject. - IMi API 05902 TH E EFFECTS OF 3 :4-BENZpTKENE ON HUMAN SKIN 187 i histologic changes in the treat! areas, fhf 0,1 var*ous spontaneous lesions involving the skin. C l a s s if ic a t io n of Pa t ie n t s following conditions were represented in the 26 patients treated with nZr,vreoe: pemphigus vulgans, 2; mycosis fungoides, 1; porokeratosis, 1; ^fyderma pigmentosum, 1; basal-cell cancer, 2; squamous cell cancer, 5; ''pu* erythematosus, 2; psoriasis, 3; syphilis in various stages, 7; ring- ,,,,rrn- 2- . Recording to age, the patients were distributed as follows: less than twenty- r f years, 5; between twenty-five and fifty, 12; more than fifty, 9. P rocedure \ 1 per cent solution of synthetic benzpyrene (Roche) in benzol was aplied to protected and unprotected surfaces of the skin, generally the thigh ind the upper extremity. In isolated instances slight friction was utilized in ` rhimscribed zone after application of a 1 per cent solution in olive oil. ft,e diameter of the treated area did not exceed 2 cm., and care was exercised | jd^successive applications to the original site. At the conclusion of this irfatment a fragment of skin was excised and examined histologically; the [fiiiainder was observed for possible late effects. Practically all patients re eved daily applications, a maximum of 120 being given in a period of four AH patients were hospitalized In several instances treatment was ,,lininistered on alternate days. The carcinogenic potency of the benzpyrene , i- ascertained by means of skin paintings in the mouse. E ffec t on X orm .u . S k in A progressive, relatively constant series of alterations developed in normal ijn following applications of benzpyrene. In chronological order they were: rvthi ina. pigmentation, desquamation, formation of verrucae. and infiltration. Ervthrma. ically perceptible erythema occurred in only 2 patients with jj-al-iell cancer, who were more than fifty years of age. In the vast majority ,nv erythematous reaction was either uansitory, ill-defined, or obscured by hr residue of yellow benzpyrene crystals deposited after evaporation of the "K# ' . ' . Pigmentation: This occurred in all patients. It wa- strictly limited to the reated area, but was more evident in the centra! than in the peripheral zone, fhc time of appearance wa- variable. 1In carlo d observation was after the wentieth application in unprotected normal 'kin of pat; nts more than -i\ty irars old with basal cell or sipiamou- ieli cancer. In patient' between ivrnty five and fifty years, with syphili'. myco-is fungoides. lupus erythema tosus or porokeratosis it developed ..iter the thirtieth application and in the lounged patient, with ringworm, after tin forty-tilth. The change consisted :naiT increase in melanin in the basal layer of the epidermis. - Rarely, small nasses of pigment granules were found in the more superficial layers. A number of factors'appeared to be of importance in this respect. I'ig- API 05903 API 05904 EFFECTS o r 3 :4 -benzpyrene o n h um a n s k in Kit;. H i c a i m M v ; M r i t \ ti. . \ >i Fit. C. S i m w i m I ' io u i i i k \ o <>\ i \ i i i i . F i ' i n m \ i i s nhins. It was more pronounced in dark skinned than in light-skinned per..n'. The nature of the primary skin ailment was without significance. Desquamation: Desquamation, when it occurred, was slight in amount and resembled that of pityriasis. It was proportional in extent to the erythema of the first stage and generally more conspicuous on the unprotected skin. In une patient of, advanced age with porokeratosis and a second with basal -cell canotr, desquamation manifested it-clf by the formation of superficial but scales which, when detached. left a bleeding surface. In some in stances desquamation was accompanied be prurilu~. Formation oj Vrrrunn ' Tin- w.i- the m o s t constant mauitestation pro (luted by benzpyrene. Verrut ,n <le\i I .[.id late and disappeared rapidly after cessation of treatment. They incurred in prat t it .illy all patients, either ac companying or directly succeeding the stage of pigmentation Ycrrucoti' lesiims were observed earliest in older individuals, but no relationship between the time of their appearance and the primary 'kin condition was detected. Of special interest was the occurrence of a network of papilliform elevations in a ruling. dark-skinned patient. The lesions, comparable with those of verruca Although flinica lh IhV lcr-uui" vmtf nut trm- v r r r m . u . t h r \ an- !>r cm u rnilTUv i n n - M r m l j' :nh. It w o u ld !* more- ai'i ura te t* tlr-triln' IIn- a ltera tio n *a~ an a m u tu a ti o n *l the relict juD.irn of the "kin x.u m lar\ to a drriH-niim *! tin* "iiUi API 05905 190 GIAN BATTISTA COTTINI AND GIAN BATTISTA MAZZONE vulgaris, regressed rapidly and disappeared completely one month after treat ment was suspended. , Infiltration: In 2 patients of advanced age with cutaneous cancer and in one with porokeratosis signs of infiltration were perceptible on unprotected skin surfaces or in parts affected by senile atrophy after eighteen treatments Infiltration did not occur in other subjects despite intensive treatment with the hydrocarbon. In one patient in whom evidence of infiltration was observed, benzpyren was applied to the thin, dry, atrophic and pigmented skin on the dorsal surface of the fourth interosseous space of the left hand. Erythema, pigmentation and verrucae developed after eight, twenty, and thirty days respectively. The lesion which ultimately resulted was a well defined, superficial and indolent Fir. 4 l > i . i \ n ' *r Ft. I V if i i- w f< \ : \ i h i m m k \ i ( i h w i h > s i M \\ m m I * -KJ V V:. placque resembling a -trip of pan linn n t . without a<!h--~ion to the deeper Inver of the curium. The histologic nature of the p r o u - a' the ;i ii of tr e a tm e n t amt fit', teen days later was identical. F.--eiitiul <h a n g e - w ere lim ited to the epiderv.i-. In the superficial layer of the d erm a tin te u a - r I;!::::' : a with p o |\ morpho- nuclear leukocytes, chiefly about 'h e d ,,,1 to r t,m u - blood ve--el-. -ur.it and sebaceous gland- an d hair -f* - ' ig. T h e e p id e r m i- -bowed evi dence of acantho-i- i Idg. ? f. u ii^ n iete form ed au irre g u la r -ucc e-sion uf prolongations into the curium . T h e 'tr u t h , u c o ru e u m a - w idened and i on- tained zones of p a r a k e r a to - i- . (Y Hular p ro life ra tio n w a- presen t in the stratum granulosum. ren d erin g t h i - - l a y e r m ore co n sp icu o u s than norm.illy (Fig. 4). The cell- of the -tratum -p in o -u m were en larg ed a n d vesicular, the intercellular b rid g e - w idened a n d the in tercellu lar -p a c e - clea rlv demarcated. API 05906 T H E E FFEC TS O F 3 :4-BENZPYRENE ON H U M A N S K IN 191 larged vacuolated cells were common. The basal layer of cells, however, k 4 ,,naffected. No evidence of nuclear abnormalities nor of an absence of clear demarcation of epidermis and derma was observed. In some areas 1 products of inflammation in the corium extended into the surface layer. ' The pronounced acanthosis and the potential invasive power of the lesion further application of benzpyrene hazardous. The benign course of the ,^5 in the following eight months, however, substantiated its non-malignant racter. .... 1 fomparable early infiltrative lesions with subsequent regression were ob- ^rVtd in 2 additional patients, but more complete observations were rendered difficult by local phenomena of intolerance and hypersensitivity. -fhe average time required for complete regression of the manifestations f-u; 5. B n . u r s L i .s m x s iv a P v u i n t w h i i P i m p ' i h . i s. P h . hu i n tt\ JO A p p i i i \ i n * \ s of Bkn/ pv k im i\ T \u m \ i m o i |) \\x ..reduced by benzpyrene was two months. The lesions generally disappeared nthe reverse order of their appearance. In summarizing the effects of benzpyrene on the normal skin, it mav be .aid that the two most constant alterations were pigmentation .and the formalinn nf verrucae. Erythema, desquamation, and infiltration, although rare, .err most common in unprotected areas of atrophic senile skin. Sex was of no ojw-quence. E ff e c t on \ iin o h \ i \ i S k in The effects of benzpyrene in several patients with pemphigus vulgaris and mum- with xeroderma pigmentosum dien d from those de~eribed in the previ ous section. It is perhaps logical to assume that the iminvolvcd -kin in irni|)higiis or xeroderma is potentially the site of future manifestations of the disorders. , : ' . In the 2 cases of pemphigus, benzpyrene, after 20'applications, precipitated jltxal bullous eruption (Fig. 5) comparable in appearance and course with the eruption of the spontaneous disease. The bullae were preceded by slight :>ignientation and pruritus. The specific effect produced by benzpyrene proh ibir is related to a characteristic sensitivity of the skin in pemphigus (Xikol- hy- sign). Pure benzol produced no reaction. API 05907 |9 2 C U N BATTISTA COTTINI AND GIAN BATTISTA MAZZONE O f special interest are the observations in a child of four years, in good gen. eral condition but having xeroderma pigmentosum, multiple squamous-cefl cancers of the face, and a melanoma of the right cheek. After 85 applies, tions of a solution of benzpyrene to the deltoid and abdominal regions, and to the unprotected skin of the dorsum of the hand, where the symptomatic triad of xeroderma was evident, only pigmentary and minimal verrucous manifesta- (ions developed. The induced lesions regressed, and the parts were unchanged after two years. In contrast to the relative innocuousness of benzpyrene was the more in. tease reaction following exposure of the lumbar region to ultraviolet irradia tion for two minutes at a distance of 0.5 meter. Intense erythema developed following which there ap[H ared raised pigmented patches alternating with flat -, M - '. \,v Vv iAv;\V 7 1 it* Fi t; f>. V m m u T s n i l m i i m \ i \ i i- \ . i \ m i Ki t i n \ m \ H \ i r F t n i j r u . i \ x I* \ I 11 \ r tt| T u t M \ I I ! 1.1 I \ I \ h -* W f i l l KI M U 11. \l . M 1 1 1 ! 4 4 AI T ! It \ T 10\ S O f . Ill \ / h M M | | I | ^ k l v " | Fltkf \M depigmented /one-. The -kin became dry and -caly. The intensity of this reaction did not dimini-li alter tun month- t orre-ponding with the grossh perceptible papular lesion-. hyperpla-ia with hyperkeratosis, parakeratosis, and dyskeratosis, and Inptrpigmi ntatiuti were visible microscopically. Less advanced change-, without dy-keratosis. were observed in areas treated with benzpyrene. It is difficult to a-irihc tIn- difference in reaction to ultraviolet light and benzpyrene in xeroderma to any known fat tor or -pecitic change in the dis ease. Theoretically it i- po-dhle that -ome metabolic disturbance (porphyrin) may be re-pon-il>le lor a relative re-i-tarue to one agent and a hypersensitivitv to the other. The effect of ultraviolet light oil skin previously treated with benzpyrene was not investigated, a- dangerous complications were feared. The two irritants, however, were combined inadvertently, since benzpyrene API 05908 U K EFFECTS OF 3 :4-b e n z ft e e n e o n h u m a n s k in 193 applied to exposed skin surfaces. No significant differences from the of benzpyrene in other portions of the body were observed. histologic changes, like the gross lesions, were closely related to the --of the patients rather than to the nature of their skin disorders, their gen- ^ condition or sex, as follows: . In patients less than twenty-five years oj age: ' 1 1 ) Minimal irregularities in the superficial strata of the epidermis, with tendency to the formation of epithelial outgrowths in the region of the hair follicles- (2) Hyperkeratosis with occasional formation of small cornified masses at jic openings of the hair follicles. tit, 7. CltANtil'.S l \ T i l t S l R V t l M l i | U \ l (MM M u t l ' \ l a|i 11. 311 SKIN SfR l' Vf l. IN \ PMILNT Ofr T w e n t y m \ k V f m s . w i t h At r i \ t aftfr 40 A p p ih \ tio\ s of B k\/i* Y k iM . in F orty i \yt> D ays . 3) Inconstant vacuolization of cells in the stratum spinosum. Increase in melanin in the basal layer. W ) Inflammatory changes in the papillary portion of the corium about ihr hair follicles and sebaceous glands. (6) Occasional cellular irregularities and giant-cell formation at the open ings of the hair follicles I l-'ig. i>>. //. In patients between twent y-firee anil fifty years oj aye: Variations from the changes in the younger group were minimal. In patients with active gum matous lesions of the skin more accentuated manifestations were observed: (1) Focal proliferative lesions in the stratum grnnulosum. The width, however, did not exceed that of five layers of cells ( Fig. 7). Nuclear abnor malities were not observed, but there was acanthosis! (2) More intense inflammation in the derma. API 05909 194 GIAN BATTISTA COTTINI AND GIAN BATTISTA MAZZONE / / / . In patients older than fifty years: All changes were more intense. Ad vanced lesions in several cases have been described. Special studies of the cutaneous nerves were not made. Basophilic in filtration was not encountered. E f f e c t o f B e n z p y r e n e o n A c t iv e S k in L e sio n s The action of benzpyrene on the following processes involving the skin was observed: ulcerated squamous-cell cancer, basal-cell cancer, tumor manifesta tions.of mycosis fungokles. desquamating form of psoriasis and porokeratosis Ulcerated Squamous-eell Cant er: An ointment containing 1 per cent benz pyrene was applied daily to an ulcerated growth in the case of multiple tumors associated with xeroderma. The initial effect appeared to be favorable, hut treatment was discontinued after 50 applications, when no definite retardation in growth was evident. The hydrocarbon did not accentuate the malignant nature of the process. Similar results were obtained in a case of roentgen-ray cancer in a man of sixty years. The benzpyrene in addition exerted a pro longed analgesic effect. The destructive action of tar on nervous tissue in ani mals may be a related phenomenon. Basal-a ll Cana r: Xo gross or histologic changes were observed in several cases of basal-cell cancer after 87 treatments with benzpyrene. Myeosis Fungoftlrs: I.eiions in the tumor stage in the iliac, thoracic and brachial regions regressed incompletely' after daily applications of benzpyrene for two months. The treated areas appeared less cyanotic, and the external surface less irregular. Further regression did not occur with continued treat ment. Associated with the changes were the pigmentary and verrucous mani festations commonly produced by the hydrocarbon. Xon-treated areas of mycosis fungoides in other portions of body progressed and ulcerated exten sively. Biopsies unfortunately were not available. . Desquamating Psoriasis anti Porokeratosis: Desquamation appeared to be hastened after 15 treatments. The residual erythematous lesions, however, remained unaffected, and the exfoliative manifestations of the primary erup tions recurred. Histologic evidence of complicating processes was not evi dent after 50 applications of the hydrocarbon. Syphilis: The accentuation of the skin reactions in patients with gum matous lesion- ha- been described above. This phenomenon may bear, a re lationship to the assumption, frequently expressed, that syphilis acts as a pre disposing factor in the -poutaneou- development of neoplasia: ' ( '( I X I I.I s i O N S 1. Dailv skin applications of a 1 per cent solution of benzpyrene in benzol in 26 human subject' for variable period', not exceeding four months, resulted indefinite manifestation- localized to the treated areas. Erythema, pigmenta tion. desquamation, the formation of verrucae and infiltration developed in chronological order. Pigmentary and verrucous lesions were the most com monly observed change'. The gross and-microscopic characteristics of each stage are described. API 05910 THE effects of 3:4-benzpyrene on human skin 195 j Following the suspension of treatment after a maximum of 120 appli e s or when evidence of infiltration appeared, the manifestations regressed vTpjetely within two to three months. Although reversible and apparently pign, the changes undoubtedly represented early stages of a process which have ultimately gone on to neoplastic proliferation. In a broad sense may be designated as prototypes of a precancerous process. 1 3 The alterations were more pronounced in older than in younger indi.jppls, and on the unprotected skin surfaces. The reaction of young pawith active gummatous lesions was comparable to that observed in the 'idee rouP0 4 In patients with pemphigus vulgaris benzpyrene produced a bullous ruptin comparable in appearance and course to the spontaneous disease. protected and unprotected skin of a patient with xeroderma pigmentosum, bile extraordinarily sensitive to ultraviolet irradiation, did not react charac*ristically to benzpyrene. 5. Treatment with benzpyrene of ulcerated squamous-cell cancer of the j.|n was followed by temporary amelioration. In one instance an analgesic was observed. Limited retardation of the progress of mycosis fungoides lesions in the tumor stage occurred in one case after local application of benz- Basal-cell cancer and desquamating lesions of psoriasis or porowere unaffected. (, The assumption appears warranted that benzpyrene, if applied to hujnan skin for protracted periods, Twould be carcinogenic as it is in animals. >'oTE: All patients were seen hv the authors at the end of April 1*13*1. In none were lunges found at. the site of the benzpyrene applications. R efehences . (.IK. J. W.. Haslkwood. (1 A. LI.. Heweit. C L I l i u m . I . Kfnnaw ay. E L . a\ d Mayneokd \V V.: Am J. Cancer 2'>: 2 to. IOAT t ,hiK. J. \Y . AND Kennaw AY. E I. Am J Cancer As. 50. I*I.?S. pycZA. G .: Boll. cl. Soc. med.-chtr . Catania. 1*153. p. h2. . Jluinow. A.: Nature lab: SOS. 1055. Ilcnnow. A., and R o binson. A \1 : I'roc Roy Soc ser It 122. 442. 1*137. . AIoHELU. E.. and G t AST.AU.A. A .: Boll, lega ital. p. 1 loti a c o n tro il can cro 14 ( 1 0 ; : 110, 1*136. . API 05911 f 4 I A r r ; . . :i:< Copy From D. V. Strocp For Information of Members of Medical Advisory Committee____________________________ __________________ July 21, 195^ ESSO LABORATORIES Standard Oil Development Company P. 0. Box 51, Linden, N. J. July 13, 195*4- Dr. George M. Saunders Socony-Vacuum Oil Co., Inc. 26 Broadway New York U, New York Dear Dr. Saunders: I am attaching the report of the Subcommittee on Carcinogenicity to the Medical Advisory Committee of the API. This report summarizes briefly the work which has thus far been done, the expenditures thus far made, the anticipated future work to be done under the auspices of this Sub committee, and the proposed budget for the period July 1, 1955-June 30, 1956. You will note that the Subcommittee suggests that funds in the amount of $5,000 be solicited for the continuation of the work of the Kettering Laboratory during the period July 1, 1955-June 30, 1956. This represents a reduction of approximately $7 ,2 5 0 in the amount already budgeted for the interval July 1, 195^-June 30, 1955- It has been suggested by one of the Subcommittee members that this report should contain a statement which would describe the benefits which this study would accomplish for the oil Industry. It is my belief that the benefits to be expected by the oil industry are contained in the brief outline of future work. However, if in your opinion this would not suf fice, then I believe the following statement might be added to this report immediately following the section on future work and Immediately preceding the estimated budget for July 1, 1955-June 30, 1956. "It is anticipated that this work will benefit the petroleum industry by providing basic information which will permit the petroleum industry to control its own potential carcinogenic hazards without gov ernmental regulation." As Chairman of the Subcommittee on Carcinogenicity, I respectfully submit this report of the Subcommittee. Very truly yours, KEE:ilk Attachment cc: Messrs. J. J. Fhair A. Wesley Horton R. A. Kehoe D. V. Stroop R. E. ECKAROT, M. D. API 05912 4 4 THE REPORT OF THE SUBCOMMITTEE ON CARCINOGENICITY TO THE MEDICAL ADVISORY COMMITTEE OF THE API The Subcommittee on Carcinogenicity of the API Medical Advisory Committee, one of the first subcommittees created, was established early in 19^5. It was formed after it became known that certain petroleum oils and fractions had been demonstrated to possess fluorescence similar to certain known carcinogens, and, when painted on mice, produced tumors and cancers. Negotiations were then begun between this Subcommittee and Dr. Kehoe of the Kettering Laboratory which culminated, early in 19^6, in an agreement between the API and the University of Cincinnati, under which the Kettering Laboratory of Applied Physiology would "investigate the toxi city and mode of action of certain petroleum products." The investigation began with the testing of 8 oils supplied by three participating companies, in addition to negative controls of water and white oil and positive controls of methylcholanthrene. Of the 8 oils tested, only 2 produced skin tumors in mice. One of these was an unflashed thermal residue, the other a clarified oil. These investigations were carried out at a cost of $6,000, and were reported in detail to the Sub committee by the Kettering Laboratory on November 5; 19^8* However, when the results were becoming apparent early in 19W , the Medical Advisory Committee, on the advice of the Subcommittee on Carcinogenicity, forwarded to the members of the Safety Committee of the Board of Directors of the API a resolution dated April 5> 19^8 This resolution called upon the Executive Committee of the Board of Directors "to establish a comprehensive research project for the development of fundamental facts, analytical techniques and related information pertain ing to cancer-causing compounds which may occur in petroleum or be produced therefrom." This resolution called for the solicitation of contributions to a special research fund in the amount of $110,000 for the first year, of which $ 50,000 would be a non-recurring capital expenditure for construc tion of extraordinary experimental facilities at the Kettering Laboratory. This resolution was presumably approved by the Executive Committee and on July 1, 19h8 a check in the amount of $65,000 was sent to the Kettering Laboratory, $40,000 of this amount as a grant for animal quarters. With the expansion of this research project in 19^8, A. W. Horton, Ph. D. was added to the staff of the Kettering Laboratory to supervise the chemical and biological work on petroleum and petroleum fractions, and the services of J. J. Phair, M. D., Professor of Preventive Medicine, University of Cincinnati, College of Medicine, were made available to the staff to develop and supervise an epidemiological survey of the incidence of cancer in the petroleum industry. From this point on, the research project can be divided into two phases, namely (l) the chemical and biological work, and (2 ) the epidemiology. API 059 4 - 2- 4 1. The Chemical and Biological Work Under Dr. Horton's supervision, an expanded program of biological testing for carcinogenicity was begun on a variety of petroleum fractions selected and provided by the technical staffs of the participating petroleum companies. It soon became evident that in order to express the relative carcinogenicity of different petroleum fractions in a quantitative fashion, it would be necessary to develop standards of reference to which the petroleum fractions could be related. For this Dr. Horton began biological work with solutions of a recognized carcinogen, methylcholanthrene, in varying con centrations in various solvents. This work clearly showed that a biological assay for carcinogenesis was a highly complicated test which was affected by a great number of variables, such as the general health of the animals used in the test, the genetic strain of the mouse used for the test, the toxicity of the material applied in the test, the presence or absence of lice or mites on the animals, fighting among the experimental animals, etc. There fore, steps were taken to control these variables insofar as possible, although it must be recognized that even after k or 5 years of research it has not been possible to eliminate or completely control these variables but only to minimize them. Where these variables were incapable of control, this early research did permit the biological experimenter to recognize their effects so that he could throw out as unsatisfactory those biological tests in which these variables have played too great a role in the final results. This bio logical research on the development of `standards of reference is summarized in the report "Carcinogenesis of the Skin, I. Basic Methods Employed in Testing Complex Hydrocarbon-Type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies" by A. Wesley Horton and Dorothy T. Denham. This report has been proposed for publication in a cancer research journal. The second achievement of this biological research has been the demonstration that solvents influence results obtained with pure carcinogens. At present two types have been recognized. The first type, represented by benzene, has no effect on the activity of pure carcinogenic hydrocarbons. This type of solvent has been called a non-accelerating solvent. When the pure hydrocarbon, methylcholanthrene, is dissolved in this type of solvent, the carcinogenic activity in animals is directly related within limits to the concentration of the carcinogen in solution. The second type of solvent, represented by dodecyl benzene, has an additive or accelerating effect on the activity of pure carcinogenic hydrocarbons. This type of solvent has been designated an accelerating solvent. When the pure hydrocarbon, methyl cholanthrene, is dissolved in this type of solvent, the carcinogenic acti vity in animals is not directly related to the concentration of the carcinogen in solution but rather appears to be more dependent on the amount of accel erating solvent applied with each application. The research work leading to this discovery is soon to be summarized in a paper which the laboratory will propose for publication, also in a cancer research journal. In addition to the work m i reference standards and solvents outlined above, the laboratory has tested biologically between 150 and 200 petroleum materials and fractions of these for their carcinogenic activity in mice. This work alone required some k,000 mice for test purposes. A summary of the results of these investigations will undoubtedly be prepared as a manuscript for publication. API 05914 0 - 3- The laboratory has also investigated the effect of hygienic measures such as washing and protective creams on the carcinogenic acti vity to mice of several petroleum materials. It is anticipated that these results will be used in the framing of practical hygienic control measures. The chemical work has encompassed the separation by distillation, chromatography, and chemical methods, of petroleum materials into various fractions in an attempt to define precisely the agents in petroleum respon sible for this carcinogenic activity in mice. This work has already led to the submission of one manuscript entitled "The Use of Catalytic Iodination on Activated Alumina for the Determination of Benzo (a) Pyrene in Complex Mixtures." This manuscript was authored by R. Tye, M. J. Graf and A. W. * Horton. It has been reviewed by the Subcommittee, the Medical Advisory !* Committee, and the Medical and Health Committee of the Board. No objections to publication were voiced and it has been submitted to the editors of the Journal of Analytical Chemistry for'publication. i In addition, the following general information has come out of the combined chemical-biological studies: a. Most petroleum materials boiling in the range 500-6509F probably contain accelerators. This includes paraffins from normal decane to hexa- or hepta-decane and naph thalenes which have at least a butyl chain attached. b. Carcinogenic petroleum materials boiling between 700725F probably contain phenanthrene-like carcinogens similar to benzo (c) phenanthrene. c. Carcinogenic petroleum materials boiling between 725800F probably contain anthracene-like carcinogens, possibly cyclopenteno benzanthracene. d. Carcinogenic petroleum materials boiling above 800F probably'contain benzo (a) pyrene. 2. Epidemiology I It was early recognized by the Subcommittee on Carcinogenicity that the chemical and biological work outlined above would give only a partial answer. It was explained that the epidemiological work would de velop information on the following two questions: a. Does work in the petroleum industry or at any specific Job within the petroleum industry subject the worker to an increased risk of developing cancer? A w b. Have newer refining processes, such as catalytic cracking, or increased severity of thermal cracking introduced any new risks of developing cancer to any segment of the petroleum workers? It is in general these newer processes which have been shown to pro duce materials which have a high degree of carcino genicity to mice. API 05915 $ 4 - 4 - A central registry for the reporting of cancer cases occurring in the petroleum industry represented by the participating companies of the API was established at the Kettering Laboratory under the direction of Dr. Phair. In addition to reporting such cases, it was deemed essential that a complete occupational history for each case also be reported so that correlations with occupation could be made. In the course of setting up this registry, it soon became apparent to Dr. Phair that the medical departments and medical records of some petroleum companies were inadequate to permit accurate reporting of cancer cases. Initially, therefore, Dr. Phair's major effort was in assisting medical directors, or refinery managers where medical directors were unavailable, in organizing their departments and/or records so that good case reports and occupational histories would be received in his office. Therefore, it has only been in the past year or two that an approach to effective reporting of all types of cancer cases and occupational histories has been achieved. Despite incomplete uniformity of reporting and partici pation, over 1,300 cancer cases have been reported to Dr. Phair from 15 com panies, and are in the pr.ocess of statistical analysis. It is believed that worthwhile statistical analysis of these cases will require maintenance of this registry for a minimum of 6 years. In the interim, Dr. Phair continues to act in an advisory capacity to the petroleum companies and their medical directors for the collection and reporting of these cases. Expenditures of the Subcommittee on Carcinogenicity 1946 19^7 1948 $ 6,000.00 0 6 5 .000. 00 (includes $40,000 grant for animal quarters) 1949 1950 1951 49,413.00 89, 963.39 59,065.12 to June 30, 1952 31,692.42 July 1, 1952 to June 30, 1953 July 1, 1953 to June 30, 1954 104,760.00 7 9 .000. 00 Budgeted July 1, 1954 to JQne 30, 1955 $484, 893.93 $ 72,250.00 API 05916 - 5 - Future Work As indicated in the above outline of expenditures, a peak of activity was reached in the period July 1 , 1952 to June 30, 1953- Since that time there has been a gradual tapering off of expenditures for this research effort. The Subcommittee recommends that future work in the chemical-biological phases should be directed towards 1. Work on physico-chemical methods for assaying the carcinogenesis of petroleum materials. These methods will be validated by a. Determining the relative importance of carcinogens and mixtures of carcinogens and accelerators in the process of carcinogenesis. b. Determining the relative carcinogenicity of additional petroleum products and materials, not adequately covered to date. c. Isolating and identifying additional carcinogens and/or accelerators in petroleum materials. d. Continuing work on the improvement of standard bio assays for carcinogenesis. 2. Effective hygienic control measures for use by the petroleum industry. 3. Work on possible carcinogenic inhibitors. In the Subcommittee's opinion, this type of research should con tinue until at least 1959- However, ejqjenditures will continue to taper off during this interval and should stabilize at levels not in excess of $25,000 after June 30, 1956. 1 The epidemiology should be continued until at least 1959* The budget for this phase of the work will probably run between $15,000 and $20,000 per year. In the opinion of the Subcommittee, if full coopera tion with Dr. Phair in the epidemiological work has not been achieved by June 30, 1955, the Subcommittee would recommend that epidemiology work be discontinued since it would be likely that full cooperation would not be expected to be achieved after this date. If cooperation has been achieved by this time, expenditures should stabilize at a somewhat lower level, since the work will consist principally of collecting and analyzing statis tically those cases reported. It is thus estimated that the anticipated annual expenditures for 1956-1959 will be in the vicinity of $40,000-$^5,000. Estimated Budget July 1, 1955 to June 3 0 1 1956 The Subcommittee recommends that the Medical Advisory Committee request funds in the amount of $6 5 ,00 for the continuation of this work during the interval July 1, 1955-June 30, 1956. A brief breakdown of expenditures is approximately as follows. API 05917 - 6- Biological Work $20,000 Technical Work 32,250 Epidemiological Work 12,750 $65,000 Another breakdown of these figures can be given as follows; Salaries Direct Indirect Histopathology Others $22,000 2,000 20,000 Miscellaneous Exp. Animals Supplies Travel Overhead $ 1,250 5,000 i+,000 10,750 $144,000 21,000 t* Composition of Subcommittee R. E. Eckardt, M. D., Chairman Standard Oil Development Company E. W. Adams Standard Oil Company (Indiana) L. C. Beard, Jr. Socony-Vacuum Oil Company, Inc. Allan E. Dooley The Texas Company F. D. Gassaway, M. D. Gulf Oil Corporation L. M. Henderson The Pure Oil Company $65,000 API 05918 REE:ilk June 21, 1951* -A 1 -7 - C. H. Hine, M. D. Shell Development Company E. K. Linder, M. D. The Atlantic Refining Company R. C. Mithoff California Research Corporation M. N. Nevquist, M. D. The Texas Company James W. Osborn, M. D. Standard Oil Company (Ohio) P. J. Sanders Standard Oil Company (Ohio) r # APPENDIX A - 2 ESSO LABORATORIES 3TANDARD OIL DEVELOPMENT COMPANY P. 0. BOX 51LINDEN, N. J. June 22, 1951*- Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New York 17, New York Dear Mr. Stroop: On June 19, 195^> the following members of the Subcommittee on Carcinogenicitymet at the Kettering Laboratory in Cincinnati to discuss the manuscript by A. Wesley Horton and Dorothy T. Denman entitled "Carcinogenesis of the Skin, I." R. E. Eckardt, M. D., Chairman C. H. Hie, M. D. W. E. Scovill, representing Dr. Osborn and Dr. Sanders L. M. Henderson A. E. Dooley, representing Dr. Newquist and himself E. K. Linder Prior to the meeting of this group, I had received communications from all the other representatives of the Subcommittee indicating no objection by any member of the Subcommittee to the publication of this manuscript from a policy standpoint. Individual members of the Subcommittee had submitted certain suggestions to Dr. Horton for his consideration, and the representatives of the Subcommittee were advised that he is taking these comments into considera tion and introducing all of those with which he is in agreement, prior to * submitting the paper to the publisher for publication. The following changes were recommended by the Subcommittee and will be incorporated into the manuscript before submission: (1) A footnote identical with that attached to the previous manuscript from Dr. Horton and his associates acknowledging support of the API will be attached to this paper. (2) The word "experimental" will be inserted between the words "basic" and "methods" in the title of the paper. (3) The second sentence of the second paragraph of the paper will be changed to read as follows: "Certain materials, such as shale oil (1,2,3)> which have induced an abnormally high incidence of cancer of the skin of men as a result of repeated exposure to their occupation, have been found capable of producing epitheliomas in the skin of mice." (Continued) API 05920 Mr. D. V. Stroop * 2 June 22, 195^ As indicated, Dr. Horton will introduce an additional three literature references in the bibliography on page 29 of the manuscript. (U) Dr. Horton was not agreeable to the rewriting of the first para graph of this paper since it is the stated purpose of the Kettering Laboratory that their interest in this problem is only insofar as it will lead to the development of information which will be of help in the estimation of the extent of the potential hazard of skin cancer in certain industries, which include not only the petroleum industry but also the coal tar industry and others. The Kettering Laboratory quite clearly recognizes and so states in the first paragraph that the determina tion of the relative carcinogenic potencies of various materials is only part of the determination of the potential hazard. It is, however, their belief that in industries or segments of industries which do not have adequate medical and industrial hygiene facilities of their own to determine adequately the extent of exposurej the Kettering Laboratory will be requested to complete the picture and make a full estimation of the potential hazard. For this reason they feel that the first paragraph of this paper as written in the manuscript accurately states their position in regard to skin cancer and they are not, therefore, willing to change the wording of this opening paragraph. By copy of this letter to each of the members of the Medical Advisory Committee, I am requesting that they be informed of the action of the Subcommittee and request that they indicate any objections to publication of this paper directly to Dr. Saunders so that he may express the opinions of the Medical Advisory Committee concerning this manuscript at the time that he submits it to the Medical and Health Comnittee of the Board for their review. Very truly yours, /s/ R. E. Eckardt R. E. ECKARDT, M. D. REE:ilk cc: Members of the Medical Advisory Committee API 05921 4 ESSO STANDARD OIL COMPANY 15 WEST 51ST STREET NEW YORK 19, N. Y. APPENDIX A - 3 July 20, 195U Re: Proposed, publication by the Kettering Laboratories of "Carcinogenesis of the Skin,I." Dr. R. E. Eckardt Medical Research Division Standard Oil Development Company Post Office Box 51 Linden, New Jersey Dear Dr. Eckardt: Please forgive me if I see* overly persistent in objecting to the opening paragraph of the above paper. The opening sentence "The investigation of skin cancer in this Laboratory has been concerned with the estimation of the extent of the potential hazard of this disease in certain industries" is simply not in keeping with established facts and contributes nothing to the meaning of the paper other than possibly abetting some already established misconceptions. The statement is erroneous in the sense that only doctors dealing with human beings exposed to the substances in question under existing working conditions can determine whether or not an actual hazard exists. I believe Mr. Horton and Miss Denman would be well advised not to read into their data more than is justified by the facts which they have established. To my way of thinking, this detracts nothing from the importance of the fine Job they have done. It would be my suggestion that the first sentence be replaced by the following: "A number of chemical substances have been suspected of being etiological agents for cancer of the skin." In addition, I would insist that the final sentence read ". . . . methods of determining the first variable for mice, the level of activity . . . . " If it is firmly established at some time in the future that the mouse data are directly transferrable to humans, there i6 no reason to doubt that this publication will have proved any less useful. Sincerely yours, /s/ Leo Wade LEO WADE, M. D. LW:HH cc: Dr. Mr. Dr. G. M. D. V. R. A. Saunders Stroop Kehoe API 05922 A .X A / Copy From D. V. Stroop For Information of Members - Medical Advisory Committee____________ __________________________ July 21, 19% B 15 July 195^ REPORT CF SUBCOMMITTEE ON FLUORIDES G. M. W. H. L. E. A. C. Saunders, M. D. Barcus Curtis, M. D. Pabst - Chairman The introduction of EF alkylation caused the petroleum industry to be confronted with the question of health hazards arising from the operation of HF alkylation units as affecting man, animals, and plant life. In January, 19^+6, considerable hysteria arose among farmers and residents in the industrial regions of Southern New Jersey relative to potential and alleged crop damage. Prior to this and in the central part of the United States, the aluminum industries had experienced great difficulty with fluorosis of cattle grazing in the vicinity of aluminum plants. Accordingly, the A.P.I. Subcommittee on Fluorides was formed to study the matter from the standpoint of the petroleum industry. One of the first efforts of the Subcommittee was to seek a scientific organization that would undertake such studies. After consider able investigation it was found that the Kettering Laboratory was already working on fluorides, the project being sponsored by five companies handling i t HF or fluorides to the extent of $17,000.00 per year. The five companies extended an invitation to A.P.I. to participate in the work and made avail able all previous data and reports on the project. A.P.I. agreed to con tribute $2 ,500.00 per year (on a year-to-year basis) and thus received the benefit of an extensive investigation at a nominal cost. Payments of $2 ,500.00 per year started in 19^7 and have been continued to the present time. During 1953-5^ the fluoride project at Kettering Laboratory, University of Cincinnati, was sponsored by ten companies or organizations, total contri butions amounting to $28,310.00. American Petroleum Institute yearly contri butions amount to $2,500.00 and payment was made in June 195^ for the twelve month period ending 30 June 1955* Investigations carried out during the past year include: 1) Urinary excretion of fluorides by subjects exposed to HF laden air. 2) Urinary excretion of fluorides ingested as sodium fluoride. Two individuals with evidence of renal dysfunction stored 76-81$ of ingested fluoride whereas three normal individuals stored only 30-35$ fluoride. API 05923 \I - 2- 3) Rabbit experiments to evaluate effectiveness of sodium pyruvate as an antidote for acute fluoride poisoning. Son inhibiting effects were noted. h) Rabbit studies on intratracheal injections of aqueous suspensions of fluoride bearing dusts. 5) Radiological studies of 50 persons residing for 20 years or more in Lake Preston, South Dakota, where drinking water contains 6 ppm of fluoride. No increase was detected in radiopacity of the bones in h8 persons, and only very slight increase in remaining two. The American Petroleum Institute has contributed to the above project for some seven years, and it is believed this has aided in the accumulation of useful knowledge on fluoride compounds. During this period, the petroleum industry has gained much practical experience in the use and handling of HF so that today studies on fluorides are of considerably less importance than they were seven years ago. In view of this, it is the recommendation of the Subcommittee that consideration be given to the termination of this activity at the close of the current period, June 1955- /s/ G. M. Saunders API 05924 oa APPENDIX C opy Frcn D. V. Strocp For Information of Members of Medical Advisory ommittee ____________________________ July 16 , 195^ ESSO LABORATORIES Standard Oil Development Company P. 0. Box 51> Linden, N. J. July lL, 195L Dr. George M. Saunders Socony-V cuum Oil Co., 26 Broadway New York L, New York Inc. Dear Dr. Saunders: ' I respectfully submit the report of the Subcommittee on the Physiology of the Skin to the Medical Advisory Committee. You will note that our Subcommittee recommends that the API solicit funds in the amount of $5,000 for the interval July 1, 1955 June 30, 1956, in continuing support of this work by the Medical Advisory Committee. Very truly yours, REE:ilk Attachment cc: Messrs. R. A. Kehoe D. V. Stroop R. E. ECKARDT, M. D. TEE REPORT CF THE SUBCOMMITTEE ON THE PHYSIOLOGY OF THE SKIN TO THE MEDICAL ADVISORY COMMITTEE OF TEE API The Subcommittee on the Physiology of the Skin was created at the 13th Meeting of the API Medical Advisory Committee in response to a request from Dr. Kehoe for support of work that his laboratory was performing on this basic problem. In the original letter from Dr. Kehoe, it was indicated that this work would require approximately $25 ,0 0 0 per year for its continuance, and that he was asking support only to the extent of $5 ,0 00, the remaining to be made up from contri butions of other interested groups. A committee appointed by B. B. Reeve, M. D., the Chairman of the Medical Advisory Committee, recommended that the API support Dr. Kehoe's work to the amount of $5,000 per year. The first contribution of $2,500 supported this research until June 30, 1952. Since that time, $5,000 annually has been given to the Kettering Laboratory in support of this work. To date, the Kettering Laboratory has worked on the following projects in connection with this work: * (1) The influence of surface active agents on skin reactions produced in experimental animals by primary irritants and sensitizers. It is believed that this work may be of value to the petroleum industry since it may suggest certain surface active agents which will be useful in the petroleum industry without increasing the effect of primary irritants or sensitizers on the skin, or, conversely, may point out certain surface active agents which should be avoided by the petroleum industry since they will en hance these primary irritating or sensitizing actions. (2) Occupational acne. It is known that occupational skin diseases constitute approximately 13$ of all occupational diseases. The factors which influence the production of occupational skin diseases, including occupational acne, may furnish leads to the petroleum industry for their adequate control. This information may be helpful to the study of oil dermatitis proposed by the Lubrication Com mittee of the API, if and when such a study is undertaken. (3) Changes in the sweating mechanism in psoriasis and other skin diseases. The wide geological distribution of the petroleum industry, from the tropics to the arctic, suggests that an understanding of the contribution of the sweating mechanism to skin disease may see wide application in the control of such diseases in the petroleum industry. (U) Work on evaluation of protective creams. A portion of this work has already been published by Dr. Suskind in the Archives of Industrial Hygiene and Occupational Medicine. It represents the most scientific work that has ever been performed on protective creams, and has already found applications within the petroleum industry. (5) Cooperative work of the API carcinogenicity project. This naturally has direct potential benefits to the petroleum industry. This cooperative work includes the effect of variables on the absorption of materials applied to the skin and the interpretation of microscopic changes produced by co-carcinogens and carcinogens. It is the recommendation of the Subcommittee on the Physiology of the Skin that the API, through the Medical Advisory Committee, continue to support the basic skin physiology work being performed at the Kettering Laboratory in the amount of $5,000 for the interval July 1, 1955-June 30, 1956. The actual budget for-195^ for this work is $33,000, contributed by 5 other parties in addition to the API. The API contribution is therefore less than l/ 6 of the total budget, and the API is one of 6 contributors. Composition of the Subcommittee on the Physiology of the Skin Robert E. Eckardt, M. D., Chairman Standard Oil Development Company F. F. Boys, M. D. Sinclair Refining Company M. N. Newquist, M. D. The Texas Company REE:ilk July lk, 195k API 05927 ,\ i I Appendix D - 1 I S e p te m b e r 1U, 19 54 Mr. M. J. Rathbone, Chairman API Medical and Health Committee c/o Standard Oil Company (New Jersey) 30 Rockefeller Plaza New York 20, New York Dear Mr. Rathbone: As requested in your letter of August 17 to David V. Stroop, the Medical Advisory Committee on September 9 reviewed, discussed, and offered comments on the project proposed and described in R. Cubicciotti's letter of July 19. The stated purpose of the proposal has been the subject of 0 repeated discussions notably at the joint meeting of representatives in New York on.July 16, 1953 when members of the Subcommittee on Cutting Fluids failed* to persuade the members of the Lubrication Committee panel that a search for causative agents might be unduly costly and lead to no practical results, from a medical viewpoint. This led to a tacit agreement on the compromise proposal for the modified project at an arbitrary maximum cost of $20,000. The Medical Advisory Committee concurred In the compromise agreement despite its persistent belief that the estimated cost of the modified proposal was high in the light of the meager results which could be anticipated on the medical aspects of the dermatoses problem. During the past year nothing has developed to change the views of the Medical Advisory Committee members. They now reiterate their belief that the project will probably contribute little or nothing to the fund of knowledge on the medical aspects of the dermatoses problem, although we are not in a position to assess any public relations value. However, it is the wish of the Medical Advisory Committee not to obstruct the project, if the Division of Marketing desires to proceed with the work. Very truly yours, SIGNED BY George M. Saunders, M. D. Chairman Medical Advisory Committee API 05928 T T APPENDIX D - 2 j A REVIEW OF MEDICAL ADVISORY COMMITTEE STOPIES RELATING TO CUTTING FLUIDS General - The Medical Advisory Committee throughout the ten years of its existence has recognized the prevalence of dermatitus incidental to exposure to oil and products containing oil. Morbidity figures show that 1G -15 percent of all industrial disease cases involve dermatitus following such exposures. 19^5 - At its organization meeting on May 7 and 8 19^5, the Medical Advisory Committee recommended a study on chlorinated compounds, particularly chlorinated cutting oils. This study resulted in tentative conclusions (November 19^5) that: O "1. Chlorinated paraffins appear to be relatively free from condemnation due to either systemic or dermatological effects. "2. The likelihood of systemic effects by the use of chlorinated aromatics in cutting oils is remote. There are claims in the literature of adverse dermatological effects. "3. Suitable precautions should be taken in the selection of workers, the working conditions and hygiene. "4. The toxicity of carbon tetrachloride is well recognized and therefore should not be used as a cutting oil additive.1' 19^6 - The conclusions reached in 19^5 were confirmed and the Medical Advisory Committee expanded its study to include all cutting fluids. 19^9 - "For information only - Not for publication" the Medical Advisory Committee had prepared two mimeographed bulletins: 1. "The Cutting Oil Dermatoses - A Critical Review" By Bertrand C. Kriete - Harvard University School of Public Health. 2. "Safeguarding the Use of Cutting Fluids and Coolants" By Roy S. Bonsib - Standard Oil Company (N. J.) 1950-195^ Medical Advisory Committee studies on cutting fluids were suspended awaiting the development of recommendations under consideration by the Lubrication Committee, Division of Marketing. -- 0 0 O0 0 - - 05929 ( APPENDIX E - 1 Ccpy From D. V. Stroop For Information of Members of Medical Advisory Committee_________ . _______________________________ July l6 , 195* SHELL DEVELOPMENT COMPANY Emeryville, California July 9, 195*+ AIR MAIL George M. Saunders, M. D. Socony-Vacuum Oil Co., Inc. 26 Broadway New York k, N. Y. Dear Dr. Saunders: Enclosed you will find reports of the Subcommittees on Economic Poisons and Cg- C^o Aromatics. These reports are submitted in accordance with the request or Mr. D. V. Stroop, as outlined in his letter of May 5, 195^. I have the concurrence of Dr. Curtis with the form and content of the report on economic poisons. Unfortunately, the Subcommittee on C9- C-j_2 Aromatics has had only one meeting, and the report reflects rather arbitrarily my conclusions as to the activities of this subcommittee. I have as of this date submitted the report to the other members of the sub committee. If these members have serious objections to the contents, I will review them and perhaps modify the report. I trust that this will not be necessary, but I would like to reserve the right to alter this second report in some measure, if necessary. Very truly yours, /s/ C. H. Hine C. H. Hine, M. D., Ph.D. Consulting Toxicologist CHH/jck Enclosures API 05930 U Report of the Subcommittee on Cg- Cj_2 Aromatics to the Medical Advisory Committee of the American Petroleum Institute July 195^ I. Objectives 1. To assess the potential health problems associated with Cg- Cy aromatics. 2. To investigate the desirability of conducting industrial hygiene and toxicological studies concerning Cg- C-^ aromatics. I I . Scope and Accomplishment 1. Past Accomplishments Consideration of possible health problems associated with the Cg- C ]_2 aromatic fractions from petroleum solvents was first given by the Subcommittee on Permissible Concentrations at the seventh meeting (19^8) of the Medical Advisory Committee. At that time it was pointed out that there was a paucity of data concerning the industrial hygiene and toxicology of these compounds that it would not be possible to prepare suitable toxi cological reviews. The Medical Advisory Committee therefore agreed to spon sor a toxicological investigation of representative compounds in this group. A sum of $10,000 was voted to support these studies, and preliminary arrange ments were made with the Department of Pharmacology at the Baylor University School of Medicine in Houston, Texas. However, at the ninth meeting of the Medical Advisory Committee, the funds allotted for this study were diverted to support of the more pressing problems of the study of carcinogenicity. It was the feeling of some members of the Medical Advisory Committee that the potential problem presented by these compounds did not warrant investigation. No further action was taken on this problem until 1952, when a survey of Committee members, conducted by Dr. R. E. Eckardt of the Standard Oil Development Coupany, indicated that eight members showed an active interest in re-examining this problem. Further interest was elicited at the seventh meeting of the Medical Advisory Committee, when several case reports of injury and death following exposure to Cg- C12 aromatics were presented. Subsequently, a subcommittee was appointed to study and report recommendations on this problem. 2. Present Activities The first meeting of the Subcommittee on Cg- C^p Aromatics was held on April 26, 195^* The subcommittee decided that in order to assess potential health problems, a review should be made of the incidence of undue exposure and intoxication allegedly caused by both individual compounds and mixtures of C9- C|2 aromatics. A questionnaire was prepared and sent to medical officers of petroleum companies in the API in order to develop this informa tion. The results of this survey are not yet available. API 05231 2 - III. Future Plans and Recommendations 1. The subcommittee reports that at this time it does not have in its possession sufficient data to adequately attain its objectives. Consequently, it recommends that it continue its effort to develop additional information. 2. Should the subcommittee uncover sufficient evidence to recommend that the Medical Advisory Committee support experimental studies, it will submit a budget requirement for the year 1955-56 in the amount of approximately $20,000. IV. Benefits to the Petroleum Industry It is the subcommittee's opinion that investigation of this problem will more adequately assess the potential health problems associated with Cq - C ^ aromatics. The increased use of these solvents and the proportionately greater quantities of varied constituents make it highly desirable for the petroleum industry to be cognizant of the problems associated with their use. V. Members of the Subcommittee C. H. Hine, M.D. (Chairman) Shell Development Company R. D. Bent Atlantic Refining Company J. W. Crookshank, M.D. Cities Service Refining Company Kieffer Davis, M.D. Phillips Petroleum Company A. E. Dooley The Texas Company R. E. Eckardt, M.D. Standard Oil Development Company API 05932 APPENDIX E - 2 C - C12 AROMATICS COMMITTEE REPORT A review was made of the incidence of undue ejqposure and intoxi cation allegedly caused by both individual compounds and mixtures of Cc) - C]_2 aromatics. A questionnaire was sent to the medical officers of the petroleum companies in the API and the opinions of experts in the field of industrial health were solicited in order to develop this informa tion. On the basis of fragmentary and inconclusive evidence that was obtained, it became necessary to classify the hazard associated with C9 - C^2 aromatics as similar to that of benzene. However, it is the feeling of the subcommittee that the actual hazard is probably of a lower order than that of benzene, and it is there fore the recommendation of this subcommittee that the Medical Advisory Committee sponsor a research program to establish the magnitude of this hazard in comparison to benzene or n-hexane. It is further recommended that monies in the amount of $20,000 be appropriated for support of this program for a period of one year. It is finally recommended that other interested groups be contacted to deter mine the desirability of entering a joint' research program in this matter. API 05933 i n >\ Sh e 'l l Development Company EMERYVILLE. CALIFORNIA August 9, 1954 TILK P H O N I OLYMPIC 3 -2 1 0 0 Messrs. E. w. Adams E. K. Linder L. c. Beard, Jr. R. c. Mithcff Allan E. Dooley M. N. Newquist ' F. D. Gassaway J. w. Osborn t. M. Henderson F. J. Sanders Gentlemen: In accordance with Dr. Eckardt's letter of June 25, a meeting of the Subcommittee on Carcinogenicity is to be held in Chicago at the Conrad Hilton Hotel, in private dining room No. 9 at 9:30 A.M. on Sep tember 10. The following is a tentative agenda for this subcommittee meeting: 1. Approval of minutes of the last meeting. 2. Progress report by Dr. A. W. Horton. 3. Progress report by Dr. J. J. Phair. 4. New items of business. 5. Time and place of the next meeting. If any of the members of the subcommittee have additional items which they would like to include in the agenda, they may submit them prior to the meeting and I will see that they are scheduled. Very truly yours, CHH/jck cc: A. w. J. J. G. W. D. V. Horton Phair Saunders S troop TM ,scofy fo* s ( M J J L x s ^ _________ C. H, Hine, M.D., Acting Chairman Subconmittee on Carcinogenicity API 05934 r - i N V'- // i; u 'e s s o L a b o r a t o r i e s STANDARD OIL DEVELOPMENT COMPANY P . O . BOX 31, LINDEN N .J . MEDICAL RESEARCH DIVISION ROBERT HORACE E. E C K A R D T . P H D .. M D. D IR E C T O R W . G E R A R D E . P H .D .. M .D . HEAD TO X IC O LO G IS T August 18, 195U INDUSTRIAL HYGIENE SECTION N A T H A N V . H E N D R IC K S . C H .E . HEAD OP SECTION F R A N K L IN W . C H U R C H . M .S . G E O R G E M . W IL K E N IN G . M .S . IN D U S TR IA L HYG IENISTS Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New York 20, New York Dear Mr. Stroop: I am sending you a' copy of a letter idiich I would appreciate your attaching as a cover letter to Dr. Horton's and Mrs. Denman's manuscript, the stencils for which you have recently received from Dr. Kehoe. Would you please see that this letter and the manuscript are distributed to each of the members of the Subcommittee on Carcinogenicity and also to each of the members of the Medical Advisory Committee. Thank you for your cooperation. Sincerely yours, REE:ilk Enel R. E. ECKARDT, M. D P.S I am also enclosing 10 copies of the revised manuscript with the alternate page 1 . ESSO L A B O R A T O R IE S STANDARD OIL DEVELOPMENT COMPANY P .O . BOX Jl, LINDEN, N.J. medical research division R O B E R T E. E C K A R D T , PH D .. M D. D IR E C T O R H O R AC E W . G E R A R D E . P H .O . M .D . HEAD TO X IC O LO G IS T Aiigust 18, 195L INDUSTRIAL HYGIENE SECTION N A T H A N V . H E N D R IC K S . C H .E . h e a d o f SECTION f r a n k l in w . c h u r c h , m .s G E O R G E M . W IL K E N IN G . M .S . 1NOUSTRIAL H Y G IENISTS Messrs. E. W. Adams L. C. Beard, Jr A. E. Dooley F. D. Gassaway L. M. Henderson C. H. Hine E. K. Linder R. C. Mithoff M. N. Newquist J. tf. Osborn F. J. Sanders Gentlemen: Attached to this letter is the revised manuscript of the paper by Dr. Horton and Mrs. Denman entitled: CARCINOGENESIS OF THE S O N . I. Basic Experimental Methods Employed in Testing Complex Hydrocarbon-Type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies for Mice. Tou will note that fr. Kehoe has suggested an alternate page 1 for this manu script. I would greatly appreciate it if each of you would communicate with Dr. Hine and express your preference either for the original page 1 or the alternate, which is attached. With your information, Dr. Hine will then be in a position to recommend to the Medical Advisory Committee that our subcommittee approves this paper for publication, either with the original page 1 or with the alternate page 1, depending upon the preferences which each of you expresses to Dr. Hine. Since the meeting of the Medical Advisory Committee is to be held one day prior to the meeting of the Subcoaaittee, Dr. Hina will need your expressions as to preference for page 1 prior to the 9th of September, 195k* It is my understand ing that the paper as a whole has the approval of the Subcommittee for publica tion and that we w i n so recommend to the Medical Advisory Committee. It is then assumed that the Medical Advisory Committee will go along with our recommenda tion and recoannd to the Medical and Health Committee of the Board that the paper be published, and that the Medical and Health Committee of the Board will review the paper at its next meeting and indicate whether or not it has any objections to the publication of the manuscript. Very truly yours, REE:ilk Attachment T T .& W * R. E. ECKARDT, M. D. cc w ith attachm ent: Members of the Medical Advisory Committee API 05936 ESSO LABORATORIES STANDARD OIL DEVELOPMENT COMPANY P. 0. BOX 51 LINDEN, N. J. E. W. Adams L. C. Beard, Jr. A. E. Dooley F. D. Gassaway L. M. Henderson C. H. Hine E. K. Linder R. C. Mithoff M. N. Nevquist J. W. Osborn F. J. Sanders . August 18, 195^ Gentlemen: Attached to this letter is the revised manuscript of the paper by Dr. Horton and Mrs. Denman entitled;. CARCINOGENESIS OF 1EE SKIN. I. Basic Experimental Methods Employed in Testing Complex Hydrocarbon-Type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies for Mice. You will note that Dr. Kehoe has suggested an alternate page 1 for this manu script. I would greatly appreciate it if each of you would conaminicate with Dr. Hine and express your preference either for the original page 1 or the alternate, which is attached. With your information, Dr. Hine will then be in a position to recommend to the Medical Advisory Committee that our subcommittee approves this paper for publication, either with the original page 1 or with the alternate page 1 , depending upon the preferences which each of you expresses to Dr. Hine. Since the meeting of the Medical Advisory Committee is to be held one day prior to the meeting of the Subcommittee, Dr. Hine will need your expressions as to preference for page 1 prior to the 9th of September, 195^. It is my understanding that the paper as a whole has the approval of the Subcommittee for publication and that we will so recommend to the Medical Advisory Committee. It is then assumed that the Medical Advisory Committee will go along with our recommendation and recommend to the Medical and Health Committee of the Board that the paper be published, and that the Medical and Health Committee of the Board will review the paper at its next meeting and indicate whether or not it has any objections to the publication of the manuscript. Very truly yours, R. E. ECKARDT, M. D. REE: ilk Attachment cc with attachment: Members of the Medical Advisory Committee API 05937 Alternate Page One C A R C I N O G E N E S I S OP T HE SKIN. I. Basic E x p e r i m e n t a l Methods Employed in Testing Complex Hydrocarbon-type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies for Mice.1 b y A. Wesley Horton and Dorothy Templeton Denman The investigation of skin cancer in this Laboratory has been directed toward the appraisal of the potential hazards which arise in Industry out of the handling of chemical substances which are known to have, or suspected of having, carcinogenic properties under appropriate conditions. The experiments which have provided the background for this paper have been concerned with the relative carcinogenic potency, for experimental animals, of hydrocarbontype oils and tars. The data of these experiments are employed h erein to the extent that they have contributed to the development of reproducible methods for measuring and expressing such potencies. Mice were chosen as the primary test species since n e o plasms of the skin, comparable in certain respects to those of man in the same tissue, can be reproduced in these animals under s u i t a b l e e x p e r i m e n t a l c o n d i t i o n s (1). C e r t a i n m a t e r i a l s s u c h as shale oil, which have induced an abnormally high incidence of cancer of the skin of m e n as a result of repeated exposure in their 1 The majority of this work was carried out as part of the American Petroleum Institute Research Project MC-1. API 05938 C A R C I N O G E N E S I S OF THE SKIN, I. Basic E x p e r i m e n t a l Methods Employed in Testing Complex Hydrocarbon-type Tars and Oils, and the Development of a Quantitative Scale to Express Their Relative Potencies for Mice. 1 b y A. W e s l e y H o r t o n a n d D o r o t h y T e m p l e t o n D e n m a n The investigation of skin cancer in this Laboratory has been concerned with the estimation of the extent of the potential hazard of this disease in certain industries. As in any other aspect of the g e n e r a l f i e l d of toxicology, it m u s t be r e c o g n i z e d that a s p e c i f i c h a z a r d is the p r o d u c t of s e veral variables, two of which assume dominant importance. These may be termed conven iently the physiological activity of the material or mixture in question, and the conditions of exposure. The latter aspect in volves the practices of personal and industrial hygiene. This paper will be devoted primarily to the development of reproducible methods of determining the first variable, the level of activity, or relative carcinogenic potency, of hydrocarbon-type oils and tars. Mice were chosen as the primary test species since neo plasms of the skin, comparable in certain respects to those of man in the same tissue, can be reproduced in these animals under suit able experimental conditions(1). Certain materials such as shale oil, which have induced an abnormally high incidence of cancer of the skin of m e n as a result of repeated exposure in their The m a j ority of this work was carried out as part of the American Petroleum Institute Research Project MC-1. API 05939 - 2 - occupation (2,3), have been found capable of producing epitheliomas in the skin of mice Conversely, in our experience, certain inbred mic e , such as the C 3 H strain, have r a r e l y d e v e l o p e d ''spon taneous" skin tumors, and have not reacted by skin tumor formation to repeated application of a number of materials, including benzene and white mineral oil, which are presumed to be non-carcinogenic to the skin of man on the basis of extensive practical experience. The specification, "suitable experimental conditions", in the previous paragraph, was used advisedly. It has become evident that the problem of excessive mortality among the experi m e n t a l animals, due- to the t o x i c i t y of the m a t e r i a l un d e r e x a m i n ation as well as to the occurrence of incidental disease, cannot be met simply by increasing the number of animals on test. Such an approach ignores the probability that the survivors of these conditions are not at all representative of the population involved, with respect to their susceptibility to carcinogenesis. Hence, stress will be laid upon the necessity for standards of selection and care of the experimental animals which will maintain their health at as h i g h a level as possible. EXPERIMENTAL METHODS Most of our work has been carried out with male mice of the C3H strain, purchased from the Jackson Memorial Laboratory, Bar Harbor, Maine. A number of experiments have also been carried out with male mice of the CFW strain obtained from Carworth Farms, but the results were not as reproducible as those derived from the C3H strain. Although the difference in genetic homogeneity API 05940 - 3- is u n d o u b t e d l y a factor, the f a c t that the C3H m i c e are m o r e docile probably contributes to the reproducibility of experiments in which they are employed. The frequent occurrence of bites and scratches of the skin of the C F W mice, i n f l icted by their companions, pro b a b l y results in occasional direct intraepithelial introduction of car cinogens, Added to this complication is the possibility that w o u n d - h e a l i n g m a y p l a y an a c c e l e r a t i n g role in c a r c i n o g e n e s i s (6), Current experimentation with Swiss mice from Jackson Memorial Laboratory indicates that, for similar reasons, the males are un satisfactory animals for quantitative tests. The females of this stock, on the other hand, do not seem particularly pugnacious and, hence, should prove quite useful for this purpose. 2he mice were received at six to eight weeks of age and placed under observation for approximately four weeks before use in any experiment. To control Infestation of the animals with mites or lice (which when uncontrolled affects adversely their health), the mice were dusted routinely with Aramite8 by the sup plier prior to shipment and then twice again with the same miticide during the period of observation in this Laboratory. Any group in which there was evidence of poor health during this period was withheld from use in tests in which quantitative results were needed. In all experiments described herein, the mice were fed on Purina Laboratory Chow and water without restriction. At six months of age, C3H mice fed on this diet and subjected to no adverse experimental procedure reach an average weight of about 29 to 3 1 g., w h i l e c o r r e s p o n d i n g C F W m i c e w e i g h about 33 to 35 6 API 05941 - ij. - Since growth of both the hosts and their tumors m i g h t vary with the t e m p e r a t u r e of their surroundings, an effort h as b e e n m a d e to maintain a relatively constant temperature of 75-2F. in the animal rooms. For a given experiment, a group of 20 to 30 mice was divided into three to five cages. The animals were assigned individual numbers for identification and marked by clipping their toes. The contact of the mice with any given material involved its r e p e a t e d a p p l i c a t i o n u p o n the i n t e r s c a p u l a r r e g i o n of their skin. When dosages of 100 mg. of a material were to be applied, the fur was left intact, but when the dosage was to be less than 100 mg., the fur was removed by means of electric clippers. A small camel-hair bru s h of a selected size to deliver the desired dosage was used to apply the material, which was then simply allowed to spread according to its individual physical characteristics. Applications were repeated in accordance with a predetermined schedule (one, two, or three times each week) until the animals died. Care was taken to keep the dosage and the location of each application as constant as possible throughout the experiment* Normal growth of the mice could not be maintained in testing certain of the materials, particularly those of high potency, w h e n the experiment involved three applications of the material per week in the dosage of 100 mg. Hence, when there was some physicochemical or biological evidence that a given sample was likely to hav e a r e l a t i v e l y h i g h c a r c i nogenic p o tency to the API 05942 - 5 - mice (equivalent to that of a solution of 0 .1 5 percent, or greater, of m e t h y l c h o l a n t h r e n e in benzene), the m a t e r i a l was ap p l i e d at a lower frequency, usually once each week. In the case of materials of l o w e r potency, m o r e f r e q u e n t a p p l i c a t i o n s were r e q u i r e d to obtain tumors in a reasonable length of time. Hence, when the toxicity of such samples p r o v e d to be a serious p r o b l e m at the 100 og. level, the dose was reduced, in some cases to as low as 5 to 10 mg. per application. Experience has shown that, even when the mortality among a t an e x p e r i m e n t a l g r o u p was low, the time of a p p e a r a n c e of tumors was sometimes delayed seriously by conditions of variable or uncertain origin which also interrupted the normal growth of the animals. As a m e a s u r e of s u c h c onditions, therefore, the m i c e were w e i g h e d at f r e q u e n t intervals throughout the experiments. Since early observations gave evidence that interruptions of growth were usually the r e s u l t of i n t e r c u r r e n t infe c t i o n s of low s e v e r i t y a f f e c t i n g most of the animals in a given cage, the weighing of individual animals was discontinued in favor of determining the average weight of the s u r v i v o r s In e a c h group. V a r i o u s precautions, s u c h as regular cleaning and sterilization of cages, feeders and water bottles, decontamination (with a bacteriostatic agent) of gloves used to handle animals f rom one cage before proceeding te another, and Isolation of sick mice, were taken to minimize Infections and to prevent their spread from one group to another. Throughout each experiment, the animals were examined at least once each week by one of us (D.T.D.), and the presence and time of appearance of any abnormal gross changes in the skin were 1 "*"`" .... API 05943 -- 6~ noted and, in the early stages of tests on solutions of greater than 0 .2 percent of methylcholanthrene or benzpyrene in benzene, large s w e llings, of the type p r e v i o u s l y d e s c r i b e d b y C r a m e r (7), and small ulcerated areas, were occasionally observed. In response to the appli c a t i o n of solutions of these synthetic carcinogens, papillomas eventually developed on the skin of practically all of the surviving animals except when the lowest level of concentration was being tested (0.01 percent). In general, the induced neoplasms progressed through the familiar pattern from papilloma to squamous cell carcinoma, with gross evidence of invasion. In the first test of a given material, sections of all epidermal tumors produced were prepared for microscopic examination. The date on whi c h each typical p apilloma was fou nd to have reached the arbitrary size of 1 mm. in diameter and elevation was r e c o r d e d as the ''time of a ppearance" of the papilloma. In addition, the date on which it was grossly apparent that the tumor was invading the subcutaneous tissues was noted (rolled border, generally accompanied by cratering necrosis of the center). When a mouse developed such an apparently malignant tumor, it was killed* Usually, the others were maintained on test until death from "natural causes" occurred. The data, thus obtained, were recorded on g r a p h s s u c h as those s h o w n in F i g u r e s 1 t h r o u g h Ij., As experience was gained in associating the physical and chemical characteristics of various types of complex materials with their relative toxicity and with their ability to induce tumors of the skin in C3H mice, certain criteria were developed for i selecting optimum conditions for any given biological test. API 05944 - 7 - The interpretation of the results of the different experiments was made possible by comparison with external standards of reference, discussed in a later section of this paper. As a general practice, a schedule of application was chosen which would lead to induction of papillomas in a n average time of not less th a n 12 weeks. This restriction was based on repeated observations that the mean time of appearance of tumors was unduly variable in experiments in which the seve r i t y of exposure r e s u l t e d in more rapid induction of tumors. On the other hand, experimental conditions which involved very long periods of time before the appearance of tumors sometimes proved to be even more unsatisfactory than those associated with very short latent periods. The longer the experiment, the greater was the possibility of the occurrence of infection or other u n identified interference with the normal growth of the mice. Even when these interruptions resulted in little or no loss of animals, the irregular effects on the growth of the induced tumors made it very difficult to interpret the results of the experiment. Therefore, as a general rule the f r equenoy and severity of the applications were scheduled to produce an average latent period as close to 10 weeks as possible. When the materials were low in potency, this obviously meant the use of as severe conditions as were compatible with the normal growth and survival of the mice. J I INTERPRETATION OP DATA OBTAINED In general, under the conditions of these experiments ! one of two results has been obtained in response to the applications I of any one of a series of materials, l.e., most of the animals API 05945 - 8 - of the a p p l i c a t i o n , or none of t h e m did so. The outcome of the experiments, therefore, has been essentially unequivocal, and the relative potencies of the different active materials could be ex pressed in terms of the relative rates of induction of tumors. The calculation of the "mean time of appearance of tumors" involved the determination, by the method of least squares, of the linear function whi c h best fitted the plot of the cumulative f r e q u e n cy of tumor response (in probits) versus the time (in weeks) after the f i r s t a p p l i c a t i o n . The l a tter coordinate is e q u i v a l e n t to cumulative dosage. Typical plots of such data are shown on the right side of F i g u r e s 1 t h r o u g h lj.. It is to be n o t e d that, for these experiments involving repeated applications of carcinogenic materials to the skin, the response varied linearly with time, and not wit h the l o g a r i t h m of time. It Is assumed that the exponentially decreasing rate of appearance of tumors usually observed following single or limited numbers of subcutaneous injections of carcinogens (8) was n o t s e e n in these e x p e r i m e n t s be c a u s e the m o r e r e s i s t a n t animals in any given group were given a larger effective dosage than were the more susceptible. CALCULATION OF CUMULATIVE FREQUENCIES OF RESPONSE The c o n v e n t i o n Is a d o p t e d (after Bliss, 9) that, at the midpoint of the period between two given observations, the n u m b e r of m i c e b e a r i n g tumors is e q u a l to the average of the numbers at the times of the observations. At the midpoint of each period during which one or more mice d e v e l o p the i r first p a pilloma, a c a l c u l a t i o n is made of API 05946 T \4 ^ # - 9 - the cumulative percentage of the "effective group of mice" which bear tumors on that date. The "effective group" at a g i v e n time is d e f i n e d as the s um of the n u m b e r of s u r vivors plus the number of mice which died previously after developing tumors of the skin, except that after the appearance of a tumor in the "average" mouse this sum is h e l d constant. The "average" mo u s e is d e s i g n a t e d by the median in a simple arithmetic array of the times of appearance of the first tumor in each animal. As an example, the calculations used to obtain the cumulative frequencies of tumor response from the data of F i g u r e 2 are shown in Table I, CALCULATION OF MEAN TIME OF APPEARANCE OF TUMORS, x Experience has shown that, If the frequency of response, r (in problts), is plotted against the time, x, a linear r e l a t i o n s h i p Is obtained, e.g., Figures 1 t h r o u g h ij.. T h e r e f o r e , the m e a n time of appearance of tumors, x, m a y be obtained graphically from the best straight line fitted by eye to such a plot. However, we have found that x m ay be obtained by algebraic methods more rapidly and without the subjective aspects Inherent in graphical analysis, so long as a suitable calculator is available. The f o l l o w i n g exam p l e shows the c a l c u l a t i o n of the m e a n time, x, f r o m the d a t a of Tab l e I. API 05947 ) ) ) y-- -- ---------------------------------------------------------------------------i ----------------------------------------------------------------------------------------------------------- TABLE I TIME OP OBSERVATION OF NEW POSITIVES MIDPOINT OP PERIOD BETWEEN OBSERVATIONS, X NUMBER OF NEW POSITIVES, R' TOTAL NUMBER OP MICE BEARING TUMORS, R* + R a CUMULATIVE RESPONSE, 2R + (zn + R) 2 EFFECTIVE NUMBER OP MICE FREQUENCY OF RESPONSE, r ( probits) 12.5 0.5 19 ,1.06 {2.6%) 13 1 1 iii.5 2.0 19 3.75 15 2 3 i ii 3.5 19 U.10 16 1 k 16.5 k-S 19 U.28 17 1 5 U iS. 7.5 19 \Ls2 1 18 5 10 1M . n.5 19 liil 19 3 13 2*1 15.0 19 5.81 20 k 17 20.5 18.0 19 6.62 21 2 19 1 Number of weeks after first application. a R is the total number of mice with tumors at the time of previous observation. API 05948 k Determine constants, m and b, by m e t h o d of least squares (10). Normal equations: r x = m z xz + b i x z r - m i x + 8b (note that the coefficient of b equals the number of points used) (Values of x and r are u n d e r l i n e d i n Table I) 656.07 = 2328.0m + 135.0b 37.62 = 135.0m + 8 b b = -2.520 m s O.i^.28 s slope of d o s e - r e s p o n s e r e l a t i o n s h i p .*. r = ,!+28x + -2 .5 2 2. x s time when r equals 5 = 17*6 weeks s, s t a n d a r d d e v i a t i o n of mean, z x minus time of i^-probit response = 2.3 weeks. CLASSIFICATION OF EXPERIMENTAL RESULTS ON THE BASIS OF THE HEALTH OF THE ANIMALS In the appraisal of data derived from these experiments, consideration has been given to the following apparent relation ships between the average growth and survival of the mice and the mean time of a p p e a r a n c e of t h e i r e p i d e r m a l tumors. 1. If, in the course of an e x p e r iment, an intercurrent infection overtakes a group of mice at about the time when papillomas would be expected to appear (on the basis of other comparable experi ments), there is usually a d e l a y in the m e a n time of appearance of API 05949 12 tumors. The infection need not be lethal to any of the mice, a change in sign of the slope of the average growth curve frequently b e i n g an i n d i c a t i o n of such a condition. It is b e l i e v e d that, in many such instances, the mice would probably be unaffected were their resistance not diminished somewhat by systemic toxic effects induced by the hydrocarbons being applied upon their skin. Thus, untreated controls may not show any symptoms of the infection. Not infrequently, as the animals recover from such dis turbances, tumors grow to measurable size in a large proportion of the animals in a relatively short time. The slope of the plot of the frequency of response versus time is then exceedingly steep (see Figure 2), In other words, the standard deviation of the mean time of appearance of tumors in the group is smaller than the normal range, 3 to 5*5 weeks, for such experiments using C3H mice. Thus, it could be assumed mistakenly, if it were not for the evidence from the weight curves of the presence of uncontrolled variables, that such an experiment had defined the fiducial limits of the mean, calculated from the standard deviation, better than another in which such disturbances had not occurred. 2. In some experiments the cumulative toxic effects of the material applied, or chronic infections, or a combination of both, result in prolonged inhibition of the growth of the mice. A number of examples have been observed in which the average weights have increased for a few weeks, became stationary at a point 10 p e r c e n t or m o r e b e l o w that of adult controls, a n d then, in m a n y instances, gradually declined (see Figure 3) Whether or not these conditions influence the timing of the initial changes resulting API 05950 - 13 - in the neoplasms, they may retard the growth of the induced tumors to v i s i b l y d i s c e r n i b l e size and, accordingly, l e n g t h e n the apparent average period of time required for the induction of tumors by the material under test. In such experiments, the standard deviation of t h i s m e a n time is f r e q u e n t l y m u c h g r e a t e r t h a n normal. This somewhat extended discussion of the apparent relation ships between the irregularities in growth due to the toxicity of applied materials and to intercurrent infections, and the rate of induction of tumors, does not derive from the originality of this observation but rather from our recognition of its importance in experiments in which various materials are being compared. Occasion al mention of the phenomenon has appeared in the literature on exper imental carcinogenesis (e.g., 1 1 , 1 2 } and, in the field of chemo therapy of cancer, recognition has been given to the necessity of considering the systemic toxicity of a material in evaluating the specificity of its apparent "antlcarcinogenlc" effects (e.g., 13). Hence, it is believed that, in the experimental assay of the relative potencies of various materials or of the susceptibility of various species or strains of one species to a given material, information on the health and survival of the groups of animals be i n g c o m p a r e d is e s s e n t i a l to a n y q u a n t i t a t i v e i n t erpretation of the data. The results of all experiments described herein have been examined, therefore, in the light of these relationships. Each one has been classified according to the estimated significance of such effects in the following manner. API 05951 - 14 - Class A. Those characterized by normal growth and sur vival of the mice at least until the time when most of the animals have developed papillomas (see F i g u r e 1;). Class B. Those in which all or m o s t of the m i c e lived long enough to develop tumors, but in which there was evidence (from average weights) of intercurrent infection at times during the period when the new growths were appearing. So long as recovery from the infection was prompt, and no obvious skewing of the distri bution of the times of appearance of tumors oc c u r r e d , it was f e l t that reli a b l e e s t i m a t e s of the mean time of appearance could reasonably be made (see Figure 1), However, when no tumors had appeared prior to the onset of an infection, and when, with recovery, tumors developed somewhat precipitously, the reli ability of the apparent mean time of appearance was not determinable, and therefore such tests were classified X (see Figure 2). Class X. Those characterized by poor growth (see Section 2 a b o v e a n d F i g u r e 3) or h i g h rate of m o r t a l i t y . The confidence limits of the apparent mean time for the induction of tumors could not be calcu lated by standard statistical methods, because of inability to assign proper weight to the effects of the uncontrolled variables. API 05952 - 15 - Experiments involving the application of m oderately toxic materials frequently failed to meet the requirements of the Class a category. On the other hand, results of the frankly unsatisfactory type (Class X) were usually found to be avoidable through the exercise of appropriate care in the initial choice and the subsequent handling of the mice. REFERENCE STANDARDS To provide a background of reference which might permit a more nearly quantitative interpretation of the mean time of appearance of tumors Anduoed in C3H mice by complex oils and tars under various experimental conditions, a number of experiments have been carried out over the past five years employing solutions of the synthetic carcinogens, 3-methylcholanthrene (MC), benzofa]pyrene (BP), and 7 , 12-dimethylbenzfajanthracene (DMBA), in various solvents. It has been found that one of the essential requirements for o b t a i n i n g r e p r o d u c i b l e a n d l o g i c a l l y c o n s i s t e n t r esults is the maintenance of the animals in a good state of health as measured b y g r o w t h curves. Thus, In Table II, the r e s u l t s of E x p e r i m e n t s 2014. show the precision of the determination of the m ean time, x, when Class B experiments are involved. In using volatile materials such as benzene as solvents, occasional checks on concentration of the carcinogens are necessary, with appropriate corrections for evap oration. Similarly, it has bee n found that even in the case of a complex material, so long as the conditions of the experiments permitted reasonably normal g r o w t h of the animals, the repeatability of the determination of the m ean time, x, has been quite satisfactory (within *10 percent). API 05953 - 16 - To determine the effect on the rate of induction of tumors of v a r i a t i o n s in the level of c o n c e n t r a t i o n a n d in the f r e q u e n c y of exposure, comparable experiments were conducted with other solutions of MC in benzene, s h o w n in Tab l e II. F r o m c ertain o f these results (Experiments 201;, 209, 238, an d 2i;3) it is apparent that v a r i a t i o n in the q u a n t i t y of s o l u t i o n u s e d f o r e a c h a p p l i c a t i o n f r o m 20 to 100 mg., o t h e r fa c t o r s r e m a i n i n g constant, h a d no s i g n i f i c a n t effect on the rate of induction of tumors. Similar results have been observed when solutions of BF in benzene were used as the carcinogenic stimulus. This constancy of potency with variation in the quantity of material applied (or area of skin exposed) also holds for some complex oils a n d tars, b u t for o t hers there is signif i c a n t change of potency with changes in the quantity used for each application. The chemical basis for these differences will be discussed in a later paper. It has b e e n d e t e r m i n e d e x p e r i m e n t a l l y that vari a t i o n s in the concentration of MO over the range, 0.05 to O .35 percent, did not affect the average area covered by a given weight of its sol utions in benzene. It was further assumed on the basis of the results of e x p e r i m e n t s 201;, 209, 238, and 2i;3 that the dosage of MC per unit area of the skin was not significantly different when 20 or 100 mg. of the same solution were applied. Therefore, the values in col u m n 5, Table II, were calculated directly f r o m the concentration of MC in the solutions and the number of applications per week. Since the amount of MC, in mg,, in 100 mg. of solution is e q u a l to the c o n c e n t r a t i o n , in p e r c e n t b y weight, the values in c o l u m n 5 ere e q u a l to the p r o d u c t s of the v a l u e s in columns 2 a nd 1;. API 05954 ) ) ) +-- ................................................... ....................................................* --------------------------------------------------------------------------* TABLE II ------------------------------------------------------------------------------------------------------------- 4 1 EXPERIMENTS I N V OLVING THE APPLICATION OP SOLUTIONS OF METHYLCHOLANTHRENE IN B E N Z E N E UPON THE SKIN OF C3H MICE EXPERIMENT TOMBER 206b 20$A 201+A 20J.B 201+C 201+D 21+3A 21+3B 203A 203B CONCENTRATION OF METHYLCHOL ANTHRENE IN SOLUTION (# b y weight) DOSAGE OF SOLUTION PER APPLICATION (mg.) 0.69 100 0.31+5 100 0 .17 2 100 0 .17 2 20 0 .17 2 20 0 .17 2 20 0 .15 100 0.15 20 0.086 100 0.086 100 NUMBER OF APPLI CATIONS PER WEEK 3 3 3 3 3 3 3 3 3 3 DOSAGE OF METHYLCHOLANTHRENE PER UNIT AREA1 PER WEEK (mg.) I.9 1.035 O .516 O .518 O .5 16 0.516 o.i+5 0.1+5 0.258 0.258 MEAN TIME OF APPEARANCE OF TUMORS, x (weeks) 6.9-I+.23 11.1-2.0 1 6 .0-1 . 7 16.ij.il. 1 16.6*1.2 15.7*1.1 18 .1 *1 .8 18.7*2.0 30.02.3 26.3*3.1+ PERCENT OF MICE WITH TUMORS cl a. 100 B 100 B 100 B 100 B 100 B 100 B 100 A 100 B 90 B 100 B API 05955 205B 0 .31+5 100 2 2 3 0A 0.23 20 2 230B 0.23 100 2 0.69 O.I4.6 0.1+6 15.1*1.5 100 B 1 8 .7*2 .3 100 B 16.1|2.0 85 B i Area of akin covered by 100 mg. of solution of raethylcholanthrene in benzene $% f i d u c i a l limits (li|.) . ) ) ) ----------- S----- --------------- TABLE II (page 2) EXPERIMENTS INVOLVING THE APPLICATION OP SOLUTIONS OF METHYLCHOLANTHRENE IN BENZENE UPON THE SKIN OF C3H MICE :p e r i m e n t NUMBER 220 209A 209B 205C CONCENTRATION OF METHYLCHOL ANTHRENE IN SOLUTION (i, by weight) 1.03k 0.517 0.517 0.3k$ DOSAGE OF SOLUTION PER APPLICATION (mg.) 100 20 100 100 NUMBER OF APPLI CATIONS PER WEEK 1 1 1 1 DOSAGE OF METHYLCHOLANTHRENE PER UNIT AREA1 PER WEEK (mg.) _ 1.03U 0.517 ' 0.517 0.3k$ MEAN TIME OF APPEARANCE OF TUMORS, X (weeks) 10.1-1.62 18.72.0 16.22.7 22.t+2.7 PERCENT OF MICE WITH CLASSIFITUMORS CATION 100 B 100 B 100 B 100 B 207 1.1$ 100 3 3.1+$ 9.1 100 X 209c 0.517 100 3 l.$$l 10.1+ 100 X 205d 0.3k$ 100 3 1.03$ 11+.8 100 X 219 0.115 100 3 201 0.012 100 3 0.31+$ 0 .0 3 6 21+.2 72.6 . 96 X 1+1+ X 268 0.287 20 2 0.$7l+ 17.6 100 X 2 0 6A 0.69 100 1 205e 0.3l+$* 100 1 0 .6 9 0.31+$ 17.1 12.9 100 X 3 100 Z 238c 0.23* 100 1 0 .2 3 ll+.U 100 1 Area of akin covered by 100 mg. of solution of methylcholanthrene in benzene. * %% f i d u c i a l l i m i t 8. 3 Sample of methylcholanthrene used in these experiments contained undesirable impurity z3 (see p.19). API 05956 - 19 - Comparison of the data in columns 5 and 6 for Class A and B experiments shows a consistent inverse relationship between the dosage of carcinogen per unit area per week and the mean time of appearance of tumors. The data are p l o t t e d in Figure 5 together with the curve of the corresponding hyperbolic function (calculated by the m e t h o d of least squares f r o m the data on Class A and B experiments), or, (x - 3 . 7 ) (d + 0.10) = 8.2 8.2 x * (d + 0.10) + 3.7 It is i n t e r e s t i n g to note the s i m i l a r i t y b e t w e e n this e q u a t i o n and that calculated f r o m the theoretical considerations of the mechanism of c a r c i n o g e n e s i s b y I v e r s e n and A r l e y (1 5 ) t tap = -i-- ko + t, where t ap is the m e a n time of a p p e a r a n c e of tumors and cQ the concentration of the carcin ogen, k and T being constants. The constant, 0.10, in the experimental equation might be inter preted as a correction required by the definite, though very small, probability of the "spontaneous" excitation of a cell of the dorsal skin of a C3H mouse to the rate of proliferation necessary for the development of a new growth. The purity of the polycyclic hydrocarbon is obviously an important consideration in obtaining consistent results. The usual chemical criteria are necessary, but not always sufficient, requirements. In one case In particular, the MC, as re c e i v e d , h a d a sharp m e l t i n g point, 1 7 6 - 7 C . ( u n c o r r . ). API 05957 20 - Yet, on repea t e d bioassay, it proved to be excessively irritating and much more rapidly carcinogenic than the normal material. The results of Experiments 205e and 238c, us i n g this sample, indicate t h a t the i m p u r i t y h a d active carcinogenic or accelerating properties. Tumors developed at a rate expected only with solutions contain ing 2 to 3 times the concentrations of MC employed. Unfortunately the quantity of this particular sample available did not permit the determination of the structure of the particular impurity responsible for this effect. . The compound apparently constituted less than 0.5 percent of the ample. Therefore, both recryatal- lization and chromatograjhy on activated alumina have been used in combination in an attempt to guard against this contamination. Bloaasay has remained the only sure w a y of a s c e r t a i n i n g that the p u r i f i e d MC is free of tv significant concentrations of this material. Similar experiments have been carried out with solutions of two other c o m m o n l y us e d eareintgenic hydrocarbons, BP an d DMBA (see Table III). As is e v i d e n t from the data, it has b e e n dif ficult to obtain satisfactory experiments with BP on C3H mice. The growth of the animals has tendei to be extremely erratic. Ev e n in tests in w h i c h the average weight event u a l l y r e a c h e d 30 g., the curve was, more often than not, interrupted at critical times by sharp downward breaks. In these experiments, recovery from these periods of poor health was sometimes accompanied by the appearance of tumors in a large percentage of the animals in a API 05958 ) ) ) RELATIVE CARCINOGENIC POTENCY, PMC DOSAGE OP CARCINOGEN PER UNIT AREA1 PER WEEK (mg.) CONCENTRATION OP CARCINOGEN IN S O L U T I O N by weight) API 05959 TABLE III EXPERIMENTS INVOLVING THE APPLICATION OP SOLUTIONS OP SYNTHETIC CARCINOGENS IN VARIOUS SOLVENTS UPON THE SKIN OP C3H MICE EXPERI M E N T CARCIN NUMBER OGEN SOLVENT 215 21k 2I44A 21*i*b 25 2A 252B DMBA DMEA BP BP BP BP Benzene Benzene Benzene Benzene Benzene Benzene 252C 261+A 26I4.B 270 BP Benzene BP White mineral oil BP White mineral oil BP White mineral oil 0.115 0.31*5 0 .17 2 0 .17 2 0.322 0.322 0.322 0 .17 5 0 .17 5 0.35 DOSAGE OP SOLUTION PER APPLI CATION (mg.) 100 100 100 20 20 20 100 20 100 20 NUMBER OP APPLI- ' CATIONS PER WEEK 3 3 3 3 l 3 3 3 3 3 0.3U5 1.035 0.516 0.516 0.322 0.966 O.966 0.525 0.525 1.05 MEAN TIME OF APPEARANCE OF TUMORS, x (weeks) CLASSIFI CATION I5.6i3.3e B 0.19 7.51.1 B 2 5 .2 2 .6 B 26.6 X 35.3 X 23.0 X 20. X 25.1* X 0.73 0.09 (0 .00) (0 .1 6 ) (0 .1 1 ) (O.1 3 ) (0.09) 25.5 X (0.09) 21.6 X (0.12) 1 Area of skin covered by 100 mg. of solution of methylcholanthrene In benzene. 8 a fiducial limits. - 22 - r e latively short time ( r e l a t i o n s h i p e x e m p l i f i e d in F i g u r e 2), it will also be noted that, even at high levels of concentration, the m e a n time of a p p e a r a n c e of tumors in d u c e d b y solutions of BP in ''n o n - a c c e l e r a t i n g " solvents, s u c h as b enzene a nd white m i n e r a l oil in the skin of C3H m i c e was i n n o case less than 20 weeks, USE OF REFERENCE STANDARDS AS A SCALE OF RELATIVE POTENCY OF COMPLEX TARS AND OILS Various methods of expressing the relative potencies of the pure polycyclic carcinogens and complex materials have been reviewed b y Badger (16). Many of these are simply rough classifi cations b a s e d u p o n a p a r t i c u l a r i n v estigator *s p e r s o n a l experience. The Ib a l l i n d e x (17) a nd a r e c e n t te c h n i q u e (18) a t t e m p t e d a more quantitative approach, involving the use of the reciprocal of the mean time of appearance of tumors. Such methods are useful in comparing results obtained in a given laboratory using a stand ardized and inflexible schedule of application of various materials. However, experimental techniques and other factors differ signifi cantly from one research organization to the next, thus rendering the comparison of data on this basis rather difficult. Further, it is s o m e t i m e s n e c e s s a r y to a l t e r e i t h e r o r b o t h the f r e q u e n c y of a p p l i c a t i o n a n d the q u a n t i t y of m a t e r i a l a p p l i e d to o b tain s a t isfactory conditions for the measurement of the potency of a sample while minimizing its toxic side effects. Hence, a scale of relative potencies which would permit such flexibility in experimental design was needed. API 05960 T - 23 - The use of the data on relative rates of induction of tumors by various lev e l s of c o n c e n t r a t i o n of solutions of synthetic carc i n ogen, MC, as e x t e r n a l s t a n d a r d s of reference, p r o v i d e d a p r a c t i c a l solution to this p r o b l e m . A l t h o u g h this h y d r o c a r b o n has not b e e n identified as such in complex tars and oils, it might be regarded as r e p r e s e n t a t i v e of the I4.- a nd 5- r i n g e d c a r c i n o g e n s present (19, 20, 21), B y use of F i g u r e 5, the m e a n time of a p p e a r a n c e of tumors induced by some complex tar or oil may be translated into wk a value of Relative Carcinogenic Potency, P MC (potency as compared to MC). The dosa g e of MC c o r r e s p o n d i n g to the g i v e n m e a n time is either read off the graph or calculated from the equation, d -- 2-- - 0 .10 . x - 3.7 Then, the value of the relative potency, PM C , for the complex material in question is PMC = d, f o r an e x p e r i m e n t i n v o l v i n g one a p p l i c a t i o n e a c h week, = , f o r an e x p e r i m e n t i n v o l v i n g two applic a t i o n s e a c h week, z for an sxperiment involving three applications each week Thus, by comparing the results of testa on complex materials with those involving MC as an external standard of reference, one may obtain the relative carcinogenic potencies of the different materials, even though the experiments m a y have involved differing frequencies of applications. For example, if the application of two 20 mg. doses of oil A each week to a group of C3H mice results API 05961 T i -2*- in the induction of skin tumors in an average time of 20 weeks, the relative potency, PM C , of oil A at the 20 mg. level of dosage is 0.1|0/2, or 0.20. It s h o u l d be noted, however, that the p otency of some complex oils varies significantly with the quantity used in each application. Hence the description of the potency of a given m a t e r i a l cannot be c o n s i d e r e d c omplete until it has b e e n t e s t e d at two appreciably different dosages, e.g., 20 mg. and 100 mg. per application, unless negative results were obtained at the higher level of dosage in a test in which the mice survived and grew at a normal rate. Because of certain differences In composition, a given oil may be much more potent than another under severe condi tions of exposure, but the former may actually have a lower potency than the latter if the comparison is carried out under mild condi tions. Chemical research on such oils has led to an understanding of this apparent paradox, and the matter will be dealt with in a later publication. The technique of r e l a t i n g results on complex materials to those on standard synthetic carcinogens should facilitate com parison of results from different laboratories. The synthetic compound chosen should, of course, be comparable to the type of carcinogens supposed to be present in the complex mixtures, under appropriate conditions of dosage and frequency of application, it m u s t be capable of inducing tumors at least as rapidly as the most potent of the samples to be tested. Por this reason, BP has not proved to be a suitable standard for work with C3H mice, since its solutions in non-accelerating solvents will not induce tumors suf ficiently rapidly e v e n at high levels of concentration. On the API 05962 - 25 - other hand, DMBA is very satisfactory from this standpoint, but has the d i s a d v a n t a g e of r e q u i r i n g sp e c i a l care in h a n d l i n g to prevent it o x i d a t i o n to the n o n - c a r c i n o g e n i c 7 * 12-ph o t o o x i d e (12), It should be reemphasized that this consideration of a quantitative scale of potencies applies only to the results of experiments which m a y be classified as satisfactory from the stand point of the g r o w t h and s u rvival of the m i c e (A and B by our criteria) Those in our Class X have only a limited value. If positive results are obtained, the material may be classified qualitatively as carcinogenic; if survival is good and most of the m i c e eventually.'develop tumors (X c l a s s i f i c a t i o n based on certain irregularities in growth) the estimation of th apparent potency may be useful as a mini m u m value. I ; ' RELATIVE POTENCIES OP POLYCYCLIC AROMATIC COMPOUNDS Arbitrarily, the relative potency of MC on the PMC scale is 100. B y c o m p a r i n g d a t a o n the rate of i n d u c t i o n of tumors by solutions of known concentration of other aromatic hydrocarbons in benzene under suitable experimental conditions, with data on MC, such as those s h o w n in Ta b l e II a n d F igure 5, it should be p ossible to arrive at l o g i c a l e s t i m a t e s of the r e l a t i v e c a r c i n o g e n i c p o t e ncies of the different compounds. Numer i c a l values for BP and DMBA m a y thus be derived f r o m the data of Table III. For example since the PMC f o r E x p e r i m e n t 211* was 0.62, i.e., 0.31*5 p e r c e n t D M B A p r o d u c e d tumors at the same rate as 0.62 percent MC under comparable con ditions, the relative potency of the former is o _ 100 rMG o .3 l*S 0.62 a l80 . API 05963 T - 26 - F r o m E x p e r i m e n t 215, the rela t i v e potency, P M C , for DfiBA is 165. The h i g h e r value, 180, is p r e f e r r e d at the p r esent time, since, if a u t o x i d a t i o n of the h y d r o c a r b o n caused a n y r e d u c t i o n in activity, it mi g h t r e a s o n a b l y be a s s u m e d that the effect w o u l d have been greater in Experiment 215. S i m ilarly, the r e l a t i v e p o t e n c y of BP on this scale is 57 to 65 (Experiments 244A and 252A). The data f r o m Experiment 252B and 252C involving three applications each week cannot be used since a pparently the m a x i m u m effective dosage of this carcinogen h a d been r e a c h e d at a lower level. Thus, as a g e n e r a l r ule the det e r mination of the relative potency of any given compound should be based u p o n the results of experiments at at least two different concentrations, which differed significantly with respect to the mean time of appearance of tumors under otherwise comparable conditions. At least one and preferably both of these should meet our classification A or B from the standpoint of growth and survival. SUMMARY Repeated applications of solutions of highly carcinogenic hydrocarbons upon the dorsal skin of male mice of the C3H strain I under suitable experimental conditions result in the induction of epitheliomas in essentially all of the exposed animals, such experiments, involving solutions of 3 -methylcholanthrene in benzene, show a consistent inverse relationship between the mean time of a p p e a r a n c e of tumors (x) a n d the dosage (d) of the c a r c i n o g e n per unit area of the skin per week. x = _ J L i --- + 3 .7 . d + 0.10 API 05964 \ T - 27 For experiments involving synthetic carcinogens in non-accelerating solvents, such as benzene and white min e r a l oil, the mean time of In d u c t i o n of tumors is independent of the q u a n t i t y of solution employed in each application, in the range 20 to 100 mg. The good h e a l t h of the mice, as m e a s u r e d b y their growth, has p r o v e d to be an essential factor in obtaining reproducible results, A quantitative scale for expressing the relative carcinogenic potencies of various synthetic carcinogens and complex tars and oils has been derived, using the experiments on solutions of methylchol- anthrene in benzene as reference standards. Utilizing the relation ship between the dosage, of methylcholanthrene and the rate of induc tion of tumors shown above, a value for the relative potency, P ^ , is obtained. The n u m e r i c a l value of the P ^ of a t a r or oil is equal to the level of concentration, in percent by weight, of the solution of methylcholanthrene in benzene which would induce papil lomas at the same rate as the material in question under comparable experimental conditions, -The potencies,P,,_, of pure compounds, relative to that of methylcholanthrene, t^ken as 100, may be derived in a similar manner. Prom the data in this paper, the relative potencies of 7l2-dlmethylbenza^anthracene and benzoJVjpyrene are 180 and 60 respectively, ACKNOWLEDGEMENTS ' The authors are indebted to their associates in the Kettering Laboratory and in the petroleum industry for their many constructive I s u g g e s t i o n s . P a r t i c u l a r than k s are d u e to Dr, P. F. H e y r o t h and ^ Dr, R. A. K e h o e f o r their v a l u e d c o u n s e l in the p l a n n i n g of the I program, to M i s s M, J. Graf f o r the t a b u l a t i o n and s t a t i s t i c a l an- 1 alyses of the data, and to Drs. Frank Cleveland and Frank Dutra, who have been responsible for the pathology. API 05965 - 28 - REFERENCES (1) Tsutsui, H. Ue b e r das K n s t l i c h erze u g t e C a n c r o i d bei der Maus. Gann. J a p a n Ztschr. f. K r e b s f o r s c h . , 12:17-21, I9I 8-I9 . (2) Scott, A. On the O c c u p a t i o n Canc e r of the P a r a f f i n and oil : - , . Workers of the S c o t t i s h Shale Oil Industry. 2 1108 9 1922 Brit. M. J., (3 ) Southern, A. H., and Wilson, S. R. Brit. M. J., 2:971-3, 1922. Cancer of the Scrotum. (I4.) Leitch, A. P a r a f f i n C a n c e r a n d its E x p e r i m e n t a l Production, Brit. M. J., 2 : IIOI4.-6 , 1922. (5) Twort, C. C., and Twort, J. M. Th e R e l a t i v e P o t e n c y of C a r c i n o g e n i c Tars and Oils. J. Hyg., 29:373-9 1929-30. (6 ) Pullinger, B. D. The L o c a l i z a t i o n of E x p e r i m e n t a l Tumors in Scars a n d Healing' Wounds. J. Path, a nd Bact., 55:301-9 19U3. (7 ) Cramer, W . , and Stowell, R. E. The E a r l y Stages of C a r c i n o genesis by 20-Methylcholanthrene in the Skin of the Mouse. I. E x p e r i m e n t a l T e c h n i q u e s and M a c r o s c o p i c Changes. J. Nat. . Cancer Inst., 2:369-78 19^2. (8) Bryan, W. R., and Shimkin, M. B. Q u a n t i t a t i v e A n a l y s i s of Dose-Response Data Obtained with Carcinogenic Hydrocarbons. J. Nat. C a n c e r Inst,, 1:807-33, 19^1. (9) Bliss, C. I The C a l c u l a t i o n of the T i m e - M o r t a l i t y Curve. Ann. Appl. Biol., 21^:815-52, 1937 (10) Whittaker, E., and Robinson, G. The Calculus of Observations, pp. 209-59 2nd Ed., London; Blackie and Sons, Ltd., 1932. (11) W a t son, A. F., and Mellanby, E. T *r c a n c e r in Mice. I. The T e c h n i q u e of the C o m p a r a t i v e E x p e r i m e n t . Brit. J. Exper. Path., 11:267-81, 1930. (12) Bradbury, J. T., Bachmann, W. Production of Cancer by Some Res., 1:685-91;, 191^. E., New and L e w isohn, M. G. Chemical Compounds. The Cancer (13) ( H 4.) Stamer, S. E f f e c t of a C a r c i n o g e n i c H y d r o c a r b o n on M a n i f e s t Malignant Tumors in Mice. Acta Path, et Microbiol. Scandinay, Supp. I4.7 , 191+3* Bliss, C. I. The S t a t i s t i c s of Bioassay, pp. I4.69- 7O. New York: Academic Press Inc., 1952. 1st Ed., API 05966 29 ' '! Iv.'-csu, 5 ., and Arley, N. O n the M e c h a n i s m of Experimental ^ r c i n o g e n e s i s . n e t a Path, et Microbiol. Scandinav., 2 7 : 1 --31, 1950. (16) (17) Badger, G. IvI. The C a r c i n o g e n i c Hydroc a r b o n s : Chem i c a l C o n s t i t u t i o n an d C a r c i n o g e n i c Activity. Brit. J. Cancer, C:309-50, l^Q. Iball, J. The R e l a t i v e P o t e n c y of C a r c i n o g e n i c Compounds. Am. J. Cancer, 35:188-90, 1939* (IS) Blanding, F. H., King, W. H . , Priestley, W . , a nd Rehner, J. Properties of High-boiling Petroleum Products. Arch. Ind. Hyg. Occ. Med., ft:335-1+5> 1951. (19) Tye, R., Graf, M. J., and Horton, A. Iodination on ,'ctivated Alumina for Benzo^a]pyrene in Complex Mixtures, W. The Use of Catalytic the Determination of Anal. Chem., (20) Ber e n b l u m , I.,and Schoental, R. C a r c i n o g e n i c C o n s t i t u e n t s of Coa l - T a r . Brit. J. Cancer, 1:157-65, 191+7 (21) Fischer, H. G. M., Priestley, tf., Eby, L.T., Wanless, G.G., and Rehner, J. Prop e r t i e s of H i g h - b o i l i n g P e t r o l e u m Products. Arch. Ind. Hyg. Occ. Med., i+:3l5-2l+, 1951. API 05967 - 30 - experiments involving the application of materials upon the skin of C3H MICE LEGEND Symbols used In connection with average weight curves! i W 0T Time of death (from disease) of Ith tumor-free mouse, Time of death (from disease) of ith tumor-bearing mouse, - Time mouse killed. Gross and microscopic pathology: (Tj Time of appearance of first papilloma in tth mouse. W h e n this s y m b o l is f i r s t u s e d f o r a g i v e n m o u s e after its death, the presence of n o n - invading c a r cinoma (intra-epithelial) or small areas of benign neoplasm was determinable only by histopathology. ft! T] or Time of appearance of gross changes indicative of malignancy in tumor of L t h mouse. Diagnosis confir med except in cases where no tissue section available. When this symbol is first used for a given mouse after its death, the invasive malignancy of the tumor was determinable only by histopathology. No tissue section available for histopathology. API 05968  EXPERIMENT 220, SOLUTION FIGURE 1 OF METHYLCHOLANTHRENE, I . O 3I4. P E R C E N T B Y W E I G H T IN BENAENE ) ) ____________ _ FIGURE 2 EXPERIMENT 268, SOLUTION OF METHYLCHOLANTHRENE, 0.287 PERCENT BY WEIGHT IN BENZENE I VjJ ro t AVERAGE WEIGHT OF SURVIVORS (grams) ) FIGURE U- ) ) ) 4 V FIGURE 5 DOSAGE OF METHYLCHOLANTHRENE (mg./Std. Area/week) 2.0 1.8 1.6 l.Il 1. 2- 1.0 1 VI 0 .8 - 1 0. 6- o.U 0.2 0.0 - X o Ul \o -I-----1-- -I - I - 1 U) 1* 8 12 16 20 2I4. 28 32 36 1*0 1+J+ 1|8 $2 56 60 6i* 68 72 Area of skin covered by x, MEAN TIME OF APPEARANCE OF TUMORS (weeks) 100 mg. of solution. Copy from D. V. S t r o o p for i n f o r m a t i o n of M e m b e r s of M e d i c a l Advis o r y Committee,_____________________________________________________ A u g u s t 23. 1954 rf ri f w , ESS0 LABORATORIES STANDARD OIL DEVELOPMENT ^ P. 0. B o x 51 Linden, N. J. COMPANY MEDICAL RESEARCH DIVISION Robert E. E c k a r d t , PH.D., M . D . , D i r e c t o r August 12, 1954 Mr. D. V. S t r o o p American Petroleum Institute 50 West 50th Street New York 20, New York Dear Mr. Stroop: I have reviewed the information which was forwarded from England and 1 am sending to you the material which I think should be duplicated for distribution to the members of the Subcommittee on Carcinogenicity. Of particular importance I believe is item 4 on the Agenda. This suggested that the British may be taking cognizance of the die sel engine exhaust potential problem. It will be interesting to follow their approach to this problem. 1 would appreciate it if you would have the attached material duplicated f o r distribution not only to the members of the Subcommittee on Carcinogenicity but to all members of the Medical Advisory Committee. Very truly yours, / s / R. E. Eckardt R. E. Eckar d t , M. D. REE :11k E n d s . Committee on the Carcinogenic Action of Mineral Oils M R C . 5 V 5 2 7 , CAMO. Ag. 16 MRC.5V528, CAMO.54/4 MRC.5V529, CAMO.54/5 MRC.54/542, CAMO.54/6 M R C .54/547, C A M O . 5 4 / 9 API 05974 MEDICAL RESEARCH COUNCIL For Specified Circulation (Committee members) M R C .54/527 CAM0.Ag.l6 COMMITTEE ON THE CARCINOGENIC ACTION OF MINERAL OILS AB E ND A for the Sixteenth Meeting to be held at 11.15 a.m. on Friday, 23rd July, 1954 at 38 Old Queen Street, London, S.W.l. 1. Minutes of previous meeting. (MRC.54/454 " already circulated^ 2. Progress Reports. Progress report by Dr. W. Carruthers and Professor J. W. Cook. (CAMO.54/4 " Enclosure A ) Progress report by Professor H. N. Green and Dr. M. J. Redcliffe. (CAMO.54/5 " Enclosure B ) 3. Future Plans. 4. Possible Carcinogenicity of Diesel exhaust fumes. There Is a continuing and growing Interest In the possible carcinogenicity of diesel exhaust fumes and It would be of assistance to the Council to have an authoritative statement on the subject. The Committee might also wish to make recommendations about possible future research. 5. A ny o t h e r business. 6 . Date of next meeting. API 05975 MEDICAL RESEARCH COUNCIL For Specified C irculation (MemDers o f Committee) MRC.5V528 CAM0.5V4 Committee on the Carcinogenic Action of Mineral Oils PROGRESS REPORT BY DR. W. CARRUTHERS AND PROFESSOR J. W. COOK We have continued our examination of the aromatic extracts ! of the two Kuwait fractions Kj,S0b (b.p. 357i - 360) and Kj.Sz-b (b.p. 377i - 380). * 2 4b' As mentioned in the last Report, anthracene hydrocarbons were removed from these* fractions by treatment with maleic anhydride, and the recovered 'anthracene-free' oil separated Into 'plcrate-form- ! lng' and 'non-plcrate-forming ' parts with picric acid in alcohol. The 'picrate-forming ' parts, recovered from the crystalline picrates by decomposing them with alkali, have now been further subdivided by J chromatography on silica gel and on alumina. From several of the small fractions thus produced small amounts of crystalline substances have been isolated. One of these compounds, from K^S^b, has been identified as 1 :8-dimethylphenanthrene. Another substance, from the same fraction, has been shown to be a derivative of fluorene, prob ably a tetramethylfluorene or its equivalent. From fraction K^Sgb a polycyclic sulphur-containing compound has been obtained. Elemen tary analysis indicates the molecular formula C^cHt ^S, and the ultra violet absorption spectrum suggests that it is a^derlvative of the tricyclic condensed thiophene compound 6 :7-benzothionaphthene, Ci2HpS. Further investigation of these and other compounds is in progress. ` API 05976 T MEDICAL RESEARCH COUNCIL For Specified Circulation (Members of Committee) MRC.5 V 529 CAM0.5V5 Committee on the Carcinogenic Action of Mineral Oils PROGRESS REPORT BY PROFESSOR H. N. GREEN AND DR. M. J. REDCLIFFE Mouse Experiment This is now in its 25th week. A large number of tumours have been obtained with 0.2$ and 0 .05$ dimethylbenzanthracene in liquid paraffin, (see mouse life sheets), but of all th oil fractions, f (Kh 4) alone has produced ' two definited tumours. ' An epidemic of haemolytic streptococcal septicaemia in May was responsible for the death of 60 mice. As the mice on e (K4Sgb) were more affected by this than any other group a second series of 50 mice was started on e on 1 1 th June, 1954. Survivors to date in the various groups are as follows: a KhSga] 34 d Kij-Sga] 2 7 g K 4 ,, 4 `> 46 j D) 13 b Kl|S3b) 23 e 19 h : S ) 8r 39 k K4s 2 ) 34 c i * 4 S 6 j 29 f 25 i k Js Jr ) ; 31 Rabbit Experiment This is in its l8th week. One rabbit died at the 10th week. So far 5 tumours have been produced. e (K^Sgb) has produced 2 tumours, both at site II (R ear) at 12 and 4 16 weeks; d (Ki^Sga) has produced 1 tumour at site I (L ear) at 15 weeks; c (K4S6) has produced 1 tumour at site II (R ear) at 15 weeks; h (Di) has produced 1 tumour at site VII (Interscapular) at 16 weeks. API 05977 MO U S E LIFE SHEETS SAMPLE J. (0.20 DMB in Liq. Paraffin) Time in Weeks 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 No. S urviving 46 46 46 45 45 45 44 44 44 40 35 30 28 23 23 19 15 13 No. w i t h Tumours C. 3 4 8 8 10 19 35 41 38 33 29 , 28 (1000) 23 23 19 15 13 SAMPLE H. (0.0556 DMB in Liq. Paraffin) 11 12 13 14 15 I 16 17 18 19 20 21 22 23 24 25 49 2 48 3 45 13 45 - 14 45 14 45 16 45 17 45 20 43 23 43 24 43 24 42 33 41 31 40 32 39 32 S A M P L E F. (Ki, I,) 12 46 0 13 46 0 14 46 0 15 46 1 16 46 0 17 46 0 18 40 0 19 35 0 20 31 1 21 30 1 22 30 1 23 28 1 24 27 2 25 25 2 API 05978 MEDICAL RESEARCH COUNCIL For Specified Circulation (Committee members) MRC.5 4 /5 4 2 C A M 0.5V 6 Committee on the Carcinogenic Action of Mineral Oils PROGRESS REPORT NO. XI. (JULY 1954) by D. L. Woodhouse < I The general observations on the tests at this centre after 44 weeks do not differ materially from those given in the previous report of April 1954 - 30 weeks. Rabbit Tests The tumours'now present on the rabbits are indicated in Table 1. It will be seen that (1) Some of the fractions which previously had produced only a few papillomas have now shown some "long term" activity e.g. K4S3A (7 cf. 1 ) and K4S3 (6 cf. 2 ). (2) The paraffinic fraction K4SP has not produced any further papillomas in this period. (3) On the contrary in one instance some small papillomas which appeared early have not developed or have actually regressed cf. K4S4A and K44. (4) The number of tumours produced by the Aromatic (SA) series still exceeds that in the (S) series. (5) The response to the 0.2# 9 :10-dimethyl-l:2-benzanthracene is now practically equal to that of the active mineral oil frac tions K4s 6a and K4S8A with 16 positive sites each. Comparison of the type, size mas in the standard carcinogens and in impression that the former are usually In all cases multiple papillomas often cation. and rate of growth of papillo the oil fractions gives the more fleshy and spreading. appear on the sites of appli Mouse Tests The tumours on the mice treated with the mineral oils are still fewer than in the rabbit series, the differences being special ly observed in K4S6A. The most active fraction is K4S8A with 7 small but definite papillomas. In many instances, papillomas about 2mm in diam eter appear to remain unchanged fo r long p erio d s. The tumours present at 44 weeks are given in Table 2. API 05979 ) ) ) Table 1. Rabbit Experiments 1953-54 Sites and Time of Appearance of Tumours (after 44 weeks) K4S1A Weeks Site 20 II 20 IV 22 I 30 III 35 I 35 III 37 VII 42 I 8 tumours K4S2A Weeks Site 18 III 25 VII 27 I 30 IV 33 I 35 VII 6 tumours K4S3A Weeks Site 13 II 27 VII 33 IV 37 I 37 III 37 VII 42 III 7 tumours K4S4A Weeks Site 13 III 13 IV 17 II 22 I 28 I 35 I 35 I 37 II 8 tumours i 1 K4S5A Weeks Site 13 III 13 II 13 VII 14 V,II 18 I 18 II 20 I 20 IV 22 III 22 IV 27 I 29 I 35 IV 42 III 14 tumors K4S6A Weeks Site 11 VII 12 IV 13 III 17 IV 17 IV 18 II 18 II 20 I 20 I 20 VII 22 IV 22 II 22 VII 27 II 40 III 42 VII T5 tumors K4S7A Weeks Site 10 VII 11 VII 17 III 17 IV 18 III 20 I 25 IV 27 I 35 III 9 tumours K4S8A Weeks Site 11 IV 11 IV 11 IV 11 VII 11 VII 13 III 17 II 18 III 20 III 22 VII 27 VII 28 I 28 I 35 IV 37 II 42 III 16 tumours API 05980 Tnhle 1 cont. Rabbit Experiments 1953-5*1 Site and Time of Appearance of Tumours (after 44 weeks) K4S1A Weeks Site 25 I 42 III 2 tumours K4S2A Weeks Site 18 I 20 VII 22 I 22 II 35 II 37 VII 6 tumours K4S3A Weeks Site 18 II 25 III 33 VII 35 IV 37 II 37 VII 6 tumours K4s 4a Weeks Site 11 I 13 VII 25 IV 35 VII 35 VII 37 VII 42 IV 7 tumours K4S5A Weeks Site 30 I 35 II 37 II 37 IV 4o II 5 tumours K4s 6A Weeks Site 18 VII 20 VII 22 II 33 h i 40 i i 40 IV 6 tumours K4S7A Weeks Site 17 III 18 VII 18 VII 37 I 37 VII 42 IV 6 tumours K4S8A Weeks Site 10 IV 11 II - 3 0 - VII 3 tumours > u M O U1 VO 00 Table 1 cont Rabbit Experiments 1953-5*1 Site and Time of Appearance of Tumours D(2) Weeks Site 17 I 17 II 18 II 18 VII 22 I 22 II 22 VII 25 IV 27 VII 33 III 35 I 37 III 42 III 42 III 42 VII 1'5 tumours D(i) Weeks Site 22 VII 27 II 30 VII 35 I 35 IV 35 IV 6 tumours Kj|4 Weeks Site 10 II 27 III 30 III 33 IV 42 i 5 itumours Ki*4(1) Weeks Site - - K4SP Weeks Site 20 VII 2^ II 2 tumours (after 44 weeks) API 05982 Table 2. K^SIA K433A K434A K4S5A K4s 6a K4S7A K4S8A % K44 K4S4 K4S5 K4s 8 Mouse Experiments lqq^-s4 (after 4* weeks) ' ' i.;o. of tumours/time In weeks 1 /22, 1/24 1/35, 1/44 1/9, 1/30, 1/34, 1/35 1/24, 1/35, 1/44 1/20, 1/40 1 /23, I/32, 2/44 1/22, 1/23, 2/28, 1/30, 2/40 1/22, 1/35 1/30, 1/40 1/34 1/36 Total Tumours 2 2 4 3 2 4 7 2 2 1 1 f I I I API 05983 MEDICAL RESEARCH COUNCIL Fc'' Spe cified Circulation t lri.bei a of C o m m i t t e e ) Copied as: MRC.5V547 CAM0.5V9 Committee on the Carcinogenic Action of Mineral Oils C O P Y O F A L E T T E R F R O M DR. W. K. T A Y L O R TO Mft. J. C. R. H U D S O N GODALMING GROUP LABORATORY Group Laboratory c/o St. Thomas's Hospital, Godaiming, Surrey. WKT/MPS. 19th July, 1954 Dear Mr. Hudson, As the Pathologist of a large group of Chest Hospitals, I am naturally very interested in the question of squamous metaplasia and bronchial carcinoma, particularly so in the relationship between this disease and the inhalation of oil and its combustion products. I feel, personally, and various items of research tend to support the view, that there is a relationship and it seems very regrettable that just at the time w h e n large scale research on the subject is being initiated, London Transport should seek to introduce 1,700 additional oil driven vehicles on the streets of London in place of electric trolley buses. It seems to be a pity that no influential group of doctors can persuade London Transport to postpone this step until more is known of the aetiology not only of carcinoma of the lung but other respiratory diseases and particularly squamous metaplasia. One important point is very clear and that is that a con siderable amount of oil is to be found in suspension in the air over our roads, as evidenced by the greasy state of the average car windscreen. It would be of interest to determine the origin of this oil -- i.e., diesel exhaust or lubricating oil and whether it is associated with substances which are likely to potentiate any carcinogenic effect. It would be of interest to examine air collected through the intake fan of a car heater and compare the findings in a centre like central London with its numerous diesel vehicles and an equally industrialised area such as might be found in Wolverhampton where passenger diesel vehicles are in the minority. A further point which should be considered is whether first class maintenance of diesel engines is likely to produce finer oil covered carbon particles which are more likely to remain in suspension in the air. The large particles in the black exhaust from badly maintained engines rapidly sink to the ground. API 05984 Mr. J. Hudson Page 2 Whilst on the question of possible toxicity of exhaust gases, attention might be paid to the desirability of using the present type of air intake for car heaters where air is collected in a vent at the front of the car about one foot from the ground. Another important factor is the increasing use of small portable 1 h.p. engines using petrol -- oil mixture, many of which are used under cover. I personally find the fumes from my rotary hoe extremely irritant. However, the problem daily becomes more confused and it may well be that many factors are responsible and that the cause of squamous carcinoma is not that of the columnar cell type. I should be delighted to place the facilities of this department at your disposal, if required, now that we have a certain amount of research capacity available following the completion of work which we have been carrying out for the Tuberculosis Research Unit. Yours sincerely, /s/ W. K. Taylor Group Pathologist Mr. J. C. R. Hudson, Medical Research Council, 38 Old Queen Street, London, S.W.l. API 05985 lH i KETTER1NO LARORATORT C0LLESE OF MEDICINE-- EDEN AVENUS CINCINNATI I * . OHIO UNIVERSITY OF CINCINNATI DEPARTMENT OF PREVENTIVE MEDICINE ANO INDUSTRIAL HEALTH S- / CABLE AODRESS: KETLAB, CINCINNATI TELEPHONE: CAPITOL M i l September 14, 1954 Mr. D. V. Stroop, Director Department of Technical Services American Petroleum Institute 50 W e s t 50t h Sfceeet New Y o r k 20, Nev Y o r k Dear Mr. Stroopj I a m sending -you h e r e w i t h a statement of expendltures made on behalf of American Petroleum Institute during the second quarter of 195^* I believe you will find this statement self-explanatory, but If you have any questions or comments, Doctor Kehoe would appreciate your bringing them to his attention. Very truly yours, t (ft A d y is E. ft. P o r t lge, Secretary to Dr. Eehoe API 05986 O' i .T=g.^ OrC<*&r i M O ' I ---------U N lVERSITYO f CINCINNATI" KETTERING LABORATORY ACCOUNT OF THE AMERICAN PETROLEUM INSTITUTE FOR 2nd Quarter 1QS4___________ SALARIES (Based on Proportion o f T im a A ctually Spent on Project) D ir e c t S a la r ie s _________________________ 3923.*33.___ Indirect Salaries H sto p s th o lo g ica l 4QQ.5Q__ P reparation___________________________ O ther .6634...43 S e r v ic e s ............................................................................... ______ 10,?pfl.9fi MISCELLANEOUS EXPENSE P u rch ase o f 367 A n im a ls.............................................................................................. . ..2Q.... S p ecia l Laboratory S u p p lies--------------------------------------------------------------- l 8 L . 4 Z -------- T ravel ........................................................... 0 1 9 1 0 2 ---------- Overhead (Proportion o f H eat, Gas, E lectricity, Steam , T elephone, G eneral Laboratory S u p p lies, P o sta g e, A n n u itie s, P en sio n s, M a in ten a n ce, e t c . - 2 1 0 9 - & 3 --------- - 3 , 9 3 ^ . 7 2 T O T A L ....................14,894.98 Balance Available for Further W ork a t End o f - J L t t j t L . Q n a r t w 3 9 5 4 ____________ 1 5 , 7 5 2 . 8 2 Balance Doe Kettering Laboratory a t End o f .------------------------------------------------------------------- R e c e i p t s ------------------------------------------------------------------------------ ---------------------------- E xp en d itu res . . . . . 2 o A ,, Q U A r . t i t X _ 1 9 5 4 ............. ............... - 1 4 , 8 9 4 ^ 9 8 Balance Available for Further W ork a t End o f . . ,, 2 B L < l . - Q i u u r i f l r _ l S 5 . 4 . ----------------- 8 5 7 .8 4 FORM mo Balance Due K ettering Laboratory a t End o f .____________________________________________ KET. LAB. M-- II-M API 05987 Co7 Ft m D. Y. Sto Tor Xteanatla* te Oaarfo M. 0 h It i . M . 1. lcterlt. M. 8.___________________________ i-- tatear ' Uml itmU tatmirT cr ubo C r T a KIWf tckt f Z JceooK cr f amdctcai c e ra i a r a m i FW ______ 2a i ftoirttr 195________ SALARUS ( u id on rotartia* af fia t Ateaally S f nt a i Frojact) Dir*t Irtte l i ............................................... J983.33 Xtelroet Salarlo Htetetholasiaal Prayoxtelaa . . . . . . . . . . fcOO.90 Otter SarriaM . . . . . . . . . . . . . . . . . . . ......663k.*3 n a e n z A K o n expense Paratew t f te la la .'.................................... SfiT. Miai Laboratory ) 1 H i ........... lB6.k7 f t w t l ..................................... 1191.te 10.958.26 (Frojartlai of --te t e , lleatrleity, l a i, talaffcoM, B tew , 3.936.72 C IB A I...................... ... Balano A it it e li for Farttei I t e te It e te Ite -- tew 19* 15,79 .82 lk ,89k.98 HftelteTM arteria lo 1 -- 0 Am ilabla far Fwrtter Work te Ite te te i teartar 195 Boloaoo ter Kottorlac LaMtetory te Ite t e ___________________ . 8 5 7 . 8k API 05988 4 Copy From D. V. Stroop t o Members of Medical Advisory Committee and Subcommittee on C arcinogenicity_____________________________________________________ October 19, 195^ ESSO LABORATORIES STANDARD OIL DEVELOPMENT COMPANY P.0. BCK 51, LINDEN, N. J. October 15, 195^ Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New York 20, New York Dear Mr. Stroop: I am attaching recent minutes of the Committee on the Carcinogenic Action of Mineral oils of the Medical Research Council of Great Britain. These have been forwarded through Dr. Newquist's office, and after review ing them I believe it would be of interest to the members of the Medical Advisory Committee to review these minutes. I would appreciate it, therefore, if you would arrange to have these minutes duplicated and distributed to all the members of the Medical Advisory Committee and, in addition, to the Kettering Laboratory group. Sincerely yours, /s/ R. E. Eckardt R. E. ECKARDT, M. D. REE:ilk Enel. API 05989 MEDICAL RESEARCH COUNCIL For Specified (Normal Committee) Circulation MRC. 5^/606 Camo.Min.l6. COMMITTEE ON THE CARCINOGENIC ACTION OF MINERAL OILS Minutes of the Sixteenth Meeting, held at 38 Old Queen Street, London, S.W.I., on Friday, 23rd July, I95U, at 11:15 a.m. Present: Professor T. Ferguson (Chairman), Colonel S.J.M. Auld, Professor J.W. Cook, Professor H.N.Green, Dr. I. Hieger, Dr. J.O.Irwin, Professor F.Morton, Professor R.D.Passey, Professor J.R.Squire, Dr. D.L.Woodhouse, and Mr.J.C.R. Hudson (Secretary). Attended: Dr. W.Carruthers, Dr. J.Faulkner and Dr. B.S.Lush. 13^. Minutes. After making the following amendments the Minutes of the previous meeting were approved and signed: (i) Minute 129, second paragraph; "originally thought that the Lagunillas" to read "originally found that the Lagunillas"; (ii) Minute 129, second paragraph; "approximately 210C" to read "approximately 310C"; (iii) Minute 130, second sentence to read: "It'was noteworthy, he said, that the potencies of the fractions relative to the standard carcinogens as judged "by mice and rabbits were different, but the response of the various fractions among themselves was similar in both types of experiment". 135 Progress Reports (i) In amplification of his report (CAMO. 5k/k) Professor Cook said that some of the compounds isolated were in minute quantities and it was accordingly difficult to analyse and identify them. Of the three compounds reported he thought that the isolation of a fluorene derivative from a mineral oil had not previously been recorded. (ii) Professor Green had nothing to add to his report (CAM0.5V5) although he hoped to be able to develop a test for carcinogenicity by tumour inhibition. (iii) The present report by Dr, Woodhouse (CAMO.5^/6) showed that the results quoted in the previous report (CAMO.5^/3) based on thirty weeks, trial gave a sufficiently close indication of levels of biological activity for the purpose of deciding on subsequent experiments although there might be inaccuracies of detail. The results indicated that the carcinogenicity 1 kPl 05990 occurred mainly in the higher (3^7-390C) boiling fractions of the aromatic portion. Prom this it appeared that the tests should be extended to even higher boiling fractions in order to make certain that no appreciable degree of carcinogenicity remained in the residue. (iv) Professor Morton reported that he had nov completed the distillation of Lagunillas up to 295C, and that modification of the Steadman still to operate under a higher vacuum would be necessary before any higher boiling fractions could be obtained. 136. Nomenclature. It was suggested that the fractions now being produced might be re-classified under single letters or figures for convenience. However, the general feeling was that the present nomenclature was known by all the people concerned and that the title of each fraction epitomised its life history. It was agreed to maintain the present nomenclature for all purposes. below. For convenience, the present system of nomenclature is summarised Three original crude oils, Kuwait (K), Lagunillas (L), and Oklahoma (0), were distilled each into four fractions and a residue, i.e. Kl-5 etc. Fractions distilled on the Steadman still (s) into 2^-C cuts, were designated lower and upper boiling fractions (a & b ) , which were then mixed to give eight 5C cuts, for convenience in the biological tests. The 5C cuts, which were given numerical suffixes, i.e. K^.j S^.g, were split into aromatic (A) and raffinate (R) portions. Subsequent chemical treatment of the aromatic extract gave a maleic anhydride reactive portion (m), i.e. an anthracene; a picrate forming portion (p) and a residue (r). Thus, K^SgsAm would be the anthracene portion of the aromatic extract from the lower boiling fraction of the sixth 5C cut in the Steadman still of the fourth fraction of the original Kuwait crude oil. The term K^.g represents the sixth cut of a laboratory distillation of K^ and is equivalent to Kl^Sg. 137. Future Plans for the year 195^-1955 It was agreed that the biological tests to be carried out at Birmingham during the next year should be on the m, p, and r extracts of the fractions K^S^A to K^SgA. The detailed plans for these tests were to be decided between Professor Squire and Professor Cook (circulated subsequently as CAM0.5h/9)* The previous proposal, recorded in Minute 131, was not to be undertaken at present. It was also agreed that Professor Morton should undertake the fractionation of another batch of Kuwait oil, and that, if possible, the aromatic portion should be separated before distillation. 2 API 05991 138. Possible Carcinogenicity of Diesel Exhaust Fuaes. At the Chairman's request Dr. Faulkner gave a brief account of the Council's position with regard to this matter. Numerous inquiries had been received and the answer given to date had been that the Council were supporting research on atmospheric pollution in general both from the point of view of its chemical and physical characteristics and of its effect on health but that apart from the work on the benz-phrene content of diesel exhaust by Mr. R.L. Cooper (in receipt of a personal grant from the Council) no specific investigation was being undertaken on the possible effect of these fumes on health. The Council would welcome the guidance of the Committee on the matter. It was then discussed and the following points raised:- (i) It was noted that visible fumes were emitted from diesel engines only if the engines were improperly maintained. The Committee felt that it would be desirable to obtain information about the nature of the pollution produced under different working conditions, (ii) In this regard Colonel Auld said that Research Group No. 2 of the Institute of Petroleum will be reviewing all the current information on this problem and he agreed to make the Group's report available to the Committee: he thought this review would be available by the end of 195b. (iii) It was noted that Professor D.H. Eey of King's College, who had expressed interest in the problem and who had diesel engines available in his laboratory for experimental work, should be invited to attend the meeting at which the above report would be discussed. London, (iv) Professor Morton said that he also had diesel engines in his laboratory and that he would be prepared to use them under varying working conditions so that the fumes produced under these conditions could be studied. (v) It was the general view of the Committee that, on the face of i t , - the question of diesel exhausts was one which seemed likely to repay study; but it was felt that the line of approach should await study of the review of existing knowledge likely to be available in time for the next meeting of the Committee. 139. Date of Next Meeting It waB agreed that the next meeting should be held on Friday, 17th December, 195^ at 2.30 p.m. 3 API 05992 MEDICAL RESEA?,CH CCHICIL MRC. 5V 607 CAMO. 5 V l O Committee on the Carcinogenic Action of Mineral Oils Basis PLANS FOR '"BIOLOGICAL EXPERIMENTS AT BIRMINGHAM 195^-5 The 1953-^ experiments at Birmingham, as reported in CAMO. 5^/6 and CAMO. 5^/3, were carried out with fractions of Kuwait 1+ (Boiling Range 350-390C) separated in Professor Morton's Steadman Still (K1*Si -8) and by subsequent extraction of aromatic-containing fractions with acetone solvent extraction (KI4.S1A-8A ) . These experiments showed that even with the Steadman apparatus little further separation was being achieved by distillation, due presumably to the formation of azeotropic mixtures of carcinogen(s) with paraffinic compounds. (Better separations might be achieved by distillation of the separated aromaticcontaining material, but this would require an even higher vacuum to keep the still temperature down to acceptable levels.) The maximum difference in concentration within the sries KkSi-8 is of the order of 2x - 3x. On the other hand, solvent extraction with acetone was shown to concentrate the carcinogen into the aromatic-containing fraction. As acetone extraction removed only 16 -22# of KLS2 -8 and 32# of K l ^ (cf. CAMO. 53/5), this effected considerable concentration, the biological results showing effects 2x - lOx greater with each KijSa fraction when ctmprd Vtl TE cOEr'efcdhlrig K^S fraction. Concurrent chemical work at Glasgow (CAMO. 5^/l, CAMO. 5^+A) shown the feasability of separating each aromatic-containing extract into three main classes of compound:- (i) "m" - maleic anhydride adduct-formers containing mainly anthracene derivatives. (ii) ' "p" - substances forming picrates from anthracene-free oils, including phenanthrenes, etc. (iii) ,"r" - remaining substances in these otbls. It was agreed therefore at the Committee meetings held on 23rd July, I95I+, and 29th April, 195U, that the biological testing at Birmingham for 195^-55 should be carried out with "m", "p" and "r" fractions derived from KkSuA " K ^ S A ( the most potent fractions so far obtained from Kuwait oil). As the quantity of material obtainable in this way was in some instances likely to be only l/lO or l/20 of the corresponding Ki^S-A fraction, it ^/as further agreed that these materials should be diluted for biological testing with liquid paraffin to the same volume as that of the KLS-A fraction from which it had been derived. 250 ml. amounts of each test material would be required for this purpose. 1 API 05993 Details cf blending, etc. Seme complication arose from the fact that each Ki^Sl-S fraction covering 5 C boiling ranges which were previously tested was derived by admixture of 2fC cuts kept separate for chemical work e.g. Kl+Sg boiling range 385c-39CcC was derived from fySga 385-387iC and K)*SSb 367-|-390C. (In each cast the subscript "a" referred to the lower boiling range subfraction.) The details of fractions to be used were therefore left to Dr. Carruthers and Proffessor Squire to work out. It is suggested that the following l8 fractions should be used:- K1+S8A m IQ+S7A m K^SgAa m w P K. SaA r k 0 K1+37A P r w K^SgAa P' K^SgAa r K^SgAb m Wb P K4s6Ab r K)+S5A m W P \SjA r h h A m Kl+Sl+A P KkSkA r It will be noted that in the ca'se of the K^Sg fractions, those derived from K^SgAa and K^SgAb have been kept separate. In the other instances blending will be used so as to maintain the same ration of a/b as in the original oil. Thus to prepare the K^SgA tests the following calculation is used Volume in Original Distillation # in 'Aromatic 1 Extract. Vol. aromatics Volume of Aromatic Fraction in 250 ml. of Blend with Same Proportion of Aromatics as in Original Oil. K. S 10.28 1. k ga 21.2 2179 ml. (KuSgAa ) 250 X 2179 - Ihk ml. 37B5 ' KUs8b 9 *13 17.6 I606 ml. (K^SgA^) 250 X 1606 : 106 ml. 3735 Total KuSp 19.^1 litres Total Volume (K^Sga) 1 3785 ml. s 250 ml. Thus for K^SpA m, K^SgA p and K^SgA r, Dr. Carruthers will supply m, p and r compounds equivalent to lM+ ml. of K^SgAa, admixed with m, p and r compounds equivalent to 106 ml. of K^SgAb. Each of these three will then be made up to 250 ml. at Birmingham with liquid paraffin for biological testing. Analogous procedures will be used for the Ki|S^A, Ki+S^A and K^S^A subfractions a and b. In the case of K^SgA and K^SgA. , tne amounts of m, p and r compounds equivalent to 250 ml. of each of these fractions will be supplied for biological testing separately. 2 API 05994 -K Hemainder o: 21 fractions for biological testing . . vo The frictions tested will be D(2) andD(|) standards SS n (-;e * Sifi-andT,*5'' 9:10 diaethyl 1:2 benzanthracene in liquid paraffin). InQaddi.ion Professor Merton will supply KkS,, a fraction boiling just above 390 C prepared irom the residue left after distillation of K.3 -A in order to see whether appreciable further amounts of carcinogen remain In this residue. Po ssib le r e s u l t s t o be a n t i c i p a t e d from "m" "p" "r" tests 1. If complete separation of carcinogen occurs, one of the "m", "p" or "r" fractions diluted in liquid paraffin should have the same potency as the corresponding K^S-, fraction in the 1953-^ tests, while the remaining two should be free from activity (e.g. in the U+ week results of CAMO. 5^/6 the K^SgA'm might show about 16/25 rabbit sites affected while K^SgA p and K^SgA r showed none). 2. If no separation occurs, each test substance should have l/3 the activity of the corresponding K S.^ fraction in the 1953-k test, (e.g. in the UU week results the K, S.A m, K^SdA p and K^SgA r fractions might each show about 8/2 5 -rabbit sites affected). 3. A result intermediate between 1. and 2. is perhaps the most likely. Anomalous results can also be envisaged. These examples are given to show that the biological tests are feasible and likely to give constructive information. Proposed date for starting 195^-5 biological testing at Birmingham. This to be 1st October. If delay in supply fractions turns out to be inevitable, Dr. Carruthers undertook to warn Dr. Woodhouse at least by 7th September. Dr. Irwin also undertook to suggest suitable allocation of 21 fractions to rabbit sites as before. 5th August, 195U 3 API 05995 96650 IdV n laww i `i /*/ JT -- m W ITT M* T W X `*ttt J* t t e w *41 CI 4 T 9 MOL M i 90S MM os t M TI M O i J M%fxn *Hn>m *A *41 w i T M i m j t t w p n m -- Trrm ^(TW ' U V M M S *d TFiT * n o m *A *g M1 J i g il irrur KI TBMDP UOCRAXGRX Aoconer cr m jm bucai n o rau o i a a rra n s ree o r o u r s r cr 195* 8ALAHXBS (Bbm A ob Proport i of T la Aetutlly 8pent ob Project) Direct S o le rla $$862.79 Indirect salarle ttatopattalaglcal Preparati 6$890 KZBCXLLABOtTS $10.777.6$ frirrlaai of A a i a l S p e d ii L IW lM T BBppli-- arerei P arteal (P re p o rti f Seat, Oea, S la c tris itjr, Maae, BeLopSeeo, 210.32 A l.23 io $3.$7 * ' * '______t s f i _____ VQZAL $16,398.87 la la e e irallafele fa r Ferther Ver* B t M t f t B i o r la r 199$ 857. 8$ H artyta A i r 199$ 36125.00 f a Ardlafcle 3^j 982.8$ r o m i t a 3ri Qpar tar 199$ 16,358.87 le i ance Avmilobla far fu rto r Ver* H M < 3rt Qwrter 199$ ,6 8 3 .9 7 ' INSTITUTE CF FETROIEUM RESEARCH GROUP No. 2. Minutes of Meeting No.lk, held at 26 Portland Place. 29.9.5^. IN THE CHAIR; ATTENDANCE P. Morton GUESTS: W. M. Catebpole P. J. M. Trollope R. B. Killingsvorth A. T. Wilford C. S. Windebank MEMBERS: APOLOGIES FORABSENCE: S. J. M. Auld W. S. Ault E. J. Boorman W. Fohl J W Cook E. J. Dunstan A. McLean J. N. Haresnape SECRETARY: E. Herbert 1. MINUTES CF MEETING No. 13. 1,11 53 These were signed as correct. It was noted that in Minute U on Page 2, line 5, "only two years ago" should read "nearly two years ago". 2. REPORT ON WORK AT BIRMINGHAM UNIVERSITY AND ON M.R.C. COMMITTEE The Chairman described the work which bad been done at the Birmingham University on carcinogenicity of mineral oils, and the progress which bad been made; he referred also to the MRC Committee Interested In this subject. He indicated the programme of future work and mentioned that a full report would shortly be circulated to members. Professor Cook added some technical details. The meeting noted the position with approval. Colonel Auld, having thanked Professor Morton for taking over the Chair of the Committee during his period of office as President, expressed the desire that there should be placed on record the high value of the work being done at Birmingham under Professor Morton's control, and a strong recommendation that this work should be carried en: and should continue to recelva the necessary financial support. This mas duly moved.and seconded, and unanimously agreed. 3. HEALTH HAZARDS ASSOCIATED WITH DIESEL EXHAUST FTOffiS The Chairman explained that the Minister of Health and the Minister of Fuel and Power, having made a joint approach to M.R.C. on the subject of health hazards associated with diesel exhaust fumes, the M.R.C. had asked Its sub committee for a report. HO read Minute 138 of this sub-committee which, although Its terms of reference covered skin cancer, and It was not anxious for additional work, evidently felt that It could not divest Itself of respon sibility towards this problem. The M.R.C. committee had asked for Information on two main points. Firstly, If any Information was available from organizations using diesel transport concerning health studies of employees over a sufficiently long period to enable API 06012 - 2- statistical analyses to be made. Secondly, did the IP. Consnittee consider that research into the composition of diesel exhaust fumes was desirable and if so, by what organization might it be initiated. I. Information concerning health records of Diesel Engine Users. Mr. Wilford, (London Transport Executive) said that such records were available in London Transport and extended over the period (about 25 years) during which diesel-engined vehicles had been in use. Since the records covered a number of garages and depots, some operating internal combustionengined vehicles, others concerned with trolleybuses, they would be expected to provide useful data of the type required. He suggested that the M.R.C. Committee should make a direct approach to the Executive's Chief Medical Officer. Mr. Ault stated that the Shell-B.P. Marketing Organizations would also have health records concerning workers in garages using diesel transport. He believed that a study of these records would be of value but agreed with Mr. Wilford that direct contact between the M.R.C. and the Medical Officers should be established. .. It was also suggested that approach should be made by M.R.C. to the Medical Officers of other transport organizations, particularly the Manchester Corpo ration Transport Department, known to have been operating diesel fleets for some 25 years. It was agreed to reconnend to the M.R.C. Committee that they should seek information from the Medical Officers of the L.T.E., Shell-Mex and B.P. Ltd., Esso Petroleum and possibly M.C.T.D. II. Information concerning Investigation of composition of diesel exhaust fumes. Several members drew attention to the desirability of considering atmos pheric pollution in general and regretted the tendency to over-emphasize the contribution made to the general problem by the diesel engine. Professor Cook gave an account of the committees of the M.R.C. concerned with atmospheric pollution, and suggested that if an investigation into fuel combustion was found to be desirable the assistance of Research Committee No. 2 of the IP. would be of the greatest value. Colonel Auld said that at the moment the Committee could be of the greatest help if the members would use their American and Continental contacts to ascertain what views were held in other countries. In particular the M.R.C. Committee would welcome guidance as to the desirability of setting up research work in this country. It was generally agreed that at this stage the examination of exhausts of individual engines, as suggested in the M.R.C. Committee Minute was not recom mended. Before any experimental work was commenced, it would be desirable to plan the investigation on a broad basis and participants in initial discussions should include representatives of engine users and manufacturers, the Motor Industry Research Association, British Internal Combustion Engine Research Association, and the Petroleum Industry. API 06013 Dr. Killingsvorth asked if investigations should be confined to diesel fuel. It was suggested that diesel fuels had been considered, because their fu.aes could be absorbed at mouth level. Dr. Killingsvorth enquired also whether information had been sought from America, and offered to check with his American colleagues if and where in America there was information on this subject. Mr. Ault felt that it would be premature to approach A.P.I. at this stage, and that it was doubtful if there was much published Information in America although some medical officers might have some interesting data. Mr. Boorman drew attention to two reports in the National Petroleum News regarding Smog in Los Angeles. (See pages 23 and 20 of N.P. News, August Uth and l8tb respectively). It was suggested that M.R.C. be referred to these articles. It was agreed that, if, as a result of their contacts with the Medical Officers of the Interested oil and transport companies, the M.R.C. Committee wish to put any questions to the IP. Committee, then the Committee would attempt to provide the answers. In the meanwhile all members were asked to trace per tinent information on both diesel fuels and petrol, not only from America but also from the Continent of Europe. The question of the. cosipetency of Research Committee 2 to handle this matter was raised. It was pointed out that the problem would eventually have to be submitted to the Research Committee for report to Council. It was con sidered that the most appropriate IP. body to deal with the subject at this stage was Committee 2, particularly as the M.R.C. regard its opinions as un biased and valuable. U. DATE OF NEXT MEETING To be arranged by the Chairman. l8.10.5fc.FMA API 06014 ADDITION TO THE MINOTES by Professor F. Morton. A Committee has recently been established in the U.S.A. under the Co ordinating Research Council, Inc., and sponsored by the American Petroleum Institute and the Automobile Manufacturers Association, to study the compo sition of eritiaust gases from internal combustion automotive engines. This committee is composed of several panels, one of vhich vill study engine variation in relation to its effect upon the composition of exhaust gases. Another panel will develop means of sampling and analyzing exhaust gases,md a third panel is interested in uncovering all possible sources of Information relating to the proper technique for sampling such gases and for obtaining accurate analyses. I have been asked by H. M. Smith of the U.S. Bureau of Mines to co operate with this committee and to send details of a nev sampling valve and techniques vhich have been developed at Birmingham, bub vhich have not yet been described in literature. In return the committee vill send to me all the details of their vork and keep me informed of progress. Do the members of IP. Committee No. 2 feel that I should ask H.M. Smith in confidence if the American Comaittee is in any vay concerned with the health hazards associated vlth diesel exhaust? API 06015 t Various agencies studying the smog problem already have agreed that ozone is linked to smog and is present in the Los Angeles atmosphere in excessive amounts* But the district report is the first flat statement that the substance can be traced to vehicle exhaust gases. 0O0- **m API 06018 \ T CONFIDENTIAL MINUTES CF THE MEDICAL RESEARCH COUNCIL 138. Possible Carcinogenicity of Diesel Exhaust Fumes, At the Chairman's request Dr. Faulkner gave a brief account of the Council's position with regard to this matter. Numerous enquiries had been received and the answer given to date had been that the Council were supporting research on atmospheric pollution in general both from the point of view of its chemical and physical characteristics and of its effect on health but that apart from the work on the benz-pyrene content of diesel exhaust by Mr. R.L. Cooper (in receipt of a personal grant from the Council) no speelflc invest gatlon vas being undertaken on the possible effect of these fumes on health. The Council would welcome the guidance of the Committee in the matter. It was then discussed and the following points raised: l) It was noted that visible fumes were emitted from diesel engines only if the engines were improperly.'maintained. The Committee felt that it would be desirable to obtain information about the nature of the pollution produced under different working conditions. ll) In this regard Colonel Auld said that Research Group No. 2 of the Institute of Petroleum will be reviewing all the current information on this problem and be agreed to make the Group's report available to the Committee: he thought this review would be available by the end of 195k. ill) It was noted that.Professor D. E. Eey of King's College, London, who had expressed Interest in the problem and who bad diesel engines available in his laboratory for experimental work, should be invited to attend the meeting at which the above report would be discussed. lv) Professor Morton said that be also had diesel engines in his laboratory t and that be would be prepared to use them under varying working conditions so that the fumes produced under these conditions could be studied. v) It was the general view of the Committee that, on the face of it, the question of diesel exhausts was one which seemed likely to repay study but it was felt that the line of appraoch should await study of the review of existing knowledge likely to be available in time for the next meeting of the Committee. API 06016 NATIONAL PET?OLEUM REVS. AUGUST U, 1951*. Page 23. MOTOR VEHICLE EXHAUST CALIED TOP SMOG CAUSE Stanford Research Institute has branded motor vehicle exhaust as a major smog contributor in its report summarizing six years of smog study. The Institute's 13^-page report, released by Western Oil and Gas Assn., estimates 3,100 tons of pollutants are emitted daily into the Los Angeles Basin atmosphere, of which 1,500 tons are hydrocarbons. Auto exhaust heads the list with 1,016 tons, followed by: * Evaporation from automobile tanks and carburetors - 16U tons * Refining - 155 tons * Marketing - 37 tons * Production - 32 tons * Filling automobile tanks - 28 tons * Filling service station tanks - 2k tons In short, exhaust gases take the blame for 8l$ of the total organic hydrocarbons emitted and nearly 95$ of the active hydrocarbons. The report notes in passing that backyard incinerators emit kOO tons of organic materials daily. The researchers say the key to smog probably is ozone, since Los Angeles has the world's highest concentration of that oxidant. Moreover, ozone concentrations Increase and diminish with the intensity of the smog. "This ozone may react in the air with harmless materials such as hydro carbons to form troublesome compounds that are responsible for many of the undesirable effects of smog", the report concludes. Copies of the document are available from Western Oil and Gas Assn., at 510, W. 6th St., Los Angeles, lU. NATIONAL PETROIEUM NEWS. August l8. 195k. Page 20. AUTOMQBIIE EXHAUST LINKED TO SMOG Automobile exhausts are responsible for a considerable amount of ozone, the "silent partner" in Los Angeles' smog problem, says Gordon P. Larson, director of the Air Pollution Control District. Larsen said three district scientists have established that unburned gasoline and nitrogen oxides emitted by exhausts create ozone in the presence of sunlight. Ozone, in turn, is believed to "trigger" other chemical reactions in the atmosphere, which causes smog's undesirable effects. So, Larson emphasized, the district now can make formal scientific indictment against exhausts as a smog contributor. As he puts it, "We are on the same firm scientific ground in demanding that automobile exhausts be cleaned up as we were in demanding two years ago that refineries clean up." API 06017 API 06019 ESSO LA B O R A T O R IE S STA N D ARD OIL DEVELOPMENT COMPANY P . O . BOX 51, L IN D E N , N . J . MEDICAL. RESEARCH DIVISION R O B E R T E. E C K A R D T . P H .D .. M .O . D IR E C T O R H O R A C E W . G E R A R O E . P H D . M .O . HEAD TO XIC OLO G IST November 22, 195k INDUSTRIAL HYGIENE SECTION N A T H A N V. H E N D R IC K S . CH .E . HE A D OF SECTION F R A N K L IN W . C H U R C H . M .S. G EO R G E M . W IL K E N IN G . M .S . IN D U S TR IA L HYG IENISTS Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New Tork 20, New Tork Dear Mr. Stroop: I am attaching some minutes received from the Institute of Petroleum's Research Group No* 2 meeting of September 29, 1951a. In view of the discussion in paragraph 3 of these minutes, I believe it would be advisable to have these minutes reproduced and distributed to all of the members of the Medical Advisory Committee. Very truly yours, R. E. ECKARDT, M. D REE:ilk Enel. - Minutes of Meeting No. li* API 06020 MEDICAL RESEARCH DIVISION R O B E R T E. E C K A R D T . P H .D .. M .D . DI RECTOR HORACE W. CERARO E. PH D . M D. HEAD TO X IC O LO SIS T Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New Tork 20, New Tork - Dear Mr. Stroop: I am attaching some minutes received from the Institute of Petroleum*s Research Group No* 2 meeting of September 29, 195U. In view of the discussion in paragraph 3 of these minutes, I believe it would be advisable to have these minutes reproduced and distributed to all of the members of the Medical Advisory Committee. Very truly yours, R. E. BCKARDC, M. D. REEsilk Enel. - Minutes of Meeting No. lit API 06021  R2S-2-14 RESEARCH CRCLT- Ho. 2 . ir.ute: cf -eetir.r 10,14. held at 26 ~ortlar.d P la c e . 2 9 . 9 .5 4 . MZMBERS; APOLOGIES FOR A35Z1CS: SECRETARY: Mort on V/.K. C a t c h p o l e F.J.-r. Trollope R.3. Killingsworth A.I. '."Uford C .S . '.Vi nde ba nk 3.J.M . Auld " .S. Ault E . J . Boorman V. Pohl JCook 3 .J . Dunstan A. McLean J.N. Hares nape 3. Herbert 1. MINUTES OF :!ZZri:TC- Ho. 1 5 . 4 . 1 1 . 5 3 These were signed ss c o r r e c t . I t was noted t h a t i n Minute 4 on Rage 2 l i n e 5, " o n ly two yea rs ago'' sh o u ld r e a d " n e a r l y two -ears ago". 2 . report o:; vork at Birmingham u n i v e r s i t y a:zo on m. r . c . committee The Chairman d e s c r i b e d t h e work which had been done a t t h e Birmir.gha- U n i v e r s it y on c a r c i n o g e n i c i t y of mineral o i l s , and th e p r o g r e s s whi ch h ad b e e n made; h e r e f e r r e d a l s o t o t h e MRC Committee m i n t e r e s t e d i n t h i s s u b j e c t . He i n d i c a t e d t h e programme o f f u t u r e work and mentioned t h a t a f u l l re p o rt would s h o r t l y be c i r c u l a t e d t o members. P r o f e s s o r Cook added some t e c h n i c a l d e t a i l s . The meeting noted the position with approval. Colonel Auld, having thanked F ro fe sso r Morton fo r tak in g over th e Chair of t h e Committee during h is period of o f f ic e as P r e s i d e n t , e x p r e s s e d t h e d e s i r e t h a t t h e r e s h o u l d be p l a c e d on reco rd the high value of the work being done a t Birmingham under P rofessor Morton's c o n tr o l, and a strong recommendation th a t th is work should be c a r r i e d on and should continue to r e c e i v e the n e c e s s a r y f i n a n c i a l s u p p o r t . Thi s was d u l y moved and s e c o n d e d , and unanimously agreed. 3 . HEALTH HAZARDS ASSOCIATED MITH DIESEL EXHAUST FUMES The Chairman e x p l a i n e d t h a t t h e M i n i s t e r of H e a l t h and t h e M i n i s t e r o f Fuel a n d Power, h a v i n g made a j o i n t a p p r o a c h t o M.R.C. on t h e s u b j e c t o f h e a l t h h a z a r d s a s s o c i a t e d w i t h d i e s e l exhaust fumes, t h e M.R.C. had asked i t s sub-committee f o r a r e p o r t . He r e a d Minute 158 o f t h i s s u b - c o m m i t t e e w h i c h , although' its terns of reference covered skin cancer, and it was n o t a n x i o u s f o r a d d i t i o n a l work, e v i d e n t l y f e l t t h a t i t could not divest i t s e l f of r e s p o n s i b i l i ty towards th is problem. The M.R.C. committee had asked f o r i n f o r m a t i o n on two main p o i n t s . F i r s t l y , i f any in fo rm atio n was a v a i l a - l e from o rg a n i sations using diesel transport concerning health studies of employees over a s u f f i c i e n t l y long period to enable s t a t i s t i c a l a n a ly s e s t o be made. S e c o n d l y , d i d ' th e IP. Committee c o n s i d e r t h a t r e s e a r c h i n t o t h e c o m p o s i t i o n o f diesel exhaust fumes was d e s i r a b l e and i f so, by what o r g a n is a ti o n might i t be i n i t i a t e d . API 06022 I . I n f o r m a t i o n c one err. i.ng h e a l t h r e c o r d s o f D i e s e l -r.sir. e I ' s e rs . Hr. H i l f - r d , ( l e u ; or. T r a n s p o r t E x e c u t i v e ) s a i l o h a t s uc h r e c o r l s were jv: - l a ' - l ; ir. London Drar.soort ar.i extended over the period (at out 25 re are ) during v/h.e.t files el-eng ir.ed v e h ic le s had 'eer. i n u s e . 3i r . c - t h e r e c o r d s c o v e r e d a number of g a r a g e s art. d e p o t s , s rrr.e o p e r a t i n g i n t e r n a l c o m b u s t i o n - e n g i n e d v e h i c l e s , o t h e r s c o n c e r n e d ith t r o l l e y b u s e s , t h e y would he e x p e c t e d t o p r o v i d e u s e f u l d a t a of t he t y p e r e q u i r e d . He s u g g e s t e d t h a t t.-.e " .r..C. Corr'l t t e e s h o u l d make a d i r e c t a p p r o a c h t o t h e E x e c u t i v e ' s Chief Medical Officer. Mr, A u l t s t a t e d t h a t t h e S h e l l - " . P. M a r k e t i n g O r g a n i s a t i o n s would also have health records concerning workers in garages u s i n g d i e s e l t r a n s p o r t . He b e l i e v e d t h a t a s t u d y o f t h e s e r e c o r d s wo ul d be o f v a l u e b u t a g r e e d w i t h Mr. V-'ilford t h a t d i r e c t c o n t a c t between t h e M.R.C. and t h e Medical O f f i c e r s s h o u l d be e s t a b l i s h e d . I t was a l s o s u g g e s t e d t h a t a p p r o a c h s h o u l d be made by M.R.C. to the Medical Officers of other tran sp o rt organisations, par t i c u l a r l y t h e M a n c h e s t e r C o r p o r a t i o n T r a n s p o r t D e p a rt m en t , known t o have b ee n o p e r a t i n g d i e s e l f l e e t s f o r some 25 y e a r s . I t was a g r e e d t o recommend t o t h e M.R.C. C o m - i tt e e t h a t they should seek information from the Medical Officers of th e L.T.E/, S hell-H ex and S . P . L t d . , Ssso Petroleu m and p o s s i b l y M.C.T.D. II. Inform&tion concerning in vestigation of composition of diesel exhaust fumes. S e v e r a l members drew a t t e n t i o n t o t h e d e s i r a b i l i t y of considering atmospheric p o llu tio n in general and re g re tte d the t e n d e n c y t o o v e r - e m p h a s i z e t h e c o n t r i b u t i o n made t o ' t h e g e n e r a l problem by t h e d i e s e l e n g i n e . P r o f e s s o r Cock gave an account of t h e Committees of t h e M.R.C. c o n c e r n e d '>vith a t m o s p h e r i c p o l l u t i o n , ana s u g g e s t e d t h a t i f a n i n v e s t i g a t i o n i n t o f u e l c om b u s t i o n was f o u n d t o b e d e s i r a b l e t h e a s s i s t a n c e o f R e s e a r c h Committee Ho. 2 of the 1?. would be of the g r e a t e s t value. C o l o n e l --iuld s a i d t h a t a t t h e moment t h e Committee c o u l d be o f t h e g r e a t e s t h e l p i f t h e members woul d u s e t h e i r American and Continental contacts to a s c e r t a i n what views were held in other c o u n t r i e s . In p a r t i c u l a r t h e M.R.C. Committee would welcome guidance as to the d e s i r a b i l i t y of setoing uc research work in this country. ' I t was g e n e r a l l y a g r e e d t h a t a t t h i s s t a g e t h e ex a m i n a ti o n o f e x h a u s t s of i n d i v i d u a l e n g i n e s , as s u g g e s t e d i n t h e M.R.C. Committee Minute was no t recommended. B e fo r e any e x p e r i m e n t a l work was commenced, i t would be d e s i r a b l e t o p l a n t h e i n v e s t i g a t i o n on a b ro a d b a s i s and p a r t i c i p a n t s in i n i t i a l d i s c u s s i o n s should include re p re se n ta tiv e s of engine users and manufacturers, the Motor Industry Research A ssociation, 3 r i t i s h In te rn a l Combustion Engine R esearch --issociation, and the Petroleum I n d u s t r y . Dr. K illia g s w o rth asked i f i n v e s t i g a t i o n s should be con f i n e d t o d i e s e l f u e l . I t was s u g g e s t e d t h a t d i e s e l f u e l s had been c o n s i d e r e d , b e c a u s e t h e i r fumes c o u l d be a b s : . e e d a t mouth l e v e l . Dr. K illingsw orth enquired also whether information had been sought from America, and offered to check with his American c o l l e a g u e s ^if and w he re i n America t h e r e was i n f o r m a t i o n o n ' t h i s s u b j e c t . Mr. A u l t f e l t t h a t i t woul d be p r e m a t u r e t o appr oach A . P . I . a t t h i s s t a g e , a n d t h a t i t was d o u b t f u l i f wherd was much p u b l i s h e d i n f o r m a t i o n i n Amer ica a l t h o u g h some m edi ca l o f f i c e r s m ig h t ha ve some i n t e r e s t i n g d a t a . Mr. Boorman drew a t t e n t i o n t o two r e p o r t s i n t h e n a t i o n a l P e t r o l e u m Hews r e g a r d i n g Smog i n Ijcs .". n g e l e s , (3e e p ag es 23 and 2C of M.P. News, August 4 t h and API 06023 lct.n respectively). these articles. ' I t 'vas sugg es ted t h a t .R.C. be r e f e r r e d zo __ In was a g r e e d t h a t , i f , as a r e s u l t of t h e i r c o n t a c t s w i t h t h e edical O f f i c e r s of the i n t e r e s t e d o i l and t r a n s p o r t companies, the M.R.C. Committee wish to put any q u e s tio n s t o th e IF. Committee, ther. the Committee would attempt to provide the answers. In the meanwhile a i l members were a s k e d t o t r a c e p e r t i n e n t i n f o r m a t i o n on : c t h d i e s e l f u e l s ar.d p e t r o l , not only from America b ut a l s o from tne Continent of Europe. * The q u e s t i o n of the competency of Research Committee 2 to handle t h i s m a t t e r was r a i s e d . I t was p o i n t e d out t h a t t h e p r ob l em woul d ev e n tu a lly have t o be submitted t o the Research Committee fo r r e p o rt to C o u n c i l . I t was c o n s i d e r e d t h a t t h e most a p p r o p r i a t e IP. body to d e a l^ .v i t h t h e s u b j e c t a t t h i s s t a g e was Committee 2 , p a r t i c u l a r l y as t h e C'-.R.C. r e g a r d i t s o p i n i o n s as u n b i a s e d a nd v a l u a b l e . 4. EAT. I'.'C- To be a r r a n g e d : j t h e C hai rman. 13.10.54.FMA. API 06024 AZ'DI. IC" IS IFF FIITl.FS '37 Professor F. Merton. Committee has re o e r .tl7 te e n e s ta b lis h e d in the U.S.A. under t h e Coordinating Research Council, I n c ., and sponsored b y t h e American P e t r o l e u m I n s t i t u t e an d tins Aut omobi le Manu facturers Association, to study the composition of exhaust rases from in te rn a l cambustion' automotive engines. This committee is composed of s ev e ral pane ls, one of which w i l l study engine v a ria tio n in r e l a t i o n to its e ffe c t upon the composition of e xh a u s t g a s e s . A no th er p a n e l w i l l d e v e lo p means of samp li ng and. a n a l y z i n g e x h a u s t r a s e s , and a t h i r d p a n e l I s I n t e r e s t e d i n uncovering a l l possible sources of Information re la tin g to the proper technique for sampling such gases and f o r obtaining accurate analyses. " I have been asked by H.I-. Smith o f t h e U. 5. Bureau of --ines to co-operate with th is committee and to send d e ta ils of a new samp li ng v a l v e and t e c h n i q u e s which have b e e n d ev e lo p e d as Birmingham, but which have not yet been described in l i t e r a t u r e . Ir. r e t u r n t h e c o m m i t t e e w i l l s e n d t o me a l l t h e d e t a i l s o f t h e i r w o r k and keep me i n fo r m e d o f p r o g r e s s . Bo t h e members o f I P . Committee tfo.2 f e e l t h a t I s h o u l d ask K." . Smith ir. confidence i f the American Committee is in any ray concerned with the health hazards associated v/ith diesel exhaust ? 06025 API g.__ -JtL " 1 3S. P o s s i b l e C a r c i n o g e n i c i t y of D ie s e l Exhaust Pmes, At t h e C h a i r m a n ' s r e q u e s t Dr . F a u l k n e r gave a b r i e f account of the Council's position with regard to this natter. Numerous e n q u i r i e s had b e e n r e c e i v e d an d t h e answo* g i v e n t o d a t e had bee n t h a t t h e Cou nci l wer e s u p p o r t i n g r e s e a r c h on atmospheric p o llu tio n in g en eral both from the point of view of it s chemical and physical c h a r a c te r is tic s and of i t s effect on h e a l t h b u t t h a t a p a r t from t h e work on t h e b e n z - p y r e n e c o n t e n t o f d i e s e l e x h a u s t by Mr. R .L. Cooper ( i n r e c e i p t o f a p e r s o n a l g r a n t from t h e C o u n c i l ) no s p e c i f i c i n v e s t i g a t i o n was b e i n g u n d e r t a k e n on t h e p o s s i b l e e f f e c t o f t h e s e fumes on h e a l t h . The C o u n c i l wo ul d welcome t h e g u i d a n c e o f t h e Committee m ri an i st eh de : m a t t e r . I t was t h e n d i s c u s s e d , and t h e f o l l o w i n g p o i n t s i i) I t was noted t h a t v i s i b l e fumes were o m itte d from d i e s e l e n g i n e s only i f t h e e n g i n e s were i m p r o p e r l y m a i n t a i n e d . The Committee f e l t t h a t i t would be d e s ira b le to o btain information about the nature of the pollution produced under diffe ren t working conditions. .* i i ) In t h i s r e g a r d Colonel Auld s a i d t h a t Research Group No,2 of th e I n s t i t u t e of Petroleum w ill be reviewing a l l the cu rren t i n f o r m a t i o n on t h i s pro bl em and he a g r e e d t o make t h e G r ou p' s re p o rt a v a ila b le to the Committee: he thought th is review would be a v a ila b le by the end of 1954. i i i ) I t was n o t ed t h a t P r o f e s s o r D.H. Hey of K i n g 's C o l l e g e , uor.don, who had e x p r e s s e d i n t e r e s t i n t h e probl em a nd who had # diesel engines available in his laboratory for experimental work, should be invited to attend the meeting at which the above rep o rt would he d iscu ssed . iv) P ro fe s s o r Morton s a i d f i a t ho a l s o had d i e s e l engines in his la b o ra to ry and th at he would be prepared t o use them under v a r y in g working c o n d i t i o n s so t h a t th e fumes produced under these conditions could be studied. v) I t was t h e g e n e r a l view o f face of i t , the question of diesel seemed l i k e l y t o repay s t u d y but approach should await study of the lik e ly to be available in tin e for Committee. the Committee t h a t , on the e x h u a s t s was one whi ch i t was f e l t t h a t t h e l i n e of review of e x is tin g knowledge the next meeting of the API 06026 p z i r c l e u :- :~-j . Auuusr 4 . 1554. ?&e e 2 3 . " CT3P. VEHICLE ZX'liL'51 CaLLZD IC? G:\C9 I.AV2E Stanford Research I n s t i t u t e has branded motor vehicle exhau.3t as a m a j o r smog c o n t r i b u t o r i n i t s r e p o r t s ummari zi ng s i x y e a r s of smog s t u d y . The I n s t i t u t e ' s 134-page r e p o r t , r e l e a s e d by Western Oil and Gas A s s n , , e s t i m a t e s 3 , 1 OG t o n s o f p o l l u t a n t s a r e e m i t t e d d a i l y i n t o t h e Los Angeles B a s i n a t m o s p h e re , of which 1,50 0 t o n s are hydrocarbons. Auto exhaust heads the l i s t with 1,016 tons, followed by: * Evaporation from automobile tanks and c a r b u r e to r s - 164 tons Refining - 155 tons * -Marketing - 37 t o n s P r o d u c t i o n - 32 t o n s P il l i n g automobile tanks - 28 tons P i l l i n g s e r v ic e s t a t i o n tanks - 24 tons In s h o rt, exhaust gases take the blame for 91$ of the t o t a l organic hydrocarbons emitted and nearly 95$ of the a c tiv e hydro carbons . The r e p o r t notes in p a s s i n g . t h a t backyard i n c i n e r a t o r s emit 400 t o n s of o r g a n i c m a t e r i a l s d a i l y . The r e s e a r c h e r s s a y t h e key t o smog p r o b a b l y i s o z o n e , s i n c e Los Angeles has t h e w o r l d ' s h i g h e s t c o n c e n t r a t i o n of t h a t o x id a n t, -Moreover, ozone c o n c e n t r a t i o n s i n c r e a s e and d i m i n i s h w i t h t h e i n t e n s i t y o f t h e smog. " T h i s ozone may r e a c t i n t h e a i r w i t h h a r m l e s s m a t e r i a l s such as hydrocarbons t o form troublesom e compounds t h a t a r e r e s p o n s i b l e r f o r many of t h e u n d e s i r a b l e e f f e c t s of smog" , t h e r e p o r t c o n c l u d e s . Copi es of t h e document a r e a v a i l a b l e from W e s t e r n Oil a nd Gas -issn., a t 10, W.eth S t . , Los Angeles, 14. ,Ia.iCuAii FZIRCLZU!' 1 3 / 3 . A u gu st 1 8 , 1 9 5 4 , AUTO."03 ILL EXHAUST LHIKED TO SMOG Page 2 0, Automobile exhausts are responsible for a considerable amount o f oz on e , t h e " s i l e n t p a r t n e r " i n Los A n g e l e s ' smog p r o b l e m , says Gordon F. Larson, d i r e c t o r of the A ir P o l l u t i o n Control D i s t r i c t Larson said three d i s t r i c t sc ie n tis ts have established that unburned gasoline and nitrogen oxides em itted by exhausts c r e a te ozone in the nresence of s u n l i g h t . Ozone, in t u r n , i s b e l i e v e d t o " t r i g g e r " other chemical re actio n s in the atmosphere, which causes smog's undesirable effects. So , L a r s o n e m p h a s i z e d , t h e d i s t r i c t now c a n make f o r ma l s c i e n t i f i c i n d i c t m e n t a g a i n s t e x h a u s t s as a smog c o n t r i b u t o r . As h e p u t s i t , "We a r e on t h e same f i r m s c i e n t i f i c g ro und i n demanding t h a t a u t o m o b i l e e x h a u s t s b e c l e a n e d up as wo we r e i n demanding two years ago th a t r e f i n e r i e s clean up." V a ri o us a g e n c i e s s t u d y i n g t h e smog p r ob l em a l r e a d y have a g r e e d t h a t ozone i s l i n k e d t o smog and i s p r e s e n t i n t h e Los Angeles atmosphere in excessive amounts. But th e d i s t r i c t r e p o r t is th e f i r s t f l a t statem ent t h a t the substance can be traced to vehicle exhaust gases. ---- o 0 o - - - API 06027 n t'" U' * cv.'CLt* ^ UNIVtHSI'H (H CINCINNATI-------- -- DEPARTMENT OF PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH TH ( UTTCRINO LAtORATORY c 0 tL C6C OF N ID IC IN C -- COIN AVCRUK CINCINNATI I t , OHIO N ovem ber 5, 1954 C A IU ADORMI KKTLAft TELEPHONE: CAPIT I s y , ii j! M r. D. V. Stroop, D irecto r D epartm ent of Technical Services A m erican P etroleum Institute 50 W est 50th S treet New Y ork C ity 20 I i D ear M r. Stroop: f I a m Bending you h e re w ith , fojr y o u r in fo rm a tio n , a s ta te m e n t of ex p en d itu res m ade on behalf of A m erican P e tro le u m In stitu te during the th ird q u a rte r of 1954. I believe you w ill find this statem en t se lf explanatory, but ii you have any questions o r com m ents, D octor Kehoe would a p p re c ia te your bringing th em to his atten tio n . V ery truly yours, ef Enc. E . R . F oar tla go e , P^ r^ccrelt a r y to D r. Kahoe ! I API 05998 UNIVERSITY OF CINCINNATI KETTERING LABORATORY ACCO UNT OF THE AMERICAN PETROLEUM INSTITUTE F O R T h i r d Q u a r t e r of 1954_________ SALARIES (Based on Proportion o f Tuna A ctually Spent on Project) D irect S a la r ie s ______________________________________________________ 4 B 6 2 . . - 7 . 9 _____ Indirect Salaries H ist o p s tb o lo g ic a l P reparation___________________________ 3 .4 8 .,,5 Q _____ O th e r S e r v ic e s .................................................................................. 5 2 6 . 6 . * J $ 5 . ____ 1 Q . 7 7 7 .64 MISCELLANEOUS EXPENSE P urchase o f A n im a ls.................................................................................................. 2 1 Q .* 3 Z . S p ecial L aboratory S u p p lies----------------------------------------------------------------- 8 4 P . T ravel _____________________________ _________________________ _______1 Q M , 4 Z . Overhead ( Proportion o f H eat, C as, E lectricity, Steam , T elephone, G eneral Laboratory S u p p lies, P o sta g e, A n n u ities, P en sion s, M a in ten a n ce, e t c ___ 3 .4 s .2 .1 ____ 5,581.25 Balance Available for Further W ork at End of.. Balance Due Kettering Laboratory at End of----------------------------- TO TAL 857.84 16,558.87 R eceipts ...J.uly...l95.4_________________ Sum Available ................. 56,125.00 56,982.84 Expenditures . . 5 i A - - Q u a f c e i i ~ 1 9 - 5 - 4 -- -- 16., 5 -5 8 .8 7 Balance Available for Further W ork at End o f.-- 3r.d Q u a r t e r -- 195.4- FORM l 0 Balance Due K ettering Laboratory a t End o f KET. LAB. 3M-- I |. |S 20,625.97 API 05999 r* Copy From D. V. Stroop to Members of Subcommittee on Carcinogenicity and Medical Advisory Committee Decetriber 1, 1951 ES80 LABBRATCRIES STANDARD OIL DEVELOPMENT COMPANY P.0. BOX 51, UNDER, N. J. November 19, 19?U Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street New York 20, New York Dear Mr. Stroop: I am attaching a copy of the minutes of the meeting of the Subcommittee on Carcinogenicity held at the Conrad Hilton Hotel in Chicago on September 10, 195^ These minutes were prepared by Allan E. Dooley of the TexaB Company and have been reviewed by Dr. Hine who was Acting Chairman at that meeting. Since I was not in attendance at the meeting, I am not in a position to offer any comments other than that Dr. Hine believes that they are a satisfactory representation of the proceedings at the meeting. I would suggest, therefore, that these minutes be duplicated and forwarded to each of the members of the Subcommittee on Carcinogenicity and the Medical Advisory Committee. Sincerely yours, f a f R. E. ECKARDT R. E. ECKARDT, M.D. REE:ilk Attachment API 06000 For Information Only Not for Publication AMERICAN FETROIEUM INSTITUTE SUBCOMMITTEE ON CARCINOGENICITY Minutes of Meeting September 10, 1954 Conrad Hilton Hotel Chicago, Illinois Members Present or Represented: C. H. E. W. L. C. Allan F. D. L. M. E. K. R. C. M. N. J. W. F. J. Hlne, M. D. - Acting Chairman Adams (by C. M. Loane) Beard Jr. E. Dooley Gassaway, M. D. _ Henderson ' Linder, M. D. MLthoff Nevquist, M. D. Osborn, M. D. Sanders (by W. E. Scovlll) Member Absent; R. E. Eckardt, M. D. Others Present: T. W. Albin J. W. Crookshank, M. D T. M. Frank, M. D. Paul D. Halley A. V. Horton R. M. London M. M. Marlslc Clark R. Miller, M. D. A. C. Pabst J. J. Fbair, M. D. B. B. Reeve, M. D. J. G. Ruddock, M. D. Shell Development Cities Service Pan American Refinery Standard oil, Indiana Kettering Laboratory Gulf Oil Pure Oil Tidewater Assoc. Oil Socony-Vacuum Kettering Laboratory Standard Oil, Indiana Richfield Oil The minutes of the Subcommittee meeting of June 19, 1954 were approved as circulated. It was the consensus of the Subcommittee that the last sentence of third paragraph in Section 1, (Chemical and Biological Work) of Dr. Eckardt's report of June 21, 1954 to the Medical Advisory Committee should read "Selected por tions of these investigations will undoubtedly be prepared as a manuscript for publication." API 06001 1 It - 2 - Dr. Horton discussed his progress report, dated September 1, 195k copies of which he had mailed to the Subcommittee at the end of August. In Dr. Horton's discussion be has brought out the following points: 1. Future work will be directed toward solidifying the qualitative findings into a good analytical tool. 2. With respect to accelerating materials, octane has been found to be a good accelerator; normal octane is not an accelerator; it appears that a C^g chain length is about the upper limit of accelerators. 3 . In uncraeked oils (paraffin distillates 650-725) the carcinogen has not been identified but it is not benzpyrene. The cholanthrene molecule may be important in low-boiling carcinogens. 4. No work is being done on inhibitors. 5. No washing experiments are being done; Dr. Horton believing that nothing more is needed in this phase of the study. 6. Six or seven oil company laboratories which are testing the analytical method for benz (a) pyrene developed by Tye, Oraf and Horton have been obtaining satisfactory reproducibility. It was voted to receive Dr. Horton's report. Dr. Phair reported on the epidemiological phases of the research project and brought out the following points: 1. Case reporting is getting better, 289 case records, most of them current ones, having been received in the past year. 2. Fifteen companies are now participating with an additional one to start shortly. 3* More detailed population figures will be needed and Dr. Pbair will write to the various medical directors advising them in detail what figures are needed. Dr. Phair stated that one oil company had mode records of skin examin ations of all its employes and that a second company had key-punched all its periodic physical examination findings for the past 10 years. He stated that an analysis of these records might be worhtwhile and asked approval of the Subcommittee to go ahead. This analysis 'would require hiring a clerk for a few weeks to copy the records of the first company. The cost involved in the second company's records would be much less; merely the cost of having IBM punch a duplicate set of cards. Dr. Phair stated that it would not require any additional funds; he could handle the costs within his present budget. After a thorough discussion, the Subcommittee concluded that it could see no objection to Dr. Phair's obtaining the epidemiological information from these two conpanies within his present budget. API 06002 - 3 - Dr. Horton commented on the statement in the Subcommittee's report of June 21, 195^ which says that "expenditures (for the chemical-biological work) - - - - should stabilize at levels not in excess of $25,000 after June 30, 1956*" Dr. Horton felt that thi6 is too great a cut since the 1955-1956 budget allotted $52,250 for these phases of the research project. After considerable discussion the Subcommittee, in executive session voted that "It is the sense of the Subcommittee that it is their Intent at this time that the budget be reduced in the order of 10 per cent annually for the next few years." It was also voted that the next meeting of the Subcommittee would be in Buffalo, New York, 12 April 1955 one day prior to the meeting of the Medical Advisory Committee. API 06003 \ API 06004 MEDICAL RESEARCH DIVISION R O e E R T E. E C K A R D T . P H .D ,. M .D . DIRECTOR HO P'AC E W . G E R A R D E P H .D . M .D HEAD TO XIC OLO G IST November 19, 19$k Mr. D. V. Stroop American Petroleum Institute t $0 West $Oth Street _ New Tork 20, New Tork " ; Dear Mr. Stroops I am attaching a copy of the minutes of the meeting of the Subcommittee on Carcinogenicity held at the Conrad Hilton Hotel in Chicago on September 10, 195U. These minutes were prepared by Allan E. Dooley of the Texas Coaqjany and have J been reviewed by Ef. Hine who was Acting Chairman at that I meeting. Since I was not in attendance at the meeting, I am not in ; a position to offer any coments other than that Ik*. Hine t believes that they are a satisfactory representation of the i proceedings at the meeting. I would suggest, therefore, t I that these minutes be duplicated and forwarded to each of r the members of the Subcommittee on Carcinogenicity*#* \ iK PffyjLWVMJL**- Sincerely yours, I. R. E. ECKARDT, M. D. REE:ilk Attachment r ( API 06005 L  r f . * , .. .. - ..ctl:w .,n rr.ar. ...`-s ~ - . >'" L . I I. Os:1 .- . It-. r.e; Jr rs -j'.- y", ;>uCfr: ~r. L in d e r, 0. Hithcff r o v p u i s t , ::. o . C s t o r r . , h . D. , J r." - r ('-y . 7. w c r : : '.; * .* ' -- e,V" ^ 1' 0 " -*vVh.-_v_--:-- J * ^_ l -rs ' r e s e n t : -- "iVJ J. ". Crookshar.V: ,IL.D. reul I. Holley H~rton ... T. X* :'..-risic 11crk Hiller,H.D. - * Ci.v _ L. J. J. Flair,li. D. ""0 -. P-eve ,/s 0, 9 -1 ***--' 1- n e .Jitlo: S :rvocs r -- :- U......... - *-* - * S t i h d ^ r d ^ * 1T) --**----5--*---~w.- I"j ^ t -"1.. T . 1 --- * Julf Col Pr^uj re C. i-lc wOccr.y-7 t>.-1 t,Vx*4"..>*'1r** *L - * b -*'* ** :ir.;tes of the Sul r.cooui t te as circu l a t e d . ..e J-ir.g - <- *-- - > - It v?s `he co n s e n sus of the ubco._ .it te ~ t:.-. A'-.s i T t Ird p a r a g r a p h in S e c t i o n 1, (Iht-r.ical =r.' 2 i o I .w iosl >ork) of j , - - 0 -- ec k eshrodutl1ds rr ee ^p do r t" S eolfe cJtuneed p21o,r t i1o9n5sU otof tthke-n*.le1c-ivcalr A i t i : 1 1 b t e d l y te p r e p a r e d os a .-.ar.uscript for ; t li c . ti or.." 1"-. H o r:t:ocnh 'J..ie : uds ercecsilneo:s1 otor o gthree s sS_h. :*.r.. ort1,j. 4. , :i ; V ~ ' r* r:e w . t 1 <r - I - ^ T the er.i of not, e 0 A API 06006 t ftuboeaartttee OB C arcin o g e n icity *2 K in u te a I 2 W ith respoet to tooo lo ra tin g a t a r ia lt , ootano haa boo found to bo a good aeeo lo rato r] normal ootano I s not an a e co lo ra to rj i t appears th a t a 0, ohain len g th la about tho upper lim it of acce le ra to rs* 3 Ib mioracked o lla (p a ra ffin d ia t illa t o a 650* -72$*) tho aarolnogoe haa not boon id o n tifio d but i t ia n o t bcnspyreae* Tho oholanthroao nolooulo a j bo im portant in Io n -b o ilin g carcinogens It* Wo work la boing dono on in h ib tora* 5 - lo washing exprin an to aro boing dono; Dr* H o rto n S bol i o Tin g th at nothing a o rt l a aoodod 1 b th la phaao o f tho nftBdpw*: 6* t in or seven o i l aompany la b o ra to rio a whioh aro tooting tho a n a ly t ic a l method fo r bona (a ) pjrono developed by Tye, O raf and Borton have boon o b tain in g a a tla fa e to ry ib i XIt j Z t waa votod to re ce iv e Dr'* Io rte n * s re p o rt. Dr* P h a ir roportod on tho opldoadologioal phaaoa of tho roaoaroh p ro je ct and brought oat tho fo llo w ing p o ia ta t 1* Caao rep o rtin g la g e ttin g betto r* 289 oaao records* oat of then w arrant ones* h arin g boos received in the paat year* 2 * P ifto o a ownponioa are now p a r tic ip a tin g w ith an ad* d it io n a l one to at a r t sh o rtly 3* More d e ta ile d pop ulation flgnzw a w ill bo aoodod and Dr* P h a ir w ill w rite to tho vario u s n o d ic a l dirootora r a d visin g thaai in d e t a il what fig u re are aoodod Dr* P h a ir ata tod th a t one o i l company had ado rooorda o f a k in exam inations o f a l l it a a p lo y e s and th at a aoeond oonpany had koy*punohod a l l it a p e rio d ic p h y s ic a l o x an iaatian fin d in g a fo r tho paat 10 yoara lo a ta tod th a t an a n a ly s is o f theao rooorda n ig h t bo worthwhile and aeked approval o f the ftubooonittoo to go ahead Thia a n a ly a la would re q u ire h ir in g a c le r k fo r a few wooka to copy tho rooorda of tho f ir a t oonpany Tho co at in vo lved in tho aoeond oonpany* a rooorda would bo aueh le a at n o ro ly tho co at of h arin g IBJf punch a d u p lica te a c t o f earda* Dr P h a ir atate d th a t i t would not re q u ire any a d d itio n a l funda) ho oould handle tho acota w ith in h is present budget A fte r a thorough d iscu ssio n * tho Buboonalttoo concluded th at i t oould see no o b je ctio n to Dr* P h air*a o b tain in g tho ep id em io lo g ical Inform ation from those two o a n lo s w ith in h is present budget 06007 SubeaMl ttee on C a ro lM g o n le lty - 3 K laut Dr* H orton eoamentad on th * s ta t a n t In tb Subcommittee r p o rt o f Jun 21, 195k which l a j i th a t "expenditure (fo r th e h B a ic a l-b io lo g i a l work) - - - - should s ta b lilM a t l v ls n o t in excess o f |25>000 a f t e r June 30, 1956.* Dr* E o rto n f e l t t h a t th is is too g re a t a out sino th 1955-1956 budget a llo tte d #52,250 \ fo r th*s phase of th reaeareh project* A fter onalderabl disuasion tha S ubeoooittee, in exeoutive aion voted th a t * I t is th n s o f th S u b o o salttee th a t i t 1 t h e i r in te n t a t th is tisM th a t th budget be rduod in th order o f 10 p r at annually fo r th nxt few y ear .* Z t was a lso ro td th a t th n x t M a tin g o f th S u b o o u itte e would b i n B u ffa lo , lew Y o rk, 12 A p r il 1955 on day p r io r to th* M tln g of th K d i a l A dvisory C o M ltt* >r API 06008 n ia : MAC Corrapood * 1 s (- o* \ V- " - / Monter 2k, 195* fir. B. E. Beksrit Standard O il fiartlupaatti Ocm/aaf Madical Itaaaarcfi U r ia l P. 0. loa n Lindan, lav Jaraay Xaar latrar of Mara ar 19tfi tea team rocolToi. ut liiimam n oa C a n la a tn la lty tea attaada* tea a n tin g te U la Chleaaa Baptateaa IM fi, Z aaatar i f it waaU te daain fila to m at coplat of tha al aataa to a ll aatean af tha tediati H f t i a ri Caaalttaa. Vary t n ly paar, 0V8ta4 i API 06009 ESSO L A B O R A T O R IE S STANDARD OIL DEVELOPMENT COMPANY P. O. BOX 51, L IN D E N , N .J . medical research division R O flE R T HORACE E. E C K A R O T . P H .D .. M .D . DIRECTOR W . G E R A R D E . P H .D .. M .D . HEAD TOXICOLOGIST November 30, 195k IN D U S TR IA L HYGIENE SECTION N A T H A N V. H E N D R IC K S . CH .E . HEAD OF SECTION F R A N K L IN W. C H U R C H . M .S. G EO R G E M . W IL K E N IN G . M .S . INDUSTRIAL HYGIENISTS Mr. D. 7. Stroop American Petroleum Institute 50 West 50th Street New York 20, New York Dear Mr. Stroop: In answer to your letter of November 2Uth, I am certainly in agreement with sending copies of the minutes of the Subcoanittee on Carcinogenicity meeting to all members of the API Medical Advisory Committee. Very truly yours, REE:ilk R. E. ECKARDT, M. D API 06010  Copy From D. V. Stroop For Information of Members of Medical Advisory Committee December lk, 1954 ESSO LABORATORIES STANDARD OIL DEVELOPMENT COMPANY P.O. BOX 51, UHDEN, H, J. November 22, 195k i Mr. D. V. Stroop American Petroleum Institute 50 West 50th Street Hev York 20, Hew York Dear Mr. Stroop: I I am attaching some minutes received from the Institute of Petroleum's Research Group Ho. 2 meeting of September 29, 195k. In viev of the discussion in paragraph 3 of these minutes, I believe it vould be advisable to have these minutes reproduced and distributed to all of the members of I the Medical Advisory Conedttee. Very truly yours, /s/ R. E. ECKARDT R. E. ECKAROT, M.D. REE:ilk Enel. - Minutes of Meeting Ho. Ik API 06011 1