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( Caeeer Leetem 9 (1980) 27T-284 a Elrwias/Nortb-ltollaod ScieadHs Pablim6ms Lad. COEiBSY.ATION OF IIYDUCD3Qd`i'Y OF ABYL BYDBOCAIflON HYDLtORYLASE W'P~H TO 3485"PSYLCHO UCBD LUNG CANCEBS H r ®0120097 RICNARD 8. KOURi, LEONARD EI. BB.LUP9°. T8O31A9 Ei. BUDS-•. CARRIS ffi. pHUMIRgsoo. BERNARD S,A88fi aod CAROL d. HENRY Dopaetment o/ &oeAamied QReologq ond Deperte++aAt of Saperimentd Oneobgr. A!lero0ido*a! Amoeiotr; 526! BiMar Road. B®thaada. MD 20018 rU.3 A.1 (Received S Febeaeey 1980) (Acaepmd 12 March 1980) 9UIMNRY C57BLl6Cnm, DBA/ZCam, fimt 91ia (F,). and baakcrm progeny from these 2 paeeatal sasiaa of mice were evaluated for their susceQtibility to 3-metbylcholanth:ene-induced Iung caacers. la, the ctossee among these mice, aryl hydrocarbon hydrosylaee ( AHH) eesponffiveness segregated as a =4e autosom®l domiaent gene (the Ah locus). AHH iespoasiae mice (Ahb allele) eapresoed 40-$0 uraib AHH activity/g W'et wt liver following intes- . periLoneai tremtment with 3-methylcbol9nthr+ene (MCa) compared to AHI3 non-eesponsve mise (Ahd allele) wbich eaptaseW 7-11 uait9 AHH activity/ g w®e wt liver after MCA el+eatmeat. lntiauachaal administration of 500 u3 MCA [or a total of 4 times ae weekly inteevals yielded a variety of pulmonary cancera, including squamous cell carainomsa. alveolar adenocarci- nom®a, and adeno-squamous cell carcioomes ainong mice that survived 1 year after the cardnogenn e4aatment. The AHH responaive C57BL/BCum, F,, and C5TBLl8Cnm x F, animals were much moes susceptible to MCA- induced lmg canceffi than the AFIIi non-responsive DBAlZCum mice. The laa5 cancem were also not randomly distributed in DBA/ZCum x F, besk- ctose progeny since significantly moes lung cancem were found in AFfH- +Pmomt mddmao: Snvieaeeitnral Patholoay 9.niaea, $09 Vi.n 3/iU tlood. Roak.qte. 3lat41.ed Y0881. U.S.A. OgPasnt oddmw: Dspeftmetst of Pathohp. 8otiod of M.dlefne, 97e. Ueiram[ey of Neeth Cuollaa as Chapel 1011. Pndiaial Sdneesioeal 5uiide8 aZ8iL Chapel = North Cnollos 27514. 0.8.A • •••PresQne add+a.: Natlond Camaer lmeltaWNaeioa.t Tosicology Propam, Bethotdl. MayLlad 20205, U.S.A. tAes.nt addiva: Catdnogeeeank 1eStlug, Di.hion of Caoast Cuw and Pre.ondoa, National Caeeer Imeitnta, Boths:da Metylond 20205, U.B.A

( c 278 HrCo1Lao9a responsive progeny than in AHH oon-respnnsive mice. Data support genetic linkage between susceptibility to MCA induced lung carcinomam and, the Ahb allele. n4'neo0vcrtoN Ary( hydrocmbon hydrosyLase ( AlDi) is one of the major multicom• poneat. microaomally bomad enzyme systema functionia8 in the biomo& formation of ineny drngs, hormones, or chemical pcllutants [1.2,13]. In '. the inbred strains of mice, conditions csn be obtaioed so tbat AFffi respon• sivenend' segi+e8ates aa a single autosomal donniaaat gene (15,18], a sai& autosomel codominaAt gene (191, or one in which non-indudbility ie domment [17]. tn each of these genetic syetem®, tbern is a corselation between the capecity of hepatic tiesue to respond to and metabolize poly- cyclic a1ometic hydrocarbons and susceptibility to subcutaneoua fmro- sataomas inducad by 3-metliylcbolant6r+aae (b4CAi [6.7.9,11,12]. Conditions which pteferestially alter pulmonary ARK activity have been estabtished (8,10]. Theae studies suggest tlmt„ as with hepatic tieene, tbeig is specific genetic regulation of AKE activity following intratt8chea[ treatment with polycydic agometic hydrocaebops: Thus, pulmonary AFIIi reaponafve. ness can be eaeily inferrsd feom determinations of AFIIi responsiveness of hepatic tiasue (12]. The metttodsv~ in these reports. coupled with those of Nettssbeira and Hammons [16]'fot producing pulmonsry carcinomee in Inbred strains of mice, suggest the possibility of an animal model syatem in which the suaceptibility, to lung carcinomas msy be speoiflcally linked to t6e capncity of that crgen to metabolin cheml'cal caccinogws. Tbis paper dpscribes such a model systeffi. biA?EA[ALv4 AND MS'i'libD8 C57$Ll6Cam (B8), DBA/ZCum (D2), and H8D2F,/G1im (F.) mice of both seaee wers purchased from Cumbedand View F'arms. Qintdn. TN. Bechcrvsm animaia were produced in our own laboratory. At 8-10 weellS of age, mice were inoculated iastratzacbeally wft 500 rs8 MCA in 0.02 ml stan7e 0.2% getstfrt-ealine eccotding to the procedures of Ho and P1amt (41 as modifted by Sourt et al. [8,10]. Immediately Prior to use, the solu• tion was sonicated for 30 s at setting No. 8 mong a Branson SonicatoL For intrazmc-beal treatment, mice were anesthetized witti the inhelation anest6atic, Metophane (P[tmen-Moore, Co., ZY+eator., NJ). Preliminary studies have abown that bletophane has neglfgbie effect on pulmonary °'Me tatm `rapoaei.aae®', aa osed ia tbb paper, demoas a relative ineesm® in rnw of do so.o synsba3in or of ensyme ettiaity from prxiaeieg maisetea. or in sate of boeti ahee eompeeed to Ata of bieakdowe. No paedadAr mneluuiom ie implied.

279 l t and/ar hepatie AEi aetlvfty (unpubiiahed obmvatioaa). Mice were origicelly scbeduted w eeceive 6 weekly iatracsscheal inatillations: the same schedule shovm by Nettesbeim and Hemmone [18] to paocloce bronchogema squamous call carciaomas (SCC) in inb2ed sUlins of mice. Howeeer, becuse oi the severe torieity obeenved. oraly 4 taocalati,ons were done. Controle conaeted of B8, 02 ®d F, mice which reaeived 0.2% geladn in sterile saline only. At necropeym luags wete 9aed in sita by intsattaclieal injection of 1.0 ml 2% gluteaaldehyde using an 11I PW needle.lhe organ was lipted at the tracltea betote removal from the eaimel. The Iuog was sectioned through a frontal pleoe. svith ser9sl 6 aun sections takm at 3levets in the lung. 3eetiona were stained with hesnatozytin and eoatn end easmined for Lung pethology. Iiepatle AFIIi ecavity vuea determined in all animals whbch aere temi- ae!!y wg+fica+ Mice wew tteesed inttaperltoneally (Lp) with 80 kg bACAlB body wt 24 h before seariBce. At sandfice. luaw were removed for tmeament described above and 11vess eaaeed and seored at - 70QC until aeeayed.'the emay tor hepaLic AHH activity [7.1a1 eae petP'osmed on all tim samp[es on the eame day. Activity was cdcuiated in taems of units (U)/B wet wt ttver. A unit is defined aa rhat eanount of enzyme causing _ the tiuomeent equivalent of I nmo13-hydroa ybeazo(a)pyt+sne (3-0fd•8P) per min at 3?°C. ne 3-OH-BP was detetmined in an Amiaco-Bovvman spectrophoto8uorometer with activation at 398 nM and emimioa at 520 abf. As deseamned previously (`8.9). tbere is en almost 10-fold diffe- tuce between non-emponeive and meponaive levele of AEIS in thee® sttaine. Z4iCA-tteaced, non-mspoadee animaie egpreseed AHH ieveis of 7-11 U!S wet wt lives. while activity in MCA-treated teaponmve aQimats was 40-W IJ/B wet waot liver. It should be poi:ited out that in this geaetic syetsm„ detqctson of hi8b levels of 3-OH-BP is indicative of the formation of high lesels of +iiRuetly all known metabotites of many polycydio aromatic tiydrocmboas, patticu[aoty metabolites formed via eazymatia activity at non-K-fton positions (12,141. RLSULT9 [ntzatachea[ instiUatfon of S00 Kg MCA to these strains of mice was very toffic. Ten weeb after tseatiment, only 20--4096 of the treated animela suzv'rmed: D2 mice appeered to be the moet Sensitive to thees to= aIDec a. The mWor cauee of death wae determined to be acat,e bronehopaeuatoni®. At monsbly iatavale, 5-10 oS the survivlog 88 animala were randomly Wed and esamined hietopathologicslly for evidence of lung lesione. An acnte iasemmafory reaction vrae o6.erred ia the bICA-tseated animals, these Iesions ansre ioitiaily locaied ptuaately around tee tetmiasl bron- chiole, oftim prawssed to bronchitis aad, in some atdmels. to broneho- pnetimonis. Of etie 40 smmele randomly seo:iSced during this tlme, only I wm obe~ to have evidence of pulmonerq patftological changes (i.e., NrC0rzou99 S

'pA9li I LIiNO LBffIONB O®BBRVfSD ApTRR 1NTRATRACiiBAL IN877LLA'PION OF MCA IN @ARBId'RB AND uFit8FR1iV(l ®ROe9 APpROpRIA'P8 CROBSBBINVOLVINU TIIS ®8 AND ®2 OY'RAINA OB MICS Parent 9ispraf• 71oat- ° No. Plo. Lr'+ng 6ielopat6obW° or aff- epriog oioo at Ab Ioena° qwnl of miee ow t°st of mice on te°t 10onnal 88A SN fiM AII ANCN BB .. Controi 46 48 44(06) •' a(4) 1 (2) B6 a MCA so° 22 g(a7) .2 (9) 718a) 84141 3(14) D2 • o IBCA 60 Ia 11(86) 1 (8) 1 (8) P, +0 60CA 60 28 4(14) 1(4) a0(71) 19(s0) P. K 6w a MCA 41 24 1(4) • a(19) 3(13) 4(17) 1B(76) 10(4a) Iz, x Da r1 BfCA 42 12 6(42) 1(®) fj(6o, 4433) ®, x oa .. eaCA 10 1(10) 4(40) ®Ieo) 4(40) Ntuaben of lung ACBA AAC A8C cancem per nnlmYle ®t r46d O/46 10) 8(98) 1 (6) 8/a2(88) 6(lfl) 8(af) O/ta (U) g!9®(R61 4(17) 8/t8) 1(4) 8/24(89) 1 (8) 2114(17) 1(10) 2(20) 6j10f80) ° 77.e pt.emyrpe eopres.ad at the Ab locus ia ranked aa: ++, fully reaponsiee or Induolhle. 40- -g011/8 wei vt 1i.er; 0, nOn-ro.poeolYe. 7- 11 UJg wet wt (irer. b I+Oee were teoo.ed intrWecheal erit4 ellher 0.02 ml 0.2% 8elmtialetine (Cantrol) pe 60006 6SCA In geletia'sdin° once per wea& for 4 conaecutlse armka. ° Data gitan ia lerom of numbera of mke with the observed lung hWopal6ology a1 18 months after obewlad tre°tmoat. Moro tMn I iedon wa ofteq obrened per rrtlniai eo tleet the percent inciAnce (given pareatbetta°uy) mey be greoter then 100%. (8ee toet for ekltreriWoue} d 8ummary of Faddsooe of fedoeo of Ihe type BCC. AAC and ABC per the number of animab at ri.k for the 12 montb observation period.'ibe percent is 8iren paranlbetic®Oy. ° A total of 40 onimale were randomly eecrirc®d from tbie group dudng the fir.t 8 munths efter ®ACA treatment. Oae 8N was obaerred in tlmae animals during Ihie time intor.at, (i.a. at 7 montlw). I I

1 sA1 H i®01 2010' C f ssquamous metaplasia). Sedai sacdfice ov^ae discontinued 8 momthe after the laot treatment. Twelve months after the laet MCA tisatment, alt rcamaiainp auimals wexe injected Lp. with b1CA, 24 h letae were sactfSted, hepadc A,1ff levels were detersined, and lung titenm were Sged and analyzed hiseopaeho- lo®ealty for presence of l:mg lesiona. Resnlts are shown in Table 1. A veitety of laag lesions was otrsetved in these animele and theee lesions ace dividl:d inot cate8oft of: (a) essentially nolmal: (b) thoae of squamoua cell od&: (c) those of alvedogeaic oc&: and (d) those of mised-ceLl odgin. Iesions of squmaoue call oai6(n an dexdbed as squamous metapleaia (SM), squamous neoplaem (SN) and squamous cell ca:ciaoms (8CC). Lesions of alveologenio oaipa were alveolae hypexplmia (AH), alveolae aon-comprmaing nodal®a (ANCN), alveolar compresmme nodules (ACN) and alveolar adanocatdno>~a (AAC). The only mtaedtell leaion observed was an atlanoequam~ Caioiuom.e (A9C). A derWl®d deacaption of thm pulmonaey lesi one will be published elsewhere [BWups, L.FIL et aL.. unpubMW]. Recent studiea in our laboratosy suggW a pxopeswon of alveolopmc lesions front Aii - ANCM p A,CN + AAC. Although lees well deBlted, the squalnou®1mons SN mtd SCC appe®e closely related (9]. Pulffionasy l®®iona SN, SCC. ACN and AAC all have beffi shown to trampl8nt into newbom syngeneic aaimela (olata not shown) and the le:4ons tenmed AAC and SCC an eapebie of inetestesis and. invaefon, with the heart, broncbial lymph nodaa, kidneys, pl8naa, oe braia the most uanel sitm. Data in the seudy we~e giv®o in tsnos of the number and p~tap of animala feam a paeticWat stolin which was observed to have a paaiculatr lung leaan. Mast acamale were observed to lm moYe than one type of lesion, thas the total pevccant of incideacea is usually gmster thmt 100%. Gelettn ealine trel>tsd contiol allhm®!e and MCA4mated D2 miae woe observed to be ®:IEentialiy no=ed with mJy 3--10% iacidence of esely aiveologenic Ieaions ( AFi and ANCN) at 12 months after tzeatment. No maligoant Uulg le:nona svme observed in these a>limel®. MCArtevated H8, B6D2F„ and F, X 88 animals (all ARR rasponive) apesmed a vaeiety of biCA-induced lung lesions, including SCC, AAC and ASC. A eotsl of 19 careinomlffi out of 74 DfC.4be&Wd ammala From these tlu+aa AMtvapanaive popalat[a:m were observed 12 moatha after chemiaal t>waemt. In the F, X D2 •.: pulacton, A98 aepepited into 2 populatlon®:10 mica asee observed to be retpoarive and 12 miae non-cespoaeive (ese Table 1). Of the AEffi aon- teeponlrive cniae, 5 had easeatially nounel iuop and only 2 ezpimed evideece of MCA-iadacxd lung carriaomas (15CC and 1 AAC). Of the 10 AHH rmponsive miae, all espnmed some evidence of bICA-iaducetd heng bfstopathology with 6 obaexvad to hava catcinomm (4 SCC mod 2 AAC). D78CV®e[ON intratrmheaL matillation of MCA was observed to cause many pulmonary cbenges in theee lebied stafm of mice. The cbmpaa obeerved in those ~

i ( 2e2 H TQ0120102 animels after 12 months on teaC wele of both alveologentc and squemoua call oagin and included not only anetaplastic or hypc•spleetic changee, but alao elveolo8enic and squamoLs c®Il tuaiom which wex+e oecadondly mixed (ASC), weJl dif,feaeaslated, invea~ve and/or metastatic. Such lesions were not observed in the gelatin-saline treated cont:ola. The Al4Y3 responeive 88, F, and F, x$6 mice espressed a much higher incidence of tactf type of lung lesion, in contrast to the AHS non-eeaponwe D2 atzaia where 85% of the mice viexe observed to have eamentially nommial palmcnery tiesue. In F, X D2 progeny, the incidence and severity of MCR ' induced lung lesions seemed to be associated with enhanced ability of theee animele to respond to, and metabolize, this chemical carcinogen (see Table 1). A total of 83% of the AHIa non-reepondvr, mice eapsersed eitfer eseenti®lly normat lungp or were observed to have only the eady alveoloenic lesaone of Al•Y or ANCN. The inciaienae of the AH or ANCN ledona waa sim8ar in the ARK-rmpoesive progeny suggesting a lar.h of genetic [inisa6e. between the presence of etiesa leffions and A18a responeiveneea (i.e., the Ahb ailele)• The incidence of 3CC and AAC, howevec, wea signiffcantly higher in the AHIi eesponsive peogeny (Mvs.17%PW0.038) =GIQootng geaeCc lialm a between the Ahb al1ele and susceptibility to MCA induced pulmonary carcinomm. Coneidedag the progreadve or inteaelatad, nobue of the 1ua81eslone, the cummulative incidence of these lang ledons in the various parental or oftring animsls makea the dUfamm that eaim between ASH-responeive anil -ANH-non-respondve animele evea mose acrikina. Total incidence of the lesdons designatad SN, SCC, ACN, AAC or ASC for the AEH-roepoasive progeny wes 38184 (46% while the AlM- non-eesponeive progeny wes 2l2B (89b). A nuha 1on81ezency pedod for the malignant lesions wae obrenied in this etudy, in contraet to the resalts of Netteaheiae and liammone [161. These anthors reported a high incidence of SCC'a within &.10 weeks after 6 weekly treemensa voith 500 ja6 b1CA The longer latency paaad (zpproa. 1 yes.-) and chR lower tumor incidence obmerved in this study could have oeeuited Oom the fact that only 4 tt+eatsnenta were givea: how- evea, other sendiea in our labonatories suggest that the me of the MCA crystula in the Vintin•ealine vehicle may be most important in the indactton of inelignent lun4 leam (B.E. Kou$, unpublished obaevatiton). Other studles tltat ata onVeing in this labowtory have also suggested that highly aonfcated MCA (used in this study) is more toxic than Wqp9artide MCA. The relative reaifftanee of the saimsis reporeed hmee to 9CC (compaeed to the eeedta of Nettesheim and Hammom [161) may reflect the selection of a patelCuledy eestetant subpopulation of mice due to the eQWo of h*ly toaic levais of MCA. Bowe.er, the selection proaese must not have been rlomely assocdated with ASIi act[vity, becavee the AHB non- iadueible D2 seiain wee the moet aeneitive to this tosic eftect and the attvivin8 population ftm. the F, X D2 bscksroaa sdll eaptemed the 50% aepegation pattem at the Ah locua (i.e.10/Z2 or 48% arere AH8 reepon- eive).

M HT®Q12(1103 i blepaC(c AM levele weee used to deteeimine the A8K ieepooweseaa of the miae used in this scvdy. sia0e the puleaonaly dasm were 8zed m eM ead were esam(ned hisWlo&ally. Howeves, hepatia and pulmon®ey AM levels ecs t+eguloed by rJie seme pene in caoaises betweean .B6 attd D2 unee (10]. Thus, measuaemant of AHIi leVeJa in bepntlc tlmntes io iadicsCEve of the AHH leeela in pulmon®eq tissiae: Data io tmi0 repoM suSps that the capacity to metabolize MCA map detemiae the sueceptlbllity of that tiestie to DdCA-emodaW camcem. These traautt® suggest that the sdtd9 of the eon>z+ol and regulatlon of eumon of epithel(sl oei®fn, the eypes of tnmooi observed maes $equenely in the hsemen lung (3l, can be iaitiaced ueing model syateme iavold* • iabad aaimel strains. It is ineereatmig to point out that one of the tleoms peopoa®d to be a detennia®nt in the suecaptt60fty of man to pnfmoeary cmc®n is the level (or inducibility) of AHR (51. Akhougb this 1efter obaetvafon requires conSrmatdon, the obeetvationm pneented in this paper tend to support this conclusion: rhet is, the capecity to reaposd to polyeyc•Jic aromatic hydrocarbons th:va* bciewed levda of A13H aotiei$y seeme to be geaedcally linked to sueceptibilfLy to certaio id'ads of caoc®m ACRMOwLSDGE6lElYTB This project has been 9appozesd in part by the Council for Tobacco Research. USA. Inc., and Contacts:NfDl-CP-43240 and NO1-Cp-63519 witts the Virve Caacer Progr®m of the Nabone! Cancee Iostftute. NIB, USP1A9. 88PE8FiCH9 1 Conaoy, A.B. (19671 Phaem=laWd impJlmitics of ®iamomd ensymo iedueNon. PbMVaatsel. $®N.,19, 317-868. 2 Conaay, AJL and Bumik J.J. (1972) Mataboiic ineereedoan emoag eowf:onmo~a( cheimmide ead drn0u. 8dsww, 178, 576- 586. 3 FOWoo.ton, J. and 141au CJ. (1973) Epdamiobgy. In: C+eces bledelna. pp. 261- 306. Editose: J. 8ollead and 8. Ptiaf. 1.u rmd B(biger, PhiladelQhia, PA. 4$o, W. aad PaZSt. a(1971) (aesetrashost ioedlladoa mathod ror Iw39L 0ftIIIhogy, ' 87, 097-701. 6 Btlleeesaan, Q. 9baw, C.R. aod LaytorSalloneean, EL (1973) &vl hYdowbad hydway(aao indaelhiiiley and beonehop.nie carelmoma, N. Eeal. L Med., 289. 9W 988. ~ 6 iGoud, RB.. Rateis, EHL and Whiumiio. CS (1973) Bddeeee oQ a g.a.dc rmladoaa4ip betvem .nbaacomow wmmn end indmcihiliey of ae9/ hydrasuboo bydtvsylua. J. Nad. CasearlaaR. 51.19q-900. 7 Eoud, R.&, 9Dlmno. LA. aed Wyd%ta(sa, C.B. (1978) Bs(at(oaAbips botwu.n aryl hfdeaoaaAm hydmzy6os indodbilk•y .nd som(d.lty to cbamia(ly-indaerd aab• Qatroaom mimoatiae in .ariooa rtaains of ad=: J. NnL Cmeo< [ms, 80. 303-888. 8 Bouei, B.8„ Damei9s, C.F. cnd Wditmin, CS (1974)'lhe,a®ef9amm of aryt hydeaeathm hydret9lme vasymn systeme; is eM .oleadoa of model % - - for tospiniory eaieiaogan.ela. in: Upafineetd L.aae Caeeer. pp. a"l. ILdleoa: E. Knebe.nd J. Paet. $prfttprVedag, Now Yoet, N.Y. t

Z84 HT00120104 9 1Loud,lt8. Ratao.lL md Whltoairs. CS (1974) Gasade eoaerol ot euoeapdbilfty :o 3-maehyl sabentasootta smaotaee. (at. J. Cemoar.. 13, 714- 720. 10 [ond, R.B., 11aft, T.. Thomas, P.B. and Whlemfte. C.$. (1976) Stodiee oa polmosaey aryl hydeomrboa hyd[osylo®s aodvtty in tabmd eaua® of mlcs. Chem•Plai. (ataaaeL. i E ~ 13. 317-831. 11 3roaei,LL. 1twda, T.H. aad Whito:,its, C.S. (19T8) Boladoaah[p bec+oom le.*of aeye hydeocaebon hydeosylma activity and weeepdbWty to 3-mathylaholaethme md h.aao(a)pyeaao-induaed oanceas in iobtad ateeiea of mke. In: Polyaudose Atomotic 8ydaoearbom, pp: 139-151. 6diW:w 1;1. Pfandeatlsel and P.W. domm Ra.on Pters. Na® York. NY. 12 Konti, ttSL and Nabst, D.W. (1977) Q.aatla e.vWaqoa of mmptlbWty to pol7- eyaLLa hydrae®rtiaadadnced tttnmw in the mots®o. Ia: Tho Crisiao of Ntmsen Caaxav, pp. 811-86. Edttois: H.g. Hatt, d.Q Watooa .ad a.A. Wtnoeaa. Cold 8p*g Naeber r+em c:dd Spaisg BaQhor. NY. 13 blavoa. D.S. (1987) btaahaoim of o;ygae m.teboli.m. Ndr. SnsymmA., 19. 79-a88. 14 Nob®a, D.tiY. and Jeaaae. N.3L (1979) Zhe Ah 100041: Ooaodo eo0telaUon af eho mmesAdiem of m[deogene, dnL% and oehe: aevfraommnml ahamieak by ayt~hroma P.480•m.d[ased fa: CRC C:iNad B®.fa®a in 111baboumm9. pP• 187-178. BdlPCS: G.D. Fmmatti CNC Pma, tae., 8oee Rsto0. Ple. '18 Nabats, D.W.. Gomdoa, P. and <Leloa, J.W. (1972) Ary1 hydtoaeeboa hyd:aayloes iudae:toa by pelyoydfa hydsocarboer: ffimpla auum®d domieaot nrnit In tho mooas. Nstnta Near Mol., Z98. 107-110. 18 Noantbem. P. aed Hammom, AS (1971) Indoeetoa ot eqtumon® es(l mrefomis of :he eapimtoey c~eet of m6ee. 3. Nad. Casa.e Iasf., 47. 807-701. 17 Robioaon, a.R,. Coosdfne, N. end Nobsst, D.W. (1974) Conoefe aayzeoion of aql hydromboa hydrmeglmo iadttstipn: 8aidmaee tor ehe iaadsoonnot othae gaaesie 1aai. J. 8ai. Chato., a49. 5881-8889. • 18 9°homee. P$. Souei, 8S md Httt).od, JJ. (1972) The ganodm of aayE hyd~owebas hydreaylam indeeefos in m6ea A dngle Qene djtfineaoa basaeon C87ffid8J and D8A/1d. l0eehoffi, Coaet., 6. 187-108. 19 T!>omm PS and Suetea, 1.J. (1973) (i.aaeice of aeyl hydrocarbon hydeoayina iaduetion io omco: /ldditlte iah.ritaeeo in cremmem hatwrm C3WHe,i med aBA/Qd. ffietbm Geera.. 8. a48-387: .