Document NeJ52JQY32ROvjqN1d2oOQR98

FILE NAME: Oil Industry and American Petroleum Institute (API) DATE: 1953 Jan-May DOC#: API030 DOCUMENT DESCRIPTION: Letters, Memos, Reports, Financial Reports & Other Relevant Documents from Jan-May, 1953 - 2- 2 P ( m c - ') The m inutes of th e F o u rth M eeting o f th e lfnil 1ri n1 A dvisory Committee were approved. Subsequently the m eeting follow ed the agenda which had been prepared by Dr. Newquist p rio r to the m eeting w ith s lig h t m od ificatio n in th at the progress re p o rt and the r e s e a r c h a c t i v i t i e s f o r th e y e a r b e g in n in g 7/ 1/53 were combined in both Dr. H o rto n 's and Dr. P h a ir 's r e p o r ts . ITEMS 2 AND U - DR. HORTON'S REPORT As th e b a s is f o r h i s d is c u s s io n s , Dr. H orton u sed th e (I a tta c h e d Memorandum from th e K e tte r in g L a b o r a to r y - F if th M eeting of 1 th e A dvisory Committee API R e se a rc h P r o j e c t MC-1 d a te d Jan u ary 16, 1953* C o n sid e ra b le d is o u s a io n was g iv e n to th e opening two p a ra graphs which ju s tif ie d the use of mice in a p p raisin g the hazards of sk in can cer in workmen. In response to a s p e c if ic q u e stio n from D r. Adams, D r. H orton s t a t e d t h a t w ashing is more e f f e c t i v e in reducing carcin o g en ic potenoy than a re d u c tio n in the dosage of c a rc in o g e n a p p lie d . As ev id e n ce he p re s e n te d th e e x p e rim e n ta l evidence w hich was o b ta in e d w ith washing experim ents w ith API-8 and 1 API-113. I t was th e n su g g e ste d t h a t c o n s id e ra tio n be g iv en to a washing experim ent w ith API-113 end API-57 in which th e se o ils would be p a in te d on a t a l e v e l o f 100 mg. two tim es a week and washed o f f a fte r 9 hours in contact w ith the akin. Perhaps the g re a te s t in te re s ts in Dr. H orton's re p o rt are his observations or th eo ries th a t the potency of a p a rtic u la r m a te ria l is dependent le s s on the amount of carcinogen than on i t s concentration; whereas, the e ffe c t of a ccelerato rs is dependent less on the c o n c e n tra tio n than on the t o t a l amount a p p lie d a t each a p p li L_c a t i o n . A lthough th is i s an i n te r e s tin g th o u g h t, i t appears somewhat API 05532 -3 d i f f i c u l t to u n d e rs ta n d how t h i s c o u ld be th e s i t u a t i o n and how th e proper d e fin itiv e experim ents could be conducted in order to con c lu s iv e ly e s t a b l i s h th e s e o b s e r v a tio n s . The co ncept t h a t a c c e le r a to rs may a c t b ecau se o f t h e i r s i m i l a r i t y i n s t r u c t u r a l c o n f ig u r a tio n w ith the carcin o g en ic agents is an in te r e s tin g one in s o fa r as i t applies to benzo(c)phenanthrene and h ig h ly branched dodecylbenzene. I t is d i f f i c u l t , how ever, to see how t h i s co n cep t co u ld a p p ly to compounds l i k e 3#^ -b en zp y ren e o r to dodecylbenzene in which th e side chain is not highly branched. The form ula p re s e n te d by Dr. Horton on page 9 o f h is re p o rt is an extrem ely in te re s tin g form ula from a th e o re tic a l standpoint, but A e th e r i t w ill ever have any p ra c tic a l ap p lica tio n seems h ig h ly p ro b le m a tic a l sin c e the d e te rm in a tio n of the e f f e c tiv e c o n c e n tra tio n o f a c c e l e r a to r in an o i l may be ex tre m e ly d i f f i c u l t , p a r tic u la r ly i f th e re is more th an one a c c e le ra to r p re s e n t in such an o i l . S im ila rly , suoh a form ula allow s fo r th e presence of only two c a rc in o g e n s , nam ely, benzpyrene and o"8 Cl^ a ro m a tic . I t seems h ig h ly u n lik e ly th a t th e re would be only two carcin o g en s p re s e n t in as complex a m ix tu re as s l u r r y o i l , and t h i s is p a r t i a l l y borne out by the observation re p o rte d by the Standard O il Development Company in whioh 6- i s o p r o p y l b en zan th ra ce n e was determ in ed in a r e l a ti v e s t a te o f p u r ity from one of th ese o i l s . I t th e re fo re seems lik e ly th a t th is form ula w ill be exceedingly com plicated by a g rea t number o f a c c e le ra to rs and in h ib ito r s , such th a t the e ff e c tiv e con c en tra tio n of a c c e le ra to r w ill have to be determ ined fo r each in d iv id u a l o i l , and a ls o by the presence of a la rg e number of carcin o g en ic compounds, which w ill h ig h ly com plicate the u se fu ln e ss of any such equation. A n oth er i n t e r e s t i n g o b s e r v a t io n made by Dr. H orton d u rin g API 05533 -il- h is d is c u s s io n was t h a t th e average l a t e n t p e rio d d ecreases more ra p id ly fo r pap illo m as, depending upon the amount of a c c e le ra to r, th an the average l a t e n t p e rio d f o r c arcin o m as. T h is may be g ra p h ic a lly ex p ressed somewhat as fo llo w s: ITEMS 2 AND lj - DR. FHAIR'S REPORT Dr. Fhair used as a b a sis of h is rep o rt the attach ed memorandum, which i s e s s e n t i a l l y a t a b u l a r summary of th e number of c a se s which he has th u s f a r c o lle c te d from the v a rio u s companies. I t appears th a t an in c re a sin g number of companies are p a rtic ip a tin g in re p o rtin g cases to Dr. Fhair b u t th a t th ere is a heavy imponderanoe o f cases being re p o rte d from companies along the e a s te rn seaboard as compared w ith companies in southw estern U nited S ta te s . I t was agreed th a t Dr. F h air would o u tlin e b r i e f l y and send to the Chairman of th e Subcommittee on C arcin o g en icity h is l a t e s t th o u g h ts on p e r t i n e n t d a twa /hniceh w ishes t o have in c lu d e d in th e c u rre n t r e p o r t i n g o f c a n c e r c a se s among employes in th e p e tro le u m in d u stry , and th a t th e Chairman of th e Subcommittee on C a rcin o g en icity would th en re a tte m p t to stim u la te a l l m edical d ire c to rs to re p o rt cu rren t cases to Dr. F h air. API 05534 -5 - ITEIM if - DR* KEHOE'S REPORT Dr. Kehoe d isc u sse d th e b u d g etary a llo c a tio n s o f funds and In d ic a te d th a t i t would be p o s s ib le fo r the K e tte rin g Labora to ry to l i v e w ith in th e p r e s e n t l y a s s ig n e d b udget of $79, 000, b u t th a t i t must be remembered th a t th is w ill r e s u lt in approxim ately o n e-q u arter le s s work being done during the year Ju ly 1, 1953June 30, 19514-. Dr. Kehoe w ished to r e s e rv e the r i g h t t o a ll o c a t e funds under any of the headings given, depending upon the K e tte rin g L a b o ra to ry 's e s tim a tio n o f how th e funds sh o u ld be a llo c a te d . Thus, fo r in sta n c e , i f f u l l co operation w ith Dr. P hair is not o b ta in e d , i t may w e ll be t h a t th e $17,500 a ll o c a t e d f o r e p id e m io lo g ic a l work may be r e - a l l o c a t e d in to o th e r p h ases o f th e R esearch P r o j e c t . There seems to be a g e n e ra l agreem ent among th e members o f th e com m ittee t h a t Dr. Kehoe sh o u ld have freedom in allo catin g the to ta l funds to any of the four phases o f the p ro je c t th a t he f e e ls th e funds should be a llo c a te d to . Dr. Kehoe a lso made a p le a t h a t i f t h e r e i s a d ete rm in e d e f f o r t t o have a g ra d u a l re d u c tio n in th e budget of t h is p r o je c t during th e coming y e a rs , he would lik e to be inform ed of t h i s as f a r i n advance as p o s s ib le so th a t he m ight make h i s p la n s a c c o rd in g ly . Thus, i f th e budget i s to be cut o n e -h a lf in the year Ju ly 1, 195if-June 30, 1955, and to a q u a rte r o f i t s p re s e n t f ig u r e in th e succeeding y e a r s , Dr. Kehoe would l ik e to know o f t h i s as soon as p o s s ib l e . ITEM 5 Dr. Horton d iscu ssed th e re p o rt from the B r itis h on the c a r c in o g e n ic ity o f th e m in e ra l o i l s . He f e l t t h a t t h e i r t e s t s had been perform ed w ith a p p lic a tio n s of o i l w hich were so small t h a t th ey are not g e ttin g tumors in the m ice, y e t th ey were observing tumors in -6- th e r a b b its due to th e e f f e c t s o f a c c e l e r a t o r s . He seemed to f e e l t h a t the f r a c t i o n a t i o n of t h e i r o i l s l e f t much to be d e s ir e d , and th a t sane f r a c tio n a tio n of o ils in h is la b o r a to r ie s , which were th en a p p lie d on h i s own anim als by h i s own te c h n iq u e s , had r e s u l te d in the appearance of a co n sid erab le number of tum ors, p a r tic u la r ly in the K-U* L-ij- and 0--U- f r a c t i o n s . He f e l t t h a t t h i s was p ro b a b ly the re s u lt p rin c ip a lly of a c c e le ra to rs, and th a t the carcinogens would be found in the h ig h e s t c o n c e n tr a tio n s in th e K-5 L-5 and 0-5 re s id u e s . I t was su g g ested a t th e m eeting th a t the workers a t the K e tte rin g L aboratory f e e l fre e to communicate on a p ro fe s s io n a l b a sis w ith the workers in England in d iscu ssin g th e ir m utual work, and furtherm ore, i t was f e l t th a t i f th e K e tte rin g L aboratory f e l t they would lik e to keep th is comm unication in the e s ta b lis h e d channels, th is could be done by having Dr. Horton prepare a c ritiq u e of the B r i ti s h work and th en having Dr. Hewquist forw ard t h is to the B ritis h workers through Colonel Auld. I t is evident th a t i t is too e a rly to draw co n clu sio n s w ith re s p e c t to the B r itis h work which has been rep o rted thus fa r. ITEM 6 Dr. Kehoe in d ic a te d t h a t he h as w r i t te n a su p p lem en tal l e t t e r to D r. K otin su g g e stin g th a t Dr. K otin more d e f in ite ly o u t lin e what in fo rm atio n he would lik e to re c e iv e in ord er th a t the K e tte rin g L ab o rato ry m ight be a b le to p ro v id e i t to him. Dr. Kehoe did not fe e l th a t any of the K etterin g Laboratory rep o rts should be sent to Dr. K otin o r anyone e lse sin ce they are a t the p re se n t time so t e n t a t i v e . I t was a lso su g g e ste d t h a t a p o s s ib le means of h a n d lin g t h i s s i t u a t i o n would be to have Dr, K o tin come in to th e K ettering L aboratory and discu ss th e ir m utual problems v e rb a lly , and that such a d isc u ssio n could get to Dr. Kotin a l l the inform ation API 05536 -7- which he m ight need and y e t not je o p a r d iz e th e p o s it io n o f th e K ettering Laboratory. ITEM 7 Dr. E ck a rd t r e p o r te d to the com m ittee t h a t 5 a d d itio n a l cases of skin cancer r e s u ltin g from c u ttin g o ils have re c e n tly come to h is a t t e n t i o n . One o f th e s e c a s e s was a s c r o t a l c a n c e r and none of the cases had p re v io u s ly been re p o rte d to the group. This simply served to emphasize the growing im portance of c u ttin g o ils as a p o ssib le carcinogenic agent in in d u stry . ITEM 8 Some d is c u s s io n was g iv en t o th e p o s s i b i l i t y o f h o ld in g the next m eeting o f the A dvisory Committee a t th e H otel Shamrock on the 29th o f Septem ber, 1953 end t h a t a t t h i s tim e i t would be a jo in t m eeting w ith the Subcommittee on C arcin o g en icity . D ecision on th is m a tte r was l e f t to th e Chairman of th e Subcom m ittee, w ith the recommendation th a t i f the m eeting is to be h e ld on th a t d ate, members sh o u ld be n o t i f i e d and h o t e l r e s e r v a tio n sh o u ld be made well in advance. M. N. N ew quist, M. D ., Chairman R. E. E c k a rd t, M. D. Acting S ecretary API 05537 MEMORANDUM FROM THE KETTERING LABORATORY 1 FIFTH MEETING OF THE ADVISORY COMMITTEE API RESEARCH PROJECT MC-I January 16, 1953 A. The use o f mice in th e e x p e rim e n ta l a p p r a is a l o f th e r e l a t i v e h azard s o f c a n c e r of th e s k in o f workmen from exposure to various petroleum products, 1, M aterials which in In d u s tr ia l experience have caused cancer o f th e s k in ,of workmen have been shown to be c a rc in o g e n ic to m ice, 2, The c o r r e l a t i o n o f th e re sp o n se s o f men and mice i s p a r t i c u l a r l y c lo s e i n th e case of s t r a i g h t ru n wax d i s t i l l a t e s , cancers having re s u lte d in both species from in te rm itte n t, sev ere exposure (100 mg, doses a p p lie d th re e tim es p e r week on th e s k in o f th e m ice) when th e c arcin o g e n ic m a te r ia l was n o t removed by w ashing. Data re p o rte d by th e New York U h iv e rs ity group i n d ic a t e t h a t mice develop v e ry few tum ors when low dosages (15 mg, p e r a p p lic a tio n ) o f wax d i s t i l l a t e s a re a p p lie d . The mouse, lik e man, seems to be a f f e c t e d a d v e rs e ly by th e s e o i l s o nly when exposure i s se v e re , A co n firm ato ry t e s t a t low dosage w i l l be c a r r i e d o u t on C3H mice w ith th e M idcontinen t p a r a f f i n d i s t i l l a t e , API-56 ( P 0, 1 a t th e 100 mg, lev el of dosage), B, R e la tio n s h ip s betw een p o te n c y of o i l s and th e l e v e l o f dosage ap p lied upon the skin per a p p lic a tio n , 1, V a r ia tio n of th e r e l a t i v e p o te n c y , PMC, w ith th e le v e l o f dosage (over c e r t a i n ran g e s o f dosage) i s J u s t what one API 05538 - 2- would e x p e c t on th e b a s is o f human e x p e rie n c e w ith such alleged carcinogens as coal ta r , shale o il or the d i s t i l l a t e s which have been h a n d led i n wax p r e s s e s in th e petroleum in d u stry , i . e . , the higher the le v e l of exposure, th e g r e a t e r th e chance of s k in c a n c e r, The r e s u l t s o f th e t e s t s on mice a t d i f f e r e n t le v e ls of dosage which have been rep o rted fo r the c a ta ly tic a lly cracked resid u a, API-8 and 71 a re th e r e f o r e n e i t h e r s u r p r i s i n g n o r d i s t r e s s i n g when c o n sid e re d from th e o v e r a l l a s p e c t o f th e program . R e a lly unexpected r e s u l ts are coming from the t e s t s on sta n d a rd s o lu tio n s and r e f in e r y stream s c o n ta in in g e ith e r very low or very h ig h le v e ls of c o n c e n tra tio n of accelerating constituents. C ertain m aterials dem onstrate a constant potency over a broad range of dosage: (1) S olutions of m ethylcholanthrene or benzpyrene in benzene (2) Some b len d e d i n d u s t r i a l f u e l o i l s , e , g . , API-113 (3) Some th e rm a l t a r s from v i r g i n f e e d s , e . g , , API-108 (Ij.) S o lu tio n s o f m e th y lc h o la n th re n e o r benzpyrene in dodeeylbenzene (5) Some c a t a l y t i c a l l y c ra ck e d gas o i l s , e . g , , A PI-102, Types (1 ), (2 ), and (3) probably c o n ta in , a t the m ost, very low c o n c e n tra tio n s of a c c e le ra tin g c o n s titu e n ts , API 05539 - 3- ty p e s (4.) and (5) r e l a t i v e l y h ig h c o n c e n tr a tio n s . The types of o ils discussed in se ctio n B .l, apparently have a c r i t i c a l l e v e l of c o n c e n tr a tio n of such com ponents. The s i t u a t i o n may b e s t be p i c tu r e d g r a p h ic a lly (see F ig u re 1 ): Figure 1 Thus, i f no a c c e le ra to rs are p re s e n t in an o i l (A), i t w ill e x h ib it only the potency due to i t s carcinogen content. S im ila r ly , o i l s c o n ta in in g a c c e l e r a to r s w i l l show only th e ir base le v e l of potency i f the dosage per ap p lica tio n is k ep t s u f f i c i e n tl y low so th a t the amount of the API 05540 - ^- a c c e le r a to r a p p lie d to th e sk in o f th e mouse each time does n o t exceed B. What dosage t h i s w i l l be f o r any g iv en o i l o b v io u sly depends on th e c o n c e n tra tio n of a c c e le r a to r s in the o il. I f the dosage per a p p lica tio n of such o ils is in creased u n t il the amount of the a c c e le ra tin g c o n s titu e n t a c tu a lly being ap p lied each time reaches the c r i t i c a l range between B and C, th e r e l a t i v e p o te n c y , as measured by a shortened la te n t p erio d fo r tumor in d u ctio n , w ill r is e . This e ff e c t w ill then Increase as h ig h er le v e ls of dosage a re t e s t e d u n t i l th e maximum re sp o n se f o r a g iv en concen t r a t i o n o f c a rc in o g e n i s o b ta in e d (upper p la te a u in F ig u re 1). P ro ceed in g in th e r e v e r s e d i r e c t i o n (from D tow ards C) i t w ill be apparent th a t the higher the le v e l of the con c e n tra tio n o f a c c e le ra to rs in an o i l , th e more the dosage p e r a p p li c a t io n may be red u c ed b e fo re a s i g n i f i c a n t red u c tio n in potency w ill be observed. This category probably in clu d es a la rg e number of the c u rre n t d i s t i l l a t e gas o i l s w ith End B o ilin g P o in ts > 7 0 0 eF. from f r a c tio n a to r s of c a ta ly tic cracking u n its . Because of the obvious importance of these re la tio n sh ip s between the amount of a c c e le ra to r ap p lied and the r a te a t which an o i l co n tain in g carcinogens w ill induce cancer in exposed anim als, step s are being taken to e sta b lis h : API 05541 - 5- (1) the ex act magnitude of le v e ls B and C (w ith blends of synthetic hydrocarbons), (2) the c la sse s and b o ilin g ranges of the hydro carbons in c a ta ly tic a lly cracked gas o ils and resid u a resp o n sib le fo r the a c c e le ra tio n observed, and (3 ) the le v e ls of co ncentration of such hydrocarbons in various refin e ry stream s. C, The n a tu re o f th e compounds c o n tr ib u tin g to th e c a rc in o g e n ic potencies of refin ery stream s. 1, P o lycyclic carcin o g en ic compounds. a. 3 .U -B e n zp y re n e A d i r e c t method o f a n a ly s is f o r the 5 -r ingd c a rc in ogen, 3 ,i|-benzpyrene, has been developed. I t s r e l i a b i l i t y has been te s te d and confirm ed w ith four of the major types of re fin e ry stream s under consideration, including cycle gas o ils and bottom s products from c a ta ly tic cracking, ta r s from therm al cracking of c a ta ly tic fee d s, and stra ig h t run d is tilla te s . By a p p li c a t io n o f t h i s m ethod, i t h as been shown t h a t about 70% o f th e base p o ten c y e x h ib ite d by th e FCC decanted o i l , A PI-8, a t -20 mg. le v e ls of dosage i s API 05542 c o n tr ib u te d by b en zp y ren e. A pproxim ately l0% o f th e c o n s ta n t p j^ v a lu e of th e No. I4. f u e l o i l b le n d , API-113, is due to benzpyrene. b. As m ight be e x p e c te d , i n view o f th e r e p o r te d b o ilin g p o in t of 3,l;-benzpyrene (860F.) , i t has been found th a t the d i s t il l a t e re fin e ry stream s analyzed thus f a r , with true end b o ilin g p oints le ss than 860, contain re la tiv e ly sm a ll amounts o f t h i s 5- r in g e d c a rc in o g e n . The a v a ila b le data from b io lo g ic a l te s ts on fra c tio n s prepared from c a t a l y t i c a l l y cra ck e d o i l s s u g g e s t s tr o n g ly t h a t th e Im portant carcinogen in such cycle gas o ils is a l;-ringed d e riv a tiv e of phenanthrene. Since the re s u lts fu rth e r in d ic a te th a t sim ple a lk y l d e riv a tiv e s of pyrene and 1 , 2-benzanthracene API 05543 - 7- are not c o n trib u tin g s ig n ific a n tly to the carcin o g en icity of these o ils , a tte n tio n is being centered upon benzo(c)- phenanthrene. This is the hydrocarbon of low est m olecular w eight which h as been shown to be c a rc in o g e n ic w ith o u t the b e n efit of m ethylation. Pure samples of benzpheuanthrene and of each of i t s s ix monomethyl d e riv a tiv e s have been o b tain ed . T heir chem ical and p h y sic a l p ro p e rtie s a re being examined in the development of a d ir e c t method of a n a ly sis ap p licab le to refin e ry stream s. 2. A ccelerating co n stitu en ts^" Lj'J's't*. 'Jo o ,t I t i s of c o n sid e ra b le i n t e r e s t th a t the number of carbon atoms ( 18 ) in th e a c c e le r a to r , dodecylbenzene, i s th e same as th e number in th e c a rc in o g e n , b e n z o (c )- p h e n an th ren e The known c a rc in o g e n ic hydrocarbons have Carbon Numbers from 16 to The approxim ate b o ilin g range of branched-chain alkylbenzenes w ith 16-25 carbon atoms i s li.75- 700F,, which i s th e v e ry ran g e t h a t has been suspected of having a c c e le ra tin g a c tiv ity from the r e s u lts of b io lo g ic a l t e s ts on re fin e ry stream s and f r a c t i o n s t h e r e o f . I t seems l i k e l y t h a t we have h e re som ething more th an j u s t co in c id e n ce . The problem o f th e i d e n t i f i c a t i o n of th e im p o rtan t a c c e le ra to rs in re fin e ry stream s Is being approached In two ways: (1) B io lo g ic a l te s tin g of c e rta in sy n th e tic liq u id hydrocarbons as so lv en ts fo r benzpyrene, to determine API 05544 - 8 - which ty p e s have a c c e l e r a ti n g p r o p e r t i e s . The r e s u l t s obtained in the p rosecution of th is phase of the program w ill fu rn ish a d e fin ite o u tlin e of the range of m olecular w eights and of the s tru c tu ra l types of possible im portance. (2) B io lo g ic a l te s tin g of fra c tio n s of c e rta in c a ta ly tic a lly cracked o ils which have dem onstrated ( in t e s t s a t th e 100 mg* l e v e l o f dosage) p o te n c ie s th re e to te n tim es as h ig h as were expected on the b a sis of th e ir content of four to six ringed arom atic hydrocarbons. These e ff o r ts are being concentrated on th e components b o ilin g below 700F. which c o n ta in n o t more th an two fu se d aro m atic r in g s . Die foregoing phase (2) of this program has already led to the concentration of a colorless, liquid fraction from the TCC bottoms product, API-71 which possesses the specific irritant properties of dodecylbenzene. This fraction is being redistilled through an 80-plate column to prepare a number of well-defined cuts to be tested as possible accel erators of the carcinogenic action of benzpyrene. In g e n e r a l, i t i s a much sim p le r m a tte r to sep arate sa tu ra te d hydrocarbons and monocyclic aro m atics from o ils and from each o th er than i t is to f r a c tio n a te any sin g le c la s s o f p o ly c y c lic compounds from an o i l . Furtherm ore, in th e se p a ra tio n of compounds b o i li n g i n th e ran g e 500- 650 a t 760 mm., API 05545 - 9 - h ig h ly e f f i c i e n t d i s t i l l a t i o n s may be c a r r i e d out a t reduced p re s s u re w ith o u t f e a r of c ra c k in g . Hence there is l i t t l e reason to doubt th a t system atic a p p lic a tio n o f a v a ila b le techniques w ill lead to an adequate knowledge of the s tr u c tu r a l types of the acceleratin g c o n stitu en ts of re fin e ry stream s in a reaso n ab le p e rio d of tim e. The s te p from th a t knowledge to a d ir e c t method of a n a ly sis fo r th e ir e ffe c tiv e co n cen tratio n in any given stream should present no g rea t d if f ic u ltie s . 3, Physicochem ical methods f o r p re d ic tin g the r e la tiv e p o ten cies of refin ery stream s. During the l a s t s ix months, the com pletion of the p ic tu re (F igure 1) o f the in te r r e la tio n s between concen tra tio n of carcinogen, concentration of accelerato r, dosage p e r a p p l i c a t i o n , and r e l a t i v e p o te n c y . PMC, has brought with i t a f in s confidence th a t a re lia b le ana ly tic a l to o l w ill r e s u lt from the fu rth e r extension of t h i s fundam ental approach to th e problem . The e q u a tio n f o r which param eters are being determ ined i s of the f o l lowing type: ^MC s * where f ( c a *d) is a fu n c tio n of the amount of the a c c e l e r a to r component a p p lie d in each a p p lic a tio n of the whole o il. ca being the e ffe c tiv e co ncentration of acc elerato r in the o il. and d the to ta l dosage of o il per ap p lica tio n ; API 05546 - 10 - and a re the c o n c e n tra tio n s of the 5-rin g e d benzpyrene and of the lj.-ringed carcinogen re sp e c tiv e ly , while and are the re sp e c tiv e p o ten cies of benzpyrene and the ^ rin g ed compound based on m eth y lch o lan th ren e . Since th e v a lu e s f o r h a lf of th e f a c to r s in th^ e q u a tio n f o r c a l c u l a t i n g F|jC a re a lr e a d y known (d , Kcj and Ccj), th e re m a in in g Job i s th e d e te rm in a tio n o f c a , and the breakpoints a t B and C in the curve of Figure 1. These phases have been d iscu ssed in S ections Bi 2 i C . l . b . and C2 I API 05547 ESTIMATED BUDGET OP THE KETTERING LABORATORY FOR THE INVESTIGATION OP POTENTIAL HEALTH PROBLEMS OF THE PETROLEUM INDUSTRY ASSOCIATED WITH THE PRESENCE OF CARCINOGENIC COMPOUNDS IN CERTAIN MATERIALS AND PRODUCTS API RESEARCH PROJECT MC-1 Ju ly 1, 1953 - Juna 30, 1954 S alaries D irect S alaries $ 30,000.00 Indirect S alaries H isto p ath o lo g ical P reparation $ 2,000.00 Other Services $ 22,000.00 M iscellaneous Expense Purchase of Animals * 1 , 500.00 Special Laboratory Supplies * 5, 500,00 Travel $ 4, 000.00 Overhead (Proportion of heat, gas, e le c tric ity , steam , telephone, g en eral lab o rato ry supplies, postage, an n u ities, pensions, m aintenance, e tc .) $ 14 . 000,00 T otal $ 54, 000.00 $ 25, 000,00 $ 79*000.00 API 05548 r V ESTIMATED BUDGET OP THE KETTERING LABORATORY FOR THE INVESTIGATION OP POTENTIAL HEALTH PROBLEMS OP THE PETROLEUM INDUSTRY ASSOCIATED WITH THE PRESENCE OP CARCINOGENIC COMPOUNDS IN CERTAIN MATERIALS AND PRODUCTS i ! API RESEARCH PROJECT MC-1 J July 1, 1953 - June 30, 1954 I F urther development of se m i-q u a n tita tiv e b io lo g ic a l * te s tin g methods f o r m easuring the r e la tiv e carcinogenic potencies of petroleum products and fra c tio n s th e re o f $ 12,000.00 I I . D eterm ination of th re la tio n s h ip s between the ' ra p id ity of induction of bnign and m alignant tumors in experim ental anim als and v ario u s exposure fa c to rs such as frequency of a p p lica tio n , dosage per a p p lic a tio n , area of exposure, number of a p p lic a tio n s , s p e c ia l i r r i t a n t or toxic p ro p erties of the o il applied, e tc . $ 14,500.00 I I I . Development of ra p id a n a ly tic a l methods fo r estim ating the re la tiv e carcinogenic potency of any given r e fin e ry in term ed iate 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 o f compounds responsible fo r observed potency of various types of o ils and determ ination of p e rtin e n t p h ysical and chem ical p ro p e rtie s of these compounds $ 30,500,00 B. Development o f se m i-e m p iric a l methods o f $ 4,500.00 estim ation of carcinogenic potencies of o ils based on the chem ical o r p h y s ic a l p ro p e rtie s o f known o r s u s p e c te d c a rc in o g e n s and a c c e l e r a to r s IV, E pidem iological Program C o lle c tio n and c o r r e la tio n of d a ta on cases of c an cer among em ployees o f the p e tro leu m in d u s tr y including any sp e c ia l in v e s tig a tio n s in p a rtic u la r plants of the industry $ 17,500.00 _____________ T otal 7 9 /0 0 0 .0 0 API 05549 r e c e iv e d by t h e c e n t r a l r e g ist r y acco rding to s e x , r a c e and MALIGNANCY SEX RACE MALIGNANT BENIGN TOTAL I White 745 23 768 C olored 26 MALE Not Given 114 - 26 38 152 TOTAL 885 61 946 White 33 FEMALE Not Given 9 TOTAL 42 1 34 5 14 6 48 NOT GIVEh 4 t TOTAL BOTH SEXES 931 - 4 67 998 * Nine reports could not be used. Six w ere not tum ors; one of these was Boeck's Sarcoid. Three gave only the occupational history. API 05550 ) ) Type of Tumor Site Buccal C avity 00-09 D igest. S ys. and P erit. 10-19 R esp iratory System 20-29 Gland. Epith. 00-09 11 247 28 B reast 30-39 1 G en ital 42 Organa 40-49 U rinary 21 System 50-59 Skin and Soft T issue 60-69 Bones 70-75 6 FIGURE 2 - MALIGNANT TUMORS IN MALES AS REPORTED ACCORDING TO TY PE OF TUMOR AND PRIMARY SITE Non Gland. Epith. 10-19 64 Leuke m ias 20-29 Lympho - Nervous Vascular m as T issues T issues 30-39 40-49 50-59 M uscle 60-69 1 NonE p ith . T issues 70-79 E m bry. - Mixed T issues 80-89 1 24 4 91 7 > 1 2 1 16 21 170 2 11 1 1 4 2 3 1 2 1 T um ors Not C lass. Total 1 90-- ' 77 3 285 3 125 2 69 44 3 181 13 B ra in 76-79 2 3 1 Lymph, and Hem. System 80-89 12 17 OS ittheesr 9 0 ---- 8 27 1 Total 364 419 12 24 5 1 13 19 29 1 4 41 17 17 26 885 API 05551 ) Type of Tumor Site Buccal Cavity 00-09 D igest. Sys. and P e r it. 10-19 R esp iratory System 20-29 Gland. Epith. 00-09 8 1 Breast 30-39 14 Genital 2 Organs 40-49 U rin ary System 50-59 Skin and Soft 1 Tissue 60-69 Bones 70-75 B rain 76-79 Lymph, and H em . System 80-89 Other Sites 9 0 ---- Total 26 FIGURE 3 - MALIGNANT TUMORS IN FEMALES AS REPORTED ACCORDING TO TY PE OF TUMOR AND PRIMARY SITE Non Gland. Epith. 10-19 Leukem ias 20-29 Lympho - Nervous Vascular mas T issu es T issu es M uscle 30-39 40-49 50-59 60-69 NonEpith. T issues 70-79 E m bry. - M ixed T i8aues 80-89 T um ors Not C lass. 9 0 --- 1 ' ' 1 1 3 1 6 1 1 1 14 1 1 Total 1 8 2 15 5 1 7 1 2 42 API 05552 ) ) FIGURE 4 - MALIGNANT TUMORS AS REPORTED BY SEX AND PRIMARY SITE COMPARED WITH U .S . A. MORBIDITY AND MORTALITY ESTIMATES SEX Site Number Reported MALES Per Cent of Total U. S. A. * U .S . A.** P er Cent P er Cent Cancer Cancer C ases Deaths Number Reported B uccal Cavity 77 8 .7 10.0 aDnidg ePsetirvietoSnyeustme m 285 3 2 .2 3 6 .0 R esp iratory System 125 14.1 8 .0 B reast 2 0 .2 4 .7 1 4 7.7 8 1 5 .4 2 0 .2 15 G enital Organs 69 7 .8 12.0 1 3 .0 5 Urinary System Skin and Soft T issue Bones 44 5 .0 181 2 0 .5 13 1.5 7 .0 17.0 6 .6 1 2 .2 7 1.3 1 B ra in 19 2.1 2 .4 ) Lym ph, and Hema- 29 3 .3 ) topoietic System ( . (.................. V _ Other Sites 41 4. 6 10.0 / 6- 5 2 TOTAL 885 100.0 100.0 100.0 42 FEMALES Per Cent of Total U .S . A.* P er Cent Cancer C ases U .S . A.** P er Cent Cancer Deaths 2 .4 2 .0 1.2 19.0 4 .8 3 5 .7 11.9 2 .4 16.7 2 .4 2 3 .0 2 .0 ) 51.0 3 .0 11.0 3 8 .7 3 .9 19. 1 2 3 .5 3 .4 1.5 1.1 1.5 4.8 ) 8.0 100.0 100.0 / 6.2 100.0 BOTH SEXES \ Number R ep orted Per Cent of T o ta l U .S.A .**I P er Cent Cancer D eath s 78 8 .4 2 .9 293 31.6 4 3 .2 127 13.7 9 .6 17 1.8 9 .7 74 ao 18.3 45 4 .9 5 .0 188 20. 3 1.8 14 1.5 0.1 19 2 .0 2 .0 29 3.1 43 4 .6 7 .4 927 100.0 100.0 * P rim ary site of developm ent of can cer among white m ales and fem ales according to Dorn, Harold F . : Illn e ss from cancer in the United S tates. Reprint No. 2537, Public H ealth R ep orts. (B ased upon m orbidity record s c o lle c te d from ten la rg e c itie s in the United States b etw een 1937 and 1939.) ** Cr.ncer deaths by site in the United S tates for 1948 as prepared by the S ta tistica l R esea rch Section, A m erica n C ancer Society, utilizing sta tistics from the National Office of Vital S ta tistics. API 05553 >t i Copy from D. V. Stroop for Information of Members and Associaies of the ftedical Advisory Conmittee ___________________________________ 1-19-53 ESSO IABORATORIES Standard Oil Development Company Medical Research D ivision P. 0. Box 51 Linden, N. J. i January 15, 1953 Mr. D. V. Stroop Aserican Petroleum In stitu te 50 West 50th S treet New York 20, New York Dear Mr. Stroop: I am attaching a rep rin t of an a rtic le recently written by Dr. Hueper. I think th at th is a rtic le should be brought to the attention of a l l of the members and associates of the Medical Advisory Conmittee of the API. Very tr u ly yours, / s / B. S. Eckardt R. E. ECKARDT, M. D. REE:ilk Enel. - Environmental Cancers: A Review by Dr. V. C. Eueper Reprinted from CANCER RESEARCH Vol. 12., No. 10, pp. 691-97, October 1952. API 05554 ) Reprinted for private circulation from CANCER RESEARCH Vol. 12, No. 10, pp. 691-97, O ctober 1952 Copyright 1952 by Cancer Research, Inc. r u m s lh u.s.a. Environmental Cancers: A Review W. C. H u eper ( Satumal Cancer Intitule, United Statee Public Health Service, Federal Security Agency, Betheeda, Ud.) although the knowledge of an environmental causation of human cancer is much older than that of the exogenous origin of infectious diseases, there exists a striking, if not alarming, contrast between the extent to which this information has been put to scientific and practical use in the study and control of these two important groups of diseases. In the attack on the infectious diseases the mam efforts were expended on the discovery of the specif ic causative agents and on their subsequent elimi nation from the external and internal human environment by appropriate preventive, prophy lactic, and therapeutic measures. Interest in the endogenous properties of the host, as represented by heredity, constitution, aging processes, race, and sex, in determining the development and course of infectious diseases did not, as a rule, assume a dominating character. These factors were recognized as having mainly a modifying, but not a causal role, by exerting a certain influence on individual susceptibility to the pathogenic micro organisms and on the course of the disease. The investigation of the anatomic and biologic proper ties on the various reaction products, particularly those of chronic granulomatous nature, remained usually a less important side issue in devising effective control measures. The investigations directed against human cancer, on the other hand, paid relatively little attention to the factual evidence of its established environmental causation, but concentrated first on the various morphological aspects of cancer and cancer cells and, more recently, on the biochemical and biophysical properties of cancer tissue. Can cer was, and still is, considered by many investiga tors a distinct disease entity and is not regarded s an atomic reaction product to a large number of Received for publication June is, 19J2. diverse chemical and physical agents. I t has been for the same reason that the rather diffuse etiologic concepts of physiologic aging and heredity have enjoyed a much greater and much more lasting appeal than the much more definite and reliable observations on the diverse exogenous causes of a considerable variety of human cancers. HxaxDirr Since, in a recent discussion of the age aspects of environmental and occupational cancers (15) the fundamental fallacy of this concept as to the majority of human cancers was discussed in detail, it remains to point out that a similar situation seems to prevail as to the validity of the heredity theory. Conclusions drawn from selectively inbred strains of mice, a notorious biologic artifact with out parallel in nature, have been instrumental in giving to hereditary factors an exaggerated and distorted significance as immediate causes of human cancer. While it may be conceded that there exist a few rare cancers which display hereditary tendencies, at least one of them (cancer of the skin in xeroderma pigmentosum) depends in its causation on the primary action of an exoge nous agent. Although xeroderma pigmentosum is due to an inherited hypersensitivity to solar radia tion prevalent among some inbred family groups, this fact would scarcely justify the conclusion that the ordinary type of solar cancer of the skin is pri marily an inherited manifestation of the host organism which is activated by an exposure to solar rays. Such an inversion or perversion of normal reasoning would have to disregard the fundamental fact that without solar radiation there is no cancer, irrespective of the constitution of the individual. The allegation that hereditary factors based on 691 API 05555 692 Cancer Research prolonged inbreeding play an important causal role in the production of the lung cancers among miners of radioactive ore in Schneeberg and Joachimsthal (Vesin [89]; Macklin [27]; Lorenz [26]) represents another example of tbe rather loose speculations which have been advanced to bolster the cause of the heredity theory. Apart from the fact that none of the various proponents has offered any concrete evidence in support of such a claim, any significant degree of inbreeding in a community of about 10,000 inhabitants (Schneeberg) having a railroad station for many years would indeed represent a most remarkable biologic and sociologic feat. This speculation ad vanced by investigators remote from these opera tions has no basis in fact according to German in vestigators.1 The occasional occurrence of lung cancers in several members of the same family working in these mines certainly cannot be regarded as proof of a hereditary liability. Similar observations of a familial appearance of skin (scrotal) cancers were reported from England during the past century in families of chimney sweeps. Such familial cancers were obviously caused by an occupational or en vironmental exposure to soot by several members of such families without necessitating the assump tion of a common hereditary susceptibility. The high rates of attack of several environmen tal cancers rather clearly indicate that heredity evidently plays a relatively insignificant role in their production (cancer of the lung in 75 per cent of miners employed in radioactive mines--Baader [S]; Hueper [16]; cancer of the bladder in almost 100 per cent of dye workers following highly exces sive and prolonged exposure to certain aromatic amines--Goldblatt [11]; Mueller [32]; Gross [12]; Hueper [17]; cancer of the skin in 100 per cent of workers with contact with pitch for more than 40 years--Sladden [35]). I t is likely, however, that such an almost com plete obliteration of individual inherited or ac quired constitutional differences in susceptibility to exogenous carcinogens occurs only in the pres ence of an overwhelming, high intensity exposure. Whenever groups of individuals become exposed to carcinogens of low potency or sustain exposures of low intensity and duration, there appears evidence indicating variations in the speed and character of the individual responses to the environmental carcinogen. A constitutional influence evidently 1None of tbe numerous German workers mentioned in their papers tbe occurrence of inbreeding in the mining population of tbe Erzgebirge and Linzbach in answer to a recent inquiry stated that nothing is known among German scientists in this respect. controls the susceptibility of different races t solar cancer of the skin, to which light-pigment individuals are much more liable than dark pigmented individuals. The effect of this racia difference in reactivity to the carcinogenic actioi of the ultraviolet radiation from solar sources i reflected not only in a lower incidence of ski] cancer in Negroes than in whites, but also in , different topographical distribution of the skii cancers observe in the two racial groups. Whi| skin cancers in whites are mainly located in th skin of the exposed parts of the bead, neck, an upper extremities, those observed in Negroes ai predominantly situated in the usually cover parts of the body and the lower extremities (31) Since arsenical cancers not infrequently involv the unexposed parts of the skin, it is reasonable t conclude from the above evidence that the varia tions in the topographical distribution of ski] cancers in whites and Negroes are, in part, due t differences in causal agents active in the produc tion of these cancers. I t is unwise, however, to generalize or overem phasize the role of racial factors in the causation c environmental cancers. Although primary hepati cancer incidence is high among West Africa Negroes, their American descendants do not dii play any such tendency (Berman [5]), indicatin thereby that not racial or hereditary factors bu environmental factors are underlying the Africa experience. Likewise, it can be considered a established that the absence of penile cancc among Jews and the high liability of Chinese t this type of malignant tumor do not depend upo race-conditioned constitutional differences, but ai mainly the result of differences in personal hj giene, especially the circumcision practiced at a early age by the Jews. There thus exist not onl "environmental familial cancers," but also "et vironmental racial cancers," which have no reli tion to heredity. Sex Differences in the sex distribution of cancers a fecting several organs (lip, mouth, larynx, bronch skin, and bladder) have given rise to the concep of a sex-conditioned susceptibility to certaii cancers. Observations on environmental cancel lend no substantial support to such a theory Cancer of the lung of unknown origin has a malt female sex ratio of from 4:1 to 24:1, according t various investigators (18). Asbestosis cancer of th lung, on the other hand, displays a male-female se ratio of 2.4:1, indicating that with an equalizatio of exposure to a carcinogenic agent there occui also an equalization of the incidence rates of lun, API 05556 H u e p e r -- Environmental Cancers: A Review 693 to iteti *ark. , aciai ction 'es is ! skin in & skin 'Vhile n th . and :s ar 1 vered (31). . ivolve j -ible t0 varia>f skin due to >rodue- -erenation of lepatic Vfrican tt dis.icating >rs but _4frican ^ d as ,ancer nese to id upon but are nal hyd at an lot only Iso "en no rela- ncers afbronchi, concept certain l cancers 1 theory, s a male ording to cer of the emale sex jalization re occurs es of lung --------- , -he tw 0 sexes (Merewether [30]). Like- canffor t improbable that the male-female wise. >t.is , j .j for Mexican men and women se* ratl0R u and Edmondson [37]) living in Los (Steiner. sex. related factors. It is note- Ange'es. j j j s re5pect that the entrance of female worth.' in .n tjje English textile plants was fol- ^ llT 'fte r a suitable latent period by the appear- |0wed a 0f tjje vulva in some of these SllCe * s w hich fact thereby demonstrates that peni o r > . rs acq Uire the anatomic equiva- feT o H h e scrotal cancer of their male counter- *en While oral cancer is rare in white Par s this tumor is rather frequent in Indian ^m en'chewing betel quids (Bashford [4]). This uv0 cancer of the oral cavity represents 12 per bU't of all cancers among Filipino women. For this ^Lson the sex ratio of oral cancer among Fili * s inverse to that found among white people P Kuhitine the temperate zones, where the male- female sex ratio is 3:1 (Maxwell [29]; Vedder [38]). These and other similar observations indicate ti8t the differences in the sex ratio of certain cancers of unknown origin are apparently not due? in the main, to hormonal factors but rather reflect variations in the types and intensity of exposure toenvironmental carcinogens affecting members of the two sexes to a different degree for reasons of occupations, habits, customs, clothing, hobbies, and general living conditions. Eptdqcoloqic P attern The type of the epidemiologic pattern of en vironmental cancers depends less on certain con stitutional factors of the population studied (heredity, race, sex, age) than on the composition, as well as the type, intensity, and duration of action of the agents constituting the environmen tal carcinogenic spectrum to which they have been exposed. The physical and chemical carcinogens exert their disease-producing effect, just like the pathogenic micro-organism, whenever and wher ever they operate under proper conditions of ex posure and irrespective of the special type of con tact (occupational, medicinal, dietary, habitual, environmental, etc.) present. The general soundness of this concept is at tested by the following observations. The appear ance of occupational cancers in the various indus trialized countries has closely followed the spread of certain industries. This sequence of events is well illustrated by the chronologic and geographic appearance of bladder cancer among dye workers in different countries following the establishment ofaniline dye industries (Table 1). Additional support for the concept proposed is derived from the demonstration of a centrifugal spread and scatter pattern of occupational cancer hazards from the focus of original production of carcinogens in basic industries through processing and consumer industries to the ultimate handler and general consumer of the finished products. This pattern becomes apparent in the occurrence of skin and lung cancer among the producers of tar and pitch in gas plants and coke ovens, the appearance of similar cancers among tar refinery workers, roofers, road construction workers, cork brick manufacturers, and other members of sec ondary industries and trades handling and using tar and pitch. An identical chain of environmental cancers among successively exposed population groups exists in regard to arsenicals, which have TABLE 1 CHRONOLOGIC AND GEOGRAPHIC APPEARANCE OF Blad der Cancer among Dye Workers in D iffer en t Countries (it) Country Germany First rear reported 1805 Switzerland Great Britain Russia Austria United States Italy Japan France 1005 1018 1025 1032 1034 1035 1040 1045 Aothor Rehn Schedler Ross Rosenbaum and Gottlieb Schueller Ferguson et al. di Maio Nagayo and Kinosita Billiard-Ducbesne Total ao. recorded op to 1951 Approxi mately 350 100 300 71 ( 250 00 * (7) 41 elicited cancers among miners of arsenic-contain ing ores, smelter workers, manufacturers of ar senical insecticides, users and consumers of food stuffs and drinking water contaminated with such arsenicals, as well as individuals receiving arseni cals for medicinal reasons (Hueper [16]; Neubauer [S3]; Arhelger and Kremen [2]; Butzengeiger [8]; HiU and Faninp [14]; Arguello, Tello, Macola, and Manzano [1]). Occupational cancer hazards also display a cancer distribution pattern which shows the highest attack rates among the population groups employed within the immediate vicinity of the carcinogenic industrial foci and gradually decreas ing attack rates among population groups increas ingly remote from it and thus less intensely and frequently in contact with carcinogenic agents. A good illustration of this concentric distribution pattern of occupational cancers is offered by the aromatic amine bladder cancers. The bladder can cer incidence rate, according to past experience, was highest among intermittently but intensely exposed maintenance and repair men who were followed in decreasing order by operators regularly 694 Cancer Research employed in the manufacture of beta-naphthylamine and benzidine, these by truck drivers, yard men, chemists, engineers, and supervisors who have irregular and mitigated exposures. Whether or not "neighborhood cases" of aromatic amine cancers actually occurred among the population residing or working within the fume and waste dis posal zone of aromatic amine operations is still doubtful and controversial (Mueller [34], Hueper [17], and Gross [14]). Since, according to Gehr mann, Foulger, and Fleming (10), workers em ployed in a building formerly used for the produc tion of beta-naphthylamine have an occupational bladder cancer hazard from contact with the minute amounts of the chemical volatilized from impregnated building material, neighborhood cases of this occupational cancer may conceivably occur among persons living near aniline dye plants with defective loading, shipping, and waste dis posal arrangements for beta-naphthylamine and benzidine. Another major factor determining the char acter of the epidemiologic pattern of occupational cancers is represented by their latent period, which in turn depends upon the relative potency of a particular carcinogen, its physicochemical proper ties (solubility, dispersion, chemical reactivity Organ Skis Lung Bladder TABLE 8 Latent Periods o r Environmental and Occupational Cancers Afcnt Anenic: median*] occupational Tar Creosote oil Mineral oil Crude paraffin oil Solar radiation X-radiation Asbestos Chromates Nickel Tar fumes Ionizing radiation Aromatic amines At. latent period (year) 18 US 0-24 US 60-54 16-18 O-SO 7 18 16 2 18 15-86 11-13 Raste of la test period (year) S-40 4-48 1-50 16-40 4-75 8-86 16-40 1-12 15-21 5-47 8-80 -28 7-60 2-40 and affinity, etc.), the physicochemical and co carcinogenic or anticarcinogenic properties of its vehicle or its associated agents, the route of con tact, the intensity of the individual exposures, their rhythm, and the total duration of exposure. These factors, which have a distinct influence upon the incidence rate of cancers among exposed popula tion groups, also exert a definite influence upon the length of the latent period. With decreasing in tensity of exposure there occurs a reduction in the incidence of cancers and a lengthening of their latent period. Apart from individual differences' susceptibility, these irregularities in the intensit of exposure to environmental carcinogens main] account for the wide range of latent periods r corded for environmental cancers (Table 4). The target organ of environmental carcinoge depends on various factors. Special tissue affinit of benzene to the fat tissue contained in liema poietic organs thus seems to account for its exclu sive leukemogenic action. The intensity of ex TABLE S Sit e of Cancer a n d Route of E xposure Typ of eoatact Cutane ous Carctnotenlc lgat. Arsenic Coal tar, pitch, soot, asphalt Creosote oil, anthracene oil Petroleum asphalt, coke, tar Petroleum oils (high boiling) Shale oil, crude paraffin oil Lignite oil and paraffin oil Ultraviolet radiation X-radiation, radioactive chemicals Benxene Site of cancer Sein Hematopoietic" sues Respira tory Arsenic dust Chromium compound dust and fumes ' Nickel dust and vapors Asbestos dust Beta-naphthylamine, benti- dine dust and vapors Coal tar fumes Petroleum oil mist Isopropyl oil vapors* Radioactive dust and gases Lung Lung Lung, nasal cavi and sinuses Lung Bladder Lung Lung Nasal sinuses, ynx, and lung Lung Inges tion Arsenic Radium, mesothorium Skin Bone * Isopropyl oil in tbs crude liquor froin which isoprop&aol is oh' by distillation (SO). posure to some environmental carcinogens seems to determine the distributory mechanism the resulting cancers. The relatively high f quency of heterotopic multiple primary can observed among dye workers with cancers of urogenous tract as well as other organs 0 stomach, intestine, prostate--Mueller [34]; H per [16]) seems to be due to an unusual hema enous spread of the carcinogenic hydrocar ordinarily mainly excreted through the ur (Bonser [7]). The principal reason for the ap; ance of cancers in various organs, however, represented by the route of exposure (Table 3). Epidemiologic and demographic studies carri out during the past two decades have demo strated the existence of marked and signific variations in the total incidence of cancer as well in the organ, sex, age, and race distribution cancers in different countries, regions and popui API 05558 lifferences in the intensity ogens mainly it periods re'able 2). il carcinogens tissue affinity id in hemato; for its excluensity of ex- r Hueper--Environmental Cancers: A Review 695 . groups. While a part of such variations is due a carcinogenic hazard, figures on incidence must ^differences in the age distribution of the various be calculated for this group only, and not for the t0 ujation groups surveyed as well as in the qual- total plant population, since the latter procedure Pfof medical care and of medical recording of results in an undue dilution of positive evidence Vlases, there remains for many of these discrep- and thereby in distorted epidemiolog'c informa cies in rates of regional cancer incidence no other tion. The limited occurrence of bladder cancer to *lusible explanation than the differences in ex relatively small and restricted groups of dye work posure to environmental carcinogenic factors ers (10), the excessive liability to scrotal cancer (T a b le 4). . limited to workers employed in paraffin pressing The striking differences in the lung cancer operations in oil refineries (34), and the increased morbidity rates in metropolitan centers located in liability to lung cancer among persons with as- tie same part of the country militate against the bestosis--and not among all persons employed in XPOSRE concept that such local discrepancies can justly be asbestos operations--provide pertinent illustra Site of ctncer fcin attributed to variations in medical care and re cording, or to differences in the biologic composi tion and hereditary properties of the populations concerned, or to the predominant action of a single etiologic environmental factor, especially cigarette smoking (Wynder and Graham [41]). The local variations as well as the consistent increase of lung cancer morbidity rates must be ascribed to a tions of this point. TABLE 4 Lung Ca n c e b M o r b id it y Ra t e , 1937 a n d 1947, for R e s id e n t s of E ig h t M etro po litan c e n t e r s p e r loo.ooo P o pu l a t io n * Cm Atlanta H alm 1*37 1*47 5 0 IS.4 Pb u l u 1*37 1*47 10 50 T otal 1*37 1*47 29 89 hematopoietic tis* sues -ung -.ung lung, nasal cavity, and sinuses Lung Bladder combination of exogenous factors affecting, to differing degrees, the various population groups as well as the two sexes surveyed. Since the known environmental lung cancers have a latent period ranging usually from 10 to 25 years, it may be concluded that the rising trend of lung cancers is attributable to changes in the environmental carcinogenic spectrum which started around the New Orleans Dallas Birmingham Denver San Francisco Chicago Pittsburgh Supplied bj 1S.1 39.1 2.8 4.2 7.6 5 9 29 0 0 5 6.4 3 1 4.5 18.9 2.1 8.9 3.8 9 1 21.9 4.2 8.1 6 6 15 6 34. S 39 81 9.8 IS .3 29.5 4.3 7.0 8 .8 9 .7 26.1 4.9 5.5 7.8 Biometrici Section. Notional Cancer Institute. 20 8 17.2 11 0 14 8 20 8 18.0 15.6 An analysis of the population "at risk" should turn of the century and the effects of which are be include not only workers presently employed for a ung iasal sinuses, lar ynx, and lung Lung coming increasingly evident (Hueper [18]). Sup porting this concept is the fact that the majority of environmental and occupational cancers dis sufficiently long period, so as to cover the minimal latent period of the average occupational cancer (5 years), but also all formerly employed workers Skin Bone opropaool ia obtained covered during the last 25 years involve cancers of the respiratory tract (cancers produced by chro mium compounds, nickel, arsenicals, asbestos, coal dead or alive who left the industry, plant, opera tion, or trade group fulfilling this condition. The inclusion of large groups of short-term employees tar, petroleum oils, isopropyl oil, radioactive sub with insufficient exposure and latent period into the rcinogens also stances) (21, 28). surveyed group also introduces a serious dilution mechanism of In the conduct of epidemiologic studies on en factor which causes misleading or inconclusive re vely high fre- vironmental cancers certain important precautions sults (23, 36). -imary cancers have to be observed, if misleading, incorrect, or in Since the occurrence of environmental cancers cancers of the conclusive results are to be avoided (Hueper [17]; rarely has the character of an epidemic, but as a organs (lung, Downing [9]). It is essential to survey as far as rule appears as an endemic, it is necessary to con Her [32]; Hue- practicable the total effectively exposed popula duct epidemiologic studies as long-term investiga lsual hematog- tion, and to exclude from membership in the tions, which means the analysis of data covering hydrocarbons actually exposed population group all those indi periods of at least 5 years. Negative results ob lgh the urine viduals who have merely an "administrative" ad tained from short-term investigations comprising for the appear- herence to it. In a survey of environmental can periods of 1 or 2 years give, at best, inconclusive is, however, is cers among oil refinery workers, for instance, it is results if not incorrect ones (6). .re (Table 3). scientifically not permissible to include office per In view of the long latent period of environmen studies carried sonnel, sales people, and other white-collar work tal cancers it is necessary to restrict occupational i have demon- ers, unless they sustain real contact within the cancer surveys to plants which have been in and significant refinery area to suspected or recognized carcino operation for at least 10 years, unless a large por ?ancer as well as genic agents for occupational reasons. Similarly, tion of the workers in their employ has previously distribution of whenever, within an industrial population to be been employed in similar operations elsewhere. ms and popula- surveyed, only a restricted group is shown to have The absence of published reports on the occur- API 05559 696 Cancer Research rence of occupational cancers in an industry hav ing known or suspected cancer hazards is no assur ance that a serious occupational cancer problem in these establishments is nonexistent. Perhaps the most notorious example that can be cited as an illustration of such occurrences is represented by the long delayed discovery of the lung cancers among uranium miners of Joachimsthal. Although the existence of a high lung cancer incidence among the Schneeberg miners was established in 1876, despite repeated inquiries made by German investigators with the various governmental au thorities under whose jurisdiction the uranium mines in Joachimsthal located nearby on the southern slope of the Erzgebirge were operated, it was not until some 50 years later that the high frequency of lung cancers among this group of similarly exposed workers was recognized. Until that time (1926), the century-old practice of mis taking lung cancers of the Joachimsthal miners for pulmonary silicosis or tuberculosis was continued. Likewise, associated pathologic conditions some times may operate in obscuring the co-existence of an occupational cancer. This danger exists, for instance, for absestosis cancer of the lung, unless a thorough necropsy study is made. CABcmooDmu An analysis of the action mechanism of the vari ous recognized, suspected, or potential human carcinogens supports the viewpoint that the ma jority of cancers develop at sites where, for some reason, the most intense or most prolonged ex posure to the carcinogen takes place. The follow ing types of mechanisms determining the distribu tion of environmental cancers may be distin guished. a) Cancers developing at sites of primary contact. --To this group belong the cancers of the skin re sulting from cutaneous exposures to substances such as coal tar, petroleum oils, creosote oil, soot, and similar combustion and high temperature dis tillation products of carbonaceous matter, as well as to ultraviolet and ionizing radiations; moreover, the cancers of the nasal cavity, nasal sinuses, larynx, and lung elicited by the inhalation of arsenicals, chromium, and nickel compounds, (beryllium?), asbestos, isopropyl oil, tarry matter, and radioactive gases and dusts. The cancers of the connective, bony, and hematopoietic tissues following exposure to penetrating ionizing radia tion may be included in this group of primary contact cancers. b) Cancers developing at sites of selective deposi tion.--Arsenical cancers of the skin, osteogenic sarcomas following ingestion of radium and/or mesothoriirm, leukemia following contact wi benzene, (thyroid carcinoma following radioactivi iodine medication, leukemia subsequent to radii active phosphorus medication, osteogenic sarco: following inhalation of beryllium compounds?)] may be included in this group. ' c) Cancers developing in organs with specialfunc tional or toxic affinityfor carcinogens.--Represen tives of this group are almost exclusively caused carcinogens of potential importance as far as hu mans are concerned and thus have been observi mainly in experimental animals. Cancers of type are the tumors of the liver developing follow, ing exposure to various azo dyes, aminofluorem compounds, chlorinated hydrocarbons and lenium, the cancers of the breast and uterus su sequent to an excessive exposure to estrogens, a: cancers of the thyroid following the prolonged ad. ministration of thiouracil derivatives. d) Cancers developing in organs of excretion oj carcinogens.--Cancers of the bladder, ureter, an< kidney observed in individuals and experimen1 animals having cutaneous, ingestive, and respi tory contact with certain aromatic amines and a: compounds due to the presence of carcinogei material in the urine belong to this group. e) Cancers developing on the basis of fu n d i abnormalities due to certain dietary deficiencies a, representing a type of indirect or secondary envin menial carcinogenesis.--Cancers of the h; pharynx and of the liver noted among populate groups subjected to a diet deficient in vitamin complex and protein as well as cancer of the livi in rats kept on a choline-deficient diet are membe: of this group. f) Cancers of the young resulting from a trai placental penetration of carcinogens.--There exi so far only experimental evidence in support such an exposure route in the demonstration of development of pulmonary tumors in the off spring of pregnant mice given urethan near te: (Larsen [25]; Klein [24]). Whether or not a simik mechanism may be active in the production a congenital cancers or cancers developing during in fancy and early childhood appears to be a worth while subject of investigation in view of thesi findings. While it is not likely that exogenous agents a: the only factors responsible for human cancers the increasing number and widening variety d environmental carcinogenic agents suggest thai they seem to be operative in a much large proportion of cancers than is recognized realized at present. It appears from the evidenci available that the study of environmental canee hazards and cancers offers not only the mos API 05560 act with dioactive to radio sarcoma pounds?) rialf une. presenta?aused by nr as huobserved rs of this lg followlofluorene and seerus sub gens, and jnged ad- cretion of eter, and erimental d respiras and azo -cinogenic ip. functional encies and y environ ne hypoiopulation ,'itamin B f the liver members n a translere exists upport of tion of the l the offnear term t a similar luction of during in e a worthof these agents are n cancers, variety of ;gest that ich larger gnized or e evidence ital cancer the most H c e p e r -- Environmental Cancers: A Review 697 promising approach to the determination of the usation of human cancers, but is also the princi p i route through which the primary prevention of ncer may be achieved. REFERENCES i \ rocello, R- A.; T ello, E. E.; M acola, B. A.: and * Manzano, L. Los cncers cutneos en el arsenicismo * [heiiomas of Medicinal Origin. Surgery. 30:977-86, 1951. BaaDEBi E. W. Der Lungenkrebs als gewerbemedizi * nisebes Problem. Verhandl. Deutsch. Gesellsch. inn. Med., 57:322-32, 1951. , n^groRD E. F. An Address on Cancer in Man and " ,,finals. Lancet, 2:691-701, 1909. , C. Primary Carcinoma af the Liver, pp. 1-18. *' London:H. K. Lewis & Co., 1951. A Bidstrct, P. L. Carcinoma of the Lung in Chromate Workers. Brit. J. Ind. Med., 8:302-5, 1951. 1 Bosses, G. M.; Clatson, D . B.; and J ull, J. W. An Experimental Inquiry into the Cause of Industrial Blad der Cancer. Lancet, 168:286-88, 1951. g Butsengeiger, K. H. Die chronische Arsenvergiftung der Winxer. Anti. Wchnschr., 4:365-69, 1949. a Downing, J. G. Cancer of Skin and Occupational Trauma. ' J.A.M.A., 148:245-52, 1952. 10. Gzhhmann, G. H.; Foulger, J. H.; and Fleming, A. J. Occupational Tumors of the Bladder. Proc. 9th Internat. Cong. Ind. Med., London, pp. 473-75, 1948. 11. Goldblatt, M- W. Vesical Tumours Induced by Chemical Compounds. Brit. J. Ind. Med., 6:65--81, 1949. 12. Gross, E. Das Carcinom vom Standpunkt des Gewerbe pathologen. Angew. Chem., 53:368, 1940. 13. Henry, S. A. Occupational Cutaneous Cancer Attribut able to Certain Chemicals in Industry. Brit. Med. Bull., 4:389-401, 1947. 14. Hq l A. B., and Fanino, E. L. Studies in the Incidence of Cancer in a Factory Handling Inorganic Compounds of Arsenic. Brit. J. Ind. Med., 6:2-6, 1948. 15. Hueper, W. C. Aging Aspects of Environmental Cancers. Pub. Health Rep., 67:773-79, 1952. 16. --------. Occupational Tumors and Allied Diseases, pp. 435-59. Springfield: C. C. Thomas, 1942. 17. ------ . Occupational Bladder Cancer. Trans. 2d Nat. Cancer Conference, Cincinnati, 1952 (in press). 18. ------ . Environmental Lung Cancer. Ind. Med. & Surg., 20:49-62, 1951. 19. ------ . A Methodology for Environmental and Occupa tional Cancer Surveys. Public Health Tech. Monograph .Vo. 1. Fed. Sec. Ag., 1950. to. Hcepeb, W. C. and Mancuso, T. F. The Role of the State Health Department in the Control of Environ mental Cancer. Pub. Health Rep., 67:644-51, 1952. 21. Hcepeb, W. C. Experimental Studies in Metal Canceri- genesis. I. Nickel Cancers in Rats. Texas Rep. Biol. & Med., 10:167-86, 1952. 8*. ---------. Experimental Studies in Metal Cancerigenesis. II. Experimental Uranium Cancers in Rats. J. Nat. Cancer Inst, (in press). 23. Inoalls, T. H. Incidence of Cancer in the Carbon Black Industry. Arch. Ind. Hyg. & Occup. Med., 1:662-78,1950. 24. Klein, M. The Transplacental Effect of Urethan on Lung Tumorigenesis in Mice. J. Nat. Cancer Inst., 12:1003-10, 1952. 25. Larsen, C. D. Pulmonary-Tumor Induction by Trans placental Exposure to Urethane. J. Nat. Cancer Inst., 8: 63-70, 1947. 26. Lorenz, E. Radioactivity and Lung Cancer: A Critical Review of Lung Cancer in the Miners of Schneeberg and Joachimsthal. J. Nat. Cancer Inst., 6:1-15, 1944. 27. Macelin, M. T., and Macelin, C. C. Does Chronic Irri tation Cause Primary Tumor of the Human Lung? Arch. Pathol., 30:924-55, 1940. 28. Mancuso, T. F., and Hceper, W. C. Occupational Can cer and Other Health Hazards in a Chromate Plant: A Medical Appraisal. Ind. Med. & Surg., 20:358-63, 1951. 29. Maxwell, J. L. Betel Chewing and Cancer. Brit. M. J., 1:729, 1924. 30. Meriwether, E. R. Annual Report of the Chief Inspector of Factories and Workshops of the Year 1947, p. 15. Lon don: H.M. Stationery Office, 1947. 31. Montpellier, J. M., and M csbini-Montfellier, J. L'pithlioma de la peau chez le nord-africain. Maroc. Md., No. 306, 1950. 32. Mueller, A. ber Blasen- und Nierenschdigungen in der Farbstoffindustrie. Helvet. chir. acta-, 18:1-41, 1951. 33. N eubauer, O. Arsenical Cancer; A Review. Brit. J. Cancer. 1:192-244, 1947. 34. Page, R. C.; Holt, J. P.; Hendricks, N. V.; Eceardt, R. E.; Stanton, C. L.; Smith, W. E.; Sunderland, D. A.; Suqiuha, K.; Fischer, H. G. M.; Priestley, W.; Ebt, L. T.; Wanlesb, G. G.; Rehneb, J.; Blandinq, F. H.; and Kino, W. H. Symposium on a Cancer Control Pro gram for High-boiling Catalytically Cracked Oils. Arch. Ind. Hyg. & Occup. Med., 4:297-345, 1951. 35. Sladden, A. F. Pitch Cancer. Rep. Intern. Conf. on Cancer, London, 1928, pp. 285-88. Bristol: John Wright & Sons, Ltd. 36. Snegibept, L. S., and Lombard, O. M. Arsenic and Can cer (Observations in the Metallurgical Industry). Arch. Hyg. & Occup. Med., 4:199-205, 1951. 37. Steiner, P. E.; Bott, E. M.; and Edmondson, H. A. Pulmonary Carcinoma Revealed at Necropsy: With Refer ence to Increasing Incidence in the Los Angeles County Hospital. J. Nat. Cancer Inst., 11:497-510, 1950. 38. Veddeb, E. R. The Incidence of Cancer in Filipinos. J.A.M.A., 88:1627-29, 1929. 39. Vesin, M. S. Cancer pulmonaire provoqu par les mana tions radioactives. Arch. mal. prof., 9:280, 1948. 40. Weil, C. S.; Smyth, H. F., J r.; and Nale, T. H. W. Quest for a Suspected Industrial Carcinogen. Arch. Ind. Hyg. h Occup. Med., 6:535-47, 1952. 41. Wtndeb, E. L., and Graham, E. A. Tobacco Smoking as a Possible Etiologie Factor in Bronchiogenic Carcinoma. J.A.M.A., 143:329-36, 1950. API 05561 mm File: guicomaittee OB Carcinogenicity t c : 2VS s o a r y 2 3 , 1953 or. A. wealey Horton University of Cincinnati 9 Kettering laboratory Collegi of Medicine Eden Arenas Cincinnati 19* Ohio Dear Or. Horton; Wo mceived today tin ste n cils of a aenorandun froa Kettering laiborataory. In accor dance e lth yonr vlabee, the eteaeile vofiltihebeAdPoI pMleldcaimtedi AanddhrdlsiosrtyriCbountendiUteoe. e*ere and aaaociatee Wary tru ly yours, Joan Biemeheek Bepartaent at Technical Barrisse API 05562 LAiOMATOtY c7*AT1II. OH'0 1 UNIVERSITY OF CINCINNATI d if a it m in t o r F U R IA T IV I MUHCINI AMO IMDUITIIAL HEALTH V CAI L I U S : KW| u . CINJ Ja n u a ry 1 9 , 1953 r ~ -/ / w> \ Mr. David V. Stroop A -V y ^ i /^ k ^ K Director, Technical Service American Petroleum I n a t i tu t e 50 W. 50th S tr e e t He* York 20, lew York Dear Mr. Stroop: to re forwarding under separate cover by Railway Express the a te n e ila o f a memorandum fo r d u p li cation and d is tr ib u tio n to members and aaaoeiatea o f the API Medical A dvisory Committee. A ll members, except Dr. T. M. Prank, o f the Research P ro ject Advisory Com m ittee rec eiv ed c o p ies a t our m eeting January 16 . . Very tru ly yours, AWH:mjg / H . A. Wesley Horton API 05563 I A merican P etroleum Institute 8 0 WEST 8 0 th STREET NEW YORK 2 0. N. Y. February 2, 1953 As requested In your le tte r of Hovember 19, the vievs of the Medical Advisory Comnlttee membership on the rep o rt and recommendations of the Working Cfroup are transmitted for your consideration, as follows: 1. That portion of the report, as well as the f i r s t recommendstion, dealing with the epidemiological phase of the Kettering project constitutes encouraging support to recommendations . which have been considered and advocated repeatedly by the committee. The members need and welcome the assistance of refining executives in the compilation of adequate standardized records for the petroleum industry. 2. The balance of the report and the recommendation for an in v esti gation of the epidemiology of cancer in other industries lack definition necessary for a c ritic a l review; however, in principle, the members deem i t unwise for the American Petroleum In stitu te to in itia te such a project. I t seems unnecessary to recite the eany d iffic u lt problems incidental to such an undertaking. For your information, the members of the Subcommittee on Carcinogenicity of the Medical Advisory Comnlttee, by le tte r b allo t in September 1952, expressed considerable doubt as to the wisdom of extending to employees of customers the epidemiological studies of cancer by ths Kettering Laboratory under the sponsor ship of the American Petroleum In stitu te . On behalf of members of the Medicad Advisory Comnlttee, X express my sincere appreciation for the in terest shown by members of the Study Comnlttee and its Working Group in th is subject. Very tru ly yours, Medicad Advisory Committee E. Kern Linder, M. D. Chairman, Medicad Advisory Committee API 05564 ^.,,T T E R IN R U A M R A T O R T ; ; : ; U . , , '* SI o f MEDICINE EDEN AVEMOE m oho UNIVERSITY OF CINCINNATI 5) - O ' S S DEPARTMENT OP PREVENTIVE MEDICINE AMD INDUETRIAL HEALTH F ebruary 1 7 , 1953 CAELE ADDRUE EETLAE. CINCINNATI TELEPHONE: CAPITOL M il Mr. D. V. S tro o p , D ir e c to r Department of T echnical S erv ices American Petroleum I n s t i t u t e 50 Vest 50th S tr e e t Hew York C ity 20 Dear Mr. S tro o p : I am sending you h e r e v ith a sta te m e n t of e x p e n d itu re s made on behalf of American Petroleum I n s t i tu t e f o r the fo u rth q u a rte r of 1952- I b e lie v e th a t th e statem en t w i l l be s e lf -e x p la n a to r y , but i f you have any q u e stio n s o r comments, Doctor Kehoe would appreciate your b rin g in g them to h is a tte n tio n . Very tr u ly y o u rs, Dr Eehoe e c r e ta r y To ef Enc. API 05565 UNIVERSITY OF CINCINNATI KETTERING LABORATORY ACCOUNT o p THB A K Z R IC jjr PBTBOLEQM J S T I TU T1 POP 4 t h Q u a r t e r 1 9 5 2 __________________ SA LA R IE S (Based on Proportion of Tim e Actually Spent on Project) Direct Salarie s........................................................................................... 9 6 S 8 ...6 4 .. Indirect Salaries Histopathological 9 9 .9 . Preparation................................................. .Q9_ Other Services......................................................................... 9.Q Z3.?.2 0 . 1 9 .7 5 1 .8 4 MISCELLANEOUS EXPENSE Purchase of Anim als.................................................................................4 4 8 .J 3 0 . Special Laboratory Supplies...................................................................... 3 S 6 . . 4 6 - Travel .......................................................................................................16.2.8.. 3 4 - Overhead (Proportion of Heat, Gas, Electricity, Steam, Telephone, General Laboratory Supplies, Postage, Annuities, Pensions, Maintenance, etc...........5 7 3 2 . 9 1 . 8 f206 .5 1 TOTAL............................. 2 7 ,9 5 8 .3 5 Balance Available for Further W ork at End o f . . . J M J i i r t f Jr _ 1 9 5 2 __________ 3 4 , 3 4 2 . 4 9 Balance Due Kettering Laboratory at End of................................ ................................. .. Receipts T o ta l S t ir a l la b l * ...... Expenditures ..4 h -Q U fc 2 l. t * r . ......................................... 4 6 ,3 7 2 .9 8 8 0 ,7 1 5 .4 7 y fjO g ft.g g Balance Available for Further W ork at End o f-.-4 .4 k A \ ^ t o r .. 1 5 3 2 - - . - - . - , - 5 2 , 7 5 7 . 1 2 Balance Due Kettering Laboratory at End of..................................................................... >tm u t la 00 7-|| API 05566 AMERICAN PETROLEUM INSTITUTE 5 0 WEST 5 0 th STREET NEW YORK 2 0 . N Y. . N T rmoor. Of TECHNICAL. EEKVICE* d .cto March 13, 1953 To Members and Associates S u b c o m m it t e e on Carceoogenicity Kieffer Davis M.D. (Chairman) Marshall Clinton M.D. L. E. Curtis M.D. R o b e rt E. Eckardt M.D, D. Gassovay M.D. C. E. Hine M.D. E. K. Linder M.D. M, N. Hevquist M.D. James V. Osborn M.D. . B. B. Reeve, M.D. L. M. R. C. L. C. R. C. R, M. R. M. L. C. R. D. Allan F. J. G. V. Henderson Cole Beard, Jr. Mithoff Shepardson Landon Burroughs Bent E. Dooley Sanders Adams Gentlemen: At the reguest of Doctor Hevquist, ve enclose agenda and minutes of F ifth Meeting - RP (MC-1) Advisory Conaaittee held January 16, 1953. Very tru ly yours, DVS: Jb me: Menfeers and Associates Medical Advisory Comittee Drs. A. Vesley Horton Robert A. Kehoe John J. Phair API 05567 A ffi TNFflBM X i a P ^ NOT FOR PUBLICATION AGENDA AND MINUTES OP THE FIFTH MEETING RP(MC-l) ADVISORY COMMITTEE rffTTERING LABORATORY, UNIVERSITY OF CINCINNATI Jan u ary 16, 1953 9 :30 A.M. E .S .T . AGENDA 1 Approval of m inutes o f F o u rth M eeting h e ld 9 /2 4 /5 2 2. progress rep o rt 3 progress re p o rt on E pidem iological Survey - D r. A. W. H orton - Dr. J . J. Phalr h Research a c tiv itie s fo r the year beginning 7/1/53 Chemical and b io lo g ic a l - D r. A. W. H orton Epidem iological' " Dr. J . J. Phalr Budgetary a llo c a tio n s - D r. R. A. Kehoe 5 Report of the Committee on the C arcinogenic a c tio n of M ineral O ils to the Medical Research Council (G reat B r ita in ) on t h e i r " F i r s t Three Y e a rs' Work" 6. Communications w ith D r. Paul K o tin , U niversity of Southern C alifo rn ia 7. Other 8. Time and p la c e o f n e x t m eeting (MAC m eets H o tel Shamrock - Houston 9 /3 0 /5 3 ) MINUTES The F i f t h M eeting o f th e RP(MC-l) A dvisory Committee was convened in Dr. K ehoe's o f f i c e a t ;he K e tte rin g L ab o ra to ry o f the U niversity of C in c in n a ti a t 9:30 A M., January 16, 1953- In attendance were the fo llo w in g : M. N. Newqulst, M.D. R. E. E ckhardt, M.D. E. K. L inder, M.D. K ieffer D avis, M.D. C. H. Hine, M.D. R* A. Kehoe, M.D. V H eyroth, M.D. J . P h a lr, M.D. A. W. H orton L. C. B eard, J r . K. G. Mackenzie R. M. Landon R. C. M ith o ff E. W. Adams R. M. Shepards on API 05568 2 2EM-1 The m in u te s o f th e F o u r th M e e tin g o f th e RP(MC-l) A d v iso r y weri e a rprp ro v ed . S u b s e q u e n tly th e m e e tin g f o ll o w e d th e enda which had been prepared by D r. N ew qulst p r io r to th e m eeting with s lig h t m o d ific a tio n in th a t the p ro g ress rep o rt and the research a c t i v i t i e s fo r th e y ea r b eg in n in g 7/1/53 were combined in both Dr. H orton's and Dr. P h a ir 's r e p o r ts . jTfMS P and ^ ~ DR. HORTON15..RSF.QRI As the b a s is f o r h i s d is c u s s io n s , Dr. H orton used the attached "Memorandum from- th e K e tte r in g L a b o ra to ry -F ifth M eeting of the Advisory Committee API R esearch P r o je c t MC-1" d a te d January 16, 1953 C o nsiderable d is c u s s io n was g iv en to the opening two paragraphs which j u s t if i e d the use of mice in a p p ra isin g the hazards of skin cancer in workmen. In response to a s p e c if ic q u e stio n from Dr. Adams, Dr. H orton s t a te d t h a t washing i s more e f f e c t i v e in reducing carcinogenic potency than a reduction in the dosage of carcinogen a p p lie d . As evidence he p re s e n te d th e ex p erim e n tal evidence which was o b ta in e d w ith washing experim ents w ith API-8 and API-113. I t was th en su g g e ste d t h a t c o n s id e ra tio n be giv en to a washing experim ent w ith API-113 and API-57 in which th ese o ils would be p a in te d on a t a le v e l o f 100 mg. two tim es e week and washed o ff a f t e r 9 hours in c o n ta c t w ith th e s k in . Perhaps the g r e a te st in te r e s ts in Dr. H orton's rep o rt are h is observations or th e o r ie s th a t the potency o f a p a r tic u la r m aterial i s dependent le s s on the amount o f carcin ogen than on i t s concentration; w hereas, the e f f e c t o f a c c e le r a to r s is dependent le s s on th e c o n c e n t r a t io n th a n on t h e t o t a l amount a p p lie d a t ea c h a p p l i ca tio n . A lthough t h is i s an I n t e r e s t in g th o u g h t, i t appears somewhat API 05569 " 3 * f f ic u lt to u n d e rstan d how t h i s could be th e s i t u a t i o n and how the oper d e fin itiv e experim ents could be conducted in order to con c lu s iv e ly e s t a b l is h th e s e o b s e rv a tio n s . The concept t h a t a c c e le r a to rs may a c t because of t h e i r s i m i l a r i t y in s t r u c t u r a l c o n fig u ra tion with the carcinogenic agents is an in te re s tin g one in so fa r as i t applies to benzo(c)phenanthrene and h ig h ly branched dodecylbenzene. I t i s d i f f i c u l t , how ever, to see how t h i s concept could apply to compounds l i k e 3 ,4 -b en zp y ren e or to dodecylbenzene in which the sid e chain i s n o t h ig h ly branched. The form ula p re s e n te d by D r. H orton on page 9 of h is report is an extrem ely in te re s tin g form ula from a th e o re tic a l standpoint, but whether i t w ill ever have any p ra c tic a l ap p lica tio n seems h ig h ly problem m atical s in c e th e d e te rm in a tio n of th e e f f e c t i v e c o n c e n tra tio n of a c c e l e r a to r in an o i l may be e x trem ely d i f f i c u l t , p a rtic u la rly i f th e re i s more than one a c c e le r a to r p re s e n t in such an o i l . S im ila r ly , such a form ula a llo w s f o r th e p resen ce of only two carcin o g en s, nam ely, benzpyrene and one C4 a ro m a tic . I t seems highly u n lik e ly t h a t th e re would be only two carcin o g en s p re s e n t in as complex a m ixture as s lu r r y o i l , and th is i s p a r t i a l l y borne out by th e o b se rv a tio n s re p o rte d by th e S tandard O il Development Company in which 6-is o p ro p y l b e n za n th ra ce n e was d eterm ined in a re la tiv e s ta te of p u r ity from one o f th ese o i l s . I t th e re fo re seems lik ely th a t th is formula w ill be exceedingly com plicated by a g reat number of a c c e le r a to r s and i n h i b i t o r s , such t h a t th e e f f e c t i v e con centration of a c c e le ra to r w ill have to be determined fo r each individual o i l , and a ls o by the presence of a la rg e number of carcinogenic compounds, which w ill h ig h ly com plicate the u sefu ln ess of any such e q u atio n . Another i n t e r e s t i n g o b s e rv a tio n made by D r. H orton during API 05570 - 4- discu ssio n vss th a t th e average l a t e n t p e rio d d e crea se s more h is 1 for papillom a3* depending upon the amount of a c c e le ra to r, average la te n t period fo r carcinom as. than the g ra p h ica lly exp ressed somewhat as fo llo w s: This may b e AVERAGE latent PERIOD AMOUNT ACCELERATOR TTCMS 2 AND 4 " DR. PHAIR'S REPORT Dr. Phair used as a b a s is of h is re p o rt the attach ed memorandum, which i s e s s e n t i a l l y a ta b u la r summary o f th e number of cases which he has thus f a r c o lle c te d from the v ario u s companies. I t appears th a t an in c re a sin g number of companies a re p a rtic ip a tin g in reporting cases to Dr. P hair b u t th a t th e re is a heavy im- ponderance of cases being re p o rte d from companies along the e a ste rn seaboard as compared w ith companies in southw estern U nited S ta te s . I t was agreed th a t Dr. P h a ir would o u tlin e b r i e f l y and send to the Chairman o f the Subcommittee on C a rc in o g e n ic ity h i s l a t e s t th o u g h ts on p e rtin e n t d a ta which he w ishes to have in c lu d e d in th e c u rre n t rep o rtin g of cancer c ase s among employes in th e p etro leu m in d u s tr y , Mid th a t the Chairman o f th e Subcommittee on C a rc in o g e n ic ity would then reattem pt to stim u la te a l l m edical d ire c to rs to re p o rt c u rre n t cases to Dr. Phair. API 05571 - 5 rff REP0RT Dr. Kehoe d isc u sse d the budgetary a llo c a tio n s of funds and in d ic a te d t h a t i t would be p o s s ib le f o r the K e tte rin g L abora tory to liv e w ith in th e p r e s e n tly a ssig n e d budget of $79, 000, b u t th at i t must be remembered th a t th is w ill r e s u l t in approxim ately one-quarter le s s work being done durin g the y ear Ju ly 1, 1953- June 30, 195^. Or. Kehoe wished to re s e rv e the r i g h t to a llo c a te funds under any o f the headings giv en , depending upon the K e tte rin g L ab o rato ry 's e s tim a tio n of how th e funds should be a ll o c a t e d . Thus, for instance, i f f u ll cooperation w ith Dr. Phair is not obtained, i t may w ell be t h a t th e $17,500 a llo c a te d f o r e p id e m io lo g ic a l work may be r e - a l lo c a t e d i n to o th e r phases o f th e R esearch P r o je c t. Biere seems to be a g e n e ra l agreem ent among th e members of th e committee th a t Dr. Kehoe should have freedom in a llo c a tin g th e t o t a l funds to any of the fo u r phases of the p r o je c t th a t he f e e ls the funds should be a llo c a te d to . D r. Kehoe a ls o made a p le a t h a t i f there is a determ ined e ffo rt to have a gradual reduction in the budget of th is p r o je c t du rin g the coming y e a rs , he would lik e to be informed of th is as f a r in advance as p o ssib le so th a t he might make h is p la n s a c c o rd in g ly . Thus, i f th e budget i s to be c u t one- half in the Year Ju ly 1, 195^"June 30, 1955, and to a q u a rte r of i t s p resen t fig u re in the succeeding y e a rs , Dr. Kehoe would lik e to know of t h i s as soon as p o s s ib le . ITEM 5 Dr. H orton discu ssed the re p o rt from the B r itis h on the c a rc in o g e n ic ity of th e m in e ral o i l s . He f e l t t h a t t h e i r t e s t s had been performed w ith a p p lic a tio n s of o i l which were so sm all th a t they are not g e ttin g tumors in the m ice, y e t they were observing ApI 05572 - 6- tumors i,rri twh1e5 r a b b its due to th e e f f e c t s of a c c e l e r a to r s . He seemed ^eei th a t th e f r a c t io n a ti o n o f t h e i r o i l s l e f t much to be d e s ire d , d th a t same f r a c t io n a ti o n o f o i l s in h i s l a b o r a t o r i e s , which were then ap p lied on h i s own an im als by h i s own te c h n iq u e s , had r e sulted in the appearance of a c o n sid erab le number o f tum ors, p a r tic u la r ly in the K-4, L-4 and 0-4 f r a c t i o n s . He f e l t t h a t t h i s vas probably the r e s u l t p r in c ip a lly of a c c e le r a to r s , and th a t the carcinogens would be found in the h ig h e s t c o n ce n tra tio n s in the K'5, L-5 and Q~5 r e s id u e s . I t was su g g e ste d a t th e m eeting t h a t the workers a t the K e tte rin g L aboratory f e e l fre e to communicate on a p ro fe s s io n a l b a s is w ith th e w orkers in England in d is c u s s in g th e ir mutual work, and fu rth e rm o re , i t was f e l t t h a t i f the K ettering L aboratory f e l t they vould lik e to keep th is communication in the e sta b lish e d chann els, th is could be done by having Dr. Horton prepare a c r itiq u e of the B r itis h work and then having Dr. Nevquist forward th is to the B r itis h vorkers through Colonel Auld. I t is evident th a t i t is too e a rly to draw conclusions w ith re sp e c t to the B ritis h work which has been re p o rte d thus f a r . item 6 Dr. Kehoe in d ic a te d t h a t he has w r itte n a supplem ental l e tte r to Dr. K otin su g g e stin g t h a t Dr. K otin more d e f i n i te l y o u t line what inform ation he would lik e to rec eiv e in o rd er th a t the K ettering L aboratory m ight be a b le to p ro v id e i t to him. Dr. Kehoe did not fe e l th a t any of the K e tte rin g L aboratory re p o rts should be sent to Dr. Kotin or anyone e lse since they are a t the p re se n t time so te n ta tiv e . I t was a ls o su g g e ste d t h a t a p o s s ib le means of handling t h is s i t u a t i o n would be to have Dr. K otin come in to th e Kettering Laboratory and d iscu ss th e ir mutual problems v e rb a lly , API 05573 - 7" and th a t such a d isc u ssio n could g e t to Dr. K otin a l l the inform ation vhic'i he m ight need and y e t n o t je o p a rd iz e the p o s itio n of the Kettering L ab o rato ry . Dr. Eckardt rep o rted to the committee th a t 5 a d d itio n a l cases of s k in can cer r e s u l t i n g from c u ttin g o i l s have r e c e n t l y come to h is a t t e n t i o n . One of th e s e c ase s vas a s c r o t a l c an cer and none of the cases had p rev io u sly been rep o rte d to the group. This simply served to emphasize the growing im portance of c u ttin g o ils as a possible carcinogenic agent in in d u stry . TTEM.B Some d is c u s s io n vas giv en to th e p o s s i b i l i t y of h o ld in g the next m eeting o f th e A dvisory Committee a t the H otel Shamrock on the 29th of Septem ber, 1953, and th a t a t th is time i t vould be a jo in t m eeting w ith th e S u b c o a n itte e on C a rc in o g e n ic ity . D ecisio n on th is m a tte r vas l e f t to th e Chairman o f th e Subcom m ittee, w ith the recommendation th a t i f the m eeting is to be h eld on th a t d a te , members should Jhe n o t i f i e d and h o te l r e s e r v a tio n s should be made veil in advance. M. N. N ev q u ist, M. D ., Chairman R. E. E ck h a rd t, M. D. Acting Secretary API 05574 ifPMOPANDUM FROM THE KETTERING LABORATORY p t w h mkpting of the advisory committee APJ RESEARCH PROJECT MC-1 January 16, 1953 . use 0f mlc e in th e e x p e rim e n ta l a p p r a is a l of th e r e l a t i v e A. hazards of c an c er of th e a k in o f workmen from exposure to v a rio u s petroleum products. 1 M aterials which in in d u s tr ia l experience have caused cancer of th e s k in of workmen have been shown to be c a rc in o g e n ic to mice. 2. The c o r r e l a t io n of th e re s p o n se s o f men and mice is p a r t i c u la r ly c lo se i n th e case of s t r a i g h t ru n wax d i s t i l l a t e s , cancers having re s u lte d in both species from in te rm itte n t, severe exposure (100 mg. doses a p p lie d th re e tim es p e r week on th e s k in o f th e m ic e ), when th e c a rc in o g e n ic m a te ria l was n o t removed by w ashing. Data re p o rte d by the New York U n iv e rs ity group I n d ic a te t h a t mice develop very few tumors when low dosages (15 mg. p e r a p p lic a tio n ) of wax d i s t i l l a t e s a re a p p lie d . The mouse, lik e man, seems to be a ff e o te d a d v e rs e ly by th e s e o i l s only when exposure ia se v e re . A co n firm ato ry t e s t a t low dosage w ill be c a r r ie d ou t on C3H mice w ith th e M idcontinent p a r a f f in d i s t i l l a t e , API-56 (PMC 0 .1 a t th e 100 mg, level of dosage). B. R elatio n sh ip s between p o ten cy of o i l s and th e l e v e l of dosage applied upon the skin per a p p lic a tio n . 1. V a ria tio n of th e r e l a t i v e p o te n c y , PMC, w ith th e l e v e l of dosage (over c e r t a i n ra n g e s o f dosage) is j u s t what one API 05575 - 2- *ould ex p ect on th e b a s is o f human e x p erien c e w ith such allagd carcinogens as coal t a r , shale o il or the d i s t i l l a t e s which have been h a n d led In wax p re s s e s In th e petroleum In d u stry , i . e . , the higher the le v e l of exposure, the g r e a te r th e chance o f s k in c a n c e r, The r e s u l t s of the te s ts on mice a t d if f e r e n t le v e ls of dosage which have been rep o rted fo r the c a ta ly tic a lly cracked re sid u a , API-8 and 71 a re th e r e f o r e n e it h e r s u r p r is in g n o r d i s tr e s s in g when c o n sid e re d from th e o v e r a ll a sp e c t o f the program. R eally unexpected r e s u l ts are coming from the t e s t s on sta n d a rd s o lu tio n s and r e f in e r y stream s c o n ta in in g e ith e r very low o r very high le v e ls of c o n ce n tra tio n of accelerating co n stitu en ts. 2. Certain materials demonstrate a constant potency over a broad range of dosage: (1) S o lu tio n s of m eth y lcholanthrene or benzpyrene in benzene (2) Some blended i n d u s t r i a l f u e l o i l s , e . g . , API-113 (3) Some th erm al t a r s from v i r g i n f e e d s , e . g . , API-108 (1*) S o lu tio n s of m e th y lch o lan th ren e or benzpyrene In dodecylbenzene (5) Some c a t a l y t i c a l l y crack ed gas o i l s , e . g . , API-102. Types (1 ) , (2 ), and (3 ) p robably c o n ta in , a t the m ost, very low c o n c e n tra tio n s of a c c e le ra tin g c o n s titu e n ts , API 05576 - 3ty p es (U) and (5) r e l a t i v e l y h ig h c o n c e n tr a tio n s . The type8 of o ils discussed in sectio n B .l. apparently have a c r i t i c a l l e v e l of c o n c e n tra tio n of such com ponents. The s i t u a t i o n may b e s t be p ic tu r e d g r a p h ic a lly (see F ig u re 1 ): Figure 1 Thus, i f no a c c e le ra to rs are p resen t in an o il (A), i t w ill ex h ib it only the potency due to i ts carcinogen content. S im ila r ly , o i l s c o n ta in in g a c c e l e r a to r s w i l l show only th e ir base le v e l of potency i f the dosage per ap p licatio n is kept s u f f ic ie n tly low so th a t the amount of the API 05577 - k- a c c e le ra to r a p p lie d to th e sk in of th e mouse each time does n o t exceed B. What dosage t h i s w i l l be f o r any given o i l obvio u sly depends on th e c o n c e n tra tio n of a c c e le r a to r s in the o il. I f the dosage per ap p lica tio n of such o ils is increased u n t il the amount of the a c c e le ra tin g c o n s titu e n t a c tu a lly being a p p lied each time reaches the c r i t i c a l range between B and C, th e r e l a t i v e p o te n c y , PMC, as m easured by a shortened la te n t p erio d fo r tumor in d u c tio n , w ill r i s e . This e ffe c t w ill then in crease as h ig h er le v e ls of dosage are t e s te d u n t i l th e maximum resp o n se f o r a g iv en concen t r a t i o n of c a rc in o g e n i s o b ta in e d (upper p la te a u in F ig u re 1). P roceeding in th e r e v e rs e d i r e c t i o n (from D tow ards C) i t w ill be apparent th a t the h igher the le v e l of the con c e n tra tio n of a o c e le ra to rs in an o i l , th e more the dosage p er a p p lic a tio n may be reduced b e fo re a s i g n i f i c a n t red u ctio n in potency w ill be observed. This category probably in clu d es a la rg e number of the c u rre n t d i s t i l l a t e gas o i ls w ith End B o llin g P o in ts > 7 0 0 ?. from f r a c tio n a to r s of o a ta ly tlc cracking u n its . Because of the obvious importance of these re la tio n sh ip s between the amount of a c c e le ra to r applied and the ra te a t which an o i l co n tain in g carcinogens w ill induce cancer in exposed anim als, step s are being taken to e sta b lis h : API 05578 - 5- (1) the exact magnitude of le v e ls B and C (w ith blends of synthetic hydrocarbons), (2) the c la sse s and b o ilin g ranges of the hydro carbons in c a ta ly tic a lly cracked gas o ils and resid u a resp o n sib le fo r the a c c e le ra tio n observed, and (3 ) the le v e ls of c o n ce n tra tio n of such hydrocarbons in various re fin e ry streams The nature of the compounds contributing to the carcinogenic potencies of refinery streams. 1 , Polycyclic carcinogenic compounds. A d ir e c t method of a n a ly s is fo r the carcin ogen, 3,^-benzpyrene, has been developed. I ts r e lia b ility has been te ste d and confirmed w ith four of the major types of refin e ry stream s under consideration, Including cycle gas o ils and bottoms products from c a ta ly tic cracking, ta rs from therm al cracking of c a ta ly tic fe e d s, and stra ig h t run d is tilla te s . By a p p lic a tio n o f t h i s m ethod, i t has been shown t h a t about 70% of th e base potency e x h ib ite d by th e FCC decanted o i l , A PI-8, a t 5-20 mg. le v e ls of dosage i s ji API 05579 c o n trib u te d by benzpyrene. A pproxim ately i+O# of the c o n sta n t Pj^ v alu e of th e No. ^ f u e l o i l b len d , API-113, is due to b en zp y ren e. b. As m ight be e x p e c te d , in view o f th e re p o rte d b o ilin g p o in t of 3 ,l|.-benzpyrene (8 6 0 P ,), i t has been found th a t the d is tilla te re fin e ry stream s analyzed thus fa r, with tru e end b o ilin g p o in ts l e s s th a n 860, c o n ta in r e l a t i v e l y sm all amounts of th is 5-rin g e d carcin o g en . Itie a v a ila b le data from b io lo g ic a l t e s ts on fr a c tio n s prepared from c a ta ly tic a lly cracked o ils suggest strongly th at the im portant carcinogen in such oycle gas o ils is a i^-ringed d eriv ativ e of phenanthrene. Since the re s u lts fu rth e r in d ic a te th a t sim ple a lk y l d e riv a tiv e s of pyrene and 1 , 2-benzanthracene API 05580 - 7- are not contributing sig n ific a n tly to the carcinogenicity of these o ils , a tte n tio n is being centered upon benzo(c)phenanthrene. This is the hydrocarbon of low est m olecular weight which has been shown to be c a rc in o g e n ic w ith o u t the b en efit of m ethylation. pure samples of benzphenanthrene and of each of i t s six monomethyl d e riv a tiv e s have been o b tain ed . T h eir chemical and p h y sic a l p ro p e rtie s are being examined in the development of a d ir e c t method of a n a ly sis a p p licab le to refinery stream s. 2, Accelerating constituents - 475 to 700 boiling range. I t i s o f co n sid erab le i n te r e s t th a t the number of carbon atoms ( 18 ) in th e a c c e le r a to r , dodecylbenzene, i s the same as th e number in th e c a rc in o g e n , b e n z o (c )phenanthrene. The known c a rc in o g e n ic hydrocarbons have Carbon Numbers from 16 to 2 5 . The approxim ate b o ilin g range of branched-chain alkylbenzenes w ith 16-25 carbon atoms i s 475-700P ., which i s th e very range th a t has been suspected of having a c c e le ra tin g a c tiv ity from the r e s u lts o f b io lo g ic a l t e s t s on r e f in e r y stream s and f r a c tio n s th e r e o f . I t seems l i k e l y t h a t we have h ere something more than J u s t c o in c id e n c e . The problem of th e i d e n t i f i c a t i o n o f th e im p o rtan t accelerato rs in re fin e ry stream s is being approached in two ways: (1) B io lo g ical te s tin g of c e rta in sy n th e tic liq u id hydrocarbons as solvents fo r benzpyrene, to determine ApI 05581 - 6- which ty p es have a c c e le r a tin g p r o p e r t i e s . The r e s u l t s obtained in the prosecution of th is phase of the program w ill fu rn ish a d e fin ite o u tlin e of the range of m olecular weights and of the s tru c tu ra l types of possible importance. (2) B io lo g ic a l te s tin g of fra c tio n s of c e rta in c a ta ly tlc a lly cracked o ils which have dem onstrated (in t e s t s a t th e 100 mg. le v e l of dosage) p o te n c ie s th re e to te n tim es a s h ig h as were expected on the basis of th e ir content of four to six ringed arom atic hydrocarbons. These e ff o r ts are being concentrated on th e components b o ilin g below 700P. which c o n ta in no t more th an two fu se d arom atic r in g s . The fo re g o in g phase (2) of t h i s program has already led to the concentration of a c o lo rless, liq u id f r a c t i o n from th e TCC bottom s p ro d u c t, API-71, which p o ssesses the s p e c ific i r r i t a n t p ro p e rtie s of dodecylbenzene. This fra c tio n is being r e d is tille d through an 80- p l a t e column to p re p a re a number of w e ll-d e fin e d c u ts to be te s te d as p o s s ib le a c c e l e ra to rs of the carcinogenic actio n of benzpyrene. In g e n e ra l, i t i s a much sim p le r m a tte r to separate sa tu ra te d hydrocarbons and monocyclic aro m atics from o ils and from each other than i t is to f r a c tio n a te any s in g le c la s s of p o ly c y c lic compounds from an o i l . Furtherm ore, in the se p a ra tio n of compounds b o ilin g in th e range 500-6$0 a t 76O a n . , API 05582 - 9- h ig h ly e f f i c i e n t d i s t i l l a t i o n s may be c a r r i e d out a t reduced p re s s u re w ith o u t f e a r of c ra c k in g . Hence there is l i t t l e reason to doubt th at system atic a p p lic a tio n of a v a ila b le techniques w ill lead to an adequate knowledge of the s tr u c tu r a l types of the acceleratin g c o n stitu en ts of re fin e ry streams in a reasonable p e rio d of tim e. The ste p from th a t knowledge to a d ir e c t method of a n a ly sis fo r th e ir e ffe c tiv e co n cen tratio n in any given stream should present no g re a t d i f f i c u l t i e s . Physicochemical methods for predicting the relative potencies of refinery streams. During the l a s t s ix months, the com pletion of the p ictu re (Figure 1) of the in te r r e la tio n s between concen tra tio n of carcinogen, concentration of accelerato r, dosage p e r a p p li c a t io n , and r e l a t i v e p o ten cy , PMC, has brought with i t a fin n confidence th a t a re lia b le ana ly tic a l to o l w ill r e s u lt from the fu rth e r extension of th is fundam ental approach to th e problem . The e q u atio n fo r which param eters are being determ ined i s of the f o l lowing ty p e : ^MC s + * where f ( c a d) i s a fu n c tio n of the amount of the a c c e l e ra to r component ap p lied in each a p p lic a tio n of the whole oil ca being the e ffe c tiv e concentration of a cc elera to r in the o il, and d the to ta l dosage of o il per ap p licatio n ; API 05583 - 10 Cg and a re th e c o n c e n tra tio n s o f the 5 -rin g e d Denzpyrene and of the 4-rln g e d carcinogen re s p e c tiv e ly , while Kg and are the re sp e c tiv e p o ten cies of benzpyrene and th e ^ -rin g e d compound based on m e th y lc h o la n th re n e . Since the values fo r h a lf of the fa c to rs in the e q u a tio n f o r c a l c u l a t i n g PMC a re a lre a d y known (d , Kg and Cg ), the rem aining Job Is the d eterm in atio n of ca , and the b reak p o in ts a t B and C in the curve of Figure 1. These phases have been discussed in S ections 5*2, C .l .b . and C2* API 05584  P I I ESTIMATED BUDGET OP THE KETTERING LABORATORY foR THE INVESTIGATION OF POTENTIAL HEALTH PROBLEMS OF THE PETROLEUM INDUSTRY ASSOCIATED WITH THE PRESENCE OF CARCINOGENIC COMPOUNDS IN CERTAIN MATERIALS AND PRODUCTS API RESEARCH PROJECT MC-1 July 1, 1953 - June 30, 19& Salaries Direct Salaries $ 30,003.00 Indirect Salaries Histopathologlcal'Preparation $ 2,000.00 Other Services $ 22,000.00 Miscellaneous Expense Purchase of Animals $ 1,500.00 Special Laboratory Supplies $ 5,500.00 Travel i l Overhead $ i^OOO.OO (Proportion of h eat, gas, e le c tric ity , steam, telephone, g en eral lab o ra to ry supplies, postage, an n u ities, pensions, m aintenance, e tc .) $ li|.,000.00 T otal $ 5 k ,000.00 $ 25, 000.00 $ 79,000.00 ESTIMATED BUDGET OP THE KETTERING LABORATORY FOR THE INVESTIGATION OF POTENTIAL HEALTH PROBLEMS OF THE PETROLEUM INDUSTRY ASSOCIATED WITH THE PRESENCE OF CARCINOGENIC COMPOUNDS IN CERTAIN MATERIALS AND PRODUCTS API RESEARCH PROJECT MC-1 J u ly 1, 1953 - June 30, 195U T Further development of se m i-q u a n tita tiv e b io lo g ic a l te stin g methods f o r m easuring the r e la tiv e carcinogenic potencies of petroleum products and f r a c tio n s th e re o f $ 12,000.00 l it Determination of the re la tio n s h ip s between the * rap id ity of induction of benign and m alignant tumors in experim ental anim als and v a rio u s exposure fa c to rs such as frequency of ap p lies tion, dosage per a p p lic a tio n , area of exposure, number of a p p lic a tio n s , s p e c ia l i r r i t a n t or toxic p ro p erties of the o il ap p lied , e tc . $ ll|,500.00 III# Development o f rap id a n a ly t ic a l methods fo r estim ating the re la tiv e carcinogenic potency of any given r e f in e r y in te rm e d ia te or p roduct A. I s o la tio n and i d e n t i f i c a t i o n o f compounds responsible fo r observed potency of various types of o ils and determ ination of p e rtin e n t physical and chem ical prope- tie s of these compounds $ 30,500.00 B. Development of se m i-e m p iric a l methods o f $ Ij., 500.00 estim ation of carcinogenic potencies of o ils based on the chem ical o r p h y s ic a l p r o p e r tie s o f known o r su sp e c te d c a rc in o g e n s and a c c e le r a to r s IV. E pidem iological Program C o lle ctio n and c o r r e l a t io n of d a ta on c a se s of cancer among employees o f the p etroleum in d u s try including any sp e cia l in v e stig a tio n s in p a rtic u la r plants of the industry $ 17,500,00 T otal $ 79*000.00 API 05586 FIGURE 1 - 1007* CASE REPORTS FROM 13 COMPANIES RECEIVED BY THE CENTRAL REGISTRY ACCORDING TO SEX, RACE AND MALIGNANCY SEX RACE MALIGNANT White 745 C olored 26 MALE Not Given 114 TOTAL 885 White 33 FEMALE Not G iven 9 TOTAL 42 NOT GIVEN 4 TOTAL BOTH SEXES 931 BENIGN 23 38 61 1 5 6 - 67 TOTAL 768 26 152 946 34 14 48 4 998 Nine reports could not be used. Six w ere not tum ors; one of these was Boeck's Sarcoid. Three gave only the occupational h istory. API 05587 ) ) I Type of 1 Tumor Site Buccal C avity 00 -09 D igest. Sye. and P e r it. 10-19 R esp iratory System 0-29 Gland. Epith. 00-09 11 247 28 Breast 30-39 1 Genital 42 Organs 40-49 SUyrsinteamry 5 0 -5 9 21 Skin and Soft Tissue 60-69 Bones 70-75 6 Non Gland. Epith. 10-19 64 24 91 1 16 21 170 3 Brain 76-79 2 Lymph, and Hem. System 80-89 Other Sites 9 0 ---- 8 27 Total 364 419 ACCORDIMO TO TTPC O f TUMOR AND PRIMARY SITE Leuke Lym pho Nervous Vasculax m ias m as T issu es T issues M uscle 20-29 30-39 40-49 50-59 60-69 1 IttO Non- \ Em bry. \ Tumors! Epith. \ - M ixed \ Not t i s s u e s i t i s s u e s v^iasB. 70-79 1 80-89 90-- 1 Total ^ 77 4 * 2 7 3 285 1 2 3 125 2 11 69 1 1 44 4 2 3 181 1 2 1 13 3 1 13 19 12 17 1 12 24 5 1 29 1 4 41 17 17 26 885 1 API 05588 ) 1 Type of | Tumor 1 Site 1 Buccal 1 Cavity 00-09 I D ig est. S y s. and I Perit. 10-19 1 R esp iratory System 20-29 Gland. Epith. 00-09 8 1 B reast 30-39 14 GO erngaitnasl 4 0 -4 9 2 U rin ary System 50-59 Skin and Soft 1 T issue 60-69 Bones 70-75 Brain 76-79 Lymph, and Hem. System 80-89 Other Sites 9 0 ---- Total 26 Mm FIGURE 3 - MALIGNANT TUMORS IN FEM ALES AS REPORTED ACCORDING TO TYPE OF TUMOR AND PRIMARY SITE Non Gland. Epith. 10-19 1 Leuke m ias 20-29 Lympho Nervous Vascular mas T issues T issues M uscle 30-39 40-49 50-59 60-69 NonEpith. T issu es 70-79 Embry. - Mixed T issues 80-89 Tumors 1 1 Not 1 \ C la ss. I Total l 90--- 1 \ 111 18 1 1 " 12 1 1 1 15 1 3 5 1 11 1 6 7 1 |1 1 1 14 1 2 1 42 API 05589 ) ) ) I T I D B t BEX AMD PR IM A R Y S IT E COM PARED WITH U .8 .A . MORBIDITY AND M ORTALITY E ST IM A T E S M SEX Site Number Reported MALES Per Cent of Total U. S. A. Per Cent Cancer C ases U .S . A.** P er Cent Cancer Deaths Buccal Cavity 77 Danidg ePsteirvietoSnyeustme m 285 R esp iratory System 125 Breast . 2 8 .7 32.2 14.1 0 .2 10.0 36.0 8 .0 4 .7 4 7.7 1 5 .4 0 .2 Genital Organs 69 7 .8 12.0 1 3 .0 Urinary System 44 Skin and Soft T issue 181 Bones 13 Brain 19 tLoypmoipehti,canSdyHsteemm a 29 Other Sites 41 5 .0 7 .0 6 .6 20.5 17.0 2 .2 1.5 1.3 2.1 2 .4 3 .3 I 4. 6 j 10.0 j 6.5 TOTAL 885 100.0 100.0 100.0 Number Reported 1 8 2 15 5 1 7 1 2 42 FEMALES P er U .S . A.* Cent P er Cent of Cancer Total C ases U .S. A.** P er Cent Cancer Deaths 2 .4 2 .0 1.2 19.0 2 3 .0 4 .8 2 .0 35.7 11.9 ) 51.0 2 .4 3 .0 16.7 11.0 2 .4 38.7 3 .9 19. 1 2 3 .5 3 .4 1.5 1.1 1.5 ( 4.8 ) 8.0 100.0 100.0 / 6.2 100.0 BOTH SEXES V Number R ep orted P er J . s . A.'^ W Cent Per Cent rl of Cancer l Total Deaths | 78 8 .4 2 .9 293 31.6 4 3 .2 127 13.7 9 .6 17 1.8 9 .7 74 a.o 18. 3 45 4 .9 5 .0 188 20.3 1.8 14 1.5 0.1 19 2.0 2 .0 29 3.1 43 4. 6 7 .4 927 100.0 100.0 * P rim ary site of developm ent of cancer among white m ales and fem ales according to Dorn, Harold F . : Illn ess from cancer in the United States. Reprint No. 2537, Public Health R eports. (B ased upon m orbidity records collected from ten large c itie s in the United States betw een 1937 and 1939.) ** Cancer deaths by site in the United States for 1948 as prepared by the S tatistical R esea rch S ection , A m erican C ancer Society, utilising sta tistics from the National O ffice of Vital S tatistics. API 05590 I' 1 / torch 17, I53 j r . r i t f f Daria _____ nnU ipe Patrola Coapaay artlo av illa, Cfclafcaaa M r Or. D * t 1* i eoMlttM oa Carainaeaatcity, aialcaa copy of tha ttatmnt af *** bjr Cattarla^ lafrciratorj la m u --*-- ito raaaarcb prajact MC-1. A cco rila to ajr raco m a ad th ia ta ta a a o t tb e ta l asv aaU tw aa by C a tta rla 19*5-1952 to v a aaoaatad to *3*3,7*9.79, lo a rla * a b a la a e a o f $3*,73T.12 valX tbla a jaaaary 1# 1953 t a a a a t aspaaaaa a ria ha f ix t tv o a a a n a ra T 1953 ta b a lita d . tbry traXr yoara, i DVSiJb ce: Or. I . M. ftaak SBelaaara API 05591 gjpy fr Hr. D. v. Stroop for Iafaraation f Dr. Slattar D*rl* ad Or. I. K. Frank v u 'iu s iS ! or c ik u ia h H a B a tte ria * Taliu r iti jtj Col l a nt e t N adiela - Id a * n m C ln rlim a ti 19, Ohio r**m ry 17, 1953 Xr. i.. V. 3treop, DlMctcr Im yertm nt ut T eeiuileal h n lO M Marie FetroLaam X a a tita ta 50 wt 50tix stntk i n Torte 90, lo Torta Dui Mr. Straopi I a M iiae / m te r a a itt ta te m t of MQaailturM m oc M te lf f A ir i-- Fatroleaa Iteti fo r th itaarth qiartor ef 1952. I teli** te i te r ta t-- n t ili te Lf-oxpLumiary, te t lf / m t e * mv uM tiow or caonenU, Scctor Kteoe moni P ir cl te /M r te la c i * te a o hi* tte a lio n . Tory raly jo u ro , / / I . B. F ortla, te c ro ta r7 te O r. Stee f tee. API 05592 m ir a g g i or c a n o t a i JBHBHIBO LABCBATCKI ACCCDIT GP Tfat Aaarlcpa Petroloaa In a tltrta FOB Fourth Oaurtur 1952___________ &tU8X** (B-- d od Proportion at firn A c tu ally fpast eu P ro je t) D ln et Salarlos .............................................. lodlract Salarlos 963.6k K ls to p a tb o lo c la s l F n p a n t i ............. 990.00 O tto r a rriem ............................................. 9073.20 jjjjgmjasccs a ? s ia F i i w n o f A sios! ..................................................... AM.0O S p o sisi la b o ra to ry S u p p lias .................................... 39^.M BwvdI ....................................................................... l& A .* Ororfeoad (P ro p o rtio n o f S o o t, Qa , Q o s t r ic i ty , M m d , T slsjfcaso, O oasral L aboratory S u p p lias, P oot a i i A M ltiM , P-- 1-- , I i I r t i M i i , te , $732.91 1 1 1 m i ra ila fe la f a r P t ft h t r Vecfc * * W t * - 195 TOZAL 3 * ,3 te.* 9 l a l a -- Dur b t t a r i a i la b o ra to ry 1 9 ,7 5 1 . 8,206.$1 ZLSS& 2 faU X Sou A vollabla *-- a l t -- o ktfo 9 ^ g a li-- A sa ilsb lo f a r Farth o r Vark a* -- af q u g ft r 1952 BdI u m Ora s o tta r lo s Lai a t sd af k 6 .-m .9 6 8 0 ,7 1 5 .* 7 2 7 .9 5 8 .3 5 5 2 ,7 5 7 .1 2 API 05593 A m erica n P etroleum Institute 8 0 WEST bo th STREET NEW YORK 20. N. Y. tech n ica l s ia v ic fs D'" CT0' March 2k, I 953 To Members and Associates Medical Advisory Committee Gentlemen: Dr. Kieffer Davis, Chairman, Subcommittee on Carcinogenicity has appointed for 1953 the subcommittee's Research Project MC-1 Advisory Committee comprising: Robert . Eckardt, M.D., C. E. Eine, M.D. E. K. Linder, M.D. M. N. Bevquist, M.D. . V. Adams L. C. Beard, J r . L. M. enderson R. C. Mithoff Chairman I t is our hope that each appointee can take an active part in the important work of th is committee, if not, we shall expect him to notify Dr. Davis promptly. Very truly yours, DVS:Jb API 05594 ,,f0,CALoe" "TMENT ,,IT" .. C T O . . h u n t in o t o w . m 0 C* - Bl . C T O . PHILLIPS PETROLEUM COMPANY BARTLESVILLE. OKLAHOMA March 18, 1953 ALBERT MERY w o A S S O C I A T A P h VSi c i AN LU CIAN C RENES iH O U T l*L MV6ICMIST Mr. D. V. Stroop American Petroleum In stitu te 50 West 50th <1* New Tork 20, New York Dear Dave: I wish to appoint the following as members of our RP*^iC-l Advisory Committee: Robert I . Sckardt, M.D., C. H. Hino, M.D. E. K. Linder, M.D, M. N. Newquist, M.D. E. W. Adams L. C. Beard, J r . L. M. Henderson R. C. Mithoff Chairman Please inform each of these men of th e ir appointment and notify me in case anyone should decline. Unless there is some reason to change our plans, the next meeting of our Subcommittee on Carcinogenicity w ill be at the Shamrock Hotel in Houston September JO, 1953* With best personal regards, I remain Sincerely yours, I KD:dt I ec: George M. Saunders, M.D L Kifer Davis, M.D. Medical Director API 05595 # n y. Stroop for Information of Merisers and Associates of Medical ___________________________________ April 3. 1953 ESSO LABORATORIES Standard Oil Development Company F. 0. Box 51 Linden, H. J. March 18, 1953 w- p. V. Stroop V^ericsn Petroleum In stitu te 50 West 50th Street 1^, York 20, Hew York ptarMr. Stroops The enclosed apparently represents a publication of the data which members of the Subcomnlttee on Carcinogenicity of tbs A.F.I. have had an opportunity to review in connection rttb the exchange of information between the B ritish and our own gubcoamittees on Carcinogenicity. I thought you might wish to have th is a rtic le reproduced and distributed to the meribers of the Medical Advisory CoHittee of the American Petroleum In stitu te . Very tru ly yours, / s / R. E. Eckardt R. E. ECKARDT, M.D. RB/hsl Enel. (The Value of the Rabbit For Carcinogenicity Testa on Petroleum Fractions, "7 Hiefer and Voodhouse *pr. from THE BRITISH JHL. CAKER, 1952, Voi. VI, Pg. 2930 API 05596 RpjiriiiWxl from fh t R ritith Journal o f Cancer, I932, Vol. VI, p. 293. the value of the rabbit for carcinogenicity tests ON PETROLEUM FRACTIONS. I. HIEGER ASD D. L. WOODHOURE. fr o m the C hester B ra ity R esearch I n s titu ir . T he B o y a l C an cer H o sp ita l, L o n d o n , S . W . 3, 15. a n d the C a n te r R esearch L aboratories, D ep a rtm en t c f P ath ology, The M edical School, B irm ingham , HerpivoU tor publication May 2H, 1052. T he experiment recorded inthisarticlewerecarriedoutinthecourseof invtMtigatioiix on behalf of the Medical Research Council, a sub-committee having iieen "ft up in 1949 to inquire into the carcinogenic properties of mineral oils and allied products. The arrangements for co-ordinating the research have been briefly describedbyAuld(1930). Theentireseriesoftestshavebeenmade induplicateattheCancerResearch laboratories, Department ofPathology, UniversityofBirmingham, and at the (,'hester Beatty Research Institute, The Royal Cancer Hospital, London. At each centre the activity of3 selected crude oils,4 fractions derived from each crude by methods of distillation specially designed to avoid high temperature cracking, and the final residues, have been tested on groups of 50 mice for each simple and alsoon a totalof 105 rabbits. The resultsappear to be ofparticular interest, since it must be concluded from them that it is unsatisfactory to exclude carcinogenicity on the basis of teats on mice only. The desirability of utilising in industry those types ofoils which are least likely to cause dermatitis or cancer of the skin of workers has been recognised for over 2<>years,andin1934 theCommitteeonCancerofManchester Corporation proposed a standard for assessing the potency of mineral oils depending on physical properties (specific gravity and refractive index) which it was believed would ensure that the lubricants used in the textile industry would be substantially free fromcarcinogenicproperties. ThisManchesterformula,whichisnotnowregarded as a satisfactory criterion, was put forward following the investigations of Twort Andhisco-workers(TwortandFulton,1929;TwortandTwort,1930,1931,1933 ; Twort.andLyth, 1939 :Twort, 1941), whotestedawiderangeofcrudemineral oils, distillates,solventextracts,spindleandshaleoils,usingmicealmostexclu sively,however, for the biological assessment of their oarcinogenicity. In the past,also,micehare forthemost partbeenemploj'ed inskintestswith "oil" fractionsderived from coal tars and with the pure chemical carcinogens either isolated or synthesised. ItistruethattheclassicalexperimentsofYamagiwa and Itchikawa (1918), in which the carcinogenicity of coal tar was first proved, utilised rabbits, and this species has also been employed in a number of early experiments on tar canoer, e.g,, Bonne (1927), Leroux (1927), Babes (1929) and Twort and Twort (1930). Also a few early worken have used other species to a limited extent, e.g., rats and guinea-pigs (Watson, 1932). The majority, however, have favoured themouse for large scale experiments, no doubt partly beoause of low maintenance costs and API 05597 294 1. HIEGEB AND D. X. WOODHOISE Availability, but probably influenced alio by the experience of early workers that theakin ofratadid notreadilyrespond tocoal tan, and that papillomata on mbbitsaoproduoedremainedamallandoftenregressedwhenapplicationaceaaed. Thua the mouse has become aooepted aa the most sensitivetest animal for such work ;thepossibilitythatotheraperiesmight provemore suitableunder certain circumstances does not appear to have been sufficiently appreciated. Schurch (1939), however, found that hit experiments with rabbits indicated that carcinogenic ingredients is addition to the reoognised carcinogen 3 : 4-benzpyrene were probably present is oertain coaltare. This was pointed out by Berenblum (1947),who demonstrated that the relative carcinogenic activity of a number of fractions, obtained by chromatographic procedures, varied greatly according to whether standardised against the akin of the mouse or ofthe rabbit. He had already abown (Berenblum, 1945a) that tumours were readily elicited on the rabbit by 9: 10-dimethyl-1:2-benzanthracene, which was found to be more potent to rabbits than to rats or guine&pigs, in which positive results were, however,obtained (Berenblum, 19456). He alsosubsequently observed (1947) that this hydrocarbon when injected subcutaneously did not induce rabbit tumours, though ratsand guinea-pigs responded by thismethod. Much earlier, Oberiing,Saanit,GuerinandGuerin(1937)hadshown thata 1percentsolution of 3:4-banzpyrene in benzene was active on rabbits' skins, and Berenblum (1945a) observed that 9: 19-dimethyl-1: 2-benzanthraoene was much more active than benzpyrene at a similar (0* 5 per cent) concentration. Experiments mung a high boiling fraction of mineral oil from an experimen catalytic cracking operation have recently been carried out by Smith, Sunder andSughm (1951),testingtheresidueafterremovingthelowerboilingnapht and light gas oil cuts, upon mice, rats, guinea-pigs, rabbits and rhesus monk Hie ratsand guinea-pigswere found tobe refractorytoskinapplications papillomata were elicitedin allofthe 6 monkeys, 2 being proved cancerous biopsy4yeanafterthestartoftheexperiment. Papillomatawereproduced theinnersurfaceoftheearsofthe21rabbitswithin100days. Thenumber siaeofsuchgrowthstendedtoincreaseduringthe2yean inwhichpainting1 continued, and in 3 of the 6 surviving animals cancerous changes in the grov were observed. For testson the type ofmaterial which they intended to sta namely, samplon containingoilswhich had been subjected to a prooess offl catalysisupto950F.inthepresenceofaluminaorsilica,theseworkersconch that white mice were the best animals. . Recently Cruikshank and Squire (1950) obtained numerous papillomata the ear and body of albino rabbits after applications ofa "cutting oil obtainedfrom a machine sump. Only Ibenign papilloma was produced by t oilinagroupof46mice,ofwhich50percentsurvived40weeks'and28per 52 weeks'treatment, and they suggested that such materials should be against both species before being regarded as non-carcinogenic to man. In the present investigation it was decided that it was of importance that properties of the mineral oils in question should be tested on both rabbits and m' C ru des a n d o il fraction*. The 3 crudes were obtained from Kuwait, Lagunillas and Oklahoma reapecti and had the following characteristics: API 05598 of early workers that that papillomata on n applications ceased, sitivetestanimal for >more suitableunder fficiently appreciated, rabbits indicated that iogen 3:4-benzpyrene tedout by Berenblum activity of a number >d greatly according to ftherabbit. He had eadily elicited on the aa found to be more positive results were, aently observed (1947) did not induce rabbit method. Much earlmr, ita 1per cent solution skins, and Berenblum atrigmuchmow frctive ,1fromanexperimental it by Smith, Sunderland ,e lower boiling naphtha its and rbesuB monkeys, bo skin applications but ng proved cancerous by omata were produced on days. The number and rs in which painting was s changes in the growths i they intended to study, ted to a process of fluid ,theseworkersconcluded lumerous papillomata on tions of a "cutting oil ma was producedby this 4aweeks'and28percent lateralsshould be tested logenic to man. ,-as of importance that the lonbothrabbitsandmice. and Oklahoma respectively CAI(C l.VUC;E.VR'lTi- UK 1'ETltUL.El'M KK.HTIU.Vi> l.V KAULUTS 295 Kuwait (KuwaitOU Co.):anextensivelyproducedMiddleEastcrudelikeltf tobemuchusedintheUnitedKingdom,ofparaffinic-asphalticbaseandcontaining lubricating oil fractions. 6 st/jiiuillas Venezuela (Shell Petroleum Co.): a typical naphthenic crude vieldiiig well-known lubricating distillates. ' O klahom a <"ity Mid-Continental (Socony Vacuum Oil Co.): yields Mid Continental distillates.brightstockandresiduallubricatingoils. * * The derived fractioas were produced by a research group appointed by the Instituteof Petroleum, which has been responsible for the choice ofthe crudes havingregard totheirtype, distributionand industrialimportance. The initial fractionation of the 3 crudes was carried out at delimited temperatures using vacuum anilsteam inan apparatusselectedtoprecludecracking.- Theseoperations were carried out at the Thornton Research centre, Shell Refining and Marketing Co. Ltd., for the Institute of Petroleum. Fulldetails regarding the physical characteristicsofthesesamples are not jwrtinent to the purpose ofthis communication, but will be presented in due course in connection with the wider aspects of the investigations. Monte test. E xperim ental arrangem ent and procedure. tor each fraction .it animals 1 to 12 weeks of age were housed in metal boxes |1 x 8.1x 41 inches.5ineachbox. At Birminghamtheywereselected,25of each sex. from an'outbred laboratory albino strain previously employed in grading aseriesofoilfractions (Woodhouse and Irwin, 195). In London the mice used were frm laboratory stocks randomised so that each group of 5 contained some ofeachcolouranilgeneticconstitution. TheywerefedonThomson'scubediet with addition of oats and a littlegreen food. Water from bottles was always available." J The oilswere applied twice weekly for52 weeks to the inter-scapular region usingapproximatelyO '2 ml.oneachoccasion. Allapplicationsweremadebythe same person throughout for each centre. Records of deaths, appearance of |apillomata. etc., were kept, and the skin from the treated area of all mice which survived 12weeks'treatmentwere preparedforhistologicalexamination. Animals surviving were killed after 52 weeks. Rabbit tests. Itwas not-possibletoobtaintherequisiteiiumlier(105)ofanimalsofone inbredstrain,and at Birmingham medium-sized males of Dutch breed witherect carsand predominantly brown or grey in colour were randomised. In London therabbits were males of mixed commercial stock, with agouti, Flemish giant, black anti albino well represented. They were housed in fairly small cages of usual type and fed on greens alternated with crushed oats and a little bran. Applications of the oils to patches of skin alunit 3 cm. square from which the hair IimIlieen removed by electric dippers,were made twice weekly. Six areas on rach of 3o animals were used for testing the 3 crudes, and 75 animals were used for the 15 ilerived fractions employing 7 areas on each animal. The 6 areas comprisedthe2ears,leftandrightthoracic,andtheleftandrightlateralabdominal. The seventh area was the intcr-sca|Milarregion. These sitesare shown inFig. 1. API 05599 1*06 1. H1ECER AM) D. L. WOODHOU8E The scheme for allocating the fraction* and crudes to the various areas was in accordance with the arrangement shown in Tables I and II, which were devised Tabu I.-- D is tr ib u tio n o f C ru d e O i k o n B a b b it S i t e t (F ig . 1). Rabbits. 1-5 6-10 11-15 16-SO 31-35 *6-50 1. s. 4. A 5. L L K K O O K K O O L L O O L L K K L L O O K K K K L L O O O O K K L L L -- LafunillM crude, 0 -- Oklahoma crude, K = Kuwait crude. T a b u I I . -- D is t r ib u t io n o f D e r iv e d F r a c tio n s a n d R e s id u e s o n R a b b it S k in s (F ig . 1). Rabbits. 51-35 15-40 41-45 45-40 51-55 66-60 61-45 66-70 71-75 76-10 51-55 56-50 51-56 56-100 101-105 Bits*. ---- 1. 4 3. 4. 6. 6. 7. L3 K3 04 LR OR L4 K4 01 03 U KS OS 04 OR OR K1 U KR 04 LI L4 03 KS K4 K1 LI LR 04 04 LR KR 01 LS K4 OS K3 04 L3 LS KR KS OS LI 01 K3 04 03 LS K1 K4 LS 03 LI L4 03 LS K1 LS KS L4 K4 01 KR KR L4 03 03 KS K1 ER L4 LI 01 K1 LR KS LS LS 03 K1 K4 01 OR K3 KS LR K4 KR OR LI 03 OR KS LS ICR LI os 01 03 OR LR LS Kt LS KS R - RttkJue. API 05600 > * t AKClNUUEXIt'n V UF 1*TKULCM FRACTIONS IK RABBITS 27 byl)T. J.0. Irwin(MedicalResearchCouncil'eStatisticalResearchUnit,Uni versity of London), so that a statistical evaluation of the effects of site variation Hnd differencesinresponse by the individual animals might be possible. It will be apparent that each crude was applied to 60 sites and each of the other fractions to 33 sites. Applications were made twice weekly using approximately 0`3 ml.foreach. It was necessary to remove hair approximately every 10 days to ensure clear areas. <pbjgwasdonewithspecialcaretoavoidscratchingtheskin. Muchlesstrouble thanhadbeenanticipatedwasencounteredfromthean im al spreadingtheoilto otberareas of the body, though some difficulty resulted because some patches becamecakedwiththicklayersofdandruff. Recordsweremadeofthecondition 0f the sites throughout the 52 weeks, and finally the appropriate skin areas were preserved for histology. &I8ULT8. The general results for all the mice and rabbit series are set out in Table III. In addition to the mouse tumours recorded in this Table, with several fractions a number wereobservedwhichregressedafterashortperiodandhadnotrecurred up to the time of death or at the fifty-second week, in spite of continued applications III BiLromnidnognha(m-- Ta b u --- R e m i t s O b ta in e d f r o m th e icith those fro m L aboratories ( -- B ) com pared L ). Kuwait Oil (Fraction* arranged in order, light-* heavy). Crude oil K Light fraction K1 I K2 i K3 H eavy fraction K4 Residue KS Rabbit te s t: Yield of tumour*. B/L. 0/0 1/0 3/1 5/4 12/8 0/0 Moueeteet Yield of tumour*. B/L. 10 10 4/1 3/0 si 1/0 l.*guiulU* ( 'rude oil L 0/0 0/0 Light fraction LI 1/0 2/0 j Li 4/2 8/0 U 13/3 8/2 Heavy fraction U 1S/K 3/1 Residue L3 0/1 0/0 Uklaliyiua t'ruih oil U 0/2 0/0 Light fraction 01 0/1 0/0 1 02 12 0/0 T 03 l* ill Heav) fractiou 0 4 4/4 2/1 Rtwidue OS 0/1 0/0 Aithough .such>|*jut'iuieoUKregressionsarenot uncommon, the frequency has been a particularfeatureofthesetestsatbothcentres. It will be observed that the total number of mouse tumours in the Birmingham series was 33 900 and only 6 Dot) in the London series, although the survival rate wasbetterin the latter. The greatestnumber observed with any fractionwas 5. Thus the activity ofall these materials was very slight in the mouse tests; the Birmingham results suggest, that the LaguniUas derived fractions were somewhat mure active than the Kuwait and that the Oklahoma set were essentially inert. API 05601 *w I. HIEUER AND V. L. WOODHOU8E In eonirast, there was a total of 01 tumour-bearing rabbit sites in the Birming hamseriesand39m theLondonaerieeoutofapossible70.r>siteineachcane,or, excluding the crude and residues, which altogether yielded only 4 tumour* (London aeries),the remaining 12 fractions produced 01 and 35 tumours in the Birmingham and London aeries respectivelyon a possible 420 sites. Thus the bin of the rabbit was distinctly more sensitive to these fractions than that of the mouse. This observation was confirmed by the type, rate ofgrowth and other characteristics of the papillomata ; for example several tumours appeared on one rabbit site, and this is also corroborated by full statistical analyses of the results with reference to survival rate of the mice, the effect of site distribution in the rabbits, etc., which have been undertaken by Dr. Irwin. DISCUSSION. Both in the rabbit and in the mouse tests s somewhat higher incidence of tumours was obtainedatBirminghamthanatLondon. Thismay possiblybe due toa differenceinthestrainofanimals, forSmith and Sunderland (1951) have observed differencesin tberesponse ofdifferentstrainsofmice. Itisimpossible to exclude thepossibilityofslightdifferencesintechnique. Consideringthat the strains of mice were different in the two laboratories, the agreement between the Birmingham and the London resultsisbetterthan might have been expected. Although tbe absolute yield of tumours is different in the two cases, the relative activity of the different fractions shows an appreciable consistency (Table III). To account forthe greater activity on rabbit skin the possibility that these mineral oilfractions may contain types ofcarcinogen differing chemically from the polycyclic hydrocarbons hithertoencountered must be entertained. In this connection itis important to emphasize that the cracked oGa used by Smith, Sunderland and Sugiura (1951) would probably constitute a series containing considerable quantities of different types of hydrocarbons from those present in the fractions utilirnd in the present investigation. From the oilsemployed by the American investigator, compounds such as isopropyl-1:2-benzanthracene and methyl chrysene were isolated, while the presence of pyrene and benzphenanthrene derivatives was also demonstrated by Fischer, Priestley, Eby, Wanlees and Rehner (1951). Tbe extraordinary complexity of mineral oilsand the minute proportion of carcinogenic components in the uncracked oils must present very considerable difficultiesinany effort to isolateor characterise them. Twort (1951) reported preliminaryattemptstodothisbuttheywerenotcontinued. Furtherexperiments using both species of animals to test fractions selected from these crude oils are being carried out which it is believed will provide further information and facilitate future attempts to identify the carcinogenic agents. - SUMMARY. Tests hsvc been carried out in two laboratories to determine the carcinogenic action to the skin ofmice and rabbits ofthree typical mineral oilcrudes and 15 fractionsderivedfrom them by proeesseeavoidingcracking. The activityon mice of all the fractions was very slight : some, particularly those with a boiling range of 325 to 375 C,, showed high activity on the rabbit ear and body skin. Bothseriesofexperimentsdemonstrated,therefore,thattherabbitwasmore API 05602 1 rARn.vnr.EXirtTY o f p e t r o l e u m fr a ct io n 's in r a b b it s 299 sensitive to these types of oil than the mouse, and it is concluded that this species should be included in carcinogenic tests on this type of material. The Chester BeRtty Research Institute receives grants from the British Empire Cancer Campaign, the Jane Coffin Childs Memorial Fund for Medical Research, the Anna Fuller Fund and the National Cancer Institute of the National Institutes of Health. I '.S. Public Health Service. REFERENCES. Aru> S. J M.--(1950)./. In st. P stm l.. 3. 235. R*B#:s A.--(1929) B ull. A ss. /ranf, C ancer. 18. 276. Rf.RF.nbu-m. I.--(1943b) C ancer R es.. S. 263.--(19456) A n n . R ep. B rit. K m p. Cancer Cnm/ ign.. 22. 62.--(1947) B ril. J . C ancer, 1, 157. Bonne. C.--(1927) Z. K rebsforsch.. 25. 1. ('rcikshank. C. N. D.. and Squire. J. R.--(1950) B ril. J . in du st. M ed .. 7, 1. Fischer. H. 0 M.. Priestley. W.. E bv. L. T.. W asless. C... and R ehnkr, J.--(1951) A rrh. indu st. H yg .. 4. 315. I. ERorx. R --(1927) B u ll. A ss. /ranf. Cancer. 18. 16. J, vth. R.--(1933) J . indu st. H yg .. 15. 226. Oberlino, C'.. Sasni. C.. Gt'ERiN. M.. and Guerin. P.--(1937) Leeu\renhoeb. vereeniging, 4. 57. ScHrRCH 0 --(1939) Z. K rebsforsch., 48. 353. Smith. W. E.. and Sunderland. D. A --{1351) C ancer R es., 11, 281. idem and SfGH RA. K.--(1931) ArfA. indu st. H yg ., 4, 299. Twort. J. M.--(1941) Ann. R ep . B rit, E m p . C ancer C am pgn .. 18. 162. Twort C. C.. and F ulton. J. D.--( 1 9 2 9 ) P ath . B a d .. 32.149. Idem and Twort. J. M --( 1 9 3 0 ) H yg .. C am b.. 28, 373.--(li>31) in du st. H yg ., 13, 294 --(1933) A m er. ./. Confer. 17. 293. Twort. J. M.. and Lyth. R.--(1939)./. H yg .. C am b.. 38. 161. W atson. A. F.-- (1932) Cnncer Ret-.. 7. 443. WiKiiiHorsE. D. 1,.. and Irwin. J. 0 .--( 1 9 3 0 ) H yg .. C am b., 41. 121. VtMAiuwA. K.. and Itchikawa. K.--(191H) J . C ancer R es., 3, 1. e d f tic 15 ice ige yttitlr eintl firintr*l in firetit Britain Jar H. K. h rn t- d ` Co. IJ/I.. inf ore 4* S(m . Hnrthoinwnr t irrnAt D arling. s 3 sr ft-*- API 05603 m STANDARD OIL DEVELOPMENT COMPANY m Ed ic a l r e s e a r c h d iv is io n M , =T E ECKARDT. PH D.. M.D rOBEP' DIRECTOR ljNDRICKS. B. E.. CH. E. N V INDUSTRIAL HYGIENE SECTION FRIAnNKrLuInN V* CHURINCDHU.STBRIASL.. H(CYHG.IEEN.I)ST P. O. BOX 51, LINDEN, N. J. March 18, 1953 Mr. D. V. Stroop American Petroleum I n s t it u t e 50 V. 50th St. Sew York, 20, B.Y. Dear Mr. Stroop: The enclosed, apparently represente a publication of the data which members of the Subcommittee on Carcinogenicity of the A.F.I. have had an opportunity to review in connection with the exchange of information between the British our own Subcommittees on Carcinogenicity. 1 thought you might wish to have this article reproduced and distributed to the members of the Medical Advisory Committee of the American Petroleum Institute. Tery truly yours, SEB/hml T T - E. B. X. SCXAEDT, M.D. S n cl. (The Value of the Babbit For Carcinogenicity Tests on Petroleum Fractions, by Eiefer & D.L.Woodhouse Bepr.fron TH3 BEITISH JHL. OF CJBCSB,l952,Tol.VI,Pg.293.) API 05604 f T I Reprinted from The British Journal o f Cancer, 1 m i , Voi. VI, p. 293. THK value of the rabbit for carcinogenicity tests ON PETROLEUM fractions. I. HIEGER AND D. L. WOODHOUSE. from thr C hester B e a tty R esearch In s titu te , T h e B o y a l C ancer H o s p ita l, L o n d o n , 8 .W . 3, 13. a n d the C a n c e r R esearch L abo ra to ries, D ep a rtm en t c j P a th o lo g y, The M edical School, B irm ingham , Keroivui for publicsion May SH, 1052. The experiments recordedinthisarticlewerecarriedout inthecourseof investigations onbehalfof theMedicalResearchCouncil,asub-committeehaving Iwensetupin1949to inquireintothecarcinogenicpropertiesofmineraloilsand idlied products. The arrangements for co-ordinating the research have been briefly describe! by Auld (1930). The entire series of tests have been made in duplicate at the Cancer Research laboratories, Department ofPathology, University of Birmingham, and at the C hester Beatty Research Institute, The Royal Cancer Hospital, London. At eachcentrethe activityof3 selectedcrude oils,4 fractionsderived from each crude bv methods ofdistillation specially designed to avoid high temperature cracking, and the final residues, have been tested on groups of *>o mioe for each ampleand alsoon a totalof 105 rabbits. The resultsappear to be ofparticular interest, since it must be concluded from them that it is unsatisfactory to exclude carrinogenicity on the basis of tests on mice only. The desirability of utilising in industry those types ofoils which are least likely to cause dermatitis or cancer of the skin of workers has been recognised for over years, and in 1934 the Committee on Cancer ofManchester Corporation proposed a standard for assessing the potency of mineral oils depending on physical properties (specific gravity and refractive index) which it was believed would ensure that the lubricants used in the textile industry would be substantially free fromcarcinogenicproperties. ThisManchesterformula,whichisnotnowregarded as a satisfactory criterion, was put forward following the investigations of Twort and his co-workers (Twort and Fulton, 1929 ; Twort and Twort, 1930,1931,1933 : Twort.and Lyth. 1939 : Twort, 1941), who tested a wide range ofcrude mineral oils, distillates, solvent extracts, spindle and shale oils, using mioe almost exclu sively, however, for the biological assessment oftheir carcinogenicity. In the past, Also, mice have for the most part been employed in skin tests with " oil " fractionsderived from coal tan and with the pure chemical Carcinogens either isolated or synthesised. ItistruethattheclassicalexperimentsofYamagiwaand Itchikawa (1913), m which thecarcinogenicityofcoal tarwas firstproved, utilisedrabbits, and this specieshasalsobeenemployed ina number ofearlyexperimentson tarcancer, s.g,, Bonne (1927), Leroux (1927), Babes (1929) and Twort and Twort (1930). Also a few early workers have used other species to a limited extunt, e.g., rats and guinea-pigs(Watson, 1932). The majority, however, have favoured themouse for large scale experiments, no doubt partly' because of low maintenance oosts and API 05605 294 1. HIEGER AVD D. X. WOODHOCSE availability, but probably influenced alio by the experience of early worker* that the akin ofrata did notreadily respond to coal tan, and that papillomata on labbita so produoed remained small and often regressed when applications'cesaed. Thus the mouse has become accepted as the most sensitivetest animal for such work;the possibilitythat otherspeciesmight provemore suitableunder certain circumstanoes does not appear to have been sufficiently appreciated. Schurch {1939),however,foundthathisexperimentswithrabbitsindicatedthat carcinogenic ingredients in addition to the recognised carcinogen 3 : 4-benzpyren* were probably presentincertaincoaltart. This was pointed out by Berenblum (1947),who demonstrated that the relative carcinogenic activity of a numb of fractions, obtained by chromatographic procedures, varied greatly according to whether standardised against the skin ofthe mouse or ofthe rabbit. He had already shown (Berenblum, 1945a) that tumours were readily elicited on the rabbit by 9: 10-dimethyl-1:2-benzanthracene, which was found to be mors potent to rabbits than to rats or guinea-pigs, in which positive results wen, however, obtained (Berenblum, 19456). He also subsequently observed (1947) that this hydrocarbon when injected subcutaneously did not induce rabbit tumours, though ratsand guinea-pigsresponded by thismethod. Much earlier, Oberiing, SaanM, Guerin and Guerin (1937)had shown that a 1per cent solution of 3:t-benzpyrene in benzene waa active on rabbits'skins, end Berenblum (1945a) observed that 9 : 10-dimethyl-1: 2-benzanthrsoene was much more activ* than benzpyrene at a similar (0-5 per cent) concentration. Experimenta mring a tgh boiljng fraction of mineral oil from an experimental catalytic cracking operation have recently been carried out by Smith, Sunder! and 8ugftira (1951), testing the residue after removing the lower boiling napht and light gaa oil cuts, upon mice, rats, guinea-pigs, rabbits and rhesus monk The rataand guinea-pigs wen found to be refractory to skin applications papillomatawen elicitedinallofthe6monkeys,2beingprovedcancerous biopsy4yeanafterthestartoftbeexperiment. Papillomatawen produced the innersurfaceoftheearsofthe 21 rabbitswithin 100days. The number m ofsuchgrowthstendedtoincreaseduringthe2yeaninwhichpainting continued, and in 3 of the 0 surviving animals cancerous changes in the gro wen observed. Fortestsonthetypeofmaterialwhichtheyintendedtost namely, samplm containing oilswhich had been subjected to a.processoffl catalysis up to 950 F. in the presence of alumina or silica, these workers ooncl that white mice were tbe best animals. Recently Qruikshank end Squire (1950) obtained numerous papillomata the ear and body m of albino rabbits after applications of a "cutting oil obtainedfrom a machinesump. Only Ibenign papilloma was produoed by t oil in a group of 46 mice, of which 50 per cent survived 4(1 weeks' and 28 per 52 weeks' treatment, and they suggested that such materials should be against both specie before being regarded as non-carcinogenic to man. In the present investigation it was decided that it was of importance that propertiesofthemineraloilsinquestionshouldbetestedonbothrabbitsandm' Crudes and oilfractions. The 3 erodes were obtained from Kuwait, LaguniUas and Oklahoma nwpecti and had the following characteristics: API 05606 (.AIUI-NUGEMUTV UK l'KTKULKL'.M KK.UTlUXS IX KAUUITS 295 fcmi'ait (Kuwait Oil Co.): an extensively produced Middle East crude likely be much used in t he United Kingdom, of paraffinic-asphaltic base and containing lubricating oil fractions. - l/vjunillas Venezuela (Shell Petroleum Co.): a typical naphthenic cnide i,.|(ling ell-known lubricating distillates. Qlcinlioiiiii city Mid-Continental (Socouy Vacuum Oil Co.): yields Mid- <_ontiaeiital distillates, bright stock and residual lubricating oil. jh e derived fractions were produced by a research group appointed by the institute of Petroleum, which has been responsible for the choice of the crudes living regard to their type, distribution and industrial importance. The initial fractionation of the 3 crudes was carried out at delimited temperatures using vacuum and steam in an apparatus selected to preclude crackingr These operations were carried out at the Thornton Research centre, Shell Refining and Marketing Vo. Ltd., for the Institute of Petroleum. Full details regarding the physical characteristics of these samples are not pertinent to the purpose of this communication, but will be presented in due course in connection with the wider aspects of the investigations. Experimental arrangement and procedure. House tests. For each fraction 5o animals 10 to 13 weeks of age were housed in inetal boxes 11 x H.1 x 41 inches. in each box. At Birmingham they were selected, 25 of each sex, from an outbred laboratory albino strain previously employed in grading a series of oil fractions (Woodhouse and Irwin, 1950). In London the mice used were from laboratory stocks randomised so th at each group of 50 contained some ,,f each colour and genetic constitution. They were fed on Thomson's cube diet with addition of oats and a little green food. M ater from bottles was always available. The oils were applied twice weekly for 52 weeks to the inter-scapular region using approximately 0 2 ml. on each occasion. All applications were made by the same person throughout for each centre. Records of deaths, appearance of liapillomata. etc., were kept, and the skin from the treated area of all mice which surv ived 12 weeks'treatment were prepared for histological examination. Animals surv iving were killed after 52 weeks. Rttbtnt tests. It was not (xissiblc to obtain the requisite numlier (105) of animals of one inbred strain, and at Birmingham medium-sized males of Dutch breed with erect cars and predominantly brown or grey in colour were randomised. In London the rabbits were males of mixed commercial stock, with agouti, Flemish giant, black and albino well represented. Tltev were housed in fairly small cages of usual typo and fed on greens alternated with crushed oats and a little bran. Applications of the oils to pate he* of skin about 3 cm. square from which the hair Imd been removed bv electric clippers, were made twice weekly. Six areas on each of 3o animals were used for testing the 3 crudes, and 75 animals were need for the 15 derived fractions employing 7 areas on each animal. The ft areas comprised t he 2 ears, left- an! right t horacic. and the left and right lateral abdominal. The seventh area was the inter-scapular region. These sites are shown in Fig. 1. API 05607 1*96 I. RIEGER AN1> D. L. WOODHOU8E The scheme for allocating the fractions and crudes to the various areas was in accordance with the arrangement shown in Tables I and II, which were devised Table I.-- D istribution o f Crude O ilt on Babbit S ites {Fig. 1). R e b b iu . 1-5 S-10 11-15 ia -io S1-S5 3 4 -3 0 - A 1. a. 4. a L L K K o K K o O L O O L L K L L O O K K K L L O O O K K L L = laguailias enids, O - O U abum crude, K - Kuwait crude. e. 0 . L K K 0 L Table II.--D istribution o f D erived Fractions and Residues on Rabbit Skins {Fig. 1). KSEgEggCBSSiEg Rabbits. 1. 31-35 . L3 SS-40 . 01 41-46 . OR 44-50 . 03 51-55 . 04 54-SO . K3 ei-45 . LI 4-70 . K4 71-75 . K1 74-40 . KR 81-44 . L4 IM 0 . L3 81-46 . K3 4-100 . LR 101-104 . OS ^----- K3 a 04 A LR a a OR L4 03 L4 KS 03 04 K1 KS L3 K4 KR 04 K1 LI LI LR LR 04 KR Ol L3 L3 Li K4 KR K3 Ol IS K3 OS 04 LI 03 L4 L3 03 L3 KS L4 K4 Ol L4 03 03 K3 K1 LI 03 Ol K! KS K4 LR KS Ol OR K4 OR 03 KS KR KR LI OR L3 OS OR LR L3 K1 "Hi R --RuUia, API 05608  9 (AKCINOUNKTIY O f I'JiTKOLKlM FRACTIONS IN RABBITS 397 jjf j.O. Irwin (Medical Research Council'sStatistical Research Unit, UniofLondon),so thatastatisticalevaluationoftheeffectsofsitevariation differ!9in response by the individual animals might be possible. Itwill beapp*"thateachcrudew" aPPliedto60to" deachoftheotherfractions 35 sites* t0Appd044*01" were`" dtwic* weeklyusingapproximately0 3ml.foreach, it was necessary to remove hair approximately every 10 days to ensure clear areas. .Ljgas done with specialcare toavoid scratching the skin. Much lasstrouble than had been anticipated was encountered from the animal spreading the oil to otherareas ofthe body, though some difficulty resulted because some patches becamecakedwiththicklayersofdandruff. Recordsweremadeofthecondition of the sites throughout the 52 weeks, and finally the appropriate hi areas were presen'e1d for histology. BXSVLTS. The general results for all the mice and rabbit series are set out in Table III. In addition to the mouse tumours recorded in this Table, with several fractions a number were observed which regressed after a short period and had not recurred uptothetimeofdeathoratthefifty-sqoondweek,m spiteofcontinuedapplications <r TtBLC IU.-- R e su lts O b ta in e d fr o m the BirminghamL a b o ra to rie s (= B ) c o m p a re d ic itk those fr o m London (asL ). KtlWMt Oil (Kraftion* arranged in order, light-rheevy). Crude oil K Light fraction K l 1 VJV 4m K3 H eavy Crartiou K4 Rmidue KS Rabbit test : Yield of tumour*. B/L. 0I//0O 3 1 ,li5/4S 0/0 HoYiweeldteosft tumour*. B/L. i/o */o 4*//10 t1i/l0 LaguuilW* Crude oil L 0/0 0/0 Light fraction LI I/O s/o Li 4/2 5/0 *t L3 H**vy frartion U 1136//36 i3/l 1i K**u<lue L5 /I 0/O Oklahoma O w l* oil O 1 0/3 0/0 Light fahrt ion Ol 0/1 0/0 Oi lit 0/0 II T 03 11* til Heavy fraction IM 44 *11 0/1 fWwadue 03 0/0 Although such spontaneous regressionsare not uncommon, the frequency has Iktii a particular feature of these tests at both centres. It will be observed thatthetotalnumberofmousetumoursintheBirmingham series was 33 900 andonly6 '9<)0 intheLondonseries,althoughthesurvivalrate as better inthelatter. Thegreatestnumberobservedwithanyfractionwas5. Thus the activity ofallthesematerialswas veryslightinthemousetests;the Birmingham results suggestthattheLagunillasderivedfractionsweresomewhat mure active thanthe Kuwaitandthat theOklahomasetwereessentiallyinert. API 05609 ** I. H IK E R AND V. L. WOODHOU8E Ineoatnmt,therewasatotalof61tumour-bearingrabbitsitesin theBinning, hamaeriesand39m theLondonaeriesoutofapossible705sitesineachcaw,or, exeluding the crudes and residues, which altogether yielded only 4 tumour (London series),the remaining 12 fractions produced 61 and 36 tumours inthe Birmingham and London aeries respectively on a possible 420 sites. Thus the fcin of the rabbit was distinctly more sensitive to these fractions than that of the mouse. This observation was confirmed by the type, rateofgrowth and other characteristics of the papillomata ; for example several tumours appeared on one rabbitrite,and thisisalsocorroboratedby{tillstatisticalanalysesoftheresults withreferencetosurvivalrateofthemice,theeffectofsitedistributionin the rabbits, etc., which have been undertaken by Dr. Irwin. DI8CT7SBIOM. Both in the rabbit and in the mouse teste a somewhat higher incidence of tumourserasobtainedatBirminghamthanatLondon. Thismaypossiblybedue toa differenceinthestrainofanimals,forSmith and Sunderland (1951) have observed differencesin the response ofdifferentstrainsofmice. Itisimpossible to exclude the possibilityofslightdifferencesintechnique. Consideringthatthe strains of mice were different in the two laboratories, the agreement between the Birmingham and the London results isbetterthan might have been expected. Although the absolute yield of tumours is different in the two cases, the relative activity of the different fractions shows an appreciable consistency (Table III). To aeeount forthe greater activityon rabbit skin the possibility that thess mineral oilfractions may oontain types ofcarcinogen differing chemically from the polycyclic hydrocarbons hithertoencountered must be entertained. In this connection H isimportant to emphasise that the cracked oOs used by Smith, Sunderland and Sugiura (1951) would probably constitute s series containing considerable quantities of different types of hydrocarbons from those present in the fractions utilised in the present investigation. From the oilsemployed by the American investigators, compounds such ss isopropyl-1:2-benzanthraceiw and methyl chrysene were isolated, while the presence of pyrene and beiuphenanthrene derivatives was also demonstrated by Fischer, Priestley, Eby, Wanfess and Rehner (1961). The extraordinary complexity of mineral oilsand the minute proportion of c a rc in o g e n ic components intheuncrackedoilsmust presentvery considerable difficulticeinanyefforttoisolateorcharacterisethem. Twort(1951)reported preliminary attemptstodothisbuttheywerenotcontinued. Furtherexperiments Mg both species of animals to test fractions selected from these crude oils are being carried out which it is believed will provide further information and facilitate future attempts to identify the carcinogenic agents. SUMMARY. Tests have been carried out in two laboratories to determine the carcinogenic action tothe skin ofmice and rabbits ofthree typical mineral oil crudes and 15 fractionsderivedfromthembyprocessesavoidingcracking. Theactivityonmice of all the fractions was very slight: some, particularly those with a boiling range of 325 to 375* C., showed high activity on the rabbit ear and body skin. Both series of experiments demonstrated, therefore, that the rabbit was more A PI 05610 rA R C lN O O E .V iriTY OF PETROLEUM FRACTIONS EN RABBITS 299 *ensiti' e to those types of oil than the mouse, and it is concluded that this species Should be included in carcinogenic tests on this type of material. The Chester Beatty Research Institute receives grants from the British Empire Cancer Campaign, the Jane Coffin ChildR Memorial Fund for Medical Research, the Anna Fuller Fund and the National Cancer Institute of the National Institutes of Health. U.S. Public Health Service. REFERENCES. Man 8. J. M.--(1939) /. Inst. Petrol.. 39. 235, juB^s A.--(1929) Bull. Aw./mnc. Cancer. 1|. 276. Rfr eSBU'M. I.--(1943a) Cancer Re*.. S. 265.--(19455) Ann. Rep. Brit. Smp. Cancer Cnmi<gn.. 22. 62.--(1947) Brit. J. Cancer, 1, 157. Rosve- C.--(1927)Z . Krebsforach.. 25.1. (uriKSHANK. ('. X. D.. and Sq h r e . J. R.--(1950) Bril. J. induct. Med.. 7. 1. F,xther. H. C. M.. Priestley. W.. E bv. L. T.. W asless. C... and Kkhnrr, J.--(1951) Arch, induct. Hyg.. 4. 315. Leboi'E. R---(1927) Bull. As, /m sf. Cancer. II. 16. Iath. R.--(1933) /. induct. Hyg., 15. 226. Obewjsc.. ('.. SAS.vit. C.. Gt'ERtx. M. and GrERix. P.--(1937) Lemycenhoek-verrmiging, 4,57. ScHfRCH. 0 .--(1939) Z . Krtbsforteh.. 49.353. Smith. W. E.. and Scnderland. D. A.--(1951) Cancer Re., 11, 281. /idem and SrcifRA. K.--(1951) -4rrA. induct. Hyg., 4,299. Twort. J. M --(1941) .4mi. Rep. Brit. Emp. Cancer Campgn., II. 162. Twort e. ('.. ANDFvlton. J. D.--(1929) J. Path. Bad., 32.149. Idem and Twort, J. M.--(1930) J. Hyg.. Comb.. 29,373 --(1931) J. induct. Hyg., 13, 204.--(1933) .4 er../. Cancer. 17. 293. Twort. J. M.. and Lvth. R.--(1939) J . Hyg.. Camb.. 39.161. Watson. A. F.--(1932) Cancer Rer.. 7. 445. WtsiiiHorsE. D. I... and Irwin. J. O.--(1950) J. Hyg.. Comb., 41. 121. YtHuuwA. K.. ash Itchikawa. K.--(1918)./. Canrer Re*., 3, 1. ih n lr n m l iriulnl in Ureal R rito in Jar H . K . / / < d' Co, Ail/,. htf A f lh tn l U` S o u , L o l,t H firth n in u iftr 1'rr**, D o rl'iiig . 05612 ) fl /TT T ) ao' CU'fii m sx& aot of aimlM r^~ & w m s m m m CARLE ADDRESS: KCTUB. CINCINNATI TRLtPHONZ: CAPITOL 1414 1 am sending you h e re w ith , f o r your in fo rm a tio n , a sta te m e n t of e x p e n d itu re s made on b e h a lf of American P etroleum I n s t i t u t e during th e f i r s t q u a rte r of 1955 I b e lie v e th a t the s t a t e ment v i l l be s e l f - e x p l a n a to r y , b u t i f you have any q u e stio n s or comments, D r. Kehoe would a p p re c ia te your b rin g in g them to his a tte n tio n . Very tr u ly y o u rs, ff. k P o rtl g e , S e c retary to D r. Kehoe API 05614 UNIVERSITY OF CINCINNATI KETTERING LABORATORY - 5- discernible but it was not anticipated that any such trends would be discernible prior to at least 5 years experience. It is hoped that the support which the General Committee, Division of Refining has given to the epidemiological study will stimulate renewed reporting of the cases with sufficient medical and occupational histories to permit some early correlations to be made about 1956. Summary 1 . The RP (MC-l) Advisory Committee believes that the Kettering Laboratory is performing important work in the field of carcinogenesis of refinery streams and materials. They believe that although a great many practical answers have already been obtained, the work should be pursued because the work of the laboratory promises to provide many fundamental answers to questions which now exist concerning industrial and occupational carcinogenesis. 2 . It is believed that the work with non-accelerating solvents which relates the carcinogenic responses of a solution of pure carcinogen to the dose applied per unit area per week to the skin is of extreme fundamental importance. The Committee urges that additional work be conducted to establish firmly the curve which expresses this relationship. 3 . It is also believed that the work with accelerating solvents is also of extreme fundamental importance and that this should be pursued vigorously in an attempt to develop a complete understanding of the relative role of carcinogens and accelerators in refinery streams and materials. 1f. Since much of the correlation of chemical analyses with biological potency is dependent upon the accuracy of values, the Com mittee laments the fact that many of the Pj^ values thus far reported are based on data which the laboratory recognizes as not having been obtained from the best biological experiment possible. The Committee suggests, there fore, that the laboratory devote considerable effort to repeating those experiments in an effort to develop P| values that are based on the soundest possible biological experiment. 5. The Comm it t e e believes that the work with washing techniques and protective creams may provide intensely practical guides to refinery hygiene practice and suggests that this work be pursued as new approaches appear to the laboratory. 6. The Committee recognizes that the epidemiological studies be expected to produce significant results until at least 1956, and, therefore, , urges that renewed efforts at adequate case and occupational history reporting | be made during the next several years. It believes that adequate medical staffs can do most to insure the success of this program, and in this connection it acknowledges the stimulus that the General Committee, Division of Refining | has given to the attainment of this objective. cannot 7. The laboratory has already determined that a number of refinery streams and materials have a relatively high degree of carcinogenicity. Although data obtained with mice cannot be directly transposed to humans, there is sufficient data in occupational cancer experience to indicate that the API 05728 petroleum industry cannot take lightly the observations of the laboratory on the carcinogenicity of these refinery streams and materials. 8. The Committee recognizes that the laboratory at best can determine from its biological and chemical work only the relative carcino genicity of various refinery streams and materials. The determination of the hazard is dependent upon factors of extent, frequency, duration, and length of exposure. As recognized by the laboratory in Dr. Kehoe's talk to the members of the General Committee, Division of Refining the determina tion of the hazard is at present a function of the individual petroleum com panies. Adequate medieal and industrial hygiene staffs are essential in these individual companies if they are adequately to appraise the hazard. The Committee supports Dr. Kehoe's plea that petroleum company managements recognize this truism and make plans for the development of such adequate aedical and industrial hygiene staffs. ' 9 It is apparent that the laboratory has accumulated sufficient data that publication of results will be deemed desirable in the near future. The Committee recognizes the right of the laboratory to publish, and urges 'that the mechanism be established so that these publications can be reviewed in a finite period of time. As a suggestion, it Is proposed that this mechanism be so established that review by API management and suggestions and criticisms for the laboratory from API management be made available within sixty (60) days, tIf no criticisms or suggestions are obtained by the laboratory at the expiration of this time, the laboratory can assume that none are forthcoming and proceed with publication. It should be recognized that this review is for the purpose of helpful criticisms and suggestions, and not for the determination of whether ior not there will be publication, since the contract with the laboratory clearly states that the laboratory determines if and when publication occurs. -- 00O00-- October 7 , 1953 gopy frca D. . Stroop far infoimtiao of Or. Qoarfu x. Saundon r-A Dr. B. B. Ickurdt___________________ Dec--b r 22, 1953 UHTVKRSXTT GP CIXCUOIATI Departn-- t of P re--a ti lto d ld M ta d Xafcirtrlal Boalth Nr. D. T. Stroap, Director Departnot af Teehnleal Serrleos jo o rlao o P e tro l* -- IarUtuta 50 VMt 50th Street - tv IM d tp Dwr H. (tra il I m o n A U f ye herew ith fo r year ix fa x w tla e , a e t e tm e t of e ^ e o d ltu re s --4o eo b e h a lf o f th e A aerleao P etro l* -- X aetltu te fo r th e tfc&i* f --r t e r at 1993* X b e lie -- th a t th i s e ta te a e e t i l l be e l f - o ^ l e n t e e p , be* t f p H b n f o r fpraofelm a o m M i M o to r Kehoe v e a l* p M l H m M i M t h n t * h i t H i o H i f Tory tru ly youre, ( / / 1 . 1 P o rtio n 1. K. P o rtio n to Or. Kehoe Inc UOTSRSITT er cueTBBSI nmSRIK LABCRATCRT Accourr cr amducai frroixcn ixsrrrors Tt. 3rd ju ir tw 1953 yjjjO lS (Butd on Proportion o t Tin Actually Spent on Project) Direct Salaries 7,961.26 U rtip tlK 1* * ^ P re p a ra tio n O ther S errlo ee 602.58 8,539-9* 1 7 .1 2 3 .7 8 jggCXLLABOOa IX R B Purchaee of tortnala artel H U rrtw y Im plies Travel *J8.60 793 1 ,1 1 1 .5 7 (P ra p c rtta a a f h e * One I le e tr ie lty , M am , Telephone, Oeaeral laboratory Stppllee, Poetape, A anaftlee, Pension*, Nelateaaaee, e tc . *,868.77 7.212.03 TOEAL............................... 335.81 Balance AeaUehla f a r fa rth e r Work at lad of a t M l e f SeM Bl f c r t e r 1953 _____ ~i -- ifi m u r i n * * l a lane* h e l ls M e f a r f a r th e r Work a t hei ar M Je S L iS a * *1** Daa K a tta rla g la b o ra to ry a t la d o f ________________ 3 ,* 5 0 .6 2 *0.000.00 56,M .S8 a h j^ 12,213.57 API 05731 A m erican P etro leum In st it u t e 50 W EST 50th STREET N E W Y O R K 2 0 . N. Y. oF T E C H N IC A L s e r v ic e s -v'O - T R O O P D I R E C T O R Daaaater 23, 1953 D r. H ater* a . K ate* U n lT w rsltj <* C la a la n fe l Ite l i t t e t Tiicw alw j coUai <* m tU im - xtn Anna C la alaa m I f , ctea V S m at 1 tea iw la a o n API 05732 y e .s o i <v m i m m * im m n i m M M CT '1 t v IW H U M T ii m *Y I --H 1 * 1 <lT HS l a s o r a t o r t , ,,.OICINE-ROEM AVENUE UNIVERSITY OF CINCINNATI DEPARTMENT OF PREVENTIVE MEDICINE AND INDUSTRIAL HEALTH J u ly 3, 1953 CAtLE ADDRESS EETLAS. CINCINNATI TELEPHONE. CAMTJI. 1414 Mr. D. V. Stroop, D irector Department of Technical Services American Petroleum Institute 50 West 50th Street New York City 20 Dear Mr. Stroop: I am returning to you herewith two copies of the executed extension of agreem ent between A m erican P etroleum Institute and the U niversity of Cincinnati, for the period June 30, 1953 to June 30, 1954. Very truly yours ef Enc. API 05735 A m e r ic a n P e t r o l e u m In st it u t e 50 WEST 50TH STREET N EW Y O R K 20. N. Y. June 30, 1953 Board, of Directors University of Cincinnati Cincinnati 19* Ohio Gentlenen: m . s earounlcation is authority to continue work on investigation of the toxicity sad rode of action of certain petrolsun products for the year ending June 30* 1954, according to the teres of the ngre-- rvrt dated January 8* 1946, and on the basis of a -- budget for operating expenses of $78*283* ' Upon receipt of your approval of the continuation of the project with this budget ee shall pay you the sua of $40,000. Die balance of $38*263* less the unexpended balance in the fund on June 30* 1993* will be paid on January 1* 1954. .* It is wa&nretee&.bhfft any balance la the fund on June 30 1994 will be returned to the Ansrleaa Petrol* Institute* toft that no txeess over the arount of the budget is to be obligated or spent without written authorisation. Very truly yours, m i s t i R X M U K M USTITOTE Accepted sad Approved: u u v n s m or cis cixk ati Through its hoard of Directors By K. R enton K. B r o d le , Chairman c B o a a i oorf Diirreecc t o r ; By. Raal1pnhh cC.. Buralek, Clerk Board of Directors API 05737 Juae 30, 1953 3oord of Directora Uaiveraity of Cincinnati Cincinnati 19, Ohio G cu tlan an s Thl in-- iluallun la authcrltp to continua werk oo inveetloatlon cf tha to sle itp ad aode of aotlco of eartaln p stro lcm producta ft tha paar andlag Juna 30, 195k, coordin to tha ta n a of tha agreanant datad Jhauary 8, 19k6, and an the baala of a aaa l i M budget fo r cperatlng aaqpanaee of 478, 283. Upen raealp t of pour approval of tha ccntlauatlcn of tha oroject Ith th la a a r iia i budaet a ahall pap peu tha awai of 4*0,000. Ute balanoa of 438,203, Lasa tha urwxpaaiad la tha fund ca Jmm 30, 1953, UL ha pald ob Jaouarp l , 195k. Xt la wadar eto aii tha* aagr telam a ln tha fand oa <Aaa 30, 195k 111 ba raturaaii to tha ,\aai'lBan Fetrolaua In stltu U , and th at ao aneas* ovar tha m n t of tha badest la to ba obligatad o r spant vltbout ritte n antherH atten . forj tn ilp paura, AMBZCAI PtTaOLB* HSTXTUTS . . Prsident Accaptad sad Approvadi ukivsrsitx e r c d c d b u t i ifcrcugh Ita Board of O lraeters .errtary API 05739 .ene 3C, D r. Rotart A. Kehoa UniTOTsitj o f u ln e in a tti Th* u u tte ria t Mlacmiury Colle.o of hsricloe - idan Arata Ciacinnafcl l j , Dntr Doctor Seno: Erfcloaod you ili fiad foar copiti of the eoptin a ttioo agraeaaat comriaa API Hedioai neeaaruh Project N- far tt flaoal yeer odine June 3 0 , 1993 Int--nrh ma Praeident Portar la out of Um c ity tod 111 aot rotara for tao aal o aacsoet tat i / yoa tato tl oca'Uaota aigned oo Whalf of tba OalTwrolty of Cinelanat i ad rotara tta to aa oo o aoqr aoal you a eeek. Va I l i aand you two oo-jdea of the agreaaant for your fila a a aooa aa they harm toea aignod ay Mr. Portar. Tory tra ly your, DVS !C Eacloaorea API 05740 A merican P etroleum Institute 30 W E S T BOTH S T R E E T N EW Y O R K 20. N. Y. *n* 30, 1958 o u rm * at in ciati-- to 19 CM* tk i* I i t u t t I T la aitoarMy to tarli-- work oa 1-- tifa tim at toa to d sltf ma a l t a t a t t a a t tartoln yv-- 1-- I-- asta f t r i f e t j a r tola i -- 30, 1953, -- rtlag to to* to-- at to* -- tom ai ato* tonar? 6, 29*6, -- on to* to* ir o jto t 150,000. to* tola-- a t $5*,70, J oa /a so, logo, to ll to rt la 30, 2993 to ll to at la 1, 1953. u rm m m * to rn a i ito toar SO-- tora Prank P. DinaBre,rC h a l man- r o - e ^ u C *LU'Vfl Ralph C. B uralak, C lerk API 05741 ,,N< J 0 WT0*Y rf^ J-E D ie lH E -W E N AVE UNIVERSITY OF CINCINNATI DEPARTMENT OF PREVENTIVE MEDICINE ANO INDUSTRIAL HEALTH D ecem ber 28, 1953 CASLE ADDRESS: EETLAB. -JJN C IN ^ tT l TELEPHONE: C A n iO L U M ' Mr. D. V. Stroop, D ir e c to r ' pepartment of Technical Services American Petroleum Institute 50 West 50th S treet jjew York City 20 Dear Mr. Stroop: I herewith acknowledge with thanks receipt of A m erican P etroleum Institute's check in the amount of $ 3 8 ,2 8 3 . 00, covering expenditures to be m ade on th eir behalf. Very truly yours, Dr. Kehoe ef API 05742