Document gDB1o47Mr70Nbkpd6o3LLbZZ3

DSRTPITTS-A6295.5, 6889.1, 6316.4 3M MEDICAL DEPARTMENT, CORPORATE TOXICOLOGY Title: Inter-Species Comparisonof Mechanisms Fluorochemical Toxicity following Intraperitoneal dosing of perfluorooctanesulfonate (PFOS, T-6295) or ammonium perfluorooctanoate (APFO, T-6859) in Rats and Guinea Pigs and oral dosing of N-cthyl perfluorooctanesulfonamido ethanol (N-EtFOSE, T-6316) in Rats. Final Report Date: May 25,2004 Study Numbers. T-6295.8, T-6889.1, 6316.4 Strategic Toxicology Study Number: DTIS-A Sponsor: Study Location: Study Director: Study Toxicologist: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul MN 55133-3220 3M Strategic Altemative Toxicology Laboratory 3M Center, Building 270-SB-181 Saint Paul, MN 55133-3220 Andrew M. Seacat Ph.D., DABT Toxicology Specialist 3M Medical Dept. Corporate Toxicology and Regulatory Services 3M Center Building 220-2E-02 Deanna Luebker M.S Senior Toxicologist 3M Medical Dept. Corporate Toxicology and Regulatory Services In-Life Start Dates Part A: In-Life End Dates PartA: In-Life Start Dates PartB: In-Life End Dates PartB: In-Life Start Date PartC: In-Life End Date PartC: 1023/1997 and 11/17/1997 11/04/1997 and 12/1/1997 02/10/1998 and 03/11/1998 03/10/1998 and 03/13/1997 09/16/1998 09/18/1998 ' SDRTPITST-A62958, T-6889.1, T-6316.4 `Tableof Contents M"ETestAMitegsprmemmrres ee emtmsm-------------- dh ProPcaerdtuA:resP.rocedures foar single intFEraperitoneal dosing ofPROS (T-.639%) and APFO (T6889) in male rats nd4 PfaorltloBw:inPgraocseidnugrleesifp.o naecTtiimoen-CoofurFsOeSofaPtFtOwSo d(o1s.s6.2.9.5.) toxicity in male and femalerats nd inca pgs 6 ResultPsaratnCdDiPrsoccuesdsuiroens.for hepatic peroxisomal gene induwctiiosn pollowing oral dosin-- gofN-EAFOS-- E in rat. 6 PaRrEtSAUSRe:suRlEtsSPoAntYheAToo xicityofPFOS and APFOe inrats nd guinpeigas. ml7 Liver samples submited for L-A FABP analysis romrats 3d uinea Pgs rated in Part Aree 8 CParot BR: RuesusltOsimofoPTAniLmAes:-CoursecofcPFOS oxic at two doses in male and female at and ina pgs. coe8d RPeasurlCtt:s:RGesusiOprigansldPearotaBs.ofiatnswigth 20mg/Kg N-EAFOSAE. --rill SiL gnatures. S --------------------------tl Guineapigs. wn nm----------_-- ""TTaabbllee21. PPaarrtt AA_. SSuummmmaarryyooffTTooxxiicciittyyooff PPFFOOSS aanndd AAPPFFOO iinn mmaallee rguaitnsea..p.igs _....................14 "`TTabalbel34e.. PPaarrttBB..SSummuarymooffBBmooddyyaWWeeirigghhttyssiinnRGautisn,eMaePaignss,aMnedaSntsdDanedvSitadtions. Deviatons. 1165 "TableS.PartBLiver 0Body Weightratios it af... ccc " 1" FFigiureg21::uaarrrtetBB::MFaelmaelReaRtaBtoBdoydWyeiWgehitghVtsVTsime...__T ____ ime...I"e FiFgiugruer4e3::PPaarrtt BB::MFaelmaelGeuGiunienaePaigPgBoBdoydWyeWighetiVsVgsTiThmiem.te... r------ 20 AFDiPgEuDrGeIX4:1.ParDtEMB:IMRaIlOeN10SGutihneePaIPOigOBGoOdLyWv eighte VsTimee . rs cool2) `APpaprentdAi:xRIfaPtardtatAa:Toxicty ofPFOSand APFO IndividualsandSr ummam ry Dm a.--..---- 23 AppPearntdAix:IG.uinPaeratPBi:gTdiamteacourse with PFOS.: Individual Dat...rm------------ ll PaPrarBtt. BI.ndLiivviedruaWleRiagthtdatoaTBiomdyeWCeOiUgThtWaitthoPiFnSm.a.l.e nd femal ats combined data. SAS anasis 22]8 F Part.Male12t LW/BE W 30SUI. arr2 PPaarttBB.,FKeimdanleey rWaetiLghWt/0BWBroadtyioWSeuigmhmtarraytio in mal and female rats combinedm daa. SAa S snalyw ss. .36 ParPtarB.tGB.uGiuinneeiagaP:igInLdiivveird0ualBDody A WeighD t Rat. io Summary Statistics... rn 39 AppenPdaitxB.IV.KPiadrnteyCOWreailgdhto0sBiowdniytghWeNig-hEtFrOatSioEiinnmaatl.e aInNdIVfGemUaRl)e0gu8in.ea.pi.s. SAS anaIyis coord]43 2 SDrRITSTA-62958, 6889.1, 6316.4 Introduction "The research presented in thisreportwas designed to elucidate the mechanism(s) that initiate peroxisomal proliferation in rats, but not necessarily in guinea pigosr primates, by some 3M fluorochemicals. The research objectives were to determineifthe fluorochemicals disrupt the function of iver faty acid binding proteins (L-FABPs) in both rats and guinea pigs andifso, to what extent. The intermediate objectives werteo investigate the hypothesis that FC compounds alter L-FABP levels andor functionality and that it i the disruption of fatty acid processing leading to higher levelsofunassociated long chain fatty acids in the cytoplasm, and that rats and `guinea pigs wil respond differently. Further objectives were to investigate and compare the `molecular mechanismsofperoxisome proliferation in rats and guinea pigs. Three fluorochemicals; perfluorooctane sulfonate (PFOS, T6295), N-ethyl perfluorooctanesulfonamido ethanol (N-EFOSE, T-6316) and perfluorooctanoate (APFO, T6889) were tested in this study that were ether previously known (Sohlenius ef al. 1994; Sohlenius ef al. 1993) or suspected to cause peroxisome proliferation in the rat. The hypothesis tested was that these fluorochemicals would induce peroxisome proliferation i the rat, but not the guinea pig, and the data supported that hypothesis. Theeffectofthese compounds on peroxisome proliferation in the rat, but not the guinea pig were presented in an abstract and poster presented at the SocietyofToxicology meeting in 2001 (Wallace ef al. 2001). Other investigators using these fluorochemicals have also subsequently shown peroxisome proliferation with someof these compounds in vivo in rats or or in-vitro (Maloney and Waxman 1999; Yang et al. 2002; Berthiaume and Wallace 2002). L-FABPs isolated from certain rats and guinea pigs dosed in the current study were used to investigate the effectof those fluorochemicals on the binding ofa fluorescently labeled probe to L-LFABP. The detailed methods, results and conclusionsofthe L-FABP analyses were published elsewhere (Nabbefeld 1998; Lucbker ef al. 2002), and are only briefly mentioned in the resultsanddiscussion sectionofthis report. Guinea pigs may serve as better surrogate models for human risk assessment than do other rodents because, ike humans, they are resistant to peroxisome proliferation. However, the `molecular and biochemical mechanisms that differentiate the responseofthese species to peroxisome proliferators in general and for the perfluorosulfonamides in particular is unclear. `Therefore, the specific aimsofthis study were to: 1. Characterize the peroxisomal enzyme and faty acid binding proteins response inthe liver to these compounds 2. Perform toxicity tests such as serum clinical chemistry and to liver histology that may indicate an explanation for any the species differences in response to these compounds. 3. Cpoerrrfelluaotreosaunlyfoonbasmeirdveesdaanldtetrhaetiironmsoetfabtohleitaebsove functions to liver and serum levels of Methods "This study, (DT-15A) was conducted in three segments, arts A, B and C undear broadly written protocol dated 9/18/1997 that had been approved by the 3M institutional animal use 3 SDRrPITST-A6205.8, T:6889., 6316.4 committee (3M IACUC Animal use application # 2088). Part A consistedofastudy designed to compare the toxicity and peroxisomal proliferation in male rats and male guinea pigs dosed via intraperitoneal (i.p.) injection with PFOS or APFO. `The results of Part A led to further testing under part B, which consisted ofa dose-response and time-course for the observed effects of PFOS in both male and female rats and guinea pigs dosed via intraperitoneal (i.p.) injection. The resultsofpartAs and B led to the conclusion that ip. dosing was not the best route of administration, therefore an exploratory oral dosing procedure i rats was performed with NEFOSE in Part C Test Materials "The T-numbers, chemical names, abbreviations used for these samples, and the chemical structuresof eachofthe compounds that were tested in this study are given below. The currently accepted abbreviations and the ones generally used in this report for each compound are in bold: 1. Vehicle control (2% Tween 80). 2. T6295: Perfluorooctane sulfonic acid, potassium salt (perfluorooctanesulfonate), PFOS, FC-95, Formula: GF S0;- K', MW = 538.1 g/mole). 3. T-6889: Ammonium perfluorooctanoate (PFOA, APFO, FC-143, Formula: CHFCOONH:', MW = 431.1 g/mole). 4. T6316: Neethylperfluorooctane sulfonamido ethanol (narrow Range N-Ethyl Perfluorooctanesulfonamido ethyl alcohol), N-EtFOSE, E(FOSE, Formula: CFRSONCHICHCHOH, MW = 571.06). 5. Wyeth-14643 (WY, MW = 323.79 g/mol) was obtained from Chemsyn Science Laboratories, Lexena, KS. Wyeth-14643 (WY),wasadded to DTISA as positive control dose group for hepatic peroxisome proliferation. "The protocol stated tht T-6868, N-ethy! perfluorooctane sulfonamide (FX-12, Sulfuramide TM), FormulaCi,SO:NHCH ) wouldbe tested, however, T-6868 was not tested in DTI5-A. Procedures Part A: Procedures for a single intraperitoneal dosing of PEOS (1-6295) and APFO (T-6889) in `male rats and guinea pigs. For part Aa t,otalof 15 male rats and 13 male guinea pigs, 6-8 weeks in age and weighing between 150 and 250 grams were purchased from Charles River. Three to four male rats or `guinea pigs per treatment group were used. Following an adaptation periodofnot less than one week after arrival at 3M, animals were administered treatments by intraperitoneal (ip. injection, a SDRIPTST-A6205.8, T6880.1, T6316.4 at equimolar doses that were less than 40% ofthe rat oral LID 50 for PFOSorthe rat ip. 24 hour LD 50 for APFO. "There were solubility problems in corn oil for the compounds. Therefore, dosesofeach test `material at 32 mM in 2% Tween-80 and using a dose volume of 5 mL/Kg were used. The doses were calculated 0 be 0 the highest dose that could be delivered at less than 40%ofthe LD 50 Ffroreschacsholtuetsitomnasteorftieasl.t sTuhbestsaenlceecstiwoenorfetphreespeardeodsfeorleevaeclshidsodsiinsgc.usTsehdedfuorstheergrinoutphse parnodtofcionlal, numberofanimals in each dose groupwereas follows: 1. `Cgounitnreoalpiggrso.upEsa.cThhaenivmeahlicline tchoentvreohlicglreocuopntcroonlsigsrtoeduopfre2cemiavleedraatssinagnlde tihpr.eeinmeaclteion 2% Tween-80. In addition, no treatment control group consistedoftwo male rats ethfaftecrtesocefitvhedenvoehtircelaetcmoennttrowleroen tihneclsuudbetdleinentdhpeositnutdsyoftgoechneeckinfdourcatinoynpprloannonuendcfeodr athnids stthuedyn.o tTrheeatbmieonltogciocnatlroalnrdatcslwineircealccohmebmiinsetdry. data from the vehicle control group 2. PcoFnOcSe.ntrationAs=i1n7g.l4e mipg. PinFjOecSt/iMonL)ofw3a2smgiMvePnFatOaSvionl2u%meTowfee5nm-L8/0K(gfin(aflinal dose ~ 3. 8A7P.F1O5.mg/KAe)sitonagletoitpa.l tihnrjeeectmioanloefra3t2s amnMd fAoPurFmOalien g2u%inTewaepeings-.80 (final c7o0ncmegn/tKragt)io0nat=o1t3.a8l mfogurAmPaFlOe/rmaLts)awnadsthgrieveemnaalteagvuoilnuemaepiogsf5. mL/Kg (final dose ~ 4. 5W2y.e4t5h-mg1/4m6L4.3 W(WYY)).wAassignigvleeniapt.ainvjoelcutimoen ooff W1 YmL/iKnecgo(rfninoailld(ofisneal~c5o2n.c3enmtgra/tKigo)n =toa total four male ras and three male guinea pigs. oEfxa35m0pl&efodrotsheecagluciunlaetaipoingftohri3s2cammMe tPo:F(O0S.3i5p.kga)t(S0.m0U0/5KLgk,ga)s(s5u3m8i.n1gga/nmoalv)e(r0a.g0e32weight mol/L)= 30.1 mg/guinea pig,orapproximately 86 mg/Kg PFOS. dAolslianngi,maatl2s4wehrouerosbasnedrvdeadilfyorthmeorretaafliietryfaorndthseidgunrsoatfitoonxoifcitthyedsutruidnyg. tBheodfyistwefiougrhhsowuersreafter erxepcoorsduerde iamnmdeduipattoelfyouprrtioirmetso tdhorsoinugg,hoduatiltyhethienr-elaiffieerp,haasnedofatthneecrsotpusdyy,. tFhoelalnoiwmianlgstwheerfeirhstoused siancdriivfiidcueadllbyyiCn mOetaanbodlgircocssagneescrooveprsnyigphetr,faonrmdeudrionneeainthderfedcaeys w1e2roercdoalylec1t4edp.ostA-ndiomsae.lsOwregraen tbiisoscuheesm(ilciavelr,ankaildynsiesy., tSesetcetsioannsdopfanicvreeraksi)dannedy,utreisnteesanadndfepcaenscrweearsewsetroreedplfarcoezdeninat1-08%0bCufffoerred pfeorrfmuasleddewhiytdhe galnudtsruablmdiethtyedde faonrdhissutbomliotgitceadl faonrahliyssteoslobgyiclailghatnamliycsriosscboypeyl.ecStoromnemliicvreorsscwoeprye. mBeltoaobdolsiatmeplfeorsmwateiroencaonlldecttoemdoantitnoercrfooprscyhbayngheesaritnpbulonocdtcuhretmeoisatnrayl.yze for est compound, 5 SDRrPITSTA.62958, 6889.1, 6316.4 Part B: `uinea piPgrsocfeodlulroewsinfgoarasiTnigmlee-iCpo.uirnsjeecotfioPnoFfOSPF(OT-S62a9t 5)twtooxdiocsietsy. in male and female rats and cBhaosseednofnorthsearmepslueltasnoaflyPsaerst wAer(edimsocduisfsieeddb.eAlowti)mtehecoduorsseeso,fdo3,se6,vo1l2uamneds,28anddaytshewtaismecoponidnutcs.ted to measure peroxisome proliferation induction in the liverofmale and female rats and guinea pigs treated with either 16 mg/Kg or 100 mg/Kg PFOS. Aguitnotcaalopfigs2,26a-t8swaenedks20inguaigneeaanpdigwse,ig12himnaglebeatnwdee1n0 f15e0maanldra2t5s,0agnrdam1s0 wmearlee paunrdch1a0sfeedmfarloem Charles animals wRievreer.adFmoilnilsotweirnegdatnreaadtampetnattsiobnypeirntiroadpoefrnitootnealles(s.tph,a)ninojnecetiwoen.ek afer arrival at 3M, dEoigshetgrraotuspasndwecrieght16gmuign/eKagpiPgFsO(4S/saenxd) w10e0remtgr/eaKtgedPiFnOeSa.chTfolluiomriotcthheemivcoalludmoeseoftgrhoeupi..p.The icnojemctoiioln,thaen10a0ddmign/gmL1..5smulsp2e%nsTiowneoefnP8F0OaSnwdacsreamtaidneg abnyedmiuslssoilvoinngina200glmasgs tPiFssOuSe girnin0d.e5r.mLA stihnegl1e00ipm.gi/nKjegctdioosneofgtrhoeup1a0n0immagl/sm,La.nPdFatOaSvsoulspuemneosifo0n.w16asmgLi/vKengattoathveoalnuimmealosf|inmtLhe/K1g6 to msagc/riKfgicdeodsoengdraouyps.3O,n6,ea1n2iamnadl2p8erpossetx-dpoesre.species per fluorochemical dose group were `oTnweovaehniicmlaelscopnetrroslexanpiemraslpeecaicehsarteceviovleudmtehoefvvehoilculemecoontf1rolmL(/2K5e%g.ccoomrroeislpionn2di%ngTtwoetehne 8100)0, amngi/mKalg dpeorsesegxropuepr sapneicmiaelss,pearnddoaste0g.r16oumpL/wKeerge, scaocrrirfeiscpeodnodinndgatyost3h,e 16 mg/Kg dose group. 6, 12 and 28 post-dose. One Taswiongmlael.ep.raitnsjreectcieoinveodfaWY100asmag/pmoLsi.tisvuescpoenntsrioolngorfouWpY. Oanteoafvtohleumpeosoitf|ivemLco/nKtrgoltoraftosrarectoetiavled dose or mg/ml. 100 mg/Kg suspension WY. The other positive ofWY ata volume of control rat 0.16 mL/Kg received a single ip. fora total dose of 16 injection ofa 100 mg/Kg dose group WY. The positive control groupraiswere sacrificed on day 3 post-dose. ParCt_ nats Procedures for hepatic peroxisomal gene induction following oral dosingof N-EtFOSE ofadmini"sTthrearteisounlt,stohfeprafrotrseAananexdpBlorlaetdotroytohrealcodnocsliunsgiopnrotcheadu.rpe. idnosraitnsgwwaassnpoerfothremebdeswtirtohutNe:- EintlFiOveSr,EainndPafrotr NC.-ETthFeOpSuErpmoesteabwoalsitteoagneanleyrsaitseisnalmipvleresanfdorspeerruomx.isAomdaolsegoefne20inmdugc/tKigon/daasysaNy-s sEtWuFdyOSwEitfhor30t0wopdpamysN-wEaXsFcOhSosEetnhaatscaanusineidtallisvecrreeefnfeicntgsdaonsde.iTnhdiusceddospealwmaitsoyblasCeod oAnoaxiddiaetsaery dacitcitvairtyysitnudliyvewraosvcearlacudl-awteedektopbeeriaopdprinoxwihmiactheltyhe2a3vmegra/gKeg/Nd-aEytFfoOrStEhecsoencsounmdptwieoenkoinfthtehe study (fRoerft.w3oMdayMse.diAca4lmDgep/tm.LT-s6u3s1p6e.n1s)i.onTohfreNe-Ema{lFeOrSatEsiwne2re% Tdowseeednw8i0thw2a0smpgr/eKpagr/eddayinN-gEl(asFsO.SE. 6 SDRrPITST-A6205.8, T6889.1, 6316.4 tissue grinder and delivered to the rats by oral gavageat a dose volume of5mL/Kg for two coofnSsemcLuItKievge dfoaryst.woTcwoonsreactustwievreeddayoss.edTwhiethantihmeavleshwicelreeceountrhoaln,iz2e%d aTnwdeenencr8o0pastaicddoosnedvaoyl3ume ppooslty-pdroosep.ylLeinveervisalwse,rpelawceeidghoenddrayndiccehaunndksthoefnisvteorrewdearte-f8r0o.zenSeirnulmiqwuiadsnoibttraoigneendthbeyn put in cleivnetrrisfaumgpaltieosnthfartomwebrleoofdlacsohlflrecotzeedn binylhieqauritdpnuintcrtougreen awterneecsreonptsoynadnrdysitcoerefdoratan8a0l.ysiTsohef~g2enge induction to: Kendall B. Wallace, Ph.D. D.A BT. Professor, Schoolof Dept of Biochemistry Medicine. and Molecular Biology 10 University Drive Duluth, MN 55812-2496 Results and Discussion Part A Results on the Toxicity ofPFOS and APFO in rats and guinea pigs. Results: Rats Part A bInodaytsw,etihgehrte rwaetrioesnwoersteatiinsctriceaalsleydsifgonlilfoiwcianntg cthreaantgmeesntinwibtohdyallwetihgrhete.coLmipvoerunwdesi,ghptaratnidculliavrelry-toAPFO; however, these changes were not statistically significant (Table 1; Appendix 1). `dTehteecsteironuomfcl4i5nimcagl/cdhLemiinsrtaryi tarneaaltyesdiswirtehvePaFleOdS.thaGtlcuhcoolessetewraosl swiagsnilfiocwaenrtelyd tloowbeerleodwitnhaetlsimit of treated with PFOS and WY. chemistry parameotferratss No statistically significant treated with APFO. changes occurred in the clinical 7"TRh4e8h4i5stoTlRo4gi8c4al7 arensdulTtsR4fo8r337)ousthoofwefdourmaractrsotsrceoapteidcimpa.swsietshof52whmigte/KsgpotWsYon(tAhnilmiavlernsuumrbfaecres covered in subchronic hin scar tissue and evidence granulomas were noted. of mononuclear infiltration. Multifocal microscopic "eTnhlearhgiesdtolliovgeircsalarnedsuKlitsdnfeoyrs.ratMsatcrraotsecdoip.ip.cwlietshioAnsPoFfOmo(iatnliemdalblnaucmkbseprosts7oRn04o8n5e1-l5iv3e)r wsehroewed noted (7R04851). Microscopically, this animal had apadofscar tissue with an imbedded `granuloma on the liver surface. "reTphaeirh.istAolllogoitchaelrraetssu,ltisnfcolrudoinnegrtahtetrceoantterdolwigtrhouPpFrOatSs,(hTaRd04n8o3s8i)gnsihfiocwaentd cshiagnnsgoefplrievseerntdaimn aligveer, kidney, or pancreas. 7 SDRTPITST-A6205.5, T6889. T6316.4 Results: Guinea pigs Part A One guinea pig (Animal number 7G0253) overnight following treatment. No tissues that was treated with were collected from 100 that mg/Kg APFO animal and the died initial body wdeeaitghhtis (u3n5k8ngo)wonf,thbuattmaaniymahlavies nboeteninccolmudpeoduinndtreelattaebdl.esTohrestgautiisnteicaalpiagnsaltyrseeast.edTwhiethcPauFsOeSof had had significantly significantly glroewaetrerweliivgehrttgoabiondsycowmeipgahrtedrattoiocsocntormopl.arTehdetogucionnteraolp.igs treated with APFO dOanmeaogfefroeupraigru.inNeoa psiiggnsitfriecaatnetdcwhiatnhge1s7.w4emreg/pKregseinp.t iPnFtOheSl(iv7eGr,02k2i5dn4e)y,shtoeswtesd, soirgpnasnocfrelaivserof anyofthe other wo outofthree guinea control pigs examined microscopically (7G:2004 through 7G2013). guinea pigs (7G2255 & 7G2256) were never analyzed and Tissues were: from disposed of. Liver samples submitted for L-FABP analysis fromratsand guinea pigs treated in Part A. iCseorltaatiinonliovfelrivsearmpflaettsyfarcoidm bcionntdrionlgapnrdotPeiFnOaSt tthreeaUtneidvreartssiatnydofguSianneaFrpaingcsiwsceore(Aunsiemdaflosr the 7PGF0O2S00r4a-t)c.onTtrhoelggouailonefatphieg,L7FGA0B2P00s5tu-dPyFwOaSsg1u0inaesasepsisg.th7eReOf4f8e4ct4-ocfonFtCrsoolnraLt-aFnAdBTPROf4u8nc4t8i-on as deivmaleutahtyeldambiynotnheapatbhilailteyonefstuhlephfolnuyo!r)es-ceunntdefcaatnyaocicidacainda(loDgAuUeD1A1 )- (t5o-bind to L-FABP isolated mfraoxmirmautsmabnidngduiingnecaappaicgisttyroeaftLed-aFnAdBnPotfrtorematPeFdOwSitthrePaFteOdSraitns vwiivtoh.ouRtesaunltisncsrheoasweedinaKdde.crTehaesed complete methods Nabbefeld 1998). and the resultsof those analyses are reported elsewhere (Lucbker ef al. 2002; Conclusions from Part A: o"fThseohmiestoalnoigmiaclals r1e2suolrtso14fdlaiyvesradfatmratgreeawtimtehntewviitdhenWcYeo,fsAcPaOr ,tisasnude/oonr tPhFeOsSurlfeadceuosfttoheconlsiivedresr nthoattntehceesdsoasreisl,ydtohseebevsotlummoedse,lrofourtetohfeaodbmjiencitsitversaotifotnheansdtutdhye.tiFmuertphoeirnatnsaclhyosseesonfftoirspsauretsAfowrere total organic fluorine, electron microscopoyr, aborted and Part B was planned. peroxisome proliferation from part A were Part B: guinea Results pigs. of Time-Course of PFOS toxicity at two doses in male and femalerats and ReAslullrtasts:RtaretastePdawrittBh either 16 mg/Kg or 100 mg/Kg PFOS gained weight following dosingand S`tmaatiinsttaiicanledanbaoldyysewseoifgbhtosdythawtewieghrtessulgigghetsltyeldestshtathatnhecroenwtreorlevnaoluseisgn(iTfaibclanet3d,iFfifegruernecses1 abnedtw2e)e.n wtreeiagthetdsawndarconitnrcorle,absuetd hinadbovtehrymalliteleanpdowfeermableecrauassetorefattheed lwoiwthN10(s0amtigs/tKisgndootseshgorwon)u.ps Liver (Appendix 3). The liver to body weight ratios inratswere significantly increased in combined 8 SDRrPITSTA-62055, T:6889.,T6316. male and female liver body weight ratio values from the 100 mg/Ke dose group versus conirol values (Table 5, Appendix 3). The female rats had moreofan increase in liver to body weight ratiosthandid the male ats. Hypertrophy and white pigmentation were noted by gross observation at Kidney to BW necropsy in ratios were nsootmseiognfitfhiceanhtilgyhafdfoescetegdroinuprartasts(A(pAppepnednidxix3).3). Kidney weights and Results: Guinea pigs Part B tPhFaOn Smawlaesgmuoirneeatpoixgisc. toThgrueineeoaftphiges ftohuarn freamtsa,laengduianpepaarpeingtsl,yamnodroenteomxiacletogfueimnaealepgiguitnreeaatpeidgs w1i2t%ho1f00tsmgin/itKiagl PbFodOySwiepi.gdhitebdyodnadya6ya2nodftwhaesshtuumdya.neTlhyeeruetmhaaniinziendgofnemdaalye 6g.uiAnlleaotphiegrlgosutinca ptihgrsouggahionuetdtwheeitgihmteacnodurmsaein(tTaaibnleed4b,oFdiyguwreei4g).hTtshtehagtuwienreea spliigghktildynleoywteorbtohdany cwoenitgrhotl rvaatliuoessfrom all animals, males and females combined, were significantly increased by treatment with 100 mg/Kg PFOS i.p. (Appendix IID). Ftheemafalcet gthuaitnctharpeiegosutinofpfarotuirculfaermwaleeregufionuenadptiogsbediveedrywistehnisnittihveeftiorsPt F2O4Shotuorxiscaitfyt,redvoisdienngcewditbhy.a p1i0g0 lmosgt/wKegigdhotseuopftoPdFaOyS6inpo2st5-%docseo.m Ionilcoanntdra7st5,%onTeweouetnof8f0oaunrd mtahleeregmuainienaipniggfdeimeadleagfutienarea ripe.madionsiengof31m0a0lemgg/uKingedaopsiegsofsuPrFviOvSedinan2d5g%aicnoemd woieligahntd f7o5l%loTwiwnegetnre8a0tmaetntth.at dose, and the The administration ofthe 100 mg/Kg PFOS dose mixed with 25% corn oil may have contributed tothe observed toxicity ofPFOS 0 guinea pigs dosed in part B. Noneofthe male guinea pigs dmiLe/dKefgo,llwohweirnegais.po. naedmmianlisetarnatditohnroeeffaem1a0l0emggui/nKega PpiFgOsSdideodsewhien n2%theTwseaemne d8o0saetwaavsodleulmieveoref5d biny2o5ra%lcgoavmagoefltaotbaevmoolruemetooxficItmhLa.n/ Kwgh.eTnhedreleiviseraepdriencaendeanqtufeoorusdesloilvuetriyono.fTPhFeOSuseinocfcoonmoiloil `iTnhteheratdo2s4ehporuerpaorraatlioLnD-ha5s0pfroervPioFuOslSyilnead 2100.:8c0leaacretdoinfef:erceonrcne oiinltshuesopreanlsiLoDn w50asofdePtFeOrmSiniendattos sbeus2p5e1nsmigon/kwgas(Ddeeatneremfailn.ed1t9o78b)e. 1I.n2c5on-t2r.a5stg,/tkhge(rGaatborriaellL1D9-756)0.foNroPdFeOaSthsinoarngraoqsuseoleussions were oabqsueerovuesds4u8spheonusrisonasfoefr dPoFsOiSng(mGaolldeeannthdafleemfaalle. r1a9t7s9w).ithThaussi,ngtlheeoursaelogafavamgiextduorseeooff2255%0 mcgo/kmg. oil in an aqueous suspension of 2% Tween 80as the vehicle to deliver PFOS may have contributed to the observed increased toxicity of PFOS to guinea pigs when compared to an 2% Tween 80 vehicle alone. `Conclusions for Part B: Intraperitoneal administrationof PFOS led to the formationofsar tissue on the liver. The findings in ths study led to the decision that .p. dosing was not the appropriate route of administration, and that subsequent studies should be conducted following oral administration of these compounds. SDRTPITST-A6205.8, T6889., T63164 Part C: Results Oral dosingofrats with 20 mg/Kg N-EIFOSE. "sTihgenrsoefwatosxnicoitayppoarrelinvterefeffefcetcotfs wterreaetampepntaroenntbboydygrwoesisgohbtsoerrvlaitveironwseiagthnte(cArpoppseyn.diTxhIeV)r.atNsoused dwaetrae. Tofhdeiflfievrerenstpeagceismeannsdwienirteialanbaoldyyzewdeifgohrtisn,duthcetrieofnoroef,msRtaNtiAsticfsowrepreernooxtispoemrefporromleidfeorantitohnese. sBpleoctifaincalgyesniess,.hTohweerveerwatshentootaaplpmarReNntAinwdaiscaptairotnoiafllgyedneegrianddeudctiinonallbysaNm-plEeFs,OtShEerebfyorNeo,rather `quantitative analysisofthe results was not aborted, and the tissues were disposed of. performed. Further analysisof these samples was Conclusions oSbttaanidnaerdd itnoxPiacrittsyAteosfttsohfissesrtuudmycilnindiiccaaltecdhethmaitstmrayleangduimniecaropsicgospwiecreexammoirneatsienosniotfivtehteoltihveertoxic eafnfdeoctnseoofuPtFoOfSfoaunrdmAalPeFgOuitnheaan pwiegrsedmieadleovreartsn.ighInt,PaarntdBt,hethrreemeaoiuntionfgfofuermafleemgauliengeuaipniega Wpiagss sIanccroinftircaesdt,onnodnaeyo6ftdhuee mtoaleexcoerssfievmealweeiagthst ldoisesd, ffoolllloowwiinngg ai1p0t0remagt/meKngtiwpi.thdo1s0e0ofmPgF/OKgS.PFOS. `toTxhiecsietydartealasthiovewteodrtahtas.t t`hTehegufiinnedianpgisgsi,n tfheimsaslteusdiynlpaerttdoicutlhaerc,ownecrleusvieornytsheantsiinttirvaepteoriPtoFnOeaSl `dcoosnidnugctweadsfnooltlotwhienagpoprraolpraidamtienirsoturtaetoifonoadfmitnhiessteractoiomnp,oaunnddsthtaottseusbtstehqeuestnattesdtuhdyipeostshheosuelsd. Aben ignriotsiasloobrsaelrdvoatsieosntsutdhyerweiftohre20thmegd/oKsegsNw-eErGeFcOoSnsEidinermeadloe rbaetslsohwoawsedthneroeawpeprareennot eafpfpeacrtesnbtyiver Nef-feEcttsF,OaSnEdwtehreegdeneeemiendducnteicoensssatruydifeosrwfeurtueraebeoxrpteerdi.mTehnetrse.fore, Oral dosing and. higher doses of 10 SDTRIPSATT.86,T2689915. T6316.4 Signatures Prepared by: Cindi J. Seat Andrew M. Seacat, Ph.D, DABT. Toxicology Specialist & Study Director 5/25/2004 Date u SDRTPITSAT6205, T6889., 6316.4 Part A. Histopathology Report. Elden Lamprecht 3M Lab Animal Research Services: Rais TR484S Macroscopic mass imbedded in liver surfacecovered in thin scar tissue. ts substance has been dissolved leaving a foamy matrix ofprotein and mononuclear cells. The interfaceofthe mass and liver consistsof foamy mononuclear cells. TR4847 ~~ Macrospopic mass protruding from liver surface covered by thin layer ofscar tissue. Is substance has been dissolved leaving a foamy matrix of protein and mononuclear cells. "The interfaceofthe mass and liver consistsof foamy mononuclear cells. R485 A padof scar tissue with an imbedded granuloma on the liver surface. R487 Multifocal microscopic subchronic granualreoprmesaenst in liver. mTiRl4d83pe8riportalPvaarciueotallatpiloenuorfahoefplaitvoercyitsemsariskepdrelsyentth.icSkeonmeed dweigtrherdeaptaiiornotifsshuee.paAtocgyetneersamliazyedbe preset All other rats had no significant change present in liver, kidney or pancreas. Guinea pigs 7G02254 -A macroscopic. padofgranulomatous inflammation present on the surface. It has a foamy appearance with pore contents being dissolved leaving. The matrix which remains is bcyoamnpoisnecdroeafsimnognloyntuhcilckealrayceerlolsf,sscoamretoisfswueh.icShcaarretiessnugeoringteedrfoarcevsacwuiotlhattehde.liTvehreppaardenicshcyomvae.red No change in liver parenchyma. A second smaller pad is present on another locationofthe liver. 762255 & 7G2256 - never analyzed and disposed of. 7G2004 - 7G2013 - No significant changes were present inthe liver, kidney, testes or pancreas. n STRIPTAT-62058, T-6889.1, T6316.4 Tables: `Table 1. Part A. Summary ofToxicity of PFOS and APFO in male rats [a Parameter Avg Jovg a] 15T0ucowsJog 150 TucowoJovg [so Jen TTTTTTT1 TTT 1 F---- TT 1 Fr Tr 7 1 fverwti@ ol sal wood] valrd]sees]sul weed] zo sal ar] [iverbodywt 1ooo]oczsl ood oom tran] ooeo] voto] tere]ood oo rere] [crfleflim emery emergence) leimcaichemisw|y T1 T 1TTT TT | ol owas | ool | a ws sind wl s3 ud |sl oa aul 02] ood anowl oodsoru o rd Bunmga1 weal od sol il ood vas im esi] wen] sel ed) forumga1 t ool oe uo] al soo ss wel cer re]sooa]d] | sol"odsolo woodwwed al ind wil ol] ed Pool od ol wal sod wl oof seed wal ml ay) | vor oll ool wl ood wlesl wool ol 12s ued] jou 1 ool ve solsl esd oulsoie]sol sulowlsq] Patmmott Tvl sel red a wore val aa oll re oss woe] fommott Toso]oe oo 7] rol sol rod od 7a]owl sq febm1msoolt sof ool a] wd oil om oor]ses]soo org] jcreamgiat Tos owl oul oo sod os ool ed ouloi] esq] rRom1g[avord mls wsdl al wl ed nl wl ed 21.. TNhAeNlotiaopfpmDliectaiebcltetion for Cholesterol (CHOL) was45 mL. *Significantly diferent from covnaluetsbyrStuodenlt's T-test 1 pSRTPITSTA-62958, T-6889.1, T6316.4 Table 2. Part A. Summary of Toxicity ofPFOS and APFO in male guinea pig ablo2GuineaPigs[ Conwol | Pros |aro | wvern | [1TTTT TTT T TT [Bodywt@)at2te| scoon] reel soson] aos ord srerlease] sod]wosor u e ood ueightgain eo |eoon| peelsoso] 16] 00] stool sss] eae] mer] rea] eos] errr essJee e Le s [veg| voloadva sa l teed vwvl oso] ew til vod l[Gimr ealGr hemir sw|y f[ri|efT eelT e| V eeLer eee reece] Prosmgat 170 22 es 1a l 72dooui sof ouowl raf mga Too oadoa or] ord ooloa] oso zw]oo] eed [BUNmga Tes orfvex sw enol iss om!sso woolaesc]ed peu 1soor ] orlmsl sor] ondsess odsss o ssn] ] ed |ssioo | sss ssaoel]s sos a sosuos n oaosolesros ol se oo] rwsodl d rsz eoceo]o zrsaun ] ] sseodd | corso]sosox]weaso]sou] ave seeocloseoro]o]mwsoes oled] | woo] sslaso]sel | ool aslsulssl sor sad suzoo]aes] soo owl tor] soo[sow]somo| valsoo sown]sown] Jer-mmoit 1so57] ose]waco]sea ood toz00]oor] seefsomo[sone]rove| eorun Toul was]nso] aml vod redzo eid vialos eed 2. TNhAeNToitnatppolfiDceatbelcetion for Cholesterol (CHOL) was 45 mg. + Significantly different from control valuesby Student' Ltst 14 SRDTPITSTA6258, T-6889.1, T6316.4 `Table 3. Part B. Summary of Body Weights in Rats, Means and Std Deviations. [Do(smegP/FgO)ST [PosDta-Dyosel [Wear Body Weigh] [F T7Tl 787e 0 [7] [F 757T e Wm TW Jo] NC NY CANN1 [Cot 28 |I = Hit [F 7Tw e 6 [4] [ TF sTwe Tw [ TF sTare 177 3] [ TF w 2%e 17 [2 [FE T 282 Tw[7] [7F 7 wee Ts Ta [FesTeTw [7] [TF sTwe T3613] [Fee 1717 [TF =7 7%o Tw[1] LC J C I 1 [o 5 WRn TWATo] [r 6120o | 11 [2 [FTeaT7e|S--T7 17] [Tr sTwe wT] [re Tew Ja) [e Tams Twa[7] [we Ts Tw 13 [F % |i =Te is resTee Tm 1] [re ese Ta [ Tm sTaee ee [Tm s12,e |12 [3] [W 72Tw wa e | 21Ja] [Wwe T728 Twa3 [1] [To TTm n Ts 12] [o TSTo NR m TWA To] [went T5120 |7[2 (roseTs Tv Cr 2[awo |W]1] is DSRIPTSTA-62955, T6889.1 T-63164 ``GTeanbdleerD4o.sePaPrFtOBS.DaSyuPomsmtaDroyseoMfeaBnoBdWy W9e)iSgihdDtsevin NGuinea Pigs, Means and Std Deviations (mag) FFo occont 31 NsOMAow 20 FFoo Ccoonntt 5 d 20 wNA 21 FFoo Cont 2 1 mon 1 dB 4 FFoo 3 us oN 1 5 mm 73 EFo os 2 w ww owx 21 E Eo o 1 3 ws 2x 4 NR NAO Foo Foo 5 2 NM oN 1 Mo MFo oocont 21 Mmw oM7 02 MMoo CCoonntt 35 2NMA0 02 MMoo CCoonntt 2 st NA on 1 MMoose 1 3 mw 4 a oN Mote Mo 5 2 as am 2s 3 a2 MMooew 2 1 0 71 ww 7s MM1100 3 5 3 NA 1 mw 2 M10 M10 22 w[ e EoNY1 16 BSRrPieTn2958, T6891 T3164 Table 5. PaBrLivter toBodyWeight ratios in rats [Cont ----"""TRat [16mgKgPFOS-- [Rat M&F [4 Jo05 M&F [8 loos Toor | Tool | T0inoReProS [Ra [Mar [5 [oes [001 | Peeler emir rr---- [Contiol TGT F TT ooe Too | [G6mgwg TGP | 18 Too ___[oo2 | [roomgwg TGP Ta Too Tooor | Scaetnfomctn viyD'essFt. et emmme ----m eel r pr es sSarena vw SRbrPITSAT6295. T:6880.1, T3164 Figures Figure 1:Part B: Female Rat Body WeiVgsThimte. "imeof-PFOCStooxiciutyatrtwosdosee, 16mg/Kgand 100mgKg. FeRmatBaWVlsTieme x0 20 f2o0 ww 0 o1 3 sn DaysPostose |n--+--Cyont| = is SDRTPTITS-A6205.8, T6880, T6316.4 Figure2:PartB: MaleRatBody Weight VsTime. `TimofePF-OStCoxiocityuattrwosdosees, 16mg/Kgand 100mg/Ke. 0 o"sWw0 100 o1 MaleRat BW VsTime ==cant | 10 3 6 2 Days Postdoso 1 S TenR 9s8, TT s, TGs Figure 3: Part B: Female Guinea Pig Body Weight Vs Time. Time-Course of PFOS toxicity at two doses, 16 mg/Kg and 100 mg/Kg. Female Guinea Pig BWVs Time 0wo 0 roorn :bi =| xwoo wo o Toss no OaysPoston 0 DSTRISPATT-T6682800. 57.63816,4 Figure 4:PartB:Male Guinea PigBody WeightVsTime. "Time-CourseofPFOStoxicityattwodoses, 16 mg/Kgand 100mg/Kg. MaleGuineaPig BW VsTime. 0 &0 S5w0 Em ES 100 o1 --o--Cont ate | = 3 6 I) Days Postdose. 2 SRDTPITSTA6205, T-6889., T6316.4 Appendix 1. Deviations to the Protocol: 1. The protocol states that there would be Five male and five female rats and guinea pigs per treatment group, with S treatment groups faototral of 50 rats and 50 guinea pigs. In Part A, only male animals were used. Three to four animals per treatment group were: used and a total of 15 male rats and 13 male guinea pigs were used in Part A. In Part B, a totalof20 rats and 20 guinea pigs, 10 malesand 10 females per species were used. "Thus, in both parts A and B, a totalof 35 rats and 33 guinea pigs were used. 2. The protocol sated that N-thyl perfluorooctane sulfonamide (FX-12) and Nethylperfluorooctane sulfonamido ethanol (FC-10, N-EFOSE, T6316) would be dosed. "These two dose groups were removed from the protocol, anda positive control, Wyeth14643 (WY), a model peroxisome proliferators, was added as a dose group. 3. The protocol sated that all treatments would be administered in corn oi. PFOS (T-6295) and APFO (T-6889) were administered in 2% Tween 80. Wyeth-14643 (WY) was administered in 25% com oil. 4. The protocol stated that Dunnett's t-test would be performed on all data. Student's t-test were performed instead where noted. 5. The protocol stated that all treatments would be given by i. dosing. Dosing by oral gavage was performed in Part C. SRDPTITSTA6295, T:6889.1, T3164 Appendix IL. Part A: Toxicityof PFOS and APFO Individual and Summary Data [pf en wn mn rp 1 f17 T 1 1 | ewarr[7 [f 7 mt fam el |8]fe l few ml[| fomn |me emm m meem ew fgoazr [om eed we we ed Fi5Joeleer 4]z==e 7 EF CM CP --LTr)r S--S r PX MGNY:IS Nr Y:M1 XMR|. [[lKeesnteey@@| | 3s32]l 3z4o] 32020saooll 33a6ooofff a333 2503 3i2 3ai2 ooid) | GinricrairGheemiestrry, ie|Spi[ tfee|es|fee |fmt |fei|feeeee|) [opormgal| 52 oo ar |Ga] wzfeas Tas es|WA|WA| JJUoNLmUmgigaaL|| ze1asl 7ai8s] gioarlo_f1270533] s 20ssaie] i foo] T1easl 1i4s033 10d JJAASTPUULL 2o7s132s6ol] stiiall27i0o0l]szasls ssoosf] ersell atsolo zsoeoll3e3t7o0 s8e2 CJL0OHULIs7 so pee]T Bsa | Soi] 7T aal rr seri) aol R daa S5529) [JNKaesmmmmoonl[T|91241 tsaesl] 7ta6d_I|8ra25l i off t5a3s stasal staaal ieses 31]7 JJoorRmEmAetmtgL|--fo[i]otsoo ooss I ooer]o1f9 oess 0oo4lo ool s soo s o10d) JesTur TooolTsooooo a so od 5 SDRTPITST-A62058, T-6889.1, T6316.4 FRaatnd:ata (cont) animal T= 5*=slavg|sof = 2 |= 3 [=a Avg][s0]| I ll FlI I FlFl llPS [EBBrogAdaMytEwAtSY.Nami21 fpmsm gms os wr|mn ame Body WG) gee JemimaYost Wt pm remap med 258 757 lutgan | sso] so] sol sro]sa]"if sun]wo! 11s] so sme ma] Jgain% |"70%[38%|37%|11%|3%|0.2]39% |43%|43%|17%| 35%| 0.1] [Lvveerriogd)y. |289] 7961236 344 266 64) 196] 186] 185] 256] 205]34] LTM 1 aos] sos]oar ous ous ao oas| eos aos] aos] aos] oo] [iidney@ | 391"35134 48]3s]oe) 37132]34] 43]37]os] [Tests |36 30]34]36]34[o3)31 34]32] 37] 34] 03] F11 1 1FF =11 1 1 [ClinicalChemisty [~~ | | [TT 1" |Choimgial. | 53.0156.0|520 7806001]2]650]630]60.0]530] 603]53] JcamgoLT1151 11411141123 117]0426 11.1]112]108]11.4]os] [Phos mgial|" 112] 72.0172.11148726]16)121 720 715] 11.5] 118]03] [remo I 011011oi]02]oTofoiloi]oi] oi oi]oo] [AbmgaL 34134139142] 37104 34 35133] 35 34] 0.1] |iPmgaL| 61] 61] 66]71] 65] 05)61]63]61] 66] 63]02] [BUNmgdI 740 730 7601 750745 1.3}77.0 720 730] 17.0]748] 26] fGLumgaLI 206[175 4s11 |254 4[75 60|210 142 139 208] 175 a0] [AKPUL1416 264]370| 415] 374| 63)332 401| 264| 245 316]67] fasTonIsa 540 960 760 880 6.9)760 80 760 384] 139]173] fAcun es esT" es 79] e7| ef sol 7a e2| eo| 67] 12] [LNOavHUL 1340| 2341 309| 451]334 | 90) 262 233| 167 |2205] 717 983| [omen | sis] sealss vr vs | es vi] saa] si] ssa] fremmo I 701891 011ool8570)73 74 65 73]71]oaf [cEmmo 7011700[701102 fo1| 1f too e6| 96] 102] sol 3f [csReEArmguisrTL--so10s4o]l s0oal] 0s5o] sooal]0eo0f)0so5]0es4]s0o4]l s0o6l]s0o5l[0o1o]] RI [me[ofvol volonlsal ofwosolsrs]sso] rs] of i Ei UU,A [ =se acrfiv cedona daCy1d o4rate her than12 EIR REE Limitofdetection for Cholesterol = 45 mgial EY SDRTTISTA2955, T6809, T3164 Far ar Guinea ra |] we ef o Pig dataoTal=STagTwfT [5 1wily [so| EEE ll eB ll cl allI [ames ----TTT TTTT raach MdWlWldd clWdddIdIO [oarae n |r| ss sr]wl3| wl& w|sr wl 1) [[hveerr@ow[t[70204]T T0o0r4] 01.7074[]0T0r4a|]5G0s0]] T0a0i3l] T0e044|] 0200s7]] nTooss] oTsrs]|G3o4r]] [[oTensetse@ || s1s2]]s16]]a1ts]] 3t8a]]ooza]] taos ossslaoas]] t41s] tcro]loods] fF Gmearc-- hemssr yr 1 rr |1 r1 1| [CChEoTimgor7P 57 P ss as] ws to] so w1s 1 a]e1 l G srl 7] [[TPoriomsamiga@t|Totoil t2e1lotir foslo2s oer1o7r1osrlowrl] oor]Io1of [[oTPmmagia ||2--a2s]T 2s8s]2d4o]]2sso]looss]| aats]]2a2s] 2a2r]2s5il] 2a3r] oosa] [[SAtUemUerLa||=iTs aasss 2277s]| sgaesr|] troer]] Tairrs]]stsoo]] eaas]]22522] aesa]] ss [[oArSTUL | s7s1 s7o1s] sess] teotstsoa]] sass]]saor]sasl| asso]l ssal soff [[LoOsHmUmLo || t7a7s711z3ro6 t2a00o]stta[Tos4o]] Teas2]] tasse]]tToral| oteass]] ttsaar] s2o] [C iemmoit|1]717]ss]28]o so] 41s]aa1]3 7s] ss Ts] l[osgtsor [f 5]=oo] 7oe oofleWel]w es]w of] owel]owfl]] _o_w]ow1]] TTT EAE Ec 7 XG x rSRiTsaT2058, T-6889., T6316.4 FGuaihne:a Pig data (cont 1 1 1 1 T --T--7" T "1 | [ animal # 1% 5 1% "Tag[so [5 1% 1 3[Ayg [s0 | EE al al I Bl I [Parameter ---- TCCCar at Gow| | wa]om]wf se]w]eww]]w) [w3igan |[6] [a| ]e]2608 78] so] so] 2 | es] deo]iro] 176]12) 62] ev] teal ieil 1s] [Testsi@15o. e] v1 o] to1 fotf oe] ds]os[ 12] os] r -- 1 t 111 FH TT 1 1 1 1 |~ ClinicalChemisty ~~| [ChoimgioTLeas T<a5 1 821s2[NAI" 53] a5] as] as] | | oil il vel nil tb wl ml el nl of [o 8 oll ol of"ss s 8] ] 1] [Abmgiia1L 2335] 222]2stoo] [24124]ot} [es sia ] a8s ]03] "a7 45]a8] 47] oz] | as el is[ as] +f 1e1 20]as 18 --sf | so iat] ssl 0 ef dar 22 212] sa sel | 323 200 367[ 32r| dof aoe so317] 3a8| 75) | ess 40] eof es]20f es] es] s5| 71 of [si ar"ss| si] af a7 20] 47[ 38] 16] LOHUL _|""aa2| 215]237] 264 68] 250] 2s2|212] 238 23] [Nasmmoit| taal tao 1 wep] SPP P ENT a [wa [7sT oil 771cefm i NA Tha 1 3 1 3 [E 2 I | 0303]os[03sloo6f oa 03 03] 033] 006] cotun Iwi] ss flwl20 wn wn 2 1 1] [|T=RFiicstmogloigya.t| 727] 771] Fes] deo] 3s) ov] Fas]Tie] ee] a] . R =sacficeR d on day 14ratthhaen1r2 % |Aes lSbpL nanLsms || ii d |stiimai)1 ub 1 1 ssa LaLedHr] i seals| gi ] se Lal nl ety iil : Ermsscdimoatiani i sz iis: LEE eh I H{ai l i nem ZZ Fi] edPaorta=eB.SALSivaermWelight to Body Weight ratio in maleand [EE -- female rats combined fou- T = YO TO | | | --_-- - ne x p = WrCor " CH aaentet Dunetrs. Rs -- en ag oFuin Re feterm Soe) bie5 [r poeI T2 sovaSonirumea MCeosmumms eGnhv A La=eJ--I--S[i--o--m sSpeEr = HCG z SrEorsTsnoorla odeotoowtpannce rc eoUna &Z P oooEm lomeens iomm D=grntentsessGarpoaoruarsS ssy smw ounls odnuoimm En 0H fe ims norJ aSansa or a2c1h50smiw inogsScn ns om DIS 11/16/95 study 110s0 o000o048t0e8s 0000004810685 000002039498 Cont 002s 00088 001143 `CoPmopsairtviesovnaslwueishshaocwopntoliousfimngesaDnusnntehtattsaMreetshiogndifcanty diferent. 242il255 Ao10s0(DALSD "000012080 C1osnt 00.000020086817 iPotsietirvae valuesshowpas of means that are signfcanty 2 WOLWW 8DosePFOS(mONG) DTIS 11/16/98study , o ODs L I |= co ~~ ~~] | = com EE bil serosa "ein msemmeonu wr | s-e E EEneem , S=E pBo O1pEEhTi EEREEER Lm"wceEeemmamae E Fd O EE eE ae: Bsusc LBOT EJ ESa eegEdaEhTE THE 5oos liEoaf nolo sgouuts i.o&S ns oE :C n0veoan ood Y DITIS 111698suady C1osnt Dooo0mcses 0000i2s8e3t 00001152611 `PCoosmiptairviesovnasluweisthsa coh nptaoilso uosfimnegwaDnusnntethftsrMeetshiogdnfcanty diferent. 270ol802 A1b0s0(DA-LSD 001C1o7n3t C1ont Zoo0t2e6s2s4 Poste valuesshowpaisofmeansthat re sinsiign fcanty 3 ons Part B. roar Female rat Liver weight summary Th I :, "17 AmyA /] -- +4 rama A-- wNnTE-- ST E-- EBEemeaem, sUbo2o pBeOTo a ER ETER mveanme uone doER smE Te Leae tAsrEEpeTEsEnerytoroeseen n agliFR iE Em wpso eJdE iE -- toasSvootme worse Py yo DIS 11/1698 sudy Cont oA72m0o2s 2s8u7s2 23eusn0 CoPomsiptiavreivaslouwenissshhaocwonptraolsuosfimnegaDnumsntehta'tsarMeetshiogdnfcanty diferent 270I0]02 Ab10S0(DILSD 164c4o1n5t C1ont Jae0s7e6 Poasfietriovnetvaluesshowpasof meansthat resignfcanty 33 F-- Part B. FeLmWalBeWryat LDoWs/epBrWosriamtgiorgS)ummary. [om | emt AN DTIS 111698 study ] ge-eNd A~N ( A) LCa ] z ZT ( aos NZ \( | = w RG f | onrosirone oo py Rsqare OnSeovamyaAyrFcata osssis RRSoaoqtuMareeaAnGSauaeErcr Q00s0is00%0i10 OHbosaernvaotfiRoenssp(oonrSseumWeis) Qos oSorlce DF2 ArSumofToavoiaoSroaiiaouniacreess MemGnSoquswe Raatvisot ECrooal 7 oooomsozs 0G0oss pooesr iLowelMefoNroaOmnbeev"rnay+oArseovMaeoan S0i0E0ro0 Cobny t 23 0aooess0 oooodo Lol S1M4NaEoramrbonreursaensdaSpeoMDeedvainaetstiiomnastSeoufDaervrvaSriua Enea Coiinwt 23ToQauoeosso 0ood0oedos oocoaoueunrss Di0c0Meanf earMl]onsComparison0s0 0000000 oo0ms60 oorcssoo Co1nt 0"0o2iss 000000000s0 0Do0c0aaezdedd Apta"Comfporeaach|praiiusiosncgsSotudnenstst Aioos00L5D 23804804 1200 0000767 0corczoannt DIS 11/1698 study C1ont 0000001027317 000001800022 000010481072 `CoPmopasrtiesovnaslwueishshaocwopntariosuosfimnegaDnunsntehtattsarMeetshgoidfcanty afferent Abs(O2A7}0aL5lS0D2 Cont 11600 "0.000000874 cont ote2t aPiotsietrievnetvaluesshowpaisofmeans thtare signifcanty 3s PartB. Kidney Weight data. SAS analysis. I to Body Weight ratio in male and femalerats combined LA A DLA WY 1 ~ WRA ~ 00 1 Cont. EmA EDfii | . DITIS 111698 study OSnuemwmaayrAynoof vFiat ReRqSquuaarreeAg MRoeoatnMoefaRnesSpqounasree. Error Observationors Sum Wets) o0o.1o0r9e132 0000000641385 2 MSooduercle OF Analys'iSsoufnVoairSiqaunacees MeanSquore Ratio 2 1000000005 258 006s ECraol 7 00000000000805855 33542447077 P0rsosweo0r LevelMeans foNruOmnbeewrayAnovaMean SwEror 11080 58 oooooiarmss 00000022 Cont 4 000280 000031 Std MEreoarnusseasndaSptodolDeevdieatiosns.toifermroravartianece L10e0w 1 Number8 00M0easn 00S0u0D1e8v 8 00a 0000s SwE0r0m0M0e1a8n 000026 Cont 4 000280 0000500 000025 OifMeani MeanM)eans Comparison1s6.0 100 0000000 00000010s 0000o1n2t5 C1o8nt 0000000001030 0000000001030 00.000000070205 Alphas`Comparisons foreach+ pair us0in0gsStudents t AL10S0(OILSD 21090100 1s Cont 000085 000065 000067 Co1snt 00000008875 000000005875 00.0000008627 CoPomsiptiavre ivaslouenssswhitha ocwonpirlauosifimngreaDsnusnthnateaMreetshiogidfcanty diferent 237ol772 As1(00DILSD 000C0o7n8t Co1nt oDooootross aPoesitrivoe values show paoifmreasns that are sigicanty Ed 1 |xa|rgL s[TWI IT it| || [deps rot|| [lead]Lal] suuidhrT L|||Ise aTssT2l 0penny efdkT LL e I bb ce 1bLLLHOOOL LL tioelh (boLo iL, IU 0 Part B. Guinea Pig Liver to Body Weight Ratio Summary Statistics: LWWBy DosePFOSImgNG) _ | smd LR | go V > Xt oo Vv { 7 NZ Say - pn oowerosrona P= Each ParA oe Wh -Cont ES DTIS 111698study OnSuemwmaayrAynoofvFat ReRaSauuaarree Ag RMoeoatneofaRnesSpqounasree. Eror Observations orSum Wets) SMooudrecle OF2Analys'iSsuofm`Vo0af0r0Si0qa0un0wc5ee0s6 ErCroarl 1i3s 000000000088053484 o0o0s8s524sa8 o00g4z146s877 is Mea0n0S0q0u0a0r3e F04R0at8i4o 0000000000s5 P0r6o7o2f3 LevelMeans oNroOmnbeevrayAncvaM.ean SwEror 110s0 G 5 oooswErs 000000102E4s Cont 4 o0wr0 o0ot2s 1d EMreoarnusseansdapSodolDeevdieatsitonas.teof ero variance L1e0v0el Numbe"r owMesan SouoD0euvs SWE0r0M0o0a6n3 C1o5nt 54 ooo0swrrso 00000as70se 0o0c0o0t8s8s Di10M0eanf}Mean(M]eans Compa0r0i0s0o0n10s000 000037156 0.001C5o0n0t C1osnt 00000001358 0000000101020 00.000010102050 Alphas`Comparisonsfor each!pai si0n0gsStudents Ab10s0(OALSD. 216037100 00038 0291 00c0o2n3t c1osnt 0000009213 0o0o06t9 00000201%6 `PCosoiimvepvaaluwreisishahsocowonpntarisorlsousfimngeDaunnsnetthaftsaMreetshgoadicanty different Abs(D2i4y62L3)S5D8 10 11080 ""000000433338 Cont "000283 fPoesrietnvte valuesshowpais of means tat are signfcanty " DIS 1/1698 study Part B. Kidney Weightto Body Weight ratio in male and female guinea pigs. SAS _ analysis KEBy DosePFOS ms) | i "N 1 _ OQ) otis oe ~ he aousrosmons Ad ZN Lm) | CA ww emovers " DIIS 111698 sudy OSnuemwmaayrAynoofwFait ReRSqquuarree Ag MRoeoatnMoefaRnesSpqounasree. Error Observationors Sun Wats) 05o2s0e3s4s87 00000040448473 1s Source OF Analys'iSsuomfoVfaSrqiuanacree.s MeanSquare FRato EMordoerl 22 000000000000323386 010906060072 P8r59o3E9 Col 1 000000573 4o%e7 000 LevelMeansfoNrOumnbeewray AnovaMean SwErmor 11080 47 o00o04ns 00000002127 Cont 4 ooo2%0 00022 Std rMoornsusaensdaSpiodoDleevdieasttomnaste ofror variance 1L0ev0el Numbe4r 000M5e2a5n0 00S0u0D5e0v0 SWE0r0M0o0a2n5 ciosnt 47 0oo0a0ns3 00000000530708 000000002154 OifeMeani}MeantM]eans Comparison1s00 Cont co10n0t 00000000010 00000010000000 00.000010110077 0001 00001 0000000 Apha=`Comparisonsfor each pair us0in0gsStudents 217t882 Av10s0(DILSD 000018080 00c00o3n1t7 c1o6nt 0o0000003s127 00000000658 00005012s 000.00005025 `PosCitveovalmwueishpsahcoowanptorauosiifinmgreDasusnnsneotthaftsnaMreetshsiogdifcanty diferent 245ol055 Ab10s0(OALSD 000017080 Coint 00000000421159. fPoesrietinvte values showpaisof meansthatare significant 2 DTI 111698sudy Appendix IV. Part COral dosing with N-EtFOSE in rats. Individual data. Anima# GendeD(omsoeKgN-oEaWyFlOoSrEtwosSpecie B(W (B@W LW(WLWB Sadcaryipfoiscte [[DNaetceropsy days. aa dose s8oRO217 M Control 68RO246 M Control 78RO246 M Control Rat 3463887173 0045 Rat 41634204201 0048 Rat 45374612219 0047 3 ones 3 ones 3 eness 68RO217 M 78RO217 M 8BRO217 M 20Rat 5237 539302 00% 20Rat 48564884238 0049 20Rat 4073 503238 0047 3 ones 3 wees 3 wees RNaottes,w2e0remdgo/sKegdiosantpwopcornosesc3i0u0tdmpiapvyamsei,t9te1h1e6ld/1i9yt9.8,and9/17/1998 with20mg/Kg/day NEIFOSE, then sacrificed on 9/18/1998, DIS 11/1698 sudy References Berthiaume, J. and Wallace, K. B. (2002). Perfluorooctanoate, Perfluorooctane Sulfonate, and N-ethy-perfluorooctanesulfonamido ethanol; Peroxisome Proliferation `and Mitochondrial Biogenenesis. Toxicology Letters 129,23 -32 Dean, W. P. Jessup, D. C., Thompson, G., Romig, G. and D, P. (1978). Fluorad Fluorochemical Surfactant FC-95 acute oral toxicity (LDS0) study in rats. International Research and Development Corporation, Mattawan, MI. Gabriel, K. L.. (1976). Acute oral toxicity in rats of T-13891. Biosearch, Inc., Philadelphia, PA. Goldenthal, E. 1, Jessup, D. C., Geil, R. G., and Jefferson, N. D. (1979). Metabolism study with FC-95 in rat Intemational Research and Development Corporation, Mattawan, MI. Luebker, D. Interactions J, of lHuaonsreonc,heKn.cJa.lsBawsist,hNr.atM.li,veBrfuattetnyahcoifdf-J,b.iLn.d,ianngd pSreoatceaitn,. AT.oxMi.co(l2o0g0y2).176. 175-85. Maloney, E. K.,.and Waxman, D. J. (1999). trans-ActivationofPPARalpha and PPARgamma by structurally diverse environmental chemicals.Toxicol Appl Pharmacol 161, 209-18. Nabbefeld, D. (1998). Investigationofthe Effects of Fluorocarbons on Liver Fatty Acid- Binding Protein. In Environmental Health, p. 50. University of Minnesota, Minneapolis. Sohlenius, A. K., Andersson, K., Bergstrand, A., Spydevold, O., and De Pierre, J. W. (1994). Effectsofperfluorooctanoic acid--a potent peroxisome prolifeator in rat--on Mors hepatoma 7800C] cells, a rat cell line. Biochim Biophys Acta 1213, 63-74. S(1o9h9l3e)n.iuPse,rAf.luKo.r,ooEcrtiaknsesosnu,lfAo.niMc., aHcoidgisstaropmo,tenC.t,iKnidmulcearnodf,pMe.roaxnidsoDmeaPlifeartrtey,acJ.idW.beta- oxidation and other activities known to be affected by peroxisome proliferators in mouse liver. Pharmacol Toxicol 72, 90-3. Wallace, K. B., Luebker, D. J, ButenhofT, J. L., and Seacat, A. M. (2001). Perfluorooctane sulfonate and 2-(N-ethylperfluorooctanesulfonamido)-ethyl alcohol are peroxisome proliferators in rats, but not guinea pigs. Toxicologist 60, 348 Abstract ID: 1657. Yang, Q., Xie, Y., Alexson, S. E., Nelson, B. D., and Deierre, J. W. (2002). Involvement ofthe peroxisome proliferator-activated receptor alphainthe immunomodulation caused by peroxisome proliferators in mice. Biochem Pharmacol 63, 1893-900. "