Document 6w6Q9ojr9Gj8JJrJdMGRQdxMg

SDRrPITST-E6295.8, T-6316.4, T-68682, T-1071.1, T1321 AR226 1903 3M MEDICAL DEPARTMENT, CORPORATE TOXICOLOGY T6295),TNit-leet:hyClomppearrflautoirvoeoMctoalneecsuullafronBaimoildoogyeotfhaPneorlfl(uNo-rEotocFtOaSnEe,suTl-f6o3n1a6t)e,(NP-FEOtSh,yl acepteartfel(uoFrOoSoAcAta,neTs-u70l7f1o)n,amaindde/o(rN-PEer{fFlOuSoAr,ooTc-t6a8n6e8s)u,lPfeornfalmuiodreo(oFctOaSnAe,suTl-f7o1n3a2m)iodfoin Rats and Guinea Pigs following Oral dosing. Final Report Date: May 25,2004 Study Numbers. T-6295.8, T-6316.4, T-6868.2, T-707L1, T-7132.1 Strategic Toxicology Study Number: DT-15-B Sponsor: Study Location(s): Study Director: Study Toxicologist: In-Life Start Date In-Life End Date In-Life Start Date In-Life End Date In-Life Start Date In-Life End Date 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul MN 55133-3220 1. 3M Strategic Alternative Toxicology Laboratory 3M Center, Building 270-SB-181 Saint Paul, MN 55133-3220 2. UniversityofMinnesota, Duluth Dept.of Biochemistry and Molecular Biology Schoolof Medicine 10 University Drive Duluth, MN 55812-2496 Andrew M. Seacat Ph.D., DABT Toxicology Specialist 3M Medical Dept. Corporate Taxicology and Regulatory Services Deanna Luebker M.S Senior Toxicologist 3M Medical Dept. Protocol: Protocol: Amendment #1: Amendment #1: Amendment #2: Amendmen#2t: 11/16/1998 1120/1998 03/01/1999 03/05/1999 02/19/2001 02/23/2001 CONTAIN NO CB! 1 "49 SDRIPSTHT2953, T-6316.4, T-68682, T0711, T7132. TaofbColnteents IDAATLmSm-BeePOnNnNCuuOmmddbbPeeRrrOmmG21BRPPeAeIIIOEOGSnSSnOEUIItEStS ove v or om oreo]7 ABYC THOS... 9 LCia lviteeraFllCuoRroEchSeTmi.ca.l.COBGERITONS so eew n e 1T132 E Palmitoy] CoAoxidise(POO) SCVIT Y..vvrrer 13 SSigUnTaatbuAlre1e.PTAavageDerI.aE.g.eComulac ive c Doses .r..nTeremLmeososm------t. 1193 19 aTabble2e3..SSuummmmaarryyooffOBrogdaynWweeiigghhttssaannddBOrWgaGnhatbogBeosdy Weight "Tabl4e.Percent TilBodyWeigh, Combined Data... Rais. . rssma--2_0d il TETaaTblbOele5..LLiiavvteertrh-PeaenrUfdVlKuiEodSrnoesyOulFtfoRoBOnoGaKdmEyiSWadEenLeid.sgMhettaRbar otliiotse,CVoalme ubeisnferdoDmaRe tast.ssndGoinrPigr s. Anar lyses 2243 A"n"TaTaballbyels7ee.sHSpeeuprmafmiocarPrmayeomdifatLtcityvh]eeCr3oPMedcEfNOlnVuioIdrtaososeunAlmcotenianvnitityad,elsCoanmdbiMLenteadbbol.ite.Va.lDa..s. rom Ratsr snd Ge uineae Figs 21 `AAAppDpPpeEenDndiidxxi3x12...-CDAunmaEulyltaVitciaIvl1e0iDAtvoHesIErePSIIOnaOdmOiNpCvlOiedLSuLalaabnIddSeunmtmairfyidcactaat.oieo.n.......................ooooon 2208 31 `AAppApppeepnneddniidxxi45.x..OLBriogvdaeynrwWFeeliigughohttrs,oInOcdrihgveiadmnuiatDolcataBaanW.ldrSaItuimonms.adrInydiiavnivdduSaiulmDadamntdauaSr.uy.adm.amtlaa.ry dots. rr3450 ooo. 45 RA. ORCatandGuincaPig B. RatandGuineaPig LiverFC LiverFC concentrations (All unitsae ut g/g).e Analyese aiin University PercentofDose Evaluations.Analy a the Universi of o4f5 C(D..A.llunRiRtaastataeLniduveGgru.PiTnFehOaeSPXSiAMgnLEainlvvyeisrreosFnCmaetPtnehtreacle3nLMtaoEbfnaDovnsiyeraEonvsnalymuzeaetnditoranastl.lAinvaerlysLesaatbte.3M.eSnamp.les).vionment]5$44 APLEbP..ETIes7cXsh:niCcUalBRCeHpoCrtH:ELIiSvIeYr.Fvluoro ocarban Metabs olites --Univee rsityof Rochester..... 5698 IAB. SnStuadttiissitiiccvssanioodnn SCCdlluiiumnnimiaccaaarlllyCChiCelnmiiinsitcaileCsRfooErrIGSrutiisn,ymeaalPeigasn,dmaelrDemaaanldavfa.esm.a.l.ecvoamlbuiensedc.o..mbined. 6713 108 AAAppppppeeennndddiiixxx1890..:HeEFipiafatlifcoREfePDaaOcclFuiEttoe0y8F]CFCaOOd-mSAiAn0(i3sTt0Tr0aIt5)io.n.o.n cataiAaEsHeVaIYn.d..a.ccy.i.CcocvAooexciodnoaosoenesxnprresion...114133350 AApAppeCnpdioxe112m1,n;EFendiodnpfilaoelifRxnafetcpstoCbroytrifrdioeforlfeaFrnteOniStto.AonrALeia(tvm7eer1ntP 1gr)o.upR s o liO verPFS OS in. at .." 86 183 B. Conoea felffecatsbtydifofernenttreatmentgroups o liver PFOS inGuineaFigs,malesandfemale 2 50 DSRTPITST.B6295.8, T6316, T-68682, T-10711 T-7132.1 Summary dAodsuelstomfavleehiacnldefceomnatlreolS(p2ra%gTuwe-eDeanwl80e)y, r4a0tsmagn/dkgH/adralyan(gmukidn)eapeprifglsuorreocoecitvaendeosruallfognaavtaege E(PtFFOOSS,E,T-T6-269351)6,),40orm4k0dmogr/k1g6/0dmakydN-NE-tEhtyhly!peprefrlfulourooroocotcatnaenseusluflofnoanmaimdied(oNe-tEha(nFoOlS(AN,-TD6a86w8l)eyforratfsourrecdeaiyvsedaonrdalwegraevagheumdaonseelsoyfsvaechriifcilceedcoonnttrhoel (fpifrtohpdyalye.neMgallyceolS)p,ra1g6u0e-mkd ppeerrfflluuoorrooooccttaannessuullffoonnaammiiddeo(aFceOtSaAt,e (T-F7O1S3A2A),foMr5f5o6u,r dTa-7y0s7a1n)d, woerr4e0hmukmdanely sacrificed on the fifth day. rAaltlsabnyim4a0lmsksudrvPiFveOdS. Tahnedp1e6r0cemnitdiniNt-iaEltbFoOdSyEwesiiggnhitfsicwaenrtleysriegdnuifciecdanbtoldyydewceriegahsted in ccoonmcpeantrreadtitooncsonotrf>ol6s0t0o u8g5g%.an4d08m7k9%diFniOtiSalAbaonddy w1e6i0gmhtk,datFaOveSrAagAe sliigvneirfPicFaOntSly lrievdeurcPeFdObSodcyonwceeingthrtattioon9s3o%fa1n4d09u5g%/gofainndit2i0a0l ubgo/dgy, rweesipgehctt,ivreelsyp.ecNteiivtehley,r 4a0t amvkedraNg-e lEitveFrOtSoEboofdy40wemikgdhtNr-aEtitoFs OwSerAehsaidgnaifsiicgannitfliycainntcrefefaescetdobnyb4o0dmykwdeiPghFtOSin arattas.verIangethleivaetr, aPtFsOfSrcoomncalelntortahteriodnossoe gf>ro6u0ps0,ubgu/tg.thIenscercehasaendgelsivwererteo bnootdysiwgeniifgihctanrtatcioosmpwaerreedftoounthdein control. a`Tnhde4g0uimnkeadpPiFgOpeSrc1e0nt91bo%daynwde9ig3h%t winaistiasligbnoidfiycwanetilgyhtd,ecrateaasveedrabgye1l6iv0ermPkFdOSN-EFOSE rcaotnicoesnwterrateiuonncshoafn4g1e9d.ugG/ugianneda 1p7i1gkuidgnge,yrtesopebcotdivyelrya.tioGsuwienreeaspiiggnilfiicvaenttrolybiondcyrewaesiegdhbty treatment in all the dose groups measured, i.e. 40 mkd N-E(FOSE, N-EtFOSA, or PFOS, aIsnpaatrst,atcehaomleisntoetrroalns(fCeHrOaLse),(AtSriTg)lywceerriedessig(nTiRfiIcGa)n,tlaylklaolwienreepdhionsprahtastatsreea(teAdLwKiPt)h,4a0nmdkd iPnFaOtSs tarnedat1e6d0wmitkhd4N0-mEktdFOPSFEO.S,Si4g0nimfikcdanNt-dEectrFeOasSeEsainnds4e0rummkpdotNa-sEsWiFumOS(AK*.)Ioncrcautrs,re4d0 amcktidviPtyF,ObSutca1u6s0eNda-EstiFgnOiSfEicadnitdinnoct.reaEsxepionsuhrepeoatficrapsalmtoit1o6y0l mCkodANo-xiEd+a-sFeO(SPECcoaAuOs)ed a tdhoeubcloinncegontfrtahteiosnpoefcimfiRcNacAtiveintcyoodfinlagurfyolr PCoCAOAoOxidinasreat(lLivCeor,AbOu)t acantdalaa2s-efaocltdiviintcyraeansde in icantcarleaasseeminRLNCAoAweOreacutinvcihtaynagnedda.3Extopo6s-uforledoifnrcarseasteo 4in0PmCdoAPOFmOSRNcaAuseexdpare2s-sfioolndin hsaetpsa.tTircecayttmoecnhtrwoimtehP4405m0kcdonFteOnSt AanodrA1c6y0lmCkodAFoOxiSdAasAecacatuivsietaydisnimgnailfeicraantts.increase in sPiFgnOiSfiwcaanstlaypipnardeuncteldyPaCmooAreO paoctteivnittpyewrhoexriesaosmeN-prEolFiOfeSraEtoirnctrheaanseNd-,EbtuFtOdSiEd.noPtFOS hsiiggnhifliicvaenrtPlyFOinSduccoencPenCtOraAtOioancstoifvgitrye,aetveerntthhaonu6g0h0bpotgh/Ktgr.eaPtCmeOntAsOacachtiievviteyd wsiamsilnaortly 3 5 SDRIPSTHT6205, T-6316.4, T-6868.2, T9071, T-7132.1 dinedtuecrtmiionnedoffoarctyhleC4o0AmkoxdidFaOseSaActainvidty16oc0cmurkrdedFaOtSreAlAatitvreelayteldomwearleavreatrsa,gbeultiave2r-fPoFldOS cforanccteinotnraofttihonesotofta1l9l3ivger/fgluaonrdoc1h4e0miucga/lg,warsescpoencttirviebluyt,edwibtyhthaenpearqenut ciovmpoaorlugnreedanotretrthe maenteaqbuoalliltye pFotOeSntApferroomxitshoemFeOprSoAliAfertarteoartetdorPatFs.OS, Tbhuetsneotdatthae sNu-gagceesttylthmaettFabOoSliAtemsaoyf be N-EtFOSE. NIn-tBhFeOgSuiAn.eaNpeiigt,hpeort4a0ssmikumd(PKF)OSvaolrueNs-wEe(rFeOsSiAg,ninfiocra4nt0lymrkedduocre1d6b0ym4k0dmNk-dEPtFFOOSSEand caused peroxisome proliferation in either genderof guinea pigs. o`fTpheesreorxeissulotmsesphroowliefdertahtaitoPnFiOnSra,tsN,-bEutFOnoStEi,n gFuOiSneAaapingds.FOThSeAsAe raellsuclatsusaerde iinndications cspoencciofircdatnocceerwtiatihn sepfefcecitess.of classical peroxisome proliferators, in that the response is 4 sz SDRTPITSTH-62958, T-6316.4, T-6868.2, T0711, T-7132.1 Introduction: `pTehreooxbijseocmteivperoofltihfeirsatsitoundiynwraatsstaonidngveusitniegaatpeigasn.dTchormepearceomtphoeumnodlescduelrairvmedecfhraonmisms of `inpietrifallulyortoeoscttead,nepesrulfflounoartoeocatnadneksnulofwonnattoec(aPuFsOeSp)e,rNo-xEitshoym!e pperorlfilfueorraotoiocntainnetshuelfraotnawmeirdee (amNe-nEd(mFeOnStA,)atnwodNot-hEetrhcyolmppeorufnlduso,ropoecrtfalnueosruolofcotnaanmeisduolfeotnhaanmoildo(Nac-eEt(aFtOeS(EF)O.SABAy) and Npe-rEf(lFuOorSoAo,ctNa-neEsFuOlfSoEn,amFidOeS(AFAOSaAn)d wFeOrSeAadwdaesdptroetshuemsetdudtyo.bTehPeFuOlSt,imhaotweemveetarbtohleirteeiosf speormoexidseboamteeparboloiuftertahtati.onThinethhyeproatt,hebsuitsntohtatthtehegsueinfelauopriogc,hweamiscbaalssewdoounldseivnedruaclelines of pervoildifeenrcaetiionn.dicHaotiwnegvtehra,t gtuhienmeoalpeicguslaarndanpdribmiaotcehsemaircearlesmiestcahnatntiospmesrotxhaitsdoimfeferentiate the runecslpeoanrs.eoTfhethsepseecisfpiecciaeismstoofptehriosxisstoumdye pwreorleifteor:ators for the perfluorosulfonamiwdaess 1. TinoduecltuicoindaotfemthReNmoAloefcguelanrersesthpaotnsaereinasbsootchiartaetds waintdhgpueirnoexaipsiogmsebpyromleifaesruartiionng. the 2. bTiondmienagsuprroitneginst.he hepatic activity ofperoxisomal enzyme systems and fatty acid 3. Tlioveprefrofrohrimstsotlaongdicaarld tcohxaincgietys twehstiscohfsmearyuimndcilciantiecaalnchexepmliasntartyiaonndfotrotehxeasmpiecnieest.he 4. Tdioffceormernecleastesaeenny ionbsreersvpeodnsaeltteortathieosnesocfotmhpoeunadbso,ve functions to liver and serum levels ofperfluorosulfonamides and their metabolites. molecul`aTrhiasnsdtubdiyocwhaesmipcaarlt omfecahsaenriiessmosfifnovresthteigeaftfiecotnssodfetshiegsneedcotompuonduenrdsstaonbdsetrhveed invWiavlol.acTeh,iUsnsitvuedrsyiwtayosfcaMrirnineed-soouttaiDnucloultlha,bowrhatoiohnaswibtehenotehnergaignveedstiingasttourdsi.esDrd.esKiegnned to upnedrefrosrtmaenddmtohleecefufleacrtsoanfdthbeisoechpeemrifclauloraonsaullyfsoensoafmtidhees ionndumcittioocnhoofndgreinaelsbiaoscsnoecrigaetteidcsw,ith o`fpterhoexi3sMomaElnvpriorloinfmeeranttiaoln Taendc/honrolceolglyreapnldicSaatfioent.y SDerr.vKirciess lHaabnpseernfaonrdmeLdisqauaAn.tiSttateivveenson squearnutimtaatnidvelilvievrepreprefrlfulouroorsousluflofnoanmaimdiedememteatbaobloiltieteanaanlaylsyesse.s Dorn.sLoimneXsuamppelrefsorimneDdr. Marion Anders lab at the Universityof Rochester. Study Timelines: `nTuhmisbesrtsud1y (anDdT125.-TBh)ewparsotcoocnodluhctaeddainnitnh-rliefeepsatratrst,dtahtee opfro1t1o/c1o6l/a9n8dapnrdotaoncionl-laimfeeennddment deantdedoafte1o1f/320//59/89.9.AAmmeennddmmeennttnnuummbbeerr12hhaaddaanninin--ilfiefestsatratrtdadtaetoefof3/21//199/90a1nadnadnainn-ilni-fe life end date of2/23/01. Amendment #3 was procedural only. 5 3 TSRPSTTE-6295.5, T-6316.4, T-6868.2, T7071, T-7132.1 Regulatory Compliance: 0`T9h5i0s,wSatsraatnegeixcplToorxaitcoorlyosgtyuLdyebanGdLtPhuPsrcolgarssaimfiPerdoacsednuorne.-GLP as explained in TOX SOP TestMaterial: d"Tohceusmpeonntsaotriopnroofvtihdeedsstaarmtpilnegsmoaftelrliafllwuaorsotchheemriecsaplosnstiobitlhietyionfvteshteigsatpoornss.orA.naAlyctihceamlical tceommppeorsaitturieo.n sTpeesctifmiactaetriioanlswhaesetswtoarsedketpitghotlnyfsileea.lCedomatporuonodmstewmeprereatsutarbel.e at room s`tTrhuecTt-urneusmobfeerasc,hcohfetmihcealconmapmoesu,ndasbbtrheavtiwateiroensteussteedd ifnorthtihsessetusdaympalreesg,ivaenndbtehleocwh.eTmihceal currently accepted abbreviations for each compound are in bold 1. Vehicle control: 2% Tween 80,orpropylene glycol. 2. T-6295: Perfluorooctane sulfonic acid, potassium salt m(poelrefl)u.orooctanesulfonate), PFOS, FC-95, Formula: CyFrS0y- K', MW = 538.1 3. ETt-h6y3l1P6e:rfluorooNeccttahnyelspuelrffolnuaomriodoocteatnheylsuallfcoohnoalm)i,dNo-eEthtaFnOolS,E,(nEaXrFrOoSwER,anFgCe-1N0-, Formula:C.F:SON(C:H:)CH,CH;OH, MW = 571.06). 4. eTt-h6y8l6a8m:ide), PFONeStAhy(laspesrtfatleudorionotchteanperostuolcfool,nabmuitden,ot(upseerdflfuoorrtohoicstcanoemspuolufnondylin ltahbi)s,rFepXo-r1t)2,,NF-oErmtuFlOaSCA,,FPyFsOSSOENAH(Cab:MbHrW,evi=a5t2i7o.n2)u.sed by the 3M Environmental 5. Ti7F0S71O:NHCH;COP0er'f,luMorWoo--ct5a5n6esgu/lmfoolnea)mido acetate, FOSAA, MSS6, (Formula 6. bTy-7t1he323M EnvirPoenrmfelnutoraoloLcatba)n,esFuolrfmonualmaidCeaF, iF7OSSOA;,NHP,F,OFSOASA(,abMbrWevi=at4i9o9n.u0s6ed mole). 7. LWaybeotrhat-o1r4i6es4,3L(eWxeYn,a,MKWS.=W3y2e3t.h7-914g/6m4o3l)(WwYa)s,obwtaasinaedddefdrotmo CDhTeImSsAynasScpioesnitcieve control dose group for hepatic peroxisome proliferation. 6 SY DSRTPITST-B6295.8, T-6316.4, T-6868.2, T-7071.1, T-7132.1 Methods ``mTehtehmoedtsahrodesgainvdendionsethgeropurpostofcoorlcaancdh apamretnodfmtehnitsss.tuDdeyvairateisonusmmtaortihzeepdrobteolcoowl.arDeetliasitleedd in Appendix 1. DTIS-B Protocol Procedures ``UThnedeprrotthoecoplrhotaodcaonl, eini-glhitfemsatlaertadnadteeiogfht1f1e/m1a6/l9e8SapnrdagauneiDn-alwifleeeynRdatdsa,te1o0-f1211w/e20e/9k8s, old HwaerilgahninLgabaoprpartoorxiiemsa,teIlncy.25E0ig-h3t00maglreamasndatcitghhettfiemmeoaflienHiatriattlieoynGwueirneeaobPtiagisne1d0-f1r2owmeeks folrdomanHdarwleainghLianbograatpoprrioexsi,mIantce.lEya6c0h0dtoos7e5g0rgourpamcsonattatihneedfi2meanoifmailnsit/isaetxi/osnpweceirees.obTtahiened mdogs/ekgg/rdoauypsN-wEerIeFOveShAi,claencdon4t0romlg/(k2g%/dTawyePeFnO8S0.),A4020mgm/gk/gm/dLaysuNs-pEetnsFiOonSoEf,e4ac0h test `fcouormpcoonusnedcwutaisveprdeapialryeddosiens2. %ThTewdeaeynof80thien afirgsltasdsosteiswsuaesgdriensdiegrn.atTehdedaanyizmearlos,rtehcuesivtheed cdoosmepsowuenrdesaudsmpiennisdteedriend2o%n dTawyese0n,18,02 aonrdth3eofvtehhieclsetucdoyn.trRoallbsyreorcaelivgeadvathgeetaetsta volume molf2/kgmlb/ykgdrboopdleytiwenitghhetb.aTchkeofgutihneemaopuitghs.recTehieveadnitmheailrsdwaeilryeohrualmadnoesleyvosalcurmifeicoedf2on day4 Amendment Number 1 Procedures 3A/m5e/9n9d.mTehnet pnuurmpboesre o1fhaamdeanndimne-lnitfe1stwaartsdtaoteadodfg3r/o1u/p9s9oafnbdoatnh imna-lliefeaennddfdeamtaeleofrats and hgiusitnoelaogpiicgasl tarnedatceldinwiictahl cvheehmicilsetrcyonrtersoull,tsNo-fE(thFeOtSrEea(tTm-en6t31g6r)ouorpsPuFnOdSer(Tth-e62p9r5o)t.ocoTlhewere wnoetreredmealrakyaebdldyudriifnfgersehnitptphianng ianndthweecronetdreoglsr,adaendd. thTehetriesfsuoerse,foarmneonrdthmeernntbl1owtaasnadleyssiisgned atcohrieepvleacaemtoherespeeffceicmteinvse tlehvaetlw.ere lost and elevate the doseofN-EtFOSE administered to p`rTowteolcvoelraatmseannddmetwnetlv#1e. guEianceha dpiogssetgortaolup(6cmoanltea,in6edfe2maalnei/msalpse/csieexs/)spweecrieesu.sed under the `The dose groups were mg/kg/day PFOS. vehicle control (2% Tween 80), 160 mg/kg/day N-Et FOSE and 40 aFovroNlu-mEeFoOfS2E,mla/ksguswpaesnsaidomnionifs8t0ermegd/bmyloNra-lEgGaFvOaSgEe itno 2th%e rTawtseaennd8b0ywdarsopplreetpianretdheand `bTahciskodfotsehewamsouctohmptaortahbelgeutionetahepicgusmoulnadtaiyvesd0ostehorofuNgh-EdatyFO3SoEftthheasttiunddyu.ced peroxisomal PCOAO Medical activity in the rat Dept. T-6316.1), in a and 4 week feeding study with 300 was less than haolfthfe LDS0. ppm N-EFOSE (Ref. 3M 7 [3 SDRIPSTHT6958, T-6316.4, T-6868.2, T-2071.1, T-7132.1 FvoorluPmFeOSo,f2amslu/skpgenwsaisonoadfmi2n0ismtge/remdlboyfPorFaOlSgaivna2g9e%toTwtheeernat8s0awnadsbyprderpoaprleedt,ianntdhea back odfosteheofmoPuFtOh todetlhievegrueidneuandpeigrsporontdocaoylzaemreontdhrmoeungth d#1ayw3asoft~h16e0smtgud/yk.g,Thasewcausmuulsaetdivien tthherepsrhootlodcoflo.r TinhdeuccuimnuglpaetriovxeidsoosmeewparoslibfeelroatwiotnheinLtDhSeOratf.or POS and well above the Amendment Number 2 Procedures pTehreflpuuorrpoooscetoafneasmulefnodnmaemindtoa2cewtaasteto(aFdOdSgArAo)uposr mpaerlfelruaotrsotorcetaatnedeswuiltfhonvaemhiicdlee(cFoOntSrAol), at daomseensdemqeunitva#l1enitntoortdheerdtoosceosmopfaNre-EthFeOeSffEectasndanPdFOmeSt,abroelsipteectpirvoefliyl,esionfptrhoetosceol `2c1o9m/p0o1unadnsd iannriant-sliafteeeqnudivdaalteenotfd2o/s2e3s/.01A.meNnindememnaltenruamtsbewrer2ehuasdedanunidn-elriftehsetaprrtotdoactoelof oalmdewnedimgehnitng#2.apEparcohxidmoasteelgyr2o5u0p-c3o0n0tagirnaemds3atmatlheetSipmreoagfueinDitaiwatlieoyn wRaetrse,o1b0t-a1i2newdeferkosm Hmagr/lkagn/dLaayboFraOtSorAiAes,, aInndc.40Tmhge/dkogs/edagyroFuOpSsAw.erTehevevheihcilcelceocnotnrtorlo(lpwraospydleelnieveglryecdolb)y, o1r6al0 glaarvgaegrevoaltaumveoolfu5memoLf5/Kmgl/wkagsbuosdeyd wtheainghitn pornotdoacyoslzearmoetnhdrmoeungth #d1aybe3ocfautsheeonsltuydrya.tsA were dosed by oral gavage, whereas the guinea pigs cannot be dosed by oral gavage. FvoorluFmOeSofAA,mla/skugspweanssiaodnmionfi3s2temrge/dmblyForOaSl AgaAvaigneprtooptyhleernaetsgloyncodlaywsas0-3p.repTahriesddaonsed a achieved a cumulative dose of640 mg/kg FOSAA. zFeorroFtOhSroAu,gha ddaoyse3ooff4t0hemgst/ukdgy.bAosduyspweeingshitownasofa8dmmign/ismtleroefdFvOiSa gAavinagpreotpoyrlaetnseognlydcaoyl dwoasseparcehpiaervede,d aancdumauvloaltuivmeedoosf5e molf/1k6g0wmags/akdgmaifntiesrtfeoruerddbayyosroafldgoasviangge. tTohtehedorastes,ofThis FprOotSoAcolw.aTshteheLsDaSm0efaosrtFhOe SdoAseisonfoPtFkOnoSwand,mibnuitsttheerecdumuunldaetriavemednosdemoefntFO#1SoAfthis `iandcmionmisotielr,eadsuandpeoirntthoifsrperfoetroecnoclew.as below the LDS0 for PFOS of251 mg/kg for PFOS Specimen Handling: dLeisvcerriabnedd isnertaheweprroetcooclolleactneddaamnedndfmreoznetns.rapiAdloynaeftteortewutohganraasmiaalaicqcuoortdoifntghteoltihveerdetails saammepnldemsensthsi.ppeTdheiniddreyntiicfeictaotitohneoafneaalycthiclailvelrabsoarmaptolreiess leistotneedaitcnhthleabpriostloicsoteldainnd Appendix 2. 8 Se SIRPSTTB-62958, T-6316.4, T-68682, T707L1, T-7132.1 Analytical methods Ccoenrtteanitn, lLiavuerrosyalmpCloeAs woxeirdeasseenatctfiovritayna(lyPsoiosscbhyapnrdevYiaomuaslzyakpiubl1i9s8h6e)danmdetphrootdesinfocronPt4en0t (Bradford Biology at 1t9he76U)nivienrtshietylaobfoMraNtorbyyotfheKefonllWoawlilnagcemeDtehpotdso:f Biochemistry and Molecular Enzyme Sample supematant of2 Preparation 10% (wt/vol) ~ The enzyme homogenateof fraction consisted of the 6,000 g 0.5-1.0 g frozen liver tissue in 300 emstMimmaatnenditaoclc-o1r0dimngMtoHtEhPeEmSe-t1homdMofEBGrTadAfo(rpdH 7.2). using Protein concentration was commercial bovine serum albumin as standard. Lsu-sCpoedndOexdidinas6e0AmssMayKH~yTPhOe,-e0q.u0i2va%lenTtriotfoncaX,1S00pg(/pmHl 7t.i4s)suceonthaoimnoignegn1atmeMwaps- hlyadurroyxly-pChoeAny(lLaCcoeAt)a.teT(hPeHPrAea)c,tio4nsunwietrs/emallpleorwoexdidtaosei,nc2u0batueMatF3A7D,Cafnodr 3600 miiMn in a 100 shaking water bath and mM sodium carbonate terminated (pH 10.5). by adding 3 volumes The concentration of of 2 mM KCN in HyO; generated dwuirtihngantheexrceiatcattiioonn wwaasveelsetnigmtathedoffr3o1m7thne mfluaonrdesceemnicsesoiofnPHatAP40a5s measured nm. The afsluonrmesoclencpeerwoaxsidcealigbernaetredatweidt/hmicno/mimmgercmiiatlocHh;onOd;riaanld the results protein. are expressed Protein was quantitated by the Bradford method. oCraitgailnaaslemAestshaoyd-puTblhieshacetdivbiytyColfaciabtoalmaesaenwdaFsriedsotviimcahte(dJ.bByioal.moCdhiefmic.at2i54o,n o42f4t5h-e 5qu2a,nt1i9f7i9e)d,swphecitcrhopihsobtaomseetdriocnaltlhyeadti2re4c0t nmme.asuInretmhiesnptroocfedHuOre;, tdhiesatpipsseuaerasnamcpelea.5 was diluted 10 27C and in SO mM potassium the reaction initiated phosphate by adding (pH 10.3 7.0). mM The medium was warmed H;O;. The progressofthe reaction was monitored at 240 nm for 5 from the initial linear rate ( E**=43.6 min. Catalase mM''cm") and activity was expressed as estimated units/mg. pcraottaeliynze(sTtahbelede1c).omOponseituinoitnoofaf1ctpivmiotlyeoisfdeHfOin;edpaesrtmhiant.amount of enzyme which N(oArCt0hAerOn) BalnodtcAatnaallaysseewser-eQpuearntfiotramteidonbyofNomrtRhNeAr blfootr abnoatlhysiascofyqlCuoiAckoxfirdoazseen lwiavser spaomwpdleersizferdomintrelaitqeudidratnsiatrnodgegnuinuesainpgigs.a Approximately mora/pestle. 1 g offrozen Total RNA liver was recovered quantified supseicntgropthhoetoPmeEtRriFcEalClTy aRt 2N6A0nTMm.isTohlaetiRonNAk.itwaasndeletchterocpohnocreensterdaotnioan 1% agarose gel, blot transferred to a cellulose membrane and hybridized to the cforrormespproinmdeirnsgtroacna.d,om3l5y0 [b*a*sPe] sleabqeuleendceoloifgotnhueclreeostpiedcetsivtehartatwleirveerPgCenRe.ampmlRifNieAd 9 S7 SDRTPITSTB-62958, T-6316.4, T-68682, T-7071.1, T-7132.1 band density software. was quantified autoradiographically using phospho-imaging `CmeerttaabionliLtievsebrystahmepl3eMs wEenrveirsoennmtefnotraalnaTleycshinsofloorgtyheapnadrSeanftectoymSpeorvuincdes(5u)sianngdptuhbelished maneatlhyosdess (thHaatnwseerneetcaol.mpl20e0t1e)d.wTehreediestsauieldsoifnatnhaenmaeltythiocdaslurseepdoratnfdrtohmetrheesu3ltMsofthe ELnIvMiSroNnom.eEn0t1a-l0L1a29b)o.nTMhaeyre8s,ul2t0so03fa(n3aMlySsteusdtyhaNtow.eFrAeCcTom-pTlOeXted~1a0r7e,in3cMluLdaebdoirnatthoirsy rtehpeo3rtManEdnviinrtoengmreatnetdawlitlahbthuseebdodtoyiwdeeintgihftyacnodmploiuvenrdwseiwgehrteddaitfa.ferTenhtetahbabnrtehveiaotnieosnslitshtaetd aabnoavley,zeadnfdorarae mfeotuanbdolaistseo,cpieartfelduwoirtohoctthaenepsruilmfaornyamdiadtoa(teatbhlyels)facreotmattehe(lPabF.OSTAhAe)l,ab that awnadsanneavleyrzdeodsefodr.inThlievearbsbraemvpilaetsioanrseugsievdenfobreleoawc,h afnoalllyotweedthbayt tthheelcahbeumsiecdalasfosrtmaunldaaradnsd the abbreviation used in this report, PFOS= PFOSA= PFOSAA = EFOSE= MSS6= PFOSEA = Perfluorooctanesulfonamide Perfluorooctanesulfonamide (Formula CyFysSOy-, PFOS) (Formula CyFpySO;NHy, FOSA) rPeefrefrlrueodrotoocatsanPeesrufllfuoonraomoicdtoa(nectshuyllf)oancaetmaitdeo.ac(eTthaitsecionmtphoeurnadwwdaastaatlasboles of FnaArCroTwTROanXge10N7)-.Et(hFyolrmPuerlfalCuosrFo1o;ct8a0n:eNs(ulCf,oHnsa)rmCiHdzoCeOtOhy"l,aNl-coEh(oFlOSAA). P(eFrofrlmuuolraooCcFt,a:nSeOs;uNl(fCo;nHa,m)iCdHoa,cCeHt;aOtHe, N(-FEoFrmOuSlEa)CyF,sSO;NHCH,COO" FOSAA) Perfuorooctane sulfonyl ethylamide EWFOSA) (Formula CF:SONHC,HN;- CfelrutoarioncaortbhoenrsbliyverLsCa-mMpSle/sMwSerbey aDnr.alLyiznedXfuorinptahreenltabcoormatpooruynodsfMa.ndW.meAtnadboelristesofthe DpuebplairsthmeednmteotfhPohdasrm(aHcaonlsoegnyetaanl.d P2h0y0s1i)ol. oTgyh,eUrneisuvletrssiatrye roefpRoorctheedsitnetrhuissirnegpoprrteavinodusly integrated with the body weight and liver weight data. pCaelrmtiaitnoyo]thCeor-lAivOerxisdaamspeleascwteivriteysaesntantoinCdoivcaatnocroefipneMraodxiissoonmeWIproalnidfewreartieonanuasliynzgead for vaaslsiadyatisedthmaettihnotdhebparseesdeonncepoufblpiaslhmeidtomyelt-hCood-sA,(Lthaezathriorwd 1st9e8p1o)f.tThheebp-roixnicdiaptlieoonfstphieral aitnv3o4l0vensm.theThreedruecstuilotns woefrNeArDepotrotNedADthHe,3wMhiocnhOccatnobbeerm2e2,as2u0r0e2disnpeacltertotperhorteopmoerttrfircoalmly `Covance Covance. andthedata is included in this report. No formal report was writen by. 10 S DSTRPITSBT6295, T-6316.4, T-68682, T907L1, T-7132.1 Results Biological Parameters Athle iann-ilimfaelpshsausreviovreadttnoetchreoepsnyd.ofthe study. No gross observations were noted during Tfrhoemcaupmpurloaxtiimvaetdeolsyes67rtaong1e2d3 fmrgomfalpuporroocxhiemmaitceally i3n3gtuoin5e3ampgigsflrueocreoicvhienmgi4ca0lmign/kragt/sd,aayn.d `(TThaebl1e601)m. gT/hKegb/oddayy wdeoisgehgtsrooufpthaenigmuailnseraepciegisvetdhaptrwopeorretidoonsaeldlyathidgifhfeerrcenutmutliamteisvweedroeses gpirgesa.tlIynddiifvfiedrueanlt,awnhdiscuhmamcacroyunctuemdulfaortitvhee wdoisdeedraatnagaerienschuomuwlnaitnivAeppdeosnediixn t3.he guinea Agivveernag4e0 bmokddyPweFiOgShtasnddec1r6e0amsekddsNig-niEfFicOanStEly(oTvaebrleth2e).dAosvienrgagpeerbiooddyfowrefiegrhntaslederactrseased s`imgkndifNi-caEntFlOy SovEerorthFeOdSosAiAng(pMe5r5i6o)d. fIonrdmiavlideuraaltsangdivseunm4m0armykdboPdFyOwSeiogrhFt OdaStAa and are 160 shown in Appendix 4. `(TThaeblmeal3)e. ratMsaglievernatNs -gEiv{eFnOPSEFOaSta4t0 4m0kmdkhdadhsaidgnaisfiigcnanitfliycainntcirnecarseeadseliivnerlwiveeirgthotsbody wsiegingihfticraanttiloys.loMwaelreedraktisdtnreeyatweediwgihtths PaFnOdS,kNi-Ed{tFnoObSeoEdyyandweNi-ghEttFraOtiSoAs.atF4e0mamlkedkialdlnheayd wvaeliugehst. aKniddnkeiydnweeyigtohtbodadtyawweaisghntotraotbitosaiwneerdefnorotthseigontihfeircadnotsley dgirfofueprse.ntIfnrdoimvicdounatlroalnd summary body weight data are shownin Appendix 5. Mdoasleegarnodupfeamnadleanpaelryczeendtotfoigneitthiearl. bTohdeycwoeimgbhitnsedonradtapyerfcoeunrtwbeordeycwoemibgihntewdafsor each sEiAgFniOfSicEan(tTlaybdleec4r)e.aTshedebcyom4b0imnekddgPuFiOneSa,pFiOgSpAer,ceanntdbFoOdSyAwAe,igahntdwbays s1i6g0nmifkicdanNt-ly decreased mid. by PFOS, N-EFOSA and N-EFOSA at 40 mkd, and by N-E(FOSE at 160 Owerrgeancowmebigihntedraftoirosm(allievseratnodbfoedmyalweesigihntdeapnedndkeindtnleyyftroobmocdaycwhediogshet wghreoruep aivnaailllapbalret)s oifntcrheeassetutdhyNaendfoarneaalcyhzeddosteoggerthoeurp. fCoormabnialnyisnigs tahnedriesljautsitviefioerdgabnecwaeuisgehttshewoarsgdanone to winecirgehatssehdalvieverbeweenignhotrmtoalbiozdeyd bweyibgohtdyrawteiiogshti,n thPeFrOatScaotm4b0inmedkdastigan(ifTiacbalnetl5y). The dciofmfebriennet dfrmoamlethaendcofnetrmoallegrgouuipnevaalpuiegslfiovreratnoyobfotdhy ewetirgehattmreanttisosgiwveerne. notSisginginfiifciacnatnltyly EinFcOreSaEs,edNk-iEdnWeFyOStoA,boodryPwFeOigSahttr4a0tiomskwde.re found in guinea pigs treated with N- n DSRTPITST-B6205.8, T-6316.4, T-68682, T-7071.1, T7132.1 Liver Fluorochemical Concentrations "The liver fluorochemical concentrations in rats and guinea pig livers are summarized from the analyses performed at the UniversityofRochester (Table 6), and at the 3M. Environmental lab (Table 7). The individual and summary liver fluorochemical concentrations data are presented in Appendix 6. The 3M Environmental lab analyzed liver samples from the PFOS 40 mkd and the N-EXFOSE 160 mkd rat dose groups and did not measure fluorochemical levels in the guinea pig livers. Aliquotsofthe livers that were analyzed by the 3M Environmental lab were also analyzed by Drag Andres lab at the Universityof Rochester. The individual rat liver data showed that one lot of the samples analyzed at Rochester had a high backgroundof FOSA in the control group. The Rochester analyses tended to have higher PFOS and FOSA determinations than the 3M Environmental lab, however the values for PFOS are listed as 35% accuracy, for FOSAA + 50% accuracy, and for FOSA, FOSAA, EGFOSE and N-E(FOSA the values are listed as qualitative only in the final report (FACT TOX 170). Therefore, the differences in the measurementof PFOS and FOSA fall within the experimental error inherent with the methods used. `The percentof the dose that wasinthe liver was calculated for each dose group from the `amountofthe total liver PFOS containing species, derived from the sumofthe concentrationsofall fluorochemical species detectedinthe liver and designated `TLPFOSX, and for the amount of PFOSitselfin the liver. The amount (mg) ofall fluorochemical species detected in the liver was derived from the TLPFOSX times the liver weight. The percentoffluorochemical dosed present in the liver was derived for boththeTL PFOSX and for PFOS itself for the values derived from both the University of Rochester lab and the 3M Environmental lab (Appendix 6, B and D). In rats dosed with PFOS, the percent of PFOS dosed present in the liver as PFOS ranged from about 17 1023 percent in both male and female rats, and was consistent between labs. In guinea pigs dosed with PFOS, the percentage ofthe total amount ofPFOS dosed that was. presenint the livers was far less than in the ra, between 3% and 5%. Guinea pig livers `were only analyzed at the UniversityofRochester so a comparison between labs for the fluorochemical content in the liversofguinea pigs cannot be determined. However, it can be seen from the data presentedin Tables B and D in Appendix 6 thattheTL PFOSX values derived at the University of Rochester and at the 3M Environmental for rats treated with 160 mg/kg/day N-EFOSE are about the same at each lab. `The UniversityofRochester analyzed both rat and guinea pig livers from animals treated with either 40 mkd PFOS, N-EFOSE or N-E{FOSA, or 160 mkd N-E(FOSE. The pTaLrtPicFuOlaSrXlyiinngtuhienmeaalpeisgsgutirenaetaedpiwgist.hT1h6e0 mpedrceNn-tEoXftFhOeSEdowsaesthlaotwwearsthparnesientntheinratth,e liver as PFOSX was approximately 2-fold the percentofthe dose present in the liver as PHoFwOeSveirts,eltf,heifnebmoatlhegruatisnetarepaitgesd awintdhin16m0osmtkrdatNs-tErteaFtOedSwEithhaNd-gEreFaOteSrEthaannd7F0O%SoAf.the TLPFOSX as PFOS in their livers. Conversely, the percentageofthe dose that was present as PFOS in the liverofras treated with 160 mkd FOSAA had high levels of FOSAAitselfand FOSA, with low levelsofPFOS. 2 DSRTPITSBT6295. 6316.4, T-68682, T-707L1, T-1132.1 Clinical Chemistry cSheermuimstcrlyinviaclaluecshewmeirsetrcioemsbfionrerdatsfraonmd mgaulienseaapnidgsfeamraelperseisnenctaecdhisnpAecpipeesnadnidx s7t.atCilsitniicscal were performed. aIsnpraarttsa,tcehaomleisntoetrroalns(fCeHrOaLse),(AtSriTg)lywceerriedessig(nTiRfiIcGa)n,tlaylklaolwienreepdhionsprhatasttarseeat(eAdLwKiPt)h,4a0nmdkd iPnFrOatSs atrnedat1e6d0wmikthd4N0-mEktFdOPSFE.OS,Si4g0nimfikcdanNt -dEec(rFeOaSseEsainnds4e0rummkpdotNa-sEs(iFumOS(AK,)woictchurthreed t`omtoasltpsriogtneiifnic(aTnPt)dewcerreeasseisgnoicfciucarnrtilnygiinncrtheeas4e0dmbykd4P0FmOkSddPoFsOeSg,roaunpd.cArlebatuimniinne(Alb) and g(rCoRupEsA.TT)hewsaescshiagnnigfeiscainntlcyliinniccraleacsheedmiinsttrhye a4r0emckondsiPsFteOnSt wiatnhdp4r0evmikodusNs-tEudtiFesOSwiAthdose dPiFffOeSreantndfrNo-mE(coFnOtSroEl.vaNlouneseoifntahtes.other clinical chemistry parameters were significantly sIingtnhiefigcuanitnecahapniggsescoinmbcihonleedstmeraolle, atrnidglfyecmearildeesc,lianlikcaallicnheepmhiosstprhyataanasley,siosr,atshpearretawteere no Pamoitnaostsriaunmsf(eKr"a)sev.alAulebsufmoirnm,atloetalanpdrofteemianlaentdhcergeuatiinneianepiwgesrceonmobtisniegdniwfeicraentsliygncihfaincgaentdl.y armeidnuocterdabnys4fe0rasmek(dAPLFT)OSwaansds4ig0nimfikcdanNtl-yEitnFcOreSaAs,edsibmyiltarreattomtehnetoraft.guAilnaenainpeigs with 40 f`mrkodmFcoOnStr.ol.vNalouneesoifntrhates.other clinical chemistry parameters were significantly different Palmitoyl CoA oxidase (PCoAO) activity dHoespeatgircopuaplmwiatsoyclomCboiAneodxiadnadsean(aPlCyozeAdO)togaecttihveirt.yPdFatOaSfoatr 4ma0lmegs/akngd/dfaeymafloersfofurrodmaeyasch c1a6u0smedkda sfiogrn4ifdicaaynstdiindcrneoatsesiignnihfeipcaatnitclyPiCnOcrAeOaseachteipvaittiycinPaCtosA(OTaibnlreat8s).. NN-oEtsiFgOniSfEicaatnt change occurred in the guinea females separaaretseholwyn pig. Individual in Appendix 8. and summary PCoAO values for males and Peroxisomal enzyme activity and gene expression PThFeOSefffeocrtsfoourf ofnoucratdaalyasseoafnodraalcdyolsCionAg ooxfid1a6s0emgge/nkege/xdparyesNs-ioEnFaOnSdEenozry4m0e magct/ikvgit/ydaiyn liver were tissue from exposed rats presented as a poster at and the 2g0u0i1neaSopciigestyaroefpTroexsienctoeldogiyn Ampepeteinndgix(W9a, lTlahceeseefdaatl.a 2001). activity Acute exposure of of LCoAO and a rats to 2-fold 160 mkd increase N-Et-FOSE caused a in the concentration doubling of the specific of mRNA encoding for 1 </ DSRTPITST-B6295.8, T-6316.4, T-6868.2, T-7071.1, T-7132.1 cPaetoalAaOse minRlNivAerwferroemubnocthhamanlgebeyadnedxpfoesmuarlee troat1s60(AmpkpednNd-ixEt9-).FOSCEatianlalsieverasctfirvoitmybaonthd bsoextehs.sexEexsp,oasunrdepoosfsriabtlsytao 4sl0igmhktdinPcrFeOasSecianucsaetdalaas2e-faocltdiviintcyreiansleivienrLfCroomAmOalaec,tivbiuttynfootr female, PCoAO rmatRs. N~AAcexuptreeessxipoonsutrheatofwraastmsotroe4p0romnkodunPceFdOSin caused female a 3- to 6-fold increase compared to male rats. in Exposure of guinea catalase activity in pigs to cither N-Et-FOSE sex. The and PFOS guinea pig did not mRNA stimulate LCoAO activity or encoding for PCoAO was undetectable, even following exposure to N-Et-FOSE or PFOS (Appendix 9). PTr4e5a0tmceonntteonftaransd wAictyhl4C0omAkodxiFdOasSeAacctaiuvsietyd(sAigpnpiefnicdainxt 1i0n)c.reaTsreesaitnmheenptaotficractystwoicthhro1m6e0 CmkodA FoxOiSdaAsAe accatuisvietdys(iAgpnipfeincdanitxi1n1c).reaTsheseiinnhdeupcaetdicthceytexopcrhersosmieonoPf4t5h0ecsoentoefntthaensde Acyl npraottse.insFoirntlhivee4r0ofmrkatds FinOdiScAataenddth1at60FmOkSdAFaOndSAFAOStAreAatwedermealpeerraotxsi,sao2m-efporlodliifnedruacttoirosn oafndac1y4l0Cgo/Ag,oxriedsapseectaicvteilvyi.tyThoeccsuerlrievderatPaFvOeSracgoenclievnetrraPtFiOonSs cwoenrceenltorwaetriotnhsanofth1e93ligve/rg concentrations in the PFO treatment group. fIrnaFctOioSnAofttrheeatetdotraaltsl,ivtehrefplauroernotccheommipcoaulnads,dFidOSthAe,FcOonStArimbeuttaebdoalniteeqPuiFvOalSenatnodregarcehatoefr ctohnetsreasftl,uotrheocFhOeSmAic-aglluscpeucrioensirdeeprperesseennttedinapFpOroSxAimtarteealtyed0r.a3ts%ocofntthreibdutoesdeoinnltyhea lsimvaerl,lIn fractionofthe TLPFOSX (Appendix 10). cIonnFtrOiSbuAtAedtarnea2tetdor3attsi,mtehse,mreetsapbeoctliivteelFy,OtShAe,amaonudntthoefptahreenttoctaolmlpiovuerndf,luForOoScAheAm,ical than did the ltihveerm,erteasbpoelcittievePlFyO(SA,papnednrdeipxre1s1)e.nted approximately 0.14% and 0.24%of the dose in pTraokliefnertaotgoerthteorP,FthOeSseindartaatss,uagngdestthatthattheFNO-SacAetmyalymebteabaonleiqtueaslolfyNp-otEetnFt OpeSrEox(iNs-ome PEFtOFSOAS.AAT,heancdoFmpOoSuAnAd)s aFrOeSwAeaaknedrFpeOrSoAxAis,omweerperonloitfetreasttoerds itnhagnuienietaheprigPsF. OS or Discussion P`TFhOeSa,nawlyhteitchaelrdaPtFaOsShowwaesdatdhmaitnFisOtSerAedwdaisreccotnlsyiostrefnotlrymeiddenatsiafimeedtaasboalimteet,abboultittheeosfource `osfatmhpeleasmsiunbomigtrtoeudpfiosrnaontarleyasdeislaytaappgairveennt.tiFmOe,SbAutwanostpirnesoetnhteringrsooumpesoofftchonetcroonltrsoalmples dsuebtmeirtmtiendedatindiaflfleorfetnht etimleisv.ersTohfeafllorPmFatOiSontroefatPeFd OaSniAmaflrsotmhaPtFwOerSetraenaatleydzaendimataltshewas UniversityofRochester, and at the 3M environmental lab for the same samples, which 14 DSRTPITSTB-62058, T-6316.4, T-68682, T-1071.1, T-7132.1 Chlaadrinfoicbaatciokngorfotuhned fPoFrOmaStioofnPoFfOtShAe.FOinStAhefcoolnlcouwrirnegnttrceoanttmreonltfwriotmh ePiFthOeSr lsahbo.uld be obtained in further studies. FOSA N-glucuronide. Whatever its routeofformation, FOSA was metabolized to cNo-uEltdFaOriSsEe aflrcoomhotlhegiNv-edseertihsyeltaot.iaornaongfeNo-fEm{aFjOoSrEaanldcomhionlo,ramnedtaNb-olEitteFsO.SAFAOScoEuladlcaorhiosle EbyAFtOheSoExiadlactoihoonlowfotuhledaglicvoehotlhetoobtsheercvaerdboNx-yEli{cFaOcSidE. alGclouhcoulroglnuicduartoinoindoefs.theFOpaSrAenAt Na`clocuolhdolb,eofrobromtehd. bFyOtSheANc-oduelctdhbyelaftoiromnoedfbNy-tEheFON-SdeAeAthoyrlabtyiotnheofoxNi-daEttiFonOSoAfFoOrSbEy.the mreamyovbaelfoofrtmheedcbayrbtohexygmleutchuyrolngirdaotuipoonfoFf OFOSSAAA. asLogslsyooxfytlhatee.caFrbOoSxyAmeN-tghlyulcgurroounpidferom FOSAA would give FOSA or lossofthe glycine moiety would give PFOS directly. c`Tohnececnotrrraetlaitoinosnwoerafefaenawolyfztehdebmyodsotsesiggnriofuipcainntrtaotxsicanedngdupionientaspitog.s (lAivpeprePnFdOixS12). P`TFhOe Sdeactre4a0semkindtahnedpeNr-cEen{tFiOnSitEialatbo16d0ymwkeidghtot 8w5as%amnosdt8s7tr%onignlityiaelffbeocdtyedwienigrhatts,baty aalvseoradgeecrleiavesredPFinOrSatcsobnyceFntOrSatAiaotns4o0 >fmk6d00angd/gF.OSTAhAe p(eMr5c5e6nt) iantit1a6l0bmokddy wteoi9gh3t%waansd. a9p5p%roxoifmiantieilayl b1o4d0tyowe2i0g0htpig,/rg.espAectgirveealtye,r fartaacvtieornaogfetlhievetrotPaFlOfSlucoornoccehnetmriactailonisn tohfe liver oFfOtShAe. FSOimSiAlaArltyr,eattheedFaOniSmAaltsrewaatsedparneismeanltsashathdeaphairgehntlicvoemr pcoonucnedntorrataisotnohefmtehtea.bolite. supnemceiteasbloilkiezleydcopnatrreinbtuctoedmptooutnhde.boTdhyuwsetihgehsteepfefrecftlutooraosgurlefaotenramdiedgerefelutohraonchtehemical `metabolite PFOS in the FOSAA and FOSA treatment groups. b`TohdeyrwanekigohrtdienrorfattshweaesffPecFtOoSfathte40difmfkerden>t FtrOeSatAmeantt4g0rmoukpsd>onNi-nEcrFeOasSesE laitve1r6w0emigkhdt >to PFFOOSSAaAnadtN1-6E0(FmOkSdE,ocacpuprrrxiinmgatatelayv1e9r3agge/lgivefrorPPOFSOScAo,ncaenndtr1a4ti0oungs/ogff>or6F00OSugA/Ag for btruetaotendlyantihmeallisv,erretsopebcotdiyvewlye.ighAtllradtoiosseogfrtohuepsPFhaOdSitnrceraetaesderdatlsivweerrteo bsiogdnyifwiecaingthlty ratios, `icnhcermeiassterdyceonmdppoairnetds toofpcootntarsosliuvmal,ucehsolbeystDeurnonleattn'dsttr-itgelstycerCiodrersetloatliiovenrofPtFhOeSclinical concentrations determination. in rats are not shown du to the limited umberof data points for each ``bTyheN-deEc{rFeaOsSeEiantth1e60pemrkcedntaninditPiaFlObSodayt 4w0eimgkhtdawnadstmoos9t1s%traonndgl9y3e%ffiencitteadlibnogduyinweeaigphitg,s fatraacvteironagoeftlhievertoPtaFlOfSlucoornoccehnetmriactailonisnotfhe41li9vearondft1h7e1 Ng-/Eg,tFrOesSpEecttrievaetleyd.guAingeraeaptiegrs was present asthe metabolites FOSAA and N-EtFOSAA than occurredinrats. Thus, these. 1s 3 DSRIPTSTH.6295.8, T-6316.4, T-68682, T-7071.1, T-7132.1 epfefrefcltuoirnogoucitnaenaespuilgfs.onTahmeisdee fdaltuaorsoucghgeemsitctahlatspreactsiemseltiakbeollyiczoentNr-ibEuFteOdStEo tthoePbFoOdSy wmeoirgeht wqueirceklsyhothwannignutihniesarpeipgosr,tpfeorrhaatpss,dbuuet twoasthneoitndduectiebrimliitnyeodfcfoyrtgoucihnreoampeigsP.450s in rats, as "wTehiepgehtrcbeynNt-iEntitiFalObSoAdyatwe4i0gmhktwdaasndalgsroodwetchrweaassedhienldgutoin9e9a%piignistitaol 9b4od%yiwneiitaglhtbobdyy40 `mkd N-EtFOSE, at average liver PFOS concentrationsof approximately 100 ug/g in the NOn-lEytPFOFSOAS tarnedatFmOenStAgrwoeurpe aanndal6y6zegd/fgorininttheheNl-iEvetroFfOtShEetNre-aEtmFenOtSgAroturpe,atreedspgeucitnievaely. pfirgasc.tiHoonowfetvheer,totthael 4f0lumokrodchNe-mEictaFlOiSnEthterelaitvmerenptregsreonutpagsutihneemaeptiagbolliivteress hNa-dEatlFaOrgSeAA aPnFdOFSOiSn AthAe lwihvierchanwdhtehnuscommabyinheadv,ewmeardeeeaqusailgntiofiocrangtrecaotnetrrtibhuatnitohnetcootnhceenotbrsaetrivoendof effects on body weight. `dTehcerelaisveerd,weailgthhtoutgohbnoodnyofwetihgehstercahtiaonsgeinsgwueirneeasipgingisf,iciannctolnytdriafsftetreonrtattsh,awnecronetraolll. The 4ra0nmkkodrd>eroNf-tEhAeFOdSecEreatas4e0dmlikvedr>toPbFoOdSyawte4ig0hmt keff>dectNi-nEgtuFiOnSeaEpaitgs16w0arsukdN.-ETthFisOSorAdeatr tishersoeudgholsyetghreouipnsv.erGsievoerndetrhoefltahcekofdtechreealsievderbroedsypownesiegshtinegfufiecnteainpgigusionefaeiptihgesrfor each of thheaptaatnomIentgraalpyeroirtoinnedaulctiinjoenocftipoenroofx1i0s0ommeg/pKrogliPfFerOaStincgaeusnezdymdeeastchoiunplgeudinweiatphitghsebfuacntot inrats (See DTLS mechanism. A), suggests that the peroxisome proliferation in the rat is a protective. oCfotrrheelaetfifeocntoofftthheecdliifnfiecraelncthtermeiasttmreyntengdrpoouipnstsonindgeucirneeaasepdigssesruhmowpeodtatshsaitutmhelervaenlks owradser oPfF1O4S8 autg4/0gamnkdd1>00Nu-gE/gt,FrOeSsApecatti4v0elmy.d,Inoccocnutrrraisntg, attreaavtemreangtoe fligvuerinPeFaOpSigcsonwcietnhtr4a0tions `mkd N-EtFOSE had no effect on the serum potassium levels at average liver PFOS `Cchoonlceesntterraotlioannsodftarpipgrlyocxeirimdaetsecloync4e5ntgr/atgioinnsfienmgaulienaenadpi8g8sgw/egreinnomtasliegngiufiincaenatlpyigs. dsitfrfoenrgenctortrhealnatcioonntrtoollivvaelruePsF,OaSndcosnhcoewnterdatnioonpsa.rticular trend between dose groups or Conclusions AwlilthtrtehaetmiennctrseacsaeusceadusiendcrbeyasePdFlOiSverbteoinbgodtyhewgeriegahttesrtataiomsoningraatlsl but the not guinea pigs, fluorochemicals ntoesttegdu.inTehaepiegvsi,decnacueserdevbeyaltehdetsheeaccluatsesiecxalpossiugrness.of Tpheersoxeidsaotmae pprroovliifdeerasttiroonnginervaitsd,enbcuet tehxaptr:ess1i)onoNf-aEcLyFlOCSoEA aonxdidaPsFeOiSn rasttsimiunlavitveo,boatnhd 2t)hethterraensicsraipmtaiornkaeldadnidffetrreanncselaitniotnhael 16 DSRTPITSTB-62958, T-6316.4, T-6868.2, T-1071.1, T-7132.1 rreessuplotsnsaereofverartys caonndsigsutiennetawpiitghstthoeisnuvgigveosteixopnostuhartetthoetsheesfelutowroocfhleumoircoaclhecmoimcaplosu.ndTsheasree "clpaesrsoicxailso"mpeerporxoilsifoemreatporrosl"ifienrartaotrs"acnhde,mimcualcsh, lgiukienweahaptighsasarebereensisdteamnotntsotrtahitsedcfffoercttohef fluorochemical exposures. 7 cs SOTRIPS8T-0295.5, 163164, 148682, 1-7071.1, 17152. Signature Page Prepared by: (nha) IW Sone? Andrew M. Seacat, Ph.D., DABT. Study Director S/z5/0f Date N cs SDRIPTSTH-6295.8, T-6316.4, T-6868.2, T-7071.1, T-7132.1 Summary Tables VarTable = Table 1. Average Cumulative Dose Cumulative Dose (ng) seorcres DcooSnEt_GRO sex u Na a N-EtOSMOmkd esoan 0ol.0 N2 0o.k0o 2 dean 3388.6 N-BeROSElGOmKd NZ 210.202.5 N-ELPOSEAOMKA Msoean 14633.4 N 2) a16s3.8 2 er0S40mkd ean 01.062.2 11.107.6 FR) 4 MSS6-160mkd Mspean 3662..74 a75l.s0 N Missing 3 cour SMDean MMiissssiinngg 11.656.1 Ne 2 F0sAdOmks dsoean 00.00 0o.k0o N missing 3 N-EtrosAomkd MSeDan MMiissssiinngg 41.03.0 N2 2 N-BtROSEiGORKd Yseoan N 2316%.6 2501..52 NepteosEdomkd dNean 212.296.3 216.739.5 erosdomka Mseoan 319a.1 0s.21.6 Ns 4 dsoean 3339.2 i3s3s 1 SRPTT-6295.8, DTISB T-6316.4, T-6868.2, T-1071.1, T-7132.1 `Ta2.bSulmmaery ofBody weights and BWchanges SPECIES --DOSE_GRO BFDO BWD4(g) BWDO HIBWDO ce cont Na(9) 4 4 4(9) Mseoan 423541 247243 1607 426496 E-tFOSA4Omkd Nz 2 2 2 sMoean 61817 25587 195 170 nBLFOSEL6Omkd. Nz 2 2 2 Mean 2653 [233 94 2 2955 NELFOSE4OmKd Nz 2 2 2 sMoean 0635 2629 909 3306 PROSG0mKd. MNeana425 43% 4 %4 a 480 rR MSS56-160mkd Nso M2i1s3sing M1i95ssing M2issing 2378 Mean Missing Missing Missing 258 cont. N SD aMissing Missing M4issing 78 Mean 2704 7211 103 2 217 13 FOSAdOmkg MNean MMiissssiinngg MMiissssiinngg MMiissssiinngg 2353 N- N SD ZMissing M2issing M2issing 26 EtFOSA4Omkd sMoean 8214 1208 927 3403 nELFOSE160mkd Nz 2 2 2 Mean 5201 1mst 3ert 2886 N= BLFOSE4OmKd Nz 2 2 2 Mean 6228 3229 140" 1 1312 PFOS4Omkd MNeana205 r1 t 4eet 2490 so 1s 19 4 12 `Significantlydifferentfromcontrol valuesby Dunnett'st-test BWD4(g) 54 03 240 2 7426 2 12033 2 7334 4 243259 320 218 297 138 235 126 1310 2 22592" 2 1316 24510 15 -- %BW_DO 4110 6 2 194 2 192 2 o100 4 8an s3et 7B 107 32 eat 24 1102 2 8ast 2 0101 g4ot 3 20 SDRTPTITS-6B295.8, T-6316.4, T6868, T-107L1, T7132.1 Ta3b.Sulmmaery of Organweights and OrtgoBaodny WeightRatios. sSPECIE GDROPSE FWV LWB KW Vv KmiWoBW LW LWoWB Ko@W WwKWB @ co ON 4 4ntio 2 2 442 Msoem 71796 NEFOSA N 2 00004s 4013 22 0000016 110917 00000359 4069 0oo000l6 2 22 2 2 omkd Men 185 002 4s 008 267 003 55 0008 NEFOSE NSD 214 160m 2000 00s 000 226 20003 001 o0o 00 MSDem 9053 NEFOSE N 2 0000003 MMiissiinngg MMiissiinngg 016 00000317 MMiisssiinngg MMiissssiinngg 22 2 2 2 2 tz "omka Men 208 003 52 0008 264 0036 55 0007 pos OsND 415 4oo 200 somkd 2000 1S 40001 201 20000 Rr sMDem 61398 0o0o35 4011 0000007 91564 00000%4 0510 00000a1 MSS ON Misng Misiog Missing Missing 3 3 0 0 160m Men Mising Missing Missing Missing 108 00 Mising Missing coNT OSND 4Mising 4Missing 2Missing 2Missing 7 00 70003 2Missing 2Missing FOSA sMDem 0815 0000m% 0119 N Mising Missing Missing 0000090 11421 0000083 2090 Mssog 3 3 0 o000o0 0 mk. SMDem MMiissssiinngg MMiisssiinngg MMiisssiinngg MiMsissiinngg 01107 00004063 MMiissssiinngg MMiisssiinngg pNEhFOSA N 2 2 2 2 2 2 2 02 Mem 80 NEFOSE NsD 024 003% 18 o2 m 002 0008 ol 141 0046 25 03 0001 01 ooo! 0000 0 22 0 0 160m Men 74 SD 01 000009 MMiissiinngg MMiissiinngg 11231 0000040 MMiisssiinngg MMiissiinngg oNmEkFdOSE N 2 2 2 2 FRE Sa Mem 95 sD 01 008 ooo 20 02 000009 i1s2ol 00008074 2017 goooot spoomsa ON 4 4 2 2 So 22 SMeDan_0872 oooor' o1i7 00.000009 Significantlydiferentfromcontolvalue byDunnett' Cit 11322 0o0p0s4! 0220 o0.o00m8" 2 7 DSTISRB ETT-6T316.- 4, T-668682.2, 9 T-1075 1.1, 3T-71, 32.1 GSoRECTES DOSE`TaGbRlOe 4. Percent Initial Bo3d5y4 WDe0ight, cont eTanF108 N-BtFosAdOmkd sNo 64 Mean gst N-BLFOSEL6OmKA X 1a ean gt N-BtFOSEdOmkd so 23 Mean got PFOSAOmkd 18 Mean gf so 5 cont Ndean u1n06 N-BLFOSEdOmKd so 3a Mseoan 2101 N-EtFOSAOmKd Mean 3100 Msse-lomka Mseoan g33 t FOSAdORkg Nso 93 Mean gst N-EROSEISORKA sNXo 44 Mean gt FOS a0nkd o s8 Mean gg! so 3 Sigificantly different fom control values by Dunnett's test Combined Data 2 DSRTPTITS-6B205.8, T-6316.4, T-6868.2, T-7071L.1, T-7132.1 SPECIETSable005s.EL_iGvReOr-andKidney-to BToad/yswWeightRFaAt/iBoRs, 3 TORT ra5 tio ratio 3 sMpean o0.o04o1 00..000016 N-ELFOSAdORKA MNean 04.034 04 .008" N-ELFOSEIGORKA sNo i0.003 o0000 sepan 000.003082 MMiissssiinngg N-EFOSBAOmKd MNean 04.034 0a.008" PPOS4OmKd. Nso 80.002 0I.000 Mean 0.036 0.008" M5S6-l60mka WsNo 30).000 00.000 con MSeDan 00..000434 11 Mmhiissssiinngg FOSAdOmkg seoan o0l.o0o430 00..000009 No3 0 SMeDan 001.000435 N-ECFOSAGORKA MN a MM4 iissssiinngg sMoean 00..000442 N-ECEOSEL60nKA N ` 00..000018 0 SMDean 000.000435 N-ECFOSEGORKA nN 4 MM4 iissssiinngg Mseoan 00l.000435 00.000018 2rOS4Omkd N Meansg.gsgt 04.009 so 0004 Significantlydifferentfromcontrol valuesbyDunnett's est 0.000 CombinedData. 5 SDRTPITST-B6205.8, T-6316.4, T-6868.2, T0711, T-7132.1 `Table 6. LiverPiPgesr.fAlnuaolryosseuslfpoenrafmoidremsedanatdtMheetaUbnoivleirtseiVtayloufeRsofcrheosmteRra.ts and Guinea (ASlPlEuC nsiaertxepsgD)OSE Es ore RFOOSC _FROOSAC _FROSAA RBICFOSAA MFOCSE ERECFOSE gWElTuReO.SE gFlOuSe [SI a Mean NA N7A WoA W7m TRoCv Mow wm N7A - sNs zWM serosa w2m om oow NA m 0 omo N2 A 40mkat Mean 106.6 0.6 A wa so 30.4 08 wm wm N MoMm oomm o0k.o2 N strosN e 2 2 2 2 22 2 2 160mkd Mean 350.2 16.2 217.9 388.2 so 338.5 7.0 1030 %.i 20.86.7 10..79 63.s1 o0.i3 s-erose N2 2 2 2 22 2 2 40mkd Mean 44.7 7.1 30.4 105.0 3.2 0.9 so 453 3s 62 27.5 1a 01 1o.i2 00.03 p20rmoksa Nn 3 3 o o oo o o M cor dseoan 11400.s8 234.53 wWma N1 o 0 WmAa o MNoNm owmm oo o nwmm 0 - Mseoan N0M.1 oNwA omm wawm Nz 2 NNA owwomom o 0 nNmA 2 s40enrkoesa - Msoean 29360.03 00.25 wwmm wwmm No 2 2 2 2 MwOoMm owmm 22 2 20o.o1 ET6toFmoksde. Msepan 5498.88.6 48.62 NN 2 2 21s2e9d.4 1226113.s6 8127 .63.0 022..23 221.10 20o.o5 E40tnFkodsE eros MseDan 888i.l40 NW 4 40.39 1 2886.2 51.46.0 31.65 013.53 oo 0o.l o00..40 40mka Mean 130.5 41.7 Ww so 131s 3 wm ow M Momm owmm mNA RF cor MnNean 10.0 WA 244 w0a w0m 04 mw Womooowm w0m mom ow 0A NA N Ecrosw e 2 2 2 2 22 2 2 T6onkd Mean 891.9 18.1 67.5 20.9 so 37.6 11 5.5 33.4 2474.62 61..45 00.12 00.02 eros wn 2 2 o oo o 0 2 DSRTPITSTB-62953, T-6316.4, T-68682,T-7071.1, T-7132.1 Toma ean 866.4 18.3 wa wa so 1300 4.3 wm wa wWmoooww oomm NnAA uo wsse no 3 3 3 o o0 o 0 160mka Mean 140.3 316.8 555.2 NA sp 85.4 1141 113.8 wa mWw omoN oomm RNYA cornMean 20.0 5109.2 W0A Noa so ole 1136 ma WA mLRo)w o0w MOM wm k0o NA 4F0omksg. wo 3 3 0 oo 3 N= Mseoan 2169.38.4 117673.9 mwam wmaa No2 2 2 2 mNoMm ooww 22 2 00.14 2 BTe6r0omsked eros Msepan 11162.41.3 91.85.5 2943.73 s29l4.s6 x2 2 0 212..51 13..28 oo 0o.l3 o0o.o1 40mka Mean 963.5 17.6 wa wa SD 88.2 105 Wa wa NMAoWw owma wNmA 2 DSRTPITSTB-6205., T-6316.4, T-6868.2, T-1071L.1, T-7132.1 `TaRbaltes7.aSndumGmuianreyaofPiLgis.veArnPaelryfsleusopreorsfuolfromneadmiatdetshean3dMMEetnavbiorlointmeeVnatlauleLsafbr.om ISEPSEC SEX oDrOeSE 3E M e 3M BLFOSAA wE=LFOSE 3 FOS ETt=FOSAaM ER a 3 z"am Mean 0.151 0.0 0.2 EY N 7 01T w7m NNsow2 Berose 200 200 N2A 2NA w1a T6omkd Mean 604.000 109.0 244.5 sD 5.657 113 24.7 310.22.5 1131.84.5 0N.A9 4er0omska 2 22 2o M cor sMoean 6870.31.15500 00.01 x2 2 o0.l1 2 nmAa o o0o1 wwmm 10 seoan 00.1239%8 00.00 00.12 NN 2 22 WMA WA0.1 wwmaa 2 22 BTe6roomskEd eros MseDan 9900.83.10000 98.95.0 1371.77.5 ow 2 22 41427..0 1106.96.5 00..14 o 2 40mkd Mean 800.000 0.1 0.3 sD 16.911 00 00 NMAw 00010 wwam 2% SDRTPITSBT-6295.8, T6316.4, T-68682, T-7071.1, T-7132.1 `Table 8 Hepatic Palmitoyl CoA Oxidase Activity, Combined Data 3 ont ean 28 N-strosMomkd Ndos esoan 21.0 N-ELROSElGONKE MNean 33 N-EtrosEomkd 0Nd0.6 2E0Sd0nkd bseoan 20.6 N71 conn esoan 31s x7 Hoan 3.2 N-StEOSElGORkd Ndean 413 eFOs40mka sNo os1s ean it Significantly diffrent rom contol values by Dun5n.et1t's test. 8 2 SDRTPITSTB6295.8, T-6316.4, T6868, T0711, T-132.1. Appendix 1. Deviations to the Protocol `pTehrefl3uMoroMoecdtiacnaelsuDlefpoanratmimdeen,tTi-d6en8t6i8f,icwaatisonnontumlibseterd fionrtNh-eEtprhoytlocol. TPhFeOSprAotaoncdolFXu-s1e2d.abPbrroetvoicaotlioanmsefnodrmNe-nEtthnyulmpbeerrfl2uolriososctthaeneasbubrlefvoinaatmiiodnes(fTo-r6868) of pabebrrfelvuioartoioocntsaunseseudlifnontahimsidreep(oTr-t7f1o3r2N)-aEsthFyOlSpAe,rfPlFuoOrSooAcatanndesFuOlSfAonmaimdied.e wTahseN-EFOSA and for perfluorooctanesulfonamide was FOSA. `stTuhdeyp"r.otTohcaotl swtaastead tthyaptogtrhaepahniicmaallesrrwoor,ulads ebveiddoesnecdedobnydtahyesf`azcetrtohtathreovuegnht4heofetxhaemple cacatlucaulladtoisoinnignptehreipordotwoacsolfowradsayfs azoe4ro-drtahyroduogshintghrpeeeroifodt,henostatudy5,-daanyddtohseianngipmearlisodw.erTehe sacrificed n day fourofthe study. Adanyimtahlreseowfertehenosttuwdeyi.gIhnetdhionsemoisnsttacnacseess,otnhedparyeovinoeousfdtahyes"stbuodyd,yawnedigihntssowmeerecausseesdofnor determinationofdosing volume. pPerortfoocromltahmeemnedtambeonltit#e2asntaaltyesdisthoeftDhre. lMi.veWresammppeleast ftrheomUnainviemraslistytorfeatReodchweitshteTr-7w0o7u1l.d1, Tp-e7r1f3o2r.m1e.d tDhueesetmoeptearbsoolninteelacnhaalnygseess,inDsr.teaXdi.n Lu at the University of Rochester CKliidnniceaylwcehiegmhitstwrayswnaostnoobttapienrefdodrumreidnognnealclrsoepsryumunsdaemrplaems.endment number 2 on 3/5/99. pHeirstfolluoogriocsaullfeovanlaumaitdieosnoafnldivmeerta,bkoildinteeyvaalnudestewsteirsewenorteonbottaipneerdf.ormed. Serum Ianndd/uocrticoenllorfemplRicNatAiofnowretrhee fnootllaonwailnygzgeednfeosraisnsloicviear:tePdewriotxhispoermoexPirsoolmiafelrpartoiloinfAercattiivoanting RNeuccelpetaorrAn(tPiPgAeRn),(PLCiNvAe)r.faty acid binding protein (L-FABP) and Proliferating Cell E 7 - SpRrIiTsTe.62058, T5164, T9692, T1071, 17152. Appendix 2 -- Analytical Liver Sample Lab Identification. [Study 7 imal [Record EI E ic [11/19/98 |-- 3|BRO4041JF RF [ee preHRRa--H PFOS, ir Me [re[mmfee] [rose s Fr [por r|e ompre yv] 5 [re re [owepe] s er emo po s dil cl A A oe [rome[owe roo] i i Fe [mp ovrep o eh, [op] o [rer [rm |owe] 7 es[rm omrepo] lo oe Fandom [ep i cdl elWc A 2 lo oe ou |p] lo 30s lM ls rs. [Dose CovancelRoche[st3o [Univ group [Necropsy[Specimen/Amt [Date [Type sentto [Amtsent [Amt [Amt to [senttofsentto. \(@) r1419-01 [Env MN fab (1- [Duluth cc CcA [1C5.0H1 (unio| rn fcont 1 v1/2008liver | 0.48919 |-1q JNA] rose [rose rose rosa eirosa mkt [21 [PFOS; [22 meBenen[[C 7"OONTRRe e[| | ar |S Susee f freoprea] FOSE; [33 1oR0049IF ioRo0ralF Cont| 5/590liver Cont | si5edliver ROCHE rNoest e| 9R00466|M [N-et | 07a7-1q | 05516119 0.812] Tafro] |-Ta 15] I~ 77 SRbTEiTssT.62958,T6316. 68682, TA07L1, T7132. al 8 el BlAL LN [[2278 JoRubdarius {pFrooss| JoRovagelw [pros| waeimseeslliivveerr ||0808t3l-7-51aJ-_Tg|[--=1T1aaa|] [[_3209 _JoJR004oTAlRF[PFoOS||0awiiismessl7liivveer3 r I|| F 067770_63-T1[g4 --p-J-1FTaaTJO--11aS q|| dc 0RL HL LIAL 5 Fp |e on op] 16 Fem er|e| omay fe 1 [fe ee epee) A F eee |e S1I960l 004[W Net er e9s600l 05[4 F Not| 3588 s lo l I lp oe o I 7s i I7l] I A A A [pro| ee) 5 GNl a l I l Ci O 0D0imailc|L |--_--[2R[1oRb00r7aan Z3 W [oCrontt|| Zp1 ErSi0lTi[lveerr [[WNAA [=TTeg J[NNAA [J-T1Tgg|| Ven" [SITRO07S0IM FOSAA|2i23i0iver NAa T=1a NL A T-T 18--]| le [eltr[oTRoor00a7rat]]w [FSA|2zotlliivveerr INnNAAt----1Tgq NA TE-T1gg] 20 rSRiPTsT6295, T6316.4, 768682, T0711, T7152. Appendix 3. Cumulative Dose Individual and Summarydata. 3 T cow aoa0oTiIiEess 00 xasca00000s5t2 00i Bdyean x0.0 N-EefosAOmkd u660c1o4ld5s0e 219061 e3an 5885.6 N-ECEOSTIGONK S9G6000000s5a3 2116606 5Hoan p16l3.4 N-EeFOSEAORKA a6cGo0iLd4e8s7 2110023 w&ean o10%2.2 e0sd0mka ssccoonnaiosnz o9n5 53560000005556 i3358 H0aan 3662.47 w co 5ss5oc00iu04rm5 00 exSca0n0045 020.0 5 alo NeEtFOSAMORKd xaGcoolisdeer 2112233 Edean a1l2o2.5 N-EeFOSEIGOMKS ~55G60000004487 2116617 5ean ilesss NeEtrosEiOmkd a6o00114s7690 117105 a -- Briss SRPT T-6295.8, T-6316.4, T-6868.2, T-7071.1, T-7132.1 TES RE. Nten G:d7T .6 SE sti srostomka sSSocoohriotensss:ss 11310s .[ScAone 3i5 eT Hs x room sSSRrRoooitooaisiesos 0 Y.Smoonoite i00 FS weEtrosadonkd 5sSRhoioodus 33:86 dwron 36.6 NeEtrossistka aSsRRo0o0tAnTzI 3113256 bweTan 203 westroskionka :SsRRoos4o0s8sz 3h400 YEoon S31 rrosdoakd sSSRhRooosoiaoonsis)s 333851 3tSeoonirs 3i32 ae 3 Wo wsse-istka mIioooonrnsss iiieess sescrss sex poseGeo NBtea 5li%es BD Gilulative pose_ieg) 5 fRoRooooorrsz RRSmoooioiesss:s 2 Yo bSRTPITST-B6295.8, T-6316.4, T68682, T0711, T1521 NSROGTET 7 Mean 0o.l0o FosadOnkg 1100000774456 3490 100747 431 sdoean 130.0 N-EFOSAGOmkd eGRR0OaA003376 S5I1 xean 251.2 5 0.8 N-BCFOSELGOmKA smSRoO0O4s6sS 117854 xdean 2179.5 5 63 N-BtFOSEAOmKA o6RR0OaA003354 5533 ven 252.6 BE) oi eros0nka RR000033 4590 ssmRo000eae6rs 1433 ven id6.8 58 3% DSRIPSTTH-62958, T-6316.4, T-68682, T707L1, T7132.1 34 8 GY oy demon Bw | o| 2 g 2 ze 22 EEge 22 2 2 PEF 2 Zr oz % ze peer p iX No PT [ER335.%3 = 8 E low ao 2 | [2382 7 ZT |[cs88 i 2 [98888228 =E 8 E 2 e 22 3 sli s gi3 2Fz HL. og) is ZH #8 < fas Ve < 3 8.54 5 2.2% 2 LE] noe 22 28 5.885, 2 2.28 8 8.23 i3 gg2 1823 Lig8 2 - 8 8 4 z9ze gf soa 22 3523 _ B.2g sane fons 3338 g 9888.3 iI o: LEga E3S 73 8% vgn. Boo. R55 a. 332d. " 5 222.31 5 4035 dedd 38.40 sEsS.a2 mn Bon gosm dnas nd. s 0 i 2l a5 Is " a9 - - dq a 55 3R3S2S2E.SSTS 2R 2ROB$Yv 2E B.EdDo &2 2d8.3=4 w2e3n5o3.3o8g LEE. BEES B 2 OBLED FEE f 4.84 Se, FE Bg EEo fillEd , ? ZPEr FE oP oP FP RP OT $P % EO PP 2EPY gr diddodd dodf f dois 4 dR.4F JE , wee . 3. ae FE 2533.85 2 81 g 3% To 58 43 moss en 3 fess gg 2 EE 3i2 g 5 2233.88 E.8s .8s 8a.in E88R.Is 8 2 : 8 i i 3 2i3 3 Eg2geg iz 18 LE 15 & 2 2 py = 8 8 ga i 55 g 2 REE4.E5 7 8.2 88 8 8.87 3842.89 38% zggP FZ 2 2 Zr oP oF FP OP PT FP PEER BP Hfdc8F # Ho88 S.88 FoF S5sj.sd 38% g EE25.i5 % E.85 & Z.ds E E.3g 2822.35 222 5 23 . I" iBaEL 1 a 3 E 7 = 95 atts am, 380 58, 33 " 43583552858.0 2 ss3.50 & 3.82 2 d.57 FE SgEaE,LEEEPEETE"IoeIRaTn.TIC2T3 SR , anI 5J 3Jt F2S7 L5nERESHRRBA2 RRE.Ed 3 3.33 8 4.H] #2 od i. 2. [ERE 23538888228.: 888.85 8 3.80 f 4.8 3 Pa 2 - - - o IFEf GeBaegfi82 5% ap 52 regrreEEze 3 BYE 3 EYE LEIERRRS494.89 ) ARR.ES d ddd 4.84 4 o 8 |.3aB22EEREd.s 222.0 EF E.ia B05 2 E 1 3 3 5 3 i : : g i 5s 48 3 2 It g ud g 15 <g i da g4 5: f6 i i 5 37 s fa2 iez sis 3 [fecEiacs g ze 2iEflRsEiiE 8228 27 288%EE330s Sass HoEE 2 pp 22 3858%3fF fgosidifis4% 588 4a;2 Sade S OF 2 JRgdB ud88d82gI8I8oEf E5 l z [lEcSs.ss So 3da8ds8s& 3 Gadd 3 3 gG8 8 addg3 Sa28y _ flpess zs 28: 5882 E 4 EEoL lsRSdiear dag 3 2.33 3 J e | [pszz gg gE I i : & pEEsE 3s B Bids i Bis 3883s, da8l 8 23 3 g 38 B.3s EEE R iLO.ERs g5 ec S| 2i 3 g ggs Zi i 153 LEE 152 , i. Tf 58 Fs . 55 n 27 i 58 228% 58 5 Badd coffadd & : rz B.35 2384.33 3 : 5 28 F 3433 zg RWS 7 er 3 E34 d.34 5 zz 4.481 3 oF 58 36.3% 3 3 33% Go L8E%EEszEsEzEN88IE= sRo E52E RE58E: mPEa: _ Sz 9%32 88s 8 Refi fgddeds CEN SEE 3 383. csv: Sadd Jedd 3 [- so Bg REGSH & fois B808.38 8 2.88 2 S.i3 8 Ge.ip E 3g g i 8 gy 3 v8 8 14% +58 13% 8 . g i 5iks :g Bdd.33 3384.33 3 3.33 8 d.38 S.dS J FEdadd IREEGA3 1 AWG3 FF dedf I LLIS g 2 3 i 3 8 44g% 2432 18I% gg 2 o i3s5 . 70 zr g gr pefgen eo: Tgp Ez 2: poep oer oz sr oer or g 233 3a8e 3 32 8soBnsssszzzz o2o8 5 8s 2en84 g 22 232.00 = 8 2 Bu mnoo9se Re rm ss.s3sbdsdndidf 5 ow 2a sss o oo ld 2 g ser 2 22 uo mmo oes wo 55. CGD. BESSCE ozE.E8. 2 g 588228 % 2 1 EZE.2QA5ZcSggfEgg RX EZ. 33 zEOE, g 42 & i 3& ls 2828 E8Hg Fi2. 25E%g Z 5 5ak3 ig 9 B 3 rT ogogg zz Sof dedf I 5.33% EEE Nef dodf dads J SELES E Es f Zf.s88 g 2 3g2 18182 g< 3 5fsa FoFE..i5 s 1882% o2 f2c2E2E..35 % s2 . 72 Ey YT TTT 3 Bossssoss S Gadd S 3.33 3 2.3% so z fy . S Feessoss 5 sass 3 RET RADE, 4 28 f Efssssns 5 sees $032 3580 3 . : Blesssss 5 goss 5 sues 3.8 5 4 Iav 5 Feesssess 5 soss cfssi.faLew ELBE 5 A030 sg 22 2883 54 } $ [lesssoss = sess 8 2.58 F.5D 2 oF 32%%FE |P g BfTe 2 REsES2 I3ds2GAs S25 TR 8 Re TgI32 s 3uEs386s4 g = 8 4 5% 289% 0 op 8 ZF 8 Eesssoss 84.32 8 8.35 6 did 8s g |p [8898.35 8 2.45 8 8.82 E.85 S 3gS gE mlueolee &Log2ie2 ) #iEtx 00b#i]i 2#i83t Es 282 Z2F E8ols 73 $ esoss ssssoss 23D EET EI EE 5.55 s555osf ssoss ssssoss 5 S035 5 best 3.37 5 es 5 sess o dens 5 0.0% 5 ag S5o85 5555.58 8 fo5f Sadt 5 G30 5 sg Efof5 585.85 8 fosf 5 3.8% o 4.37 5 sg $5055 5555.55 8 S085 2.8% 3 dus) 5 ss oo FREE sess 3 RE8 D333 383 2H z E BT EE oo PRo%3 ssis.ds %38 182 588 ec883 Fo s.ss 8 $.85 4 5.85 3 of 2 28 gigs El 1} LE 232 BS-is 8382.82 % 8.35 3.85 B.05 B Ef :: I3 M 33 E3 s 3 g 8 if i i: ia = 5 9 BoSS 55855055 S058 J Jedd So55 S55 BoS5 5555085 S058 5 Jade 5 S55 5 55 Bo55 S555055 Soff deci 5 S055 5 55% BoSE 5555055 S055 & ISI 5 SoE5 8 S85 Boss s555.5% 5 fos 8.87% 5 sos5 5 sss. Sof5 S355055 5 S055 FaS. S055 5 S55 a Fed8 $8400d0 8 S088 5 S481 5 S085 5 sds Toke, 28 sy 83 g S-3% 505.32 % goss 8 2.35 5 sossz sz ski. Z 2332 Ig zg $2 2 52% g BB-.f8s5 2B3X8E2..3030 %2 22..38302Z 28..85305E E3..8003 2 2528%, i2 iigf i3s iits g2s gis " 5: . 55 5 ssosssssssssoss Goats 8 goss Gd 55 2 S5.3ESSSSsSs.ss 5 5053 8 fos 5 = ssosssssssssoss 5 S508 f fos 4 oa 55 5 ssosssssssssoss 5 ss.53 5 s.3s 8 a. B55 55.555588588055 5 55085 goss Nd sx 38.8%sssss85055 5 s5038 5 goss or. oo RDF ER.EE8deeR0.80 4 F9R0 go ses 8 A EOR% 2st ae es 55 8% 3 Ei = 3 8 8.%85s52535.23 8 38.588 5 5.55 8 Z |g 2E5358 .97I8 3Re3B0s 22g5% s .s 2225s .=r [3= fs 2 E2.a2REEEEd5s Z 22.85 .85 2. 3 3 g 3 Egs 2iwsi 58 g2fzg i8Bs2y i8E2Hs g = 52k ES 2 so35 2 sss.ss SS 5 goss 5 sssosg Gi og fo3% 3 sss. Hi f gos8 5 sss.3s B23 5 go55 5 ses.%s 58 2 gosf sss. - 53s sess a la gzsF zos ozoss 3= 5, 5i:g5. is .82 a 2z 21s 5 shaun 2 83 24 os 5B33E.N3E5 2Eg 325%%: EF 238.:83 23 8 52 97 g2rE EF gz ' TTEEEE0EE SadS & Sada ~ dwoe o Ha |. EuB2- |efrEe2sE p2r 22 zg gzyr RCEaZ_f[5e58i8iii38 i2 c s8iz0 avwcwyn raoaann i& ifo z il, Sl $sR[EEgEo [le3p8e%r oz 58 RH 5f 2. o 32 -nt. s ew iifhy Z2 | EfEFoESS|eEhzdediroEizdr n2 io.oid$i98 .d8.13i5 i8 i5.n8 iInzEd 2 2 2238 28 23 58 a8 3 fle [s88i.is 5.5 8 8.35 B.3s EBB i008 z 25 Jo " " b 530k gE OE ge iz gs 3 if is if 1 .i g g SHE :i &gg,f &4T3ilo 5 Eb 9p zee gp gegee gY ge grr 2 zg x g 8 2 o fg 2 z2 = bE 2 gree 2% 1 o gees Ge EEEE0SE 4 deac EF 89.87 I J3.038d:d) zpes 2 [Fo EEEECEE Jade I: 772] EF EETWNS Jono EERE 2g [i+ Ec506S & dadd ge zg 2 FE Fevers I SSeSdE4EE Re EEdsaves 0 DaRR FE ESR.83 DT I8WSRE0FE a #3 2523.8 5.83 88-85 E EE ggeExy &3 5 2: 8 i < = is is i8 53 iz 8 i13 22 = 99a. Eifodd I dune # Gerd FERSWE] Bod I 23.33 8 Iosd 454.83 Foc-33 3 33.33 4 2.87 dedd.d] Eoc.dd # BR.E: B.BE 453.3% 2 :: 82y i3i Z3 fg iSsE : i: zn a ii 8 is 5 J00 , = : ss El sssgoss 3s.is ssssoss sssssssoss fl ossfts 25.20 0a3R0 cmeneided = E 22 8. El ssssoss 85.85 ssssoss sssssssoss & BE an gx BE 233.32 38 Be 8R.EF se zm 850.30 zs asl ssseslfEY EH 2s 2 Ble zz2 5s Z Ei dddnds 22 2m 3Ra8d 22 =o 2855030 an =o 2s8833033 E 2 ow 825885, 828s 83 a 22 ss _ g SE 5555.55 85.85 ssBP.EC sssssii id IO fe Eee 85.5. EEEELLS 2222255,3 3 f< g 3i FEgs 23 B2 C5H3. =i : 3i g2 g8 J0/ _ : Eo ao : : 5Ea2 Co 5% 38 Sonco S3d8 a3l FRE 3ss2s% ARenY g5ss.55 dsdr l23 g8sgoss 2% 3a sg 2 REAR 2% 8308s es 5. 3250.8253 S225 88% 83 fdccase sud fEccass| J 252i8q.8283 SgEeSsE | iz ig 3: WE | vere oe CBoeuel 85388%3 33%% 18d EEE EE oo oof "a Bas 2s FE 3|g || fog c23o2o%r BREE 2eef EE... g || EEEEEE & daaq 2 iPEl. EE Fs fie 00 Ff [FF E6Sass 2 2 2 og 2 2.84 Tl |. FEY fd.. a5 F[2Y53 #3d833%5.3<34 38 suas Fg f | EEEEER.iHIEn 2 3 zl Filf | H8|g 22 3EEEGEEe rw IE.is 8gy iHi=Hg i Zl i a E[8 l. 702 2 Iz ? 57 PEEEIPIEE PT PT IF II E8n.80 F dEofdEddffdidd f did J Iz TEZEIIr II sddedd 3 E.33ddddd48.8 2 23.50 4 Iz F gadecd 3 32.230333333.3Y 7 33.3% Tas I 7 28 2 #58.8c 3 FR.ERE3R3355.q8 8 B8.Fd g $88.33 % 25,gEEE88%8.8p 2 22.3a 2 4 i Pia2]]3 4i1212 gu = 5g2Zf7 2z#g iBEe 143 sas EEE S83 FEY s.r f dada 8.80 fiz I | 8.ds g g i 5 E E[2 i 5 5E3 ERR.IS i8s% Jo { i 70S DSRTPITSTB-62958,T-63164, T-6868.2, T-1071.1, T7132.1 REo.cTheecshtneirc.al Report: Liver Fluorocarbon Metabolites -- University of Title: Summaryof Quantitative AnalysisofFluorocarbon Metabolites in Rat Liver Samples LinXuand M.W. Anders "The paren"tThainsdismaebtraibefolistuemsmoafrthyeofftluhoroacnaarlbysoinsswoeflrievdeertsearmmpilneesdfirnolmvaetsagimvpelnesabryanLgeCo-flMuSor.ocaTrhboens results re presented below. `control at"sT,hrferogmroruatpssgoiflvevne40smamgp/ledsawyerFeOanSaAlyozreadl:lyT, ahndfifrsrtogmrorautps g(iGvreonup1610) mcogn/skigs/tdeadoyfFliOvSerAsAfom ((TMa5bl6e)1o.ralTlhy.e dTahae csohnocwentthartatPiFoOnSofpwaarsetnhtecmoamjoproumnetdasboalnidtethfeoirunmdetianbohleitleisveirnsvoferrtsswgeirveemneFaOsSurAe;d oFfOrSaAtsNg-igvleuncuFrOoSniAdAe (wMaSsS6i)d:entPiFfiOedSaasnadmFiOnSoAr.metlaibsolintoet.ewoTrwtohymetthaabtotlhietceosntwreorleadneirmatlisfe(dIRi0n0t7h4e2l,ivers F1iRsOc0h7e4r33,3a4nrdatsIRm0a0i7n4a3i)necdonitnatihneedUsniigvneirfsiictayntofcRooncchenetsrtaetiroVnsiovafrFiOuSmAdi.d nAotpasrhaolweldaentaelcyasbisloef vers from concentroaftFiOoSnAs. from contTrholeassec,orndogmroruapt(gGivreonup402)moaf/rkig/sdwayasPtRrOeaSteodrailnlyN,ofvreommbrea,t g19i9v8e.n 4G0rmogup/2dcaonytaNi.nEedGFliOvSerEs calocmopboolunordasllayn,danddenfsirfoimerdamsetgaibvoelnit4e0smagre/ksgh/odwany Nin-TEaWbFlOeS2A. oTrhaelyd.aTthsehcoownctehnattrvaetrisonroofmparratesntgiven siaPnvFdeOnFSONc-SoEnAtAAaFiOn(SeMEdSSpa6al)ro.enbtoTlhceocmomnpitonauoinrnedmdesPteFavObeoSrla;iltFemOseSwteAarbweoiaFtsOs.aSlAs,oThfFeoOumSnadEjioanrlchmoehsoeel,stVam-paEltewbsF.eOorSTEehlNae-liclEioFvhteoOrlsSehfAirAcsou,mrorPnaFisOdSe,, aanndd FFOOSSAA NN--gglluuccuurroonniiddee. wePrFeOiSdewnatisfitehdeamsamjionromretmaebtoalbiotleiic.nvIenscofnrtoramstrttso GgirvoeunpV1-,EtIhFOvSeAr;sFfOrSomA control rats did nok contain detectable coneentations ofparent compounds or metabolis. at, fom"Tahestgiinvdegnr4o0ump (gGrodupay3)PwFaOsStroeraatleldy,iannMdarfcrho,m r1a9t9s9.givGernou1p603mcogn/tsdianeyd lNi-veErGsFfOrSoEm acolnctorhooll Naorn-adEllWlyF.oOwTSchEoenaccleocnonthcroealnt,tiroNan-tsiEooInfFsFoOfOStSAhAAe,wcPehFreOemSii,dceaanlntdiafniFedOdmSienAtlaAibvo(elrMisSt5efSsr6ao)rmwreasrthseogiwidnveennntiPfTiFaebOdlSae.sm3.aInjAotshremwelititvahebrGoslriooftrueapst,3s,agnPidFvOeSn. sthhuemciurnoonirdmee.tabSiomliiltaerstoidGenrtoifuiped1,wceornetFoOlSaAn,imFalOsSsEhaolwceohdold,et-ecEtIabFlOeSheEpaatliccohcoolncgelnutcruartoinoindseo,faFnOdSFAO.SA NSDicshceumses)i,onb:utThdeosneodaatlalaolwlionwfceornecncleussiaobnsouabtohuetetxhaectovreoruatlelsmoefmteabtoalbiocliftte foofrtmhaetifoino.roFcOarSbAonwsasstudied (sce F`comOnestSiaAsbtoelwniatcsl,ymbeuitdteantbthoielfsiieozduerdacsteooafFmteOtShabAoalNmi-itgnelouocfguPrrFoounOpiSd,iesw.nhoettrheeardiPlyFOapSpawreanst.adWmhiantisetveerreidsdirrocutteyoforfofromramteidona,sa aarlicsoehoflrotVmot-thEheeIcFNa-OrdSbeoEextyahlyilcltakicodng.oifvGel-uEicsXuerFo1On0iSadEartaailnocgnoeohoofftl,hmeaapnjadoreNea-nnEtd(NmF-iOEnSFoAOrASmEectoaubaloldlcioathieossl.ewboFyuOltShdEgoilxvoiedbatthoeioocnboosufelrtdvheed reNxe-imdEoavFtaiOloSonoEfftahFlecOocShaoErlbagollxucycomuheroot,nhiyodrlebsgo.rtoh.uFpOoFSfOAFSAOASccoAouAullddabsbegeflyfooorxrmymelaedtdeb.bytythFheOeNSN-A-ddeNec-etghhlyyallcatutiriooonnonifdoefmNN-a-EyELbCFeFOOSfSoAArAmoeorrdbbbyyytthhee the glucuronidationof FOSA. 5 [06 DSTRITSBT-62958, T-6316.4, T-68682, T-1071.1, T-7132.1 giveFOSTAhoerfaltoesosoffFtOheSAglAycaipnpeemaorsierteylawtiovuelldy sgtirvaeigPhFtfOoSrwdairrde.ctlLyo.ss ofthe carboxymethyl group would 6 167 : 32 2 g : 5 588 Po aE po aT bp} S999 i Z Jd I{HAil s LH see ed ee gz moib EHH yop Ld s ul gE [H ERE T 2329358 3g NE [3] FT 3g | ETT 2 ETT | FER {Een i3z :i d2a5a 77 i [ Aefelfelmlefale alfa] ie 3dl HHH i Reess FiR [ETT | em TT fe TEE 5 & EE 3 3 AEs EE =a sista T {8 [E TTT CEeETiE 2 SITE TaTIE TEI OE oh bh 1 Hi fHAli. 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B 9RZ.B.40 F 3.3. 5 2 suf.E- a] ggz 2.2 3 = raf.8 2 g fa.5 s sens da 3 2 s.82 saeedz| og . ~3 2 3 mangda| 3B 2.28 arn] 25 3.33 sesed i) 3 5 2.83 $338.33 8 2.2 3 2 s#2.3 22%2 2 3gs ous : j 3 Fd 3 foro 2 i Leo PHLi. 2 "3 i g g 2 Ex 8 = V/A &E 147 Tor0s71F1R A. Statistics on Clinical Chemistries for rats, male and female values combined. 'CHOL mg/dL By Dose Group (mg/kg/day) Rsqure OSnoivamAarooFnaa omorrst RRNoeooikndoanfaReneysoratrsoErr CTiaasseataosrrss Ghianatons (or Sum Ws) = Soouoe OFs ArSuomfaeoVrrdSnsiacmees MenESaRerEe oFrsio ooyan7 ai0s%e 8or5s hoemre Loovner Mansfor nowNaoyArrteeTva uMmean SeSremr NNNeEEErFroOosSstnEiiiooonomkmasa P1I Ssom osamn Frosiond SR ie Lovet S44 ror ssesarapsNon lmoesr tBS aortao"Mreaonr variSanucDeoy Sn Tum Tem Sd Erphieoasn NNNEEEFFFOoOSsSEnEiolononomimsss PHoOTdBmE]mD dnemt; siseNr2 Prosionia io hae in ONtoEnFeolsnoenaant) scooonot Mea"nNsCEaArpgSarirsm]oans nerosr simiaiPro] sionks Sore bt] Sooo nase NeFosESSieihroaimnoa n tz priman LenseorEeocsecAus oumme wn oss E gaome E swowe ameww osuings oWxrenowoema ASBCoN onnesrhsMiE\stroOeMonyewSersotstweiromssoand UE atm rossroisimnis nerossoonms TMEooSgeaE, mhmwme aeemds EIaEEmm oum TLhmGEe Poste vanshow rr vsttron. aisieees Sr jHzweoolomJdeo]a 5 1i1co5gu8r Tporeoscoums , sWeEnR SCet n iston coPA onssiet01ss Srooxmesctrrerstsesnt | 79 rnin " Corgis Bs xao) FwImNVTA A SY AL R "s v \/ CONT NEFOSAOmIG. Prosigs | ahPar DoneGrup ESSE gu Wil Gonirol aes B 120 TT0I7S11 FR OSnuemwmaayryAnoof vFiat RsRSqquuaarreeAd) Root Mean Square Error 0012081195672 04t8s28 `MOebasenrvoaftRieonsspo(onrsSeum Was) 1975 1 MSoodusrc!e OF AnalyssuiomsfofVaSrqiuanrcees 3 08250000 Mea0nzS1q5u0a0r0e 1F 5R6a7o7 ErCoorta 215 2219035000000000 001785S33T3 P02r4o8b4 CLOovNeTl Means for OnewNaoymbAenrova Mean 4 121500 S0id20Er0o1r NN-EEFFOOSSAEMdOommkkdd PFOSAmKd 44 1111698500 002200004411 4 122000 020041 Lovel 51dErrorus`eMseaanspoaonldedSteDseviattoiofinas.omravartianece Number Mean SDev SUE Mean CNO-NETFOSALOmd NEFOSEAOmKI 44 11129120500 oos0uieser0i7 4 1es0 ossrast 000207426354 028305 PFOS40mkd 4 122000 0216025 010801 PFDOiMSeAa0nmiK}dMoanl] CONT! PFOMSeIaOnmsKdCompar|isConOsN.T 0000000 0.050000 N.EFO0S2M7O5m0k0d0 N-EFO0S5E7d5m0k0d0 NNE-IEFFOOSSAEMAOImmKK]Y 0020755 00000202050 0575 oss 00.202050000000 23 00532050000000 0000000 Aphas `Comparisons fo0r0e5ach par using Student's t APFsOOS4LomSkDd 217t882 PFO0S8AO8M2K7S 0s"oCsO2N7T N-EFOSAWOMK N-EFOSEdOmkd CNOENFTOSAMOmKd NEIFOSEAOmKI 00397052 00.d4s20z0r7 007027 012027 ""003472002277 "003644852277 0".0.1027002277 "osaszr "osis2r Positive values`sChomopwapraiisrosnosfwmietahnasctohnattroalruessignngiDfucannnty'sdifMeeretnhto,d 268dl201 PAFbOsS(IOIONmLiSdD 07C4o4n87t CNOENFTOSAdOmkd NEFOSEdOmK 007536495577 0.26987 dPioseirteinvte.values show pairsofmeans that are significantly PHOS mg/dl. By Dose Group (mg/Kglday) 2 ToTisLR SN -- ws/=\ A / -- ] $ Aos VA ANFAA3>]{ v VI. w \/ \/ rosea. Sa oe oa EE `DoseGroup (myKakday) .- RRoquqrueeSAOsinemwmayAnaorvtaa Ro eanSaar Eror GQouerotresrst HGeheaannovratRoenssiroresSumWis) are%s ENSrowreso oAF5es SuomfoVwraSanucrmoss 2 Sirs MeaTanirswgoe pGFeROnuiSee Con 1 ween liam same LcoNoveenrtosaioMonoinas forOravNaarAoraeT I rv|a eMean SIGerere]sr NEFFroosSitoinoidmts Dfom in ooai cLoovnet 4Erosaesana2 pNcmbie0 rtDnoavSoMfernor vriSanucDeev ome ome SSGieenm NKEPErFFoOosSisimoonma I i40h0e EoomoA doase ofurT-- NEFOShOmG PrCMOeoNoTnsCaNmEpFeOrSsoHrO2ns0A0 ars 00cano NEFOS7iMO0n00k0d PROriSIsOnmoid Prosi Srreaimn praendo 00000000000 GGoooeoy Apnan Compofocraeacrhtpiir ssingoStenntss csoensso 2SCeOaNtT 21"7N0E0FoSSeEiUnemnA NEFOSoMsIsTaksd |rOSoIsosnaiea 7 122 rTiIs07LFIR PrNNOEESFFOOASSOEMmUOaOmMkKd s0t35d5s0 035509 313.6630080000111 TA033a609e01 f3s0s0e0e1n ese Male andPfoesmiailvatvsalcuoesmisnheowdpGaaisanodfmPehaonssctrhiactaalrcshsagmitftceasr.tyNodiifegrennat.ng rom coil, 7% JX3 Torson os os ~ ss g > TBIL moe. B DoseGroup (mgr A & i= OOOO) V7 = PA 02 :: y ET Sw soompi re" 4 Veo Co " 2d TrIi0s7L1 FR OSnuemwmayarAonyfovFiat RRSsqquuaarreeAdj Root Mean Square Error 0o347a7s2s0t1s 0083491 MeObasnervoaftRieosnspo(norseSum Wats) 03583323 HSooduarc!e OF AnalSysuismooffVaSrqiuaanrcees 4 008658333 Mea0n0S2aiuoairse F 4Raaat2iso EnCoort 210 000197187353030303 00007872s9 Poroobtozr cLoovnett Means orOnewNaoymAbneorva Mean Std Ermor 8 037500 002s N-NE-IEFFOOSES1A6O0mmK NEFOSEdDmKG 43 0042050000000 0O0od7d1s2 4 015000 00u7s PFOSAOmd 5 042000 003108 Level StdErorusMeesaanpsooaineddSetsdtiDmeavitaeotifonrsor variance Number Mean StDev suErmMean N-cEonFtOSAdOmd NEFOSET6OmKd 84 0023775500000 0.000530m0s00s 3 0400000 0.000000 000021520500 000000 PFN-OESFiOOSEmddomAa 45 00422105000000 00.010050784435 000044788579 PFDOicSMHeOamnfKldMoanl] N-EIFOSE160m PF0O0S0M0m00k0iMeansNCEoFmOpaSr0Ei0Is2oG0nm0sk0i0 0.0C32O5N0T0 0.02 0000000 0012500 NEFO0S.1A4A50O0m0 NECFONOTSAdOmct NEFOSEdOmkd 000134255 ois 000112255 000010102080 025 ons 00..111225500000 00000000000000 Apha= `Compa0r0i5sfoonrs eachpairusing Studentts PFAbOsS(OiiOnLmSKD PFO"S0I0O0M7K9G0 208t301 NEFOSE0I0G86O2M2K 00C5O0N42T N-EFO0S0M4O7m4k3d3 CN-OENITFOSET60mkd NNEEFFOOSSAEddOOmmd "00085602422 00004477443333 081585175 0000875m7 0013915 0.02343 00002133493145 0.10284 0013315 0.023434 "0.1028 Posiive Vass show pars`oCfommpeaarnissotnhsawiarhesaigcnoinftircolanutsindgifDeruenet,ts Method 27)043 PAFbOsSDdiOYmLkSaD NEFOSEl6Omkd 00C7O6NsT 0.11473 CNONETF"OSMOMKS NEFOSEMmkd 00.0000309568 0ous8 NEFOS0E.A14O5m0K0S0 00..111225500000 00000000000000 N-EIFO0S0E4A7O4m3K3d 00002133493145 0".o1t0o2z844 E 125 ToiIsTLIR Abisoor avtTBeIL, maln feemal Poasiifeirvoenvaluesshow paisof combine data nosg changes. means hat aosigifcanty 7 126 TonAsLLER " A A mis. By Dose Group (gpd \/ 1 A a [N\A 2 wi S VT AN QO Da LS NN OO) I'd ) " A\ V /) con weroi\maroteim | BE Dosen rg NEFOSE16PRDES oon 4 -- Ge Query rd 0 T0711 FR TIS OSnuemwmaayryAonfoFviat RsRqSuquaarreeAd] RMoeoatnMofeaRnesSpqounasree. Error Observations (or Sum Wats) 00550058807772 30735125167344 2 Source OF AnalysSiusmooffVSaqriuainrceos EMrooderl 1+8 1276943421540103 CTotal 2 44373913 Level Means forOnowNauymAbneorva CNO-NEITFOSAOmKY 48 NN--EEFFOOSSEE1d6O0mmikd 24 PFOS40mKd 5 Mea0n8S6g1u0a3r5e F66R3a8t2i 00200010760205 P0r0o0b1o8F 36M1e2a5n0 S0td11E1r5o9r 337205000000 0o212s371E92 433580000 o01t4r1a8 Std Error usMeesaanspoaonldedStedstDiamvaitaeotofnsa.or variance. LeCvOeNlT NEFOSAtOmKd Number8 36M1e2a5n0 03S8D70o6v8 4 37000 0316228 NNE-EFFOOSSEEA1O6m0KmAkd PFOSOmKd 42 3352750000 00125102010302 5 43600 0.288097 SWE0m.M13e6a8n5 00.1155801010 000172500804 DPiFfO:SMAe0amn(k-dMean] PFOSOmAdMeansNC-oEmFpaOrSiAsoOnMsK.I CONT 000000 088000 074750 CNOENITFOSAOmK) NEFOSEdOmid 007646705000 "078500 000080705000 00.0080705000 012500 003750 NEFOSE160mkd "11000 045000 036250 Alphas Compar0i0sosns for eachpair using Studant'ts AbS(DILSD 2.10t091 PFOSAOMkS N-EFOSAOMKI CONT PFNOESFAOSIAmtKOAmKd CONT 002.1451196389 0360464 0215416688 00.33160848684 031858 033156 NENFEFOOSSEE41O6m0kmkdd 003384505119656. 0012344238 001366137584 Positive values show pairs`oCfommepanasritshwtoinrthsesaigconniifrcoalnutsyindgifDeruenntn.ett Method 272dl184 APbrsO(SOOImLkSDd. 025C7O7N3T5 CNOENITFOSAMOmKY 0"0..4432805895 NNEEFFOOSSEE146OOmmKkad "00.4381686589 N.EFOS0E47O8m5k0d0 00.0132755000 003020500000 N-EWO0S3E4d0O1m6k8i 0."03.63845380 0.r24e528 N.EEOSE11.61100m0k0 003465205000 003020500000 N.EFOS0E.1565051m9k5d. 001126412784 ""00s2e:30122 8 TomLR 10P8o)psiDtorisvsevalGuroeups sh(omwipagrsof meansthataosigfanty ar ON A FINN | O O A = 7 XN wd =< ANY T SY ~~ JA { cow \/y Werosiomarotcions i \\__/J a NEFOSEteRROS) DoseGroup (myKjday) Swienrst os Buudiy w= Rsqare SOnuevmamyaanrovFyait ose RRSMoaeouataMnrateaRAnedsSpaounarsseErr S0`3aD6oro1iz0ss5e Onsanations aSumWats) = VSEoneudrac OF" AnaSulomfyoVSfasarSivaianomscsress Me"nSZeee oFRssieo Zaes0 0ino1 owt Clot 2 Tze ames ods Lovner MeorOaneNnomAbrserca esMeiasn Saurreo NNNEEEFFFOOoSSsEEndoGmmoiimaaa i42 GaSs70s0 o0Olr3seossn Prosiomia 5 760 Greer Love 510 ErruseesarpNoaolnmebdSeetrtainoavtaetiooMfnaasaonrvariaSnuceey Son Tesi ones SEraMneoann NENNEFEFOFOoSSsEEn1AaDBmmoiimaas Prosiomia 244 ocemaomomo ooOimsmomees 0oo2dm0e0es0 5 70 030r4 ores PNOfrEeortSOeiaSorA}moimioaan] PFO5S0A0T0I0S0MeansCNoEmFpaOrS0iMs7oO5n0Ks0S0 279000 200000 NEFOSE0GA0oOm7sk0sd 01.sC1aO3r7Ns5To0 NeNCroEouFrsOeStEeOommkd SDaisreoiiono -"D2o5sr7os50o00 S202500072080000 o0o6so28eo0 NEFOSE10a66d50o0m00k00 002530710530080000 2 129 Ajpha= psoonneo raommsasi0.0n5 Nnsere2po1b'aimm apasonevonsetrousmosn ccoonn soMTesoecmmemema.s SSfScEESumREm sE smmmmnnSEedTToamgma smoemmmmny estevos sptSfTrT tr Ey at Iren D i mone e gcoounr iW a s 2Sie TL.reA--r--ie raiorens weevrostoBUoRsomS GSmasw La= a [A2 AANALSNTAN LRVICT AV ROA) I hee ome{Eroscieoroses Stdest owoery / J E2 r_te-- ,na---- ER ggrSooeamvt 2 a S BpEaE Sg0--A --TfSm--amiimmm , mvTeEmsE aoromre oas ess orm wn 4u%e wUmm " 130 TTII0S7L1FR NNE-EIFFOOSSEA1O6M0Kmdkd PNFEOISFOOSmEKAdOMKd 24 2a020s0000 433014303 54 w28r75e0 z304z90 Std Error usMeesaanspoaonldedSetsdtDiemvaitaetoifonrs.orvariance. LCoOveNlT Number8 20M3e5a0n0 Sst7.D8e67v9 NN--EEFIOFSOES1A6O0mmKk)d 42 221510000 4169664980 PFNO-SEF4O0SmEHAAOmka 45 awTresdeo s2mesest SWE 31M.e0a7n3 3S3907010 2T2881260 PDFiOtS=M4o0amnifdMoanl] NCEOFNTOSAOmK NENFEFOOSSEEA1O6m0Kmkdd PFOSAOKSMeansNC-oEmFpaOrSiAsoOnMsi.d CONT NEFOSEAOmkd 250.010000 20501000 2594010000 63278.5705 5642.815000 136.600 279705000 80705000 1500 82500 08.705000 73750 Appa PAFbOsS(DdIoAmLkSdD NCEOFNOTSAOmK) NNEEFFOOSSEET46O0mmkkdd Compar0i0s5o.ns for each pai using Students t 2.10t091 PFO"S8A1O.M05K1S N-EFOSA0M8Om6kd7-189CO5N8T 5108896578 049841177 449047787 2208318107 20065717-1869871237 NEIFOSE2A3O1m1k7d 8592786277 S21082137 Posie values show pairs`oCfommepanasrithsawotintarhseasicgonnitfrioclanutlsyindgifDeruenntn.ett Method 272dl184 PAFbOSS(dOoNmLkSdD 40C5o5n0t7 CN-OENITFOSA4OmK] T83206174111 NNEEIFFOOSSEEfd6OOmmKkdd 2480.2715619 Pdiofseirienvte.values showpairsof means thatar significantly N-EFOSE113660.m6k0d0 18112.550000 703070500 N-EEOSE126003m8K0 180851137 12387.125%2 8 Towoo aptyncp mgs SO TOTO " ya YY f1: hilE SaS KAO AA Ta N = (IZ F >)\, " e\ e Em DoseGroup(mo/giday) os oeaos " 1/32 TriIsLER OSnuemwmaayryAnofoFviat RRsSqquuaarree Ad] ooss0o6s0e7r2 RMoeoatnMoefaRnesSpqounasree Eror 0373512613744 `Observations (or Sum Wots) 2 SMooudrecle ErCorwl OF AnalysSiusomfoViarSiqaancmes MeanSquae Rao 148 1276943421540103 O00o9l9o6s2s5 P6r6ob3osF 2 4473013 0010 000i Level MeansforOnewNauymAbneorva Mean Su Emor NC-OENFTOSAOm 45 3376010200 0oi1s1r1esz NENFEOFOSSEE4lOOmmKKdd PFOSiOmd 42 3a2s5s0o0 o02s23m10 5 aa oats 51d Eorus`eMseaanpsooalneddSetsdtiDmeavtieaotifonesor variance Locvoent NEFOSAOmd Numbe5r se Mean ozsSruoDeeev 4 370000 o3tezzs N-NE-IEFIOFOSSEE1A6O0mmkkdd PROSHOmIA 24 332550000 00.211520103020 5 436000 0288097 SuE0r1M3e6a8n5 00.1155801010 00.0172580804 PFDOiMSeAsOnmiK}dMeanl] ~~ PFOSAOmi MeansNCEoWmpOaSrMisOomnksd 000000 066000 07C4O7N5T0 NCNETF.OSAOmKd N-EIFOSEdOmit 007645705000 "o78500 000080705000 000008070500 D250 00070 NEWFOSE 160m A100 45000 036250 Apha= `Compar0i0s5ons for each pair using Students t Abs(DifLSD 2.10t001 PROS N-EFOSMOmkd CONT PFNO-SE#FOOSmAk4dOmd 002415119638 Cont 0360464 02"1o5d1s6e6e 03-6030t4s6s4e 031658 03158 NENF-EOISFEO1S6Ed0OmmKad 00535450119658 00132403298 001366187548 Paste values showpais`oCfommpeaarnissohnsawairheasicgonntirfoilcaunstindgifDeruenetts Method Avs(Oi2f7L2aS1D84 cont PRNOESF4OOSmMKOmAkS 002.5473783580 CNOENFTOSEdomd Z0o.a4z80885s0 NEFOSEfGOmkd 0.31668 N-EFOSE0A7O8m5k0d0 00.0132755000 00.0302050000 NEFO0S3E40O1k6d5 "0033848380 002.44062880 N-EIFOSE1T.6101m0i0d0 004356020500 00.3020500000 NEIFOS0E5T5605m1k9d5 0D11274248 Dozeas53228 3% 133 TotiIs ER Ptosoirvoe.values showpairofmeanstataresignificant TPBy DoseGroup (mg/Kg: = . OA A NY 0: A EVA ~~ 0 wo \Jy/ CONT Nerosomerossions Dose oup mato |O = FN | 74 { SCY \ \/ Ceo a 08 on Rsquare SOnuevmamyAaorrrorFyait ose RRoeqoautnaMorfeeaARneSdsqpuoanrseeEror 0fO3s6o1rz0s5e0 Ohsanatons (or Sum Wats) = VSotwere OFo AnSuaomfoSViiaarSoiuazsanscme0s Mem"SqFurae oFRsaieo ECrlrow 2 2f35e025y00 000316 P0ra0wo0r CLoovnet Means forOneANnimobevrTa esMeiasn SouEtmoer NNEEFFOoSsEaoomoima 42 B2a0o0 O02l00e4 NP-rEoFsOiSoEmHidamia i5 e76T0 Doirmo Love S14Enoruse es apoNa oaolnmedbdSeterr sdtmaavtisas toifMoaonaonrvariaSndceDov suEmhioan NEcoFnOSAomia NEFoSE1Boma T 3 2 Geseiso0 oodosrs a0 ozmea ooozr ozo NPrEoFSOiSoEmdomia i5 sTmioo a0m3o0n4 ooorasoe PDircoMseioonmliJaoanl] PFO2S0H0OT0K0S0MeansNCEomFpOaSrA0iMs7Oo0Mn0sK0I0 NEFOSE0AOM0KS 1.C4O77N5T0 SNNoeENrFToOsSnEimmkiad 0R87575000000 202300570750500000 2o020r020s%0 00O23o632o765o00 NeFosEtsomka Fadoc0 eso SEE 0sres NEFOSE1164O4m0i0d0 0005537552000%00 30000 uw 13 Tbrsor Alpha= Go0.0m5. porea2s1e0fpr0i0r0ussinogSunest AsPNreoosnsoiLmiSvmDeama PrOo3S4iraoennkssds NEFSOasrSeaeMmsOi NEFOSSDEaiAOsemniG oosCiOeNrT NCNoeEuvrFooscsitemoama O0oasaatssaaireeesz pprSri:am] D2D0as%y oDoz2s Postvs valuesshow asCofmeoanstmetvapirensgSafocanrtsyinaigfareonnte.tnsMiahod AFsrosiib2Ln5id5laDh osicsoensts NSNoEeuvrroosstniioomnaa 0oD3di08ss8 Nevosereoma ose LUmtPGorsotevnBe:evsaleueGsroshowp(amspiofmeans atare sift NEFOSGE"Daooms0akesdse DDDaoimseasss @ A /A2 fa* 2 AA ANNU RJ VI AY TRO) NI\ ES U | CONT WeErOSwFOSEiona SPO adr oRsn E ewdentst WnCoral / ouerrs / que GSriomvmayaArryeovFiia oatazr JrRRoouosknMyeeearAnicSbaaunareErr GgS5oaarasoastsr Cheanatons (or Sum igs) 2 NSoudec OAF`as SuomfoVSraoSiangwceaesmns MeameSeuraiee oFRamio oora 2s CGormiess aammoess moewre SLoouvret Means or OnewNaoyAwreTiaomMeaan SuaEirsoer = TTIISL FR NENFEOFSOSAEMTO6m0KmYd NEFOSEdOmKd 24 2215100000 4330143%3 4 284750 304 PFOS40mkd 5 urs z1zm0 Std Error usMeesaanspoaoneddStedstDiemvaitaetiofonesror variance LCeOvNelT NEIFOSAOmK Number8 2M5ea0n sSrWDeeev StEm3M1e0a7n3 4 a2s0 1984 S971 NENF-OESFOESdEO16m0imdkd PFOSAOmKd 42 2m111000 4s6e.5a6r0 2338.10060 5 wre 2ssert 2820 PDFiOfMSoAaOn(m}K-AMean(] PFOSAOMKS MeansNC-oEmFpOaSriAsMoOnMsK.d 0000 10 5C4O1N0T0 N-EWOSE6d2O8m5k0d CNOENFTOSALOmKd NEIFOSEAOmKd 2554.110000 62850 290000000 209000000 7750 8750 378.77550 0000 NEFOSET60mkd 138.600 411500 82500 73750 Apa `Compar0i0s5ons for each par sing Student's t Abs(DILSD PFOSAOmKG 210t091 N.EFOSALOMKI CONT NEIFOSEAOMK PFNOESF4O0SAmLkOdmKd CONT" "e"801.80871 18988 00881677 1489945781 a94mr saors 2532181677 9727 NNEEFFOOSSEEd1O6m0imdkd 220131807 20058177 918782173 s70283187 Positive values show pairso`CfommepaarnissotnhsatwiatrhesaigcniofnircalntulsyidnigfDeurennntetts Method 272)184 PAFbOSS(4OOLmSKdD 40C5O5N07T CNO-NETFOSAtOmKd T302164711 NN-EEFFOFSOESET46OOmMiktd 4289.7251691. dPiofseitrievnetvalues show pair of means that re significantly N.EEOSE113660.m6i0d0 18112.550000 703070500 N-EIFOSE12800.m3k8d0 180851137 12782.13532 8 rwnim BUNmg/dL ByDoseGroup (mg/Kg/day) a iy A [ I ) AN A / I 1: WVY ANA) PO Ov NIN HoanconwcerloleEeSrcSleESmea |Sh juin oats " ys TT0I7S11 FR Rsquare O`nSeuwmamyaAronyfovFiat RRoSoqtuaMreeaAndSjauareError OMbosaenrvoaftiRoensspo(onrsSeum Was) 00121604082042 30118131785 2% MSooduercle OF Analys'iSsuomfoVfaSriqaunrcees 4" 7457500 Mean1S8q6u4i8e FR1a7t1i0o8 ECroort 129 220871.602050000 11028290784 P0r8ob0o2F LeCvOeNlT Means for OnowNauymbAneorva Mean 8 187500 Sid1iEar7o1r NNEEFFOOSSEM1O6m0mKiAd NEIFOSEAOmKd 43 2160000000 118950088 4 167500 1es08 PFOS40mKd 5 188000 14763 StdErrorus`eMseaanspoaonldedSetsdtDiemvaitaeotifonesror variance CLOovNetT NEFOSAdOmKd Numbe8r 18M7e5a0n0 44S9i6D0e3v StEr1M.e58a5n6 4 160000 244940 12267 NNEEFFOOSSEE1A6O0mmkKd PFOSiOmKd 43 2e070000 22624187735 5 18800 216795 11..51201857 09695 DiNfE=IMFeOaSnE{T-6M0emaird] NEFOSEIEOMeMaknds~ComPpaFrOiSsAonOsM.Kd 0.00000 320000 3.C2O50N0T0 N-EIFOS5E2A5O0mk0d0 PCFOONSTA0mKd NEFOSE4Omid 33225000000 525000 000050000000 00..0005000000 205000 200000 220005000000 000000 NEFOSAOmKA 600000 280000 275000 075000 Apha= (Comparisons0fo0r5each pair sing Students t Abs(DILSD 200t301 NEFOSEISOMK PFOSIOMKS CONT N-EFOSE4Omkd PFNO-ESFAO0SmEK1d60mkd CONT 6B6S43S9 148346898810 31.8432378600 42160 38880 3dsdsa 20.5002478084 223104 N-NEEFIOFSOESdMOOmmiKcdt 000722720046 s2e5ss 212438110044 -Z4a'3sasss5a0 Posiive values showpairs`oCfommpeaarnissotnhsatwiatrhesaigcniofnircoalntulsyidnigfeDruennnte.tt's Method 270dl438 AbNsEF(OOSLEfS6Domkd 27C9o3n0t2 PFCOOSNATOmId 45.4060397434 NNEEIIFFOOSSEAdOommKiAd 32.7416668833 N-EIFOS6A.I0O0m0K0d0 227850000000 000705000000 N-EIFOS0A7WO2m2K9S "118.348418088 4"1e3s8s800 91 r - m A URSortonve,aecsrosphow pals of monstat gn rsa =.= TN sm= " V/A [& O--) + cont F NeroiY_saA rolum\a/||O TTT NVG )J DoseGenimi. Oo0s 00s aA g uyrSemgese o[S ERA Eamer Sorren evrieg) YSoamr7 Esowwee oorOgF -- ENSgNmaulImmeeSmds saMTmenses arcodomme Toa BEEZ me we woissoearroanTemwmmw sEntmen NNEperorsduaiseomme, {{j oommwmes ameeems iLoow Era[n sRNpdrieirtiernoe ataSreosm a suErien NNN[ Eeorrnocgeme-- aaenes, {PD$ooommmme pnnaemmwe um mpaamms a D Rhoweiven Roscioma neroP smabnsCoT mpasrViEsroTns NErosfstiiioomnnd Bm A tid NEFOSEaBNIsnEema hEerroes, FeLa be =2 Aoe prromnosve=eems 2 TorAsLLER Aphas Comp0f0o5rasacrh|paii ussingoSundss sCNNEFTiO,SLAsODmka NEFOSSMOimakmS 22001C0O31N%T 06 anaes NEFOSPEaArgmiidd NEFOSEIZsSoeiomntiisd |PReOaaSriOe1mn0kt8d NNePErFFoosOsiSoEEmdi8ma0ima isSeassm: 3m4i7os2oe%ss S1Si4a1a0r8 ESBrsaree tai desir Postevaluesshowpars CofmeoansmthawtipiarhoaasicgorirnfvcaunsitiydngfsfDruonnotasnMotshod N5EF0O27SD0IdM4i3O8m 35c0o7n21t CNNOEENFFTOOSSEEAInGKmSG S31319255221010 Prosiom 450 ASTUPNoGBisyrDioones,veaGlruoeussphow(pmaasg ofmeanstat ro signify ps 0 oo 2 wo = rIoK, Ea N Aa L = VES TV] 2a =2 | = ) Cor aroluoVrmerotems | BE NEIFOSE16OMROSS 0080Group (moKoidey). ox fSeoi" ary on Rquare OSneuvmamyAaonfrovFyia oseo10 RoReoSaqtnuMroeafaARnegSsapuonasres Err arsTas35ri7ys5 Observations orSum Wigs) Wn MSEoneaurre OFs AnaSulomfyZoVisafroSia7ansscusoss MemeSqiuwse sFiRasiio Sasi Tse poor Clas = Stazesso mis oor Lovner Meansfor OnowNoymAbreoTrra oMmean Siiomeo 10 Ki E feomoy ews iP4R m8Ea8Em s3=em [EaomL -- LAarT LLYYoemt l PThHy weCURE eRrEcoRosEinn, Pt{ooIgmmmm ooemdse aBam oHMdweoetirnioeans he A mGcoeonu eV"sEScSEomrsSepwEns ercnsnieClroHvonsEsiaRo rrors3Tooasm5giasn Em dm om fm Te. by BCEoRsortaEeses fm a Ssoroom Ec3omr0r neacwte,rrocerdam em nerosdcweirommstaams rosTperieoenn ieoss =32 Iumm dTmE mar Ae -- s get-- admm nMm2 eoiiu1nd n "28 IH riee . =iE siispr sestr--siy ALT UL.ByDoseGroup (mg/Kg/day) erosions pi1i2aa2n nesosc8iieo8hms JAE A rJama 7 Ea . A.A | :Aa VYa-&Nye conor CONT NEFOSMOMMLEFOSEANmig NEFOSE160nB0SH =o S00POF Rr bd Bsoqm afrenme rmos SE es oyweyn LFLr ums son2e Tsfpoohomree o2rOfrVsETSaesmdEmmya cMoenwoeum eerromme OB Bum gm er to3t---rs orrTmEs> LIwnEy sipi E RRnsn H PgT mA R boo s Err r itE E ay ttRnsIrrd FF eHrreAEsi o--n i T{{ooggmeEeeR a pEawRsr Cnreems sue fbpoiarrers ,i veroseioE da imn: TCUHEE R aconw nerofscinieroemad ia am um 2as 8e3rorsiotns = pum Ezem = oom fade po Com00scpn"sotsnotnnt neroifnsoinines Cfeid 0 Twwrim oippvoaeirseesAnia eroStesen -m--Twasosmh hx ume aogcmooneww werossosiouoaemngs Fl PBosote anhwSsmrsogrEsfuartmytongm mm fnolhn=ys pas s3c8own0r BAEErgsmemngs 3ouEr w 100mg Pfateoheeevncspowsrct areresty . HooA a, A A LO [CH) L) Pe gm VY A = |) AH7 S N fe" TaATEVL A) cor relp i ere 7G Ssqrm cmrommgsoos DoseGroup (mg/Kg/day) os le ofsrs TEfEoiSLeApes r fooee2 [ e Fg B Ia A mmCmeRme WmEpR oe PpTigA a, TE1 P T oE 2wfy uai"rIEZ Sr Pi BE w 143 nevosSsoFoienees] HsEe Tborais ni prosionks 5 aem Ero s[s vldetirmatei vrnce ame tSRotorssiomis NNEErvoossctiaommies NieiTooom"nVnon FRc mRSAueamoeey sarSpeeeien pal Frosionts H oBEnS wes aSo OFfarNueant ean Nerosomd 5Sc%oor Wa"aWnEsFCGoSmEpSiamrsson.s dos ma hid NeFoaspmrMkem TrPorcgos]i HfoscioneNs E Lm psiedm aSSReoenw a soaemm Appar o00ms por saa:choinnScensts ANooeefsoos5ei0oma BNrEosrioonssosmd B25CiO5oNT NEFOS2rEoisaounm:a asl 1 Soe NEFOieSkMnOem Gen ProOsaihmeimd oom Nerostioma hJrai BSoA STemw Poi ausshow areof moana ht ar scantyaifrn: snaamm Coimop n rBoat,Mr et N5i0vos2bee1isdkoma = Secaorr BNNEerrvoooosskmtiisoammaa ovaumoe CSombiened asItaAsLoKrPmAesSanTmd TeHrOeG40omnkT FOS aT nd 180 mks--EFO-- SE r-- s, -- nEros5tSar5ao%sn apSrsoesoc NerosEShrsioemnss a h1e5m% " 144 top tan - ALE LIA ANNI WNYC) - 0 [ly con er aot J ye " fe rSEoee,mgi.ees tm L= ssooom FBaro 50A worEatTHEveiEoTmnOBeRssEn uaenee gipttroeeEmrnpel rA "111TaEBBmRE,LATuEEEe tBtoortowSns aTm1Ty raMcaiPEhmaErtEgEooEmnEy Ee sTwUepes oiperms, CEDHREE EDEeRERdSnR IeiRSErammtEenme:.et roZgAsS mmsEpIEiS EEgoEEEmm 28 3B o4TcAonEBn OBE veerrosseiEoGSnRERs -- A ow & oMm fEzrmmesEe.w LTAaTaEmmaE 2k rem3ewm EZ aEEommmn EB 8% a3ewemwere I am 2m wpolpilsan atnu mA =e, 2bi eveeenuiroesismnosns fe v>erso agmm " TLi m w LoPnSBosoytee essho sofas coup mrt at ar scm Z--z >- = 3BS meol 2 ON wiNe" AA AA LAN 6) AT \ = HI AN VE) on Weshshreatose Row Cd_7F |// DoseGroupV(moKiJciaryo)sercorasis 0r05y 7Dauvsis by Raospr OSnorvaeroeo ay RL eee oosmeeesr LmnT Sore to) Za7 T i Esoem EAraUsrRvAmmaeErmleaimcSeeenm rgm eemugsa:eee eerearsreo tLaowosuVasa Nerossome, OrvWsAe{Tfoeumw sewSeeeg NMEorroosnsism PPPoooemgme mmmeee F[ ir" -- 40Erorseas apNnidr7aamtlooiifermev sSoaDe% sutSreeminn NhNeeErrrooessiomsts, PSfFI omommmmmeeEm mmowse ge ENGSNeeivrieon,s NERSSCETS, fecuSoeenotMeErnosasCEooormpr crosSeifoowanmeedsd neostemoroemns NERS mgeear mmeew rSio aaor maro[Hoao2ssn aHioe wo | ww aoe BOHO ie EmgI se.n Pes es x w 2 ros m,an con mond eros nescstors 8GoG%hm @BomEEe BEEpEn S tm tt erm ty to 2mBRmR e-e.rw...1d, EuEn IrJMtetesesornso tdmoentomy Pwl E2 S< [l= AA LOY VE AUT 2 " mo O) pr Te "EachPar WahContra eed = NEFOSEiomARss Shdenfst Dunvarty. E Bor=efa Rg-- rA Eaw I nnof0oSorA mtee eeuornermnmTenaJss=m.msiseis Level Number Mean Std Error 14% nesomon 2 2gmS | || | TonsLR NNEENFEFOIFSoOESSiEAoBomonka PrOSdamia 42Dolumesmsw ooteammns 5 men oan cLoowur St Bor usWeesarpsNlauenmddbSestrtaDaetveiotMfneesarnr variaSn4ceDev ues ren SuiE5r5Me7n NNNEEEIFFFoOOSSSEEAtioeomamkmeas PrOSdomid i2 sulsetsmeo JoSsesiseesm ir25e0e0r0 5 Tran ress dere: CONoEfuFMrOoSwAlLdOemGl] Neosesonks NEFOSaMMoOeMannGs CampraCrcOsaNsnTs 00 domo NEFOSOTETmskEd PFOsSiIermasinsd PrNOESFIOoSmEkra mk S9S70a800o0 fSaretrraea 3S07000000000 Sgroineoenao Apha Comp0f0osaeacrh\pair ssingoStundensts coN5Euo0rS0a1d5o0nk Neoseionia NEFOZSMoOoT 2o1g0C0e0Oi1NssT Dams 20m NEFO2TSauEiAasM PRoOoSsHsmesKY PNrEOFSodSoEmriBaomi ToTuonssts aGosassmeess 2GGo0ee1s SsSaaogaawns Postve valuesshowpalsCofommepaansrithsaotwnaisrnsgGfnckoaalntsyonigfGourmtntts Method ANvEsF0iO2rS7LA55O010ns 16c3o2n5t7 PrCNOoEnSFTIOaSmEaOna Sp31ar3ie53td7 NevosEioona oa Ke molPlop.srBiovoBeeoysvealGureosuspho(wpmaasdoef)meansthatarsigfcanty NEFOSE35T58S07O00M00G0 S237700300000000 NEFOSEaTfg5Ga5Oe5m02d80 2L0o23a00s5i20s0 0 149 Torsor v . = ~~ A SN i ie BA ON LN CY . v iV OANl | a- | re ounroheirtosreemens Sons Q=) h TCd) KH OTveres on rsqure OSriemvmaayrAyonrcFaa ReauaroAd Ooarssaaorssss ROboesaternvaetoRinoonsSspqoounrrsoSguEmrWras) 0Sseinao2ss HSoooommse OF" AnSiaomfoGviaSoranssmcess MemaSdawoe hFiamo 20 orien Pane Coa 2 ame dworm eer [Lorouvrre-- Mears or OnewNaoymAbreoTDroraeiamMmemaon SoourErheomr NPNrEEoFFsOOiSSoEEmrgsaomnsa L25 odSaieonw Goomseisn Loouvre S14Err useesa2npNo3ul0me0bdeSsr1t3iDmaetvoiMfonearnovariaSnucebev eso Toons StdErhoireaen NNNEEEFFFOOoSSsEEAnIoongmwasa Prosi i32 iSSshmooeewo oovusnns 5 Shoo cea 0cTai2rc0to [4 OcfNotEeRuaonsteysHaenakrt) NEroscona 00CO0N0T0 MeNaEnsFCOoSmpTaI1rG3isO0o0mn0As0 T3000 000000 NEFOoS3SE0o0Ao0Ko0 NEFOSG2MoOarmikmnd NPErFootsohnion p3sr5oio%]o 2203900700000000 2060r0e000s00 2oo0oo0oo% Apres [i00s ---- | 103 150 PF3sOoSasAnmnadn Girorosaioen T5amm sHMmBerEEoeSEmwSRs BaGMciEEomgRn CEI neso2l1MSmlmOiuLmmfEehS oe weroaagsvsemmmurepse nneresagrumom am roaTorsoinrdsna ERES IITEIE AesA swhieomo2smIde|es J 2o1c5oa0r2 iRamee aoian Sa -- OFmma ByDose Group (ngfotey) oonl 108 ps LA soil AA aI N LAN AL 7 NA NA A AZN l ~~ iva ()(>) / - J T i= Vt Roireme gsomenr ne = asoaea S gmBre E o Obseravatoionus Es(orStVugmvgSWEtaaetmissRm)g 2 wGaveemmm onooeoee thi aHorosensiEorsreyRrAH$Ir oomemoawm vere os TbrAiLLER PROSADMKA 5 10200 tomo Level S10 Ero usMeesaanspoaonldedSea Dsovitaoifiornsomravartianece Number Mean SWDev cor 10062 2ceerr SidEr0m3h4e3a7n NKEEtIFFOOSSAEoommikaa PNFEOISFAOOSmE4KOdmka 24 4 180450000 101250 orodoooso 220% 000507040 13150 5 02000 send o7o7t ONiEcFMOeSsEnIMGeOamnKlS] NEFOSAIOmKd NEFOSEIMEeaOnMs CompaNriEsFoOnSsMOmkd 000000 050000 PROS2I0O0m0k0d0 050000 000000 150000 NEFOSE2d7O5m0k0d PFNEOFSOASOEAmOdnkd Cont 220705000000 Sars00 S22050000 Zams00 00705000000 arse0 207255000000 00600205000 Apha= Compa0for0resiacshpaornusisng Students ANbEsI(F0OSLES1DB0mkd NEFOSEIOmKS 2.10?001 ~ NEFOSMOM "473000 Seis PFOSI0O6N8KsS NEFOSaEMsOmikd PNFEOFSOASOAmOdmka NC-OENFTOSEOmkd "1ST3es5oe4is0s76ee G3aeseiess Totes Zoraorsesd zee Zarzoetss 33st Sanz Sores Aznar 2218s Positive values show pairsCofommeapnsatnhtiareasiagnifcclan1t2y0dgifCereuntt. s Motd Abs(DIY2L7S21D|84 cont NNEEFFOOSSEAIOGmOKmIS PROSAOM 01.4878858502 212506 CNoEnFtOSEAOmK S51o3r5o5s5 `CREA mg/aPdoLfsiBriyv,eDovsaleuGesrosuhpow(mpoa/ikslodfemye)ans that re significant 33C7O5N0T0 2137780000 000602080000 07CO2N2T 00sar7aens 22527107060 108 1S2 TosLR ; MA fe 2h A L8I a<Pn N=NV VVAr vW (E=) LID) 5\T /17/ <n Cot -- warotenrmoesriootanrss w|o B os TTse ars Rquare OSnoemvamyrAyrooFrlia RSauareAd oGasgsaans NGRehoiooatnrHieatsRoenSssqpuooarnrSsesuEmrWroe) QC0o7r0sae0=tnrs oSErdceso OF4 AnSuamoTfovfaiiSrimahasnrcsses Mea"mCSoquiree Rsaetsse Bm oso coosm Penk Clam 2 ome cose aoe LSoovuert MeorOanewNraoymAbrseorTra osoMaoamn SGuorses NNEENFFEOOrSSoEEs1AGsooommnisaa ProSiome 12 OoChssooom ocoooossosres 5 Como cos coLouvert S14Eror use es pNa ooalnmedbdSesirr tdmDaotvs itooMfreermovariSandcbeev soso oorssm SuEcmoiessen NNNEEeFrrOooSssEaE1sigooommnikaaa Prosdomia 2 ooosseemm Oooooomros cbooossses 5 camo ose Oommes NEPOFfrOsoSMsoAtmdaiotlnaeiaral] PR2O0S0i0O0n0iMeansNCoEmFpOa0rS0sA5aD5r0Ms0.G0 "05s 5000000 NEFO2S0EI17S35O00m00k00d NEFOSTG0E07A0emD0k00d0 00118C20O60N00T00 SNEoNFuEIorFsOSEEiteoima 0ai81s "22D05o1r 000000F000r0000 00000000000000500 O0G.oo0os5oa0on0oa0s0p Agha Co00ms pfoaasc |prarisinogStundensts 106 1S3 Theamm MjfsrogeelseDr NNEERrSSSiEEiiioownss poSsoi wef2d0foSk0hviama one pi weros( SSEnooamari NeroSCse touemua cour Seowm ae aSmaemt pori SSimm Postevansshow sofmaar tar spar torr, She ssemm Camoea lSn1o ors Nfieejers1ydmo pocoroumer NNoEEervGossEtiimaam S"oGoioes oo Cuint chn taaitorersbyt0otmrlPEsEOSSa iGPi irSpeoS.oEceop srtshiowLpaOSi1R5oEf.mo Sohnoswsrsato rerctsa ees no ms POR a 9 ne(CE wr 154 spian B. Statistics on Clinical Chemistries for `Guinea Pigs, male and female values `combined. = A -- in" VAN E"LA\M/ A AL[--] LNA~U STEN 2 neon bE SL CONT = N-EwOsasNoEnRyOdSEePOrmoEsi weSar =p Stutdhontst--" Sa ~ DJuhneas. 7 os oo 155 ToAi7sLLER OSnuomwmayaAronyfovFial RsRqSuauaarree Adj Root Maan Square Eror 0ol360i3s7i8z5 23.365 OMbesaenrovfaRdeosnspo(norsSeum Wots) waaTro SVoouder!s OF3 AnalysSiusomfo3iV8Saq7riu3aa1nrc0ee2s Mean1S1q9u1a0r3e 2F1R6a1i7o ETnoow 15 s7i661a.s94407 eSs5a5s2 porovxor Lecvoenlt Moan for OnewNauymAbneorva Mean 008m ud9Ersor PrNNoEESFF4OOoSmSAkEOdOmmkdd 445 ssTirosen rd104iecee Level 14 ErusNeosaonpsoaornldedSied Dsevitsoioifnasrmoravartianece Nomber Mean Sev cont 5 amen mers Sidr 5M.e1a5n PNNrEEOFSIOAFSOOEmSKAAdOOmmccdl 44 5 sTuisw700 0427073007 1181281775 9260 DNiEMFoOaSnAADMMoAaGn] PrOSAOmd N-EMFeaOnSsA0C0do0mO0pA0ari|sonFsFO1SA6O8m5K0d 16.2500 00000 31C6O0N8T 143167 N.EIFOS3E7d.o3m7k5d0 205280 CNOENFTOSE4Omd SSureasms 2as5te%r 407000080 057000803 Aha Comparisons or each0p0asirusing Students NAEsFsOSiAsOma NEFOSM2O13N!144 PFOSAIMKd 52147 A6sSTe 0C4O7N9T0 NEFOSEd20.m10k0d3 PCrOONST4Omd NEFOSEdDmd qeaSrTsEe 21603 5ta3g Czas a-ivsseeord zssses 2162.8403226 Sear Positive valuess`hCoowmppaariirssoonfsmwiehan tchoantralroussiinggnifBiucnanntolrytdsifMeeretnet.d Abe(Di26L2oSl0D08 cor CONPrNEOTFSOAOSMAKddOm 27203878878 Ssdeas NEFOSEmKA 33.6021 Chests, Malo an femaeguinea pig aPiofsoirteinvte. valuesshowpair combed. of means tha are signicanty 109 156 ToToisn Im 157 se PA wo 1ASAP | we r Tem TF B obdmar RnseyS--om--me RE re ceoaspn:s Re em t Sourswo weis sfg oamre d00 raEItGSmNEissEEwm nWoeemnea orrodee E-Frr W-oo--rrTrIT{rnpSaaEmn Aeros {opm sat aomwa Aeron {Bue 116 frepsiiviuxrirthinrrr sw 2ESr7esrsopsne He RaDTDooommme omcabmmme suoSemunesmre Does ome cue ALDeeoenceiel oe EP FSiToW-- wo erosCoioholnnm 02 Woe FromTmsrioeosmmomansa |oscdmcoooenn Apres 2 Sp Comorpnorias srSetsr comm sme AeAwrraioEDsnsso werosTeiobominyTM"m wesosesioonmsa me fmm proesosseiommwae accsoonmwt = SWWaEv Pose aes ho ofmrs vot mgenm cary attr msawm oonme LSas ster st, rRKiarcSoihsdt?lu.me La-no_e M=orionama 3 aone ostih aka showsota rssty PHOSmg/dL.ByDoseGroup (mg/Kg/day) Cs? roma - x H 85A N /N A VA EX E.v YY . V PN h| | i CONT N-EIFOSA4OmLS Prosiy |Seeh Par SoCo N-EFOSE4OmKI. Er _-o sgT oommioesn DoseGroup (myKgiday) Suter 00s Dues 00s ocessr Eiict BE J Ti: BC BrEEG4 TEiEmR hwaemn drma oooe rerTLeE3 erny ssuge ai. a iHEgSeenare s1ProNaEmR i20 TneeSte steerren fFfraeoewn:PCHoaEmEE IooEERmE io"Uem iS Saeoveeen A Toi= HhhEaeu geonoGwmnm UGMeHmr5oRan oS"sroaosmomuns -- coteepoBoreg s a NPR Mere en mn neon rose TrIi0s7L1 FR NcEoFntOSAOmt PFOSdOmia o0ami1s008s 4408 v1.i1z96i0o8 072108 1192610088 079608 007792810088 1.19608 Pusiive values sChomopwapraiissoonsfmwietahnasctohnattroalreussiinggniDfucmannettytdsifMeertenhto.d 266ul204 AbNsE(FOOiSfELdSODmkY CONT 1.1c0o7n8t2 tare PFNOESF4OOSMAKOMK "10390778822 he two analyses directly Above ar or guaiPofnsfeieatriepvnietgvCaaluaensdsPhHoOwSp,aimraolfemaenadnfsemtahlteacroembsiingendi.ficNaont sgchangesoccured ne 16/ wrpm ? "S>IA eS Cre oO ~~ A I:5 SS < <>< 3 pI rEis - =EachPar F=ic "WithControl us TIL FR Is `OnSeuwamymAoanforFviyat RRsSqquuaarreeAd) Root Mean Square Error o0s3a0s0e7s678 1.140108 MObesaenrvoaftiRoensspo(nosreS.um Wats) 88235279 SMooudrecle OF AnalysSiusmooffVSaqruiaanrcees 3 21359250 MeanTSqiuearse FsaRraios ErTootal 138 126380459800509 1144209587 P<r0o0b0o1F LCoOvNelT Means forOnewNauymAbnecrva Mean 4 12800 S0t5d7E0r0ro5r NN--EEFFOOSSEAAMOOmmkKdd PFOSA0mKd 44 162260000 00877000055 5 4bs0 050087 Lovel 1dError usMeesaanpsoaonldedSetsdtDiemvaitaetoiofnse.rorvariance: Number Mean SDev SErMean CNOENFTOSAOmKd NEFOSEAOmKd 44 1G28e00o0 o10p1e6s8z3 4 122000 096264 003540786217 odstsz PFOS40mKd 5 4800 1sde 069166 DCiOfeNMTeanf}Moanf] NEIFOSEdOmKd CMOeNaTns Com~pNa-rEiFsOoSnsEdOmkd 000000 0.60000 NEIFOS6A1O5M0K0A0 PFO8S.A1O2m0K0a0 PNF-OESFiO0SmAKOdmKd 06105000000 8.12000 505050000000 52000 05.5050000000 17.9572000000 15700 0.00000 Alphas `Comparisonsf0o0r5each pair using Studentts AbCSON(TDiLSD 2.16t037 1.7C4O1N6T4 "NEFOSEdOMK NEFOSAOmd PFOSAOmKG NNEEIFFOOSSAEOdmmKdY PFOSA0mKd 14.410418834 646773 7ER4160 538860787336 34.840088336 64g6e7r7r3y 174164 031773 03s assy Posiive values s`Choomwpapraiirssonosf mwietahnasctonhtaroalruessiinggniDfuicnanntetdtifMeeretnhto.d 264i5d45 AbCSO(NDTILSD. NEFOSEdOMG 2.c1o3n6t% 153593 PFNO-ESFiOOSmAiddOmi 64.0010430678 bN-aerteFOisSfAoalg4u0inmeoaKpgiigccaoym.bined male andPdiofsfeierteimnvate.pvoatlauessisuhmolwevpealsi.rsSogfrmeodauncesdthbaytPaFeOsSigainfdcantly 6 Tfiaim SN | w Tg. yous cup mn Coen on - Ba=d E AANSAA E [ AT LNNTN NN hy VV oe RomAniF r=ow Par Win 161 wr | T1IsOTLIER OSnuemwmaayrAynoofvFiat RsRqSuquiarreo Ady Root Mean Square Eror 00.0152988419 0.106758 MObasnarovfatRoensspo(norseSum Wats) 0.17108130 MSooduracle Eror OF3 AnalSysuismoo0ff0V1aS0rqi6ua2an0rc6ee1s Mea0n0S0q3u5a4r0e FR0a3t1i0o 5 0.17085833 001137 Cot 018157895 0010088 P0r8o1w7osF cLoonvet! Means or OnowNauymbAenrova Mean 6 015833 NEFOSAOms OSuoEmmosr PFNOEFSOASOEmddomk 44 5 001156005000 0210000 00005s%: ooerrd LeCvOeNlT StdErmorusMeesaanpsaacneddSetsdtiDemvaitaetoonfse.ror variance Nomber Mean SiDev StdErMean NEN-FEOFSOASEddOOmmKkda PFOS4Omd 45 4 0011550803030 0162500 000057973s8 0110888 000023878475 005843 5 0210000 01darms 006403 PFDOiSMieOanmioanl] N-EFOSEdOmct PFOMSeIaOnmsidComparNiEsFonOsSEAOmKd 0000000 007500 0.0C5O16N8T7 N-EFOS00A6W0O0m0K0 0.0475 0000000 0004167 CN-OENFTOSAOmKS <.0051867 00.000142157 000.000008030 00000182353030 0000000 Alphas Comparisons fo0r0e5achpai using Studentts PAbFsO(SOiinmLSaD 213t144 PFO0S1I4O3M0K1G NEFOS0E1O0m5k1d4 00C8O6N1T2 N.EFOSMOmkd NECFOONTSE4Omics NEFOSAOmKY "2o.o1s0e511z4 "000286 0.t1a60n9 0.1484 01g40s7r1 013858 0.00.61246844 00..113680595 Positve valuess`hCoomwppaariissonosfmweiahnasctohnattroaleussiignngiDfiucaenttdiMfeertehnatd Abs(Oi2n6L2dS8lD08 cont NPEFOFSO4SOEmdkoami CONT. 0.1167278 016208 NEFOSMOmkd 0.17288 Alb middPioBsfyietriDevoe,svealGureosusph(omwgpoaiidsaoyf)means that are signifcanty ns 16S Toros on NZ 7 a A a 2 AA Al - ANN CNE YWW A~Z N\ T | 22 NV v \ Y wl No " 1 cobn oeprroiersgoseirmeo.m| "ESaachhiPaur ow7res "WahConval square -- So Rare Ag oSaosseys jRChorsranatoonSs oorn SEumrWrt) OSSiiroen%na joStoeweyre CT oBro3 AnEofmaVTGaaronsecmmos ehompisem oFhorewue ees opmm Por LNoovnoss[ a --No-- onToadmwmeean SoeunEremesr PNrEroossEeitme HPIR SLRonFvossSiuarEiraor uNsesonpNloamenoandr5T'ssbaiotmvo"iNfeesrenrvaorSlieeesnw sEreesmn e NEroa sEiom 8i DoBghdo coowapmemm ooLeiesss fPDrrcocitotont] Nrostiamia MNEoFaOnSSoMCooOomMnpaoEris|onPs RoomOenSes 06s om NeFOSCpEairsGomendrcoccsloneet pase Aphae oblrs osm 00s GToeosnss Sboonma fAvrsoienLesD [S 21\ d R ---- NEFOSMim Prosimis NEFoSEioms cour So oa Cams Soa TM Tbison PNroonsoi ma oGTomoeoesr Soimeemr r oDxiemne a oaesme Poi vues sC howpisofmeansthta ai r siannkfren Jr 2oeo5v1dyk| Aos cont oFNoreorsioosniama SDoreSaams = 1P05yiGbroonsenGrvouepssh(onwprpisofmaar ht-- ar sian (a NINAVA NN CN TN2 = [ANA ul N2S V&y / a I~ 7 NX NeNo 7 " pr od oT a tot wa Ounnars DoseGroup (moKgiday) 00s oo Risque -- ftori] Rare Ag o0i34s65 OHRbesoaanmaoarinRoonSsgaoorrnSssurmoirg) So5s0s%8 soHooeamre Coa oAFSTva noofVtarec:o imum weomMsoweamns gFGrieamees 8 sae gum Loovn [---- er NoonSamvsan SSuEreer NPENrreotosrsotsiaoiomma 5DiiiSmamem ooampemen LoSvert 54 roruseanNoalnmoged5nbmotvio"Vfsaoanor variSauncbeer as odiom suEiSrUeEaenn 120 Twoim hjAearro--osn -- PD oOegmEeo Soommmaem ooommmee LReSi ereooslitn 5 NMeSsSiCmETaornsoatnosSanawmys sus CGH erosuMLtEomOaememese oo| csmcooeewn ar odie ofan CanbpraayorsstngoSntss om caer RfDAren0ro8s0 erTorEs"s proTsimmia Sams mA nerosanserimoosoennse occouonwn uF eSamm humm Posteaka sm howarsofmeattaan rnSstgornyattrre,s, mdmr oowmeed HNuacPilsonSdm1dae. rsI 1 pooceTotys Fsoutehne aka showpasEom+estrssry = NONRT MHiSsAVAA VAALVANA :" \v4 " \/ Vv \v/ i7 N f/ * EONTocerwerecEtEaT Prosis | aEsnhta hoaun "WilhControl at 168 TDI0S7.0LLFR `OSnuemwmaayryAonfoFviat RSRqSuquaarree Ad) Root Mean Square Error 00..01178748827 2382882 `MOebasenrovfatRieosnspo(onrsSeum Was) 145263129 SMooudrecle Ero OF Analys`iSsoufmVoafrSiqaunrcees 153 1.603509 Mean0S5q3u4a5r0e F0R0at3i2o CTotal i 8865713363834323 458617857516 P0r9o6b2o1F LeCvoenlt Means for OnewNauymAbneorva Mean 6 148667 StodgErr2osr NENW-FEOFSOES4AOOmmkKdd PFOS4Omid 445 1144050000 148000 t11i9e1n2 10854 LeCvNeTl Std Emor usMeesanaspoaonldedStedstDiemvaiatteioonfsr. or variance. Number Mean StDev Std ErrMean NN--EEFIOFSOASEd4OOmmKkdd PFOS40mkd 464 1t4a5s0e0r0 140000 213281014088 316228 d1.s19s02 15811 5 14800 277480 12410 DPiFtO=MSe4a0nm[k}dMean] CoNT PFOSMAeOamnKsdComparis"oCnOsNT 000.0103030330 0.133333 N-EFO0S3A0O0M0K0I0 N-EIFO0S.E840O0m0k0d0 NEN-FEOFSOASdEdOOmmIkdd 03 00..01060606070 08 086667 00.60600606070 05 005.60605060607 0.000000 Apha= `Comparisonsfor0e0a5chpair using Students APbFsO(SD4iOLmSkdD PFOSAOMKd 213t 14CONT 321150 284144 NEFOSMOmkd N.EFOSEd0mkd CNoENFTOSAdOmId NEIFOSE4Omkd $210046114 260831 2331116085 261105 23.1110180351 359056 226610180351 3.0906 Positivevalues sChoomwpapariirssoonfsmweiathnasctohnattroaleussiignngifDiucnannteltyt'isfeMreetnhto.2d.09056 359086 26d2l85 APbFSO(SD4IOfImLKSaD CONT 26C5o9n19t 361602 NNEEFFOOSSAEMdOOMMKKSG 33877661177 dPiofseirteinvte.valuesshowpairsofmeans tht are significantly `GLU mg/dL By Dose Group (mg/day) 122 | rpodman =w - - A A AA F $= NmVLNATNNN }| " - NN./Y \/ o J I/ eeos TTroen EE "0 DoseGroup (mp/Kpiday) "Each Par 005 "With Control 00s fESHoioRgioe ,OSmneewiaeoyeAinoenva mp_ oieee oBar TreRRresnE es L 0p Sediment Eg ame aR orma Fw wnns 8 mm wereUnNt dl et ww um of i SI nrti iPt EBgE=f OESBEE a- rS imsepsSorsr eTtrrioalme tyorsmoSnoEyw SEs stesanlne Sel {oommmomowowmes agnw REpesnoessteiet Jes eos2 dTScmoerprrersroosgsioman srowTsEoaogomEssa scpcaooeuwur arn Comao;sps irySatEge oe GEE fpooins, erToarnsste werosNeuroovmemse srowossuummss ccoonn = 170 oTmisao oPJ roog qi asan GaEwseese memesene mRmeueeer Paste values hCowpoimofmwooiannssoamatatroosiigcBaoonmty ndirmsey AsNNEEORFOo2SsdLEdaiSdloooDsmnaa a1y1coe7nrst Poronsi Spoare]s = Auk utP'JoBsyBoieovGarosupsho(wgpatrsao)fmoans htassifcanty - 371 - A RE NalNd Fag: AAA NZA AL NK A i - con WVRAIcTrrsNoy Em -- DoseGroup (moKiday) 00s. ; Voge 08 Rare S-- imo Rare Al ooseos RWoesarasRessuonneesEror soSaivtegs THsoooamrre ObserovaF5tAionsrG(oraoSfuSfvmboaWranvi.tcsio)r 0 sews 19 Moiemrwae Faso Clon 18 amew meh Peer LcNooeveFntosao[mis----Norb-- erbT oeMmyean SehuErmeor PNrEorsoistiomd 1somkm mgemn oLevra 54EroruosasnNpcomkmgeed5nbsiomnoteor"NfsaranorvariSncee v ie somm ta SUeEaYn 1 Tor0s71ER FNNrEEoFFsOoiSSoAtmdiaOoma sP5 oOisrEon mTroaerensn 38s13s% coDPuRiOreSsHOm ean) McCeOoaNnoTs ComPpRarOi3Ss2io6On0ms6d7 sew Go NEFOsS23A102O13T607Y0 NEFOS2SEs7adno0edr NNEEFrOOSSEAdOOnm SBeaewr amgee 05000000 a50o00 Aphas CompaoorsaaacrtaiirussingoScnentss EArceoossniinoinso aCiONsT 2`1P3R0OasSMnITeAS Lary sae NEFpSOnSriMrOinI NEFOSGEEaoanirkad NNeEvFoOsStEiioOmmaas osrneess ssaidsrse Speeei paeaaeat PosievaluessChowpois omf mweianpnsctohaantrlrve ssingfDcouannaynsdifMerseentt.s fcoens2ds1id|se scoonot PrNNOEFSeOaSmtriiaoomd 74p62350d888 ASTUnPGoBysoDtovesevaGlrueosupshow(pmespoifmeanstatare sifcanty KK 3 p. = 3in \ AA wo ] E N A L e ANVA AZN o c<o>r r almV Troe DoseGa (rR) as ON Vi Nalh oo ns TorsTL Rqure GSreovmayaonv:a Rqvare Ad oaoosososssy GNRheosaamnmeattaRonenssSsi(anorerSs mErWrits) pErre% Esorodrrae oF5 AnSiomraVowarriSsnascmoes 5 crn Mems"Smsuiwsie pForaiisso To 18 Tals dss onan Loovr MossforOneNnearybAerrS eva nMen NEosnona Dom stosEremonr PNroEsFdooSmEiiaoma DF osa< loss LSoot SHErruseesnspNioamnioteDrsotvisnitoMonfesamor vrSacDeov im oun Shepisd NEPNFrroOosSistouinoosmna 5Po ooZsuRam0nn alareem aSSienit PNOrEforesosisnEiiloenaknal] PRVoaOsmnSsaCopaNraEcFnsOS0E0A0m 000 sooo raCiOsNasT NEFOSATTOraznsak0ms0 coNEuFrosaomia S1a2tem a23nm eooem p15r0 Apron Compor0asuscrh\piir ussingoSndst [ANeSerOoDsIetSiOoma Cont prEosrumi'2131N40EOTSHoO 23308 Sam eoCraONesT MErOSprAooenmktsa fears Sore eee EH De rs Tereiaes Posive values sChowpoarsmofwmeipannscaotanttarursyiaaDntaoymdeinfseocamars,d fAe052ed10idss Nerostioma PZacesoindets CNoenrosnoma rShiem Pposriievaluesshowpasofmeastat resicanty ALTUL ByDoseGroup (mg/Kg/day) 126 173 Tbios ni = WKY .=LabLo2N a | 217 NN JN . = | ) >--a con vosote Trrooom S|Sr DoseGroup(mg/day) os a fed "- square pPat--e_]-- RareAd Poraressds CRoeaonrassthoRnosssressouEmrevris) Sasosoo frsEeortwsn OF5 ArSiomtaaviSaSnsgocaeans io Stes MemRoSIasmTsn rFGereeaer Chom 15 teres ses oe cLoovn MeforOanevNnaoyosonTvaanMasan RF ossiomia Dake Sui 9E8romr PNrEorsoisotmisom 1H oREeA LSoov 5Eroruees apNoonimadar$4nibmaots veoe"flnienrrvariSanocew Su Ean I PNNreEorrsooissaEoinoimad LSooRGlRme%e dEegEmmm wirmme PNDreiorseomer] PROeSSnA0Is0S0CompaWriEoFnOsSMOWmkRd 275 Soon ssCeuOrNmeT NEFOSETAi Domoknd SRoenFoseioma SareAe Sp 50 < Sam S1o3o5o% Appar Gonpor0a0asch\ptesusinog ndensts frAMOISD PRrOMvO"21NdEFOSTMOemd oCOnNT NFosEpoomei]a 2 swim uaBomscmomma a 1e2y soeeuRmss ogsTmEmen aaimmse Post vnsho seftmatarstetr adrtte., oolnois1ed J2weot yleoEscuma 13525 ona, coup ret Positivevalues showpairsofmeansthataresignificantly -" oo- Aa Lin=t] AAIP LN WAVA AN NN]I| = NN IW NAVL AVTeArry ros7a TSRe AL{ .Nm N)WVI osoGroup roNEFaOSEMmIA sub0u0sret O0u0sted] R_-- yScrmoeree Lworme TEtReTEmSSourestrwei) #S2e2 Sse oarosgo0p weRrImSmmoalEmmam ha 8 mw paMemenrrs srreooemwmee -ot a-- mis -- TWtS ferns EOmemEmn sist eawe Repose {Em osm LEott Ero saas ropnotnaigrTstsoratmersarqmebeyr statsiuosne - TBroisner \jwEereroonssaeaiioomnida P1oomfEaaE mmsemse aosmmm DDNeoriosttRsionia] PFOJSeASaG5nm0Es5SCompNaErFeOrSEMaOnMiAa pd Soo NEFOSwSMoOnToA roCoaONmnT NEsFe smi pr5 pSoodm 8So%om sem Aprae Comof0p0sch:ap urigSetensts BAj seODea SD roisoinns' 2sNEFOSShaEmG Tease NEFOaSoMneIsm 9 eCOmNT pNeeroswomd a ao: pPoE] ilseke anea Posts vesshow epreof maartht rosigan fren ffroasi0Rs5hLiSoalSn pceoer pJ NeEevosima ipvres Noo moPtopsiBiyeBe)ovseaGtrsousphowp(ansgofmeans tht asian EARN NYA ( . < IiH = oS 7 I. v NN ) hs -- CONT N-EFOSAWmid PROSIS NEFOSEMITHG oo | SONPU Rtvit A CO oxVi Col Sweeny TprIi0s7LIR RsquareSOunmomwaaryyAonfoFvita 079w3o274 MRRoeSoqatunaMorefoaRAnedsSpqounasree Error 3107742527118972168 Observations (or Sum Wgts) 19 SMooudrecle Eror OF3 Analyssiusmooff7VSa1rq8iua4an8cre2e:5s MeaSn1S2q8u2r7e 1R0a1t8i6o8 15 4478333 20856 ProboF Coal 18 21653158 20351 <000r LoCvOeNtT Meansfor OnewNauymbAenrova Mean 6 14183 NEFOSAdOmKd 4 1750 S0d70Er8o0r osed NPFEOFSOSOEmdkodm 45 1144040000 Oosrema Level S14ErrorusMeesanspoaonldedSetsdtiDemvaitaetoifonas orvariance Number Mean SidDev cont 6 116% 172260 Std0E7m0M3e1a7n NPFNE-OFESOFOOSmSAEK4OAOmmiK 445 11480070%0 104400 t1a5r0a0z0t0 zoraed 007705701010 09273 OiCfO-NMTeanii Mean] N-EFOSEdOmt 00CM0Oe0Na0Tn0s Com"pNaEriFsOoSns1E.d8m33k3d3 183338 000000 N-EIFO5S0M8O3m5k3d 325000 PFO7S4A3O3m3K8d 560000 NP-FEOFSOASOAmOdmd TS048333383 53.2650000000 203060000000 020305000000 Apha= Comparisons0f0or5eachtpairusing Students t AoCOsN(TD.inLSD NEFOSEAOmKS 2.C1O2N6T% 21N3E14F4OSEOmK "054305 054305 260418 NEFOSA2M7O0m8K05I O64se2 PFO5Si2O0mk5d 320d PFNOEFSOOSmAKOamKS 522700382055 30.412508462 C26r0o4s1s8 203122000884 Positive values `sChoomwpapraiissoonfsmweianasctohnaitralruessiinggiDcuannntory'dsifMeeretnhto.d Avs(Dif2H6L2aS6lD98 cont CNNOEENFFTOOSSAEMAOOMMKGd 21.6020286857 218113 PFOSOmKG 0a268 CH mmolid.PiofsBeiyrteiDnvtoe.svealGureosusph(omwpoakiasdaoyf)means that are significant 130 177 Tfeom = wl {ewwl : aK--N> A I ALT N N A WA Aw a () >< 1 PX \ M -tC a DoseGroup(mpKpiday) os. oo Rare S ST e Rota fosrsesa1d9 TRoConoatemmrateRonotsosarreeSumroirg) Sgae% fisaeusc or? ArNofaOvSaeiIbri 2 per ensSuIe rGiamoe Chom 1 dime gam beer LoRovFnosaMieoamnasforOnevoeyaAnrH T cva wMomc Sumy NrEaroistniond i ER mJ e - 54 Eror te es Nlon wdaontooig ma eotorn vrncs Subey sie PNrEorsoiososrmtitiosommis OooNmaaEEm remommEs daomee Ootrosaeionniia Prosi S0oC5ooO6NoT%s" Co"pNiEeFrOsSrSEeOransa PrOSSoKOnnek So Sdimo oss NEFOSpTMorOmmia NEE a i Time o%% aSso00o0o0 Apa Co0o0m 5ch:ppr inag Scen nts sposyioirisD 26co9n%t 2*3N3islfoTsocimmsy proOsiovma erosiSonss 1 oJma ABweargomsas snoms ootslmsm aIsmoRmmEoe smzmeme a c--ows Post viesS howersofn aasstattao nedeirtyt tateos, Jomo4sha 1de 1 +conr i Se. a PJsrt os towsofettrst w AA 3" /\ A /\ oN / 3 PTA IVAY YIRX -> v (OO) or on le J----Neroscuma Souwdnst Duwmwas BsamTJ orion oSmaesn HEfueEsTeESTetervei) Sibsr3 sfphrarwe o3eJoEmTrEoimsymEmeam cwTeonnsmmeer arenoee Bo mmm gome wer ai in-- ts -- TE*" ww oan wv co HSer i{oEgmeEoEme sEoma S4TEpaaronarsoTtatrotasOsnSrm,orovoanytoSsm SESeeTmn I. peri piBgoEommemss 44ihwmmmmo gsoammmeeen oooommr 5f ooyvRere. WpEwTsw0cTEoomprssiondcuommm werossoeecurwoomgmeeenss oo| cwcooonnnr sme Za ps on CWoerosrEexlo;pSseoogpSmors wero cour fone fBE c a spsoemmms eemrognysywe wUbDermeonstnwasma cSoSoogeubim esmienE ptest t reagosodt tecmers aattas, J gBirhpe1nd . ST. gJiI d J Znaa, -. A AESA a I: ~~ IN YNNIN omTare Te o 2g VN Vv NEIFOSEAOm EachPar Stain ="With Control Suey T0711 FR DTIS OnewayAnova RSquare. `Summary of Fit RSRaooqtuaMreeaAndS)quare Error OMbesaenrovaftRieosnpso(nosreS.um Wots) 00.01430289935 13243822190665 19 MSooudrecle ErCrTootral OF Analys'iSsoumfoVfaSrqiuaanrcees MeanSquare FRatio 153 182179406.558735 19211331512 P0r75o3b0 18 20937158 116318 05375 Level Means forOneNwuamybAenrova. Mean CNO-NEFTOSA4OmKd 46 11885170 PFNO-SE4FOOSmEk4dOmKd 45 110253020500 Std14E2rr1o9r 1177441155 15576 Std ErrorusMeesanapsoaonldedSetsdtiDmevaitaetoiofnss. orvariance LCeOvNelT Number 6 18M1e6a7n 4S2t6dD4e7v NN--EEWFFOOSSEA4OOmmKKdd 44 113283520500 42450348638 PROSA0MKd 5 105000 172482 SIErMean 2176.94120 172701283 ND-ifEeFMOeSanA(4iDHMMeKadn() CNO-NETFOSEd0mkd PrOS4omid N-EMFeOaSnAs4COoMmKpdarisons. CONT 4000303030 10003030303 3"3552050000 234.91166677 N-EFOSE4OMKd 1469.1265070 "108..02500000 Aphaz. ANb-SE(FDOISAL4SODmid NC-EOFNOTSE40mKd PROSAOMK `Comparisonsforeach00p5ai using Students t 2.13t 144 N-EFOSA4OMKA 524938 37C.5O8N6T7 N-EIFOSE4OMKI 37.2438 337725483687 4432.80601304 4532.40903384 163000 217863 31.6500 Positive valuess`hCoomwppaariissoonfsmweitahnasctohnattroalreussiignnigfDicuannnteltyt'disfMereetnhto.d 262a8l98 AbN-SE(WDFIOSLAS4DOmKd 48C7O7N2T1 CNO-NEITFOSE4OmKd 5542..18869630 PFOS40mkd 322197 PFO3S305m0k0d0 2183.12656070 00000 PFO"1S64.O3m0K0]0 3211.57580603 46.9518 Pdiofsoirteinvte. values show pairs of means that are significantly Above is for themaleand female guinea pigs combined clinical chemistry statistics. 134 TTO7LLER Is Appendix 8. Hepatic Palmitoyl Co-A oxidase activity & F con 88660011448856 12..0000 396G0000005s21 33.0000 Mean 32.250 so 0.957 N-SLFOSAMOmKA 88060011448380 13..0000 dean 22.000 s fot N-EFOSE160mkd 99660000005543 33..0000 Mxean 23.000 sp 0.000 N-BLFOSEAORKA 88060011448875 22..0000 Mean 22.000 s 0.000 PrOS4ORKd 88660011449921 39660000005556 xean sp M2.i0s0sing 36..0000 33.667 2.082 cow 88650011447787 11..0000 99560000004456 22..0000 Mean 11.500 so 0.5717 N-ECFOSAOmKA 8860011448821 11..0000 xean 21.000 s 0.000 N-ERFOSEL60mkd 93660000004487 uvsean M2i.s0s0ing 21.000 Missing N-ELFOSEdOWKA 8800011448709 111.0000 135 TorsLLER SPECTES seciEs SEX DOSELGRO uTs o5an Prosiomka saecoonieees3 u33660000004550 M5oan sco S8RO06M04A0 9S8R0O00446750 d5ean N-ECFOSMORKA 86RR00I400644S e3an N-BLFOSSIGONK S9RR0O0044772I vien NeECROSEAOmKA S6RROOIN004ES2 e5 an srosdonka 0860044074 xs5R80000447743 M5ean M MSSe-lomka 1180000774S9 e1Ra0n0750 sex Cboonste_cro ES5 1R00742 1e1RR80000i0707434423 sSRRO0Dii0E333 2ORO 152.000000 231...000000 33.00 251.525570 46i..a0000sing 733..60607 Tis MM0iissssiinngg MHiissssiinngg 825..0000 25020 MMoiissssiinngg MMiissssiinngg MM1ii0ss:ss00iinngg 2114..00000 sles? mMuiiissssssiiinnnggg Moissing M2ciosns0i.n0g_ MMiiisssessiiinnnggg 045...000000 136 TTIIOS7LIFR FosAdomkg N-ELFOSA(Omkd N-ECFOSELGORKA N-ECFOSE4Omkd PFOS40mkd NSRO0TET Mseoan 11R80000774465 N1R00747 Msoean 8RROO044003367 NMean s R90RO40466S Mean FE 88RRO044003354 Mxean so 88R80044003398 SRR0000446687 Nsean sp 45.00 54.355090 MMiissssiinngg Moissing Missing Missing MMiissssiinngg M0issing Missing 2109..0000 219.500 0.707 MMiissssiinngg M0issing Missing 1M1i.s0s0ing 2201..0000 317.333 5.508 137 TT0I7SL1 FR 138 TriAsLIR Appendix 9. expression Effect of acute FC administration on catalase and acylCo oxidase TPhFeOSfololnowciantgalraesepoartndsuamcmyalrCiozAesoxthiedaesfefegctesneofexi prveisvsoioandmainndisternaztyiomne oafctNi-viEtIy FiOnSlEivoerr StoiscsiueetyfroofmTeoxxpicoosleodgyratmseeatnidnggu(iWnaelalapciges,eraanld. was presented as a poster at the 2001). Tissues analyzed in this 2001 study wfeemraelederraitsveadndfrgouminaepappriogxsimtahattelryece1 ivgedofeiftrheorze4n0 lmigve/rkgf/rdoamy ePaFcOh Sofortw1o60mmagl/ekga/nddaytw2o- (eqNu-ievtahlyelnptervfolulourmoeocotfan2es%ultfwoenaemnid8o0)-(cvtehhyilclaelccoohnotlrol()Nf-rEotmFOMSaEr)chora1l,ly1f9o9r94todaMysa,rcohr a4n, 1999. All Portions of animals were the respective killed livers on March were quick S, one frozen idnalyiqaufitdernitthreogleanstancdonssheicpupteidveondodsre,y ice to Duluth for analysis. oEfnzay1me0%Sa(mwptl/veolP)rehpoamroatgieonnat~eTohfe0e.5n-z1y.m0egffrarcotzieonn lciovnesritsitsesduoeftinhe3060,0m00Mgmsaunpneirtnoalt-a1n0t tmheMmeHtEhPoEdSo-f1BrmadMfoErdGTusAin(gpcHom7m.2e)r.ciParlotbeoivninceonsceernutmraatlibonumwiansasessttiamndaatredd. according to Lsu-sCpoedndeOdxidinas6e0AsmsMay K~H:TPhOe:-0e.qu0i2va%lentTriotfoncaX,1050p(gp/mHl7.t4i)ssuceontahionmionggen1atmeMwaps- ChyodAro(xyLpChoeAn)y.lacTehteatreea(cPtHioPnAs),wer4euanlitlso/wmeldpteoroixnicduabsaet,e 2at0 3u7MCFfAoDr,30anmdin60inuaMslhaaukriynlg. water bath and carbonate (pH terminated 10.5). The bcyonacdednitnrgati3onvoolfumHe;s0;ofge2nermaMtedKdCuNrinign 100 the mM sodium reaction was e3s1t7imnamteadnfdreommitshseiofnluaotr4e0s5cenncme.oTfhPeHfAluPoraesscmeenacseuwraesd cwailtihbraanteedxcwiittahticoonmwmaevreclieanlgtHhOo;f and the results are (Table 1). Protein weaxsprqeusasnetditaatsendmboylthpeerBorxaiddfeorgdenmeertahtoedd./min/mg mitochondrial protein CmaettahloadsepAusbsliasyh-edThbye aCcltiaviibtyoomfe caantdalaFsreidwoavsicehst(iIm.atBeidolb.yCahmeomd.ifi2c5a4t,io4n2o4f5t-h5e2,or1i9g7i9n)a,l swpheiccthropishotboamseetdrioanllythate 2d4i0rencmt.meInastuhirsepmreoncteduorfe,HtOhe,tisdsiuseapspaemaprlaencweasasdilquutaendtiinfi5e0d miniMtiaptoetdabsysiaudmdipnhgos1p0h.a3tem(MpHH,7.00;).. The The medium was warmed to progressofthe reaction 27C and the reaction was monitored at 240 om for 5 min. Catalase tem") and expressed as activity was estimated from units/mg protein (Table 1). the initial linear rate One unitofactivity ( is E*=43.6 mM" defined as that `amountofenzyme which catalyzes the decomposition of 1 moleofH;0; per min. 19 TIAI0S7LIR Northern Blot Analyses - Quantitation of mRNA for both acylCoA oxidase (ACoAO) and catalase were performed by Northern blot analysisof quick frozen liver samples from treated rats and guinea pigs. Approximately 1 goffrozen liver was powderized in liquid insiotlraotgieonn uksiitnganadmotrhtearc/opnecsetnlter.atTiootnalquRanNtAifiweadssrpeecctorvoeprheodtoumseitnrgictahlelPy EaRtF2E6C0TnmR. NTAhTMe RNA was electrophoresed on a 1% agarose gel, blot transferred to a cellulose membrane PanCdRhaymbprliidfiizeedd ftorotmheprciomrerressptoondcai.n,g3r5a0ndbaosmelyse[q*'uPe]ncleaboefletdheolriegsopneucctlievoetirdaetslitvheart gweenree.. mRNA band density was quantified autoradiographically using. phospho-imaging software. RESULTS: `Rats -- Both male and female rats express LCoAO and catalase activities, and there is no substantial difference in enzyme activities between the sexes. Thesame is true for the constitutive expression ofmRNA for both enzymes, with the exception that unexposed `male rats do not express the message for PCOAO. Acute exposure to N-E-FOSE causes a doubling of the specific activity of LCOAO in liver from both male and female rats, but catalase activity is unchanged. Associated with this is a proportionate 2-fold increase in the concentration of mRNA encoding for PCOAO, but no change in catalase mRNA, in livers from both sexes. Again, there is no remarkable sex difference in response to N-Et-FOSE exposure, at-least with this limited number ofanimals tested (n=2). Exposure of rats to PFOS elicits a similar doubling of LCoAO activity for both sexes, `and possibly a slight increase in catalase activity in liver from male rats. Hepatic catalase activity in female rats does not appear to be affected by PFOS exposure. Unlike N-EtFOSE, which caused a doubling of mRNA band intensity for pCoAO, acute exposure to PFOS caused a 3-6 fold increase in PCOAO mRNA expression. This enhanced expression of message was more pronounced in female compared to male rats, and in both sexes resulted in no greater LCOAO enzyme activity than was observed athalfthe mRNA level for rats exposed to N-Et-FOSE. Guinea Pigs - The response to guinea pigs to exposure to N-E-FOSE and PFOS was dramatically different from that observed for rats. The constitutive activityofLCoAO in unexposed guinea pigs of either sex was very low (near LOD values) whereas catalase activity was 1.5-3 times higher. However, the activityofneither enzyme was stimulated dfioflfleorweinncge abceuttweeeexnpsopseucrieestoisNt-hEe-fFacOtStEhatorguPinFeOaSpiign mviRvoN. APeernhcaopdsintghefomroPsCtOdAraOmawtaisc undetectable, even following exposure to N-EG-FOSE or PFOS. Part of the explanation for this observation may be that we were using hybridization probes developed against 140 187 TbTIIOSLIER the sequence for rat liver ACOAO because the sequenceofthe guinea pig enzyme is not Known at this time. It may be that the guinea pig sequence is sufficiently different that the PCR product lacks complementarity for adequate hybridization to the probe for the rat sequence. Although this may indeed be an artifact, the absence of constitutively expressed message and the inability to stimulate expression of PCOAO mRNA arc consistent with the low and uninducible enzyme activity in guinea pigs. CONCLUSIONS: These data provide strong evidence that: 1) N-Et-FOSE and PFOS stimulate both the transcriptional and translational expressionofacylCoA oxidase in rats in vivo, and 2) there is a marked difference in the response of rats and guinea pigs to in vivo exposure to these two fluorochemicals. These results are very consistent with the suggestion that these fluorochemical compounds are "peroxisome proliferators" in rats and, much like what has been demonstrated for the classical "peroxisome proliferator" chemicals, guinea pigs are resistant to this effect of fluorochemical exposures. We were not given the data for organ weights or necropsy findings and thus cannot comment on whether exposure 10 cither agent caused hepatomegaly or other indications that are associated with peroxisome proliferation. However, based on our data, we suspect that the evidence will reveal the classical signs of peroxisome proliferation in rats, but not guinea pigs, caused by these acute exposures. 191 V2:14 TAOLIFR TIS `ASnuimmmaalryofScSrpeeecnifeorSHeepaTticrPeeroBxaiosdomymeLiPervoelrnifLeritavteiron P450 LauylCoA Camlase ug pCoAO Catalase sox ww Protin (ng/g) content Oxidase (molime)Activity activity RNAi mRNA mRNA roe (amolimin/mg)pr(oUtnietisni)mg. Control Control yo) SROOME) Rat M Veh SRROODOME6)H RRaat FM VVeehn ROTO Rat F Ven RODE Rat M NE. RODE Rat M NF-OBSt-E ROOT Rat F NFOESLE OROMT2 Rat EF NFOES-E FOSE 298 12 ISLS 0086 321130 17316 11848536 00015647 201 71 1553 005s 22 13 Isis 0331 231 ILI 1349 0284 174 75 1658 004 17S 73 2158 0.88 4 53 2199 29u 185 265d 1267 73 190222014471 219 338 100 100 91s 1085 2 2195 35 290 1401 n7 m3 26 2m ois 78 1861 193 253 178 629 1 30 wa 9ROO6T RODE Rat Rat M PROS M PFOS OROMTS 9ROOT4 Rat Rat F PROS F PFOS 257 229 119 125 11S4 153 0423 0241 17S 153 83 73 1431 2386 0077 0099 139 12 22 2369 39 208 am ssi 2m 1s7 11397915250461 198 2 60 1504 404 1413 9G004S 9GO046 99GG000000SSI2 Gpig Gpig M Veh M Veh GGppiigg FF VVeehh 296 208 113 119 1752 2527 0.23 0089 227849 114129 2132013 001.8187 9000047 9G0004S 9G00S3 9G00S4 Gpig M NE: Gpig M NF-OES-E Gpig F NF-OBS-E Gpig F NFOESLE FOSE 240 87 2127- 26 85 2601 0013 230 96 1764 0126 236 89 2183 0195 9G0004 9GONSO Gpig M PFOS Gpig M PFOS 215 72 1s32240 10 2221 007 - 9GO00SS 9G000S6 Gpig Gpig F F PFOS PFOS 221 237 84 74 1218 1716 0031 0016 (a)di)nnsouftfdiectieecnttasbalmepleforanalysis L347 133907 Low 163nd 103 97 0192550468 165nd 23nd 91 109 M93. ow - 07 393 17m 2 07 4168 1770 105 IS 4781 128nd 104 a3. ow 09 09 4608 6129 1270 Lsnd 97 103 14 $16 12nd 9% 192 TorAsLLER Appendix 10: Final Report for FOSA (T-7132.1). Comparative Molecular BiologyofPTeirtflel:uorooctanesulfonamide (FOSA, T7132) in Rats following four consecutive daysofdosing. Final Report February 18, 2004 Study Number: DTI1S ProtocolAmendment Number: 2. 3M Medical Department Study Number: T-7132.1 Study Director: Andrew Seacat Ph.D. Analytical laboratories: L KendalBl.Wallace, Ph.D. D.A.B.T. Professor, Dept of Biochemistry and Molecular Biology Schoolof Medicine 10 University Drive Duluth, MN 55812-2496 2. Dr. Xin Lu Departmentof Pharmacology and Physiology 601 Elmwood Ave, box 711 Rochester, New York 14642 Study Protocol Title: Rats and Guinea Pigs Comparative Molecular Biologyof Peroxisome Proliferation in Study Initiated: In-Life Start Date: November 16, 1998 In-Life End Date: December 21, 1998 15 bTiaso Purpose: `The purposeofthis study was to treat three male rats treated with cither no.compound (vehicle control) or 40 `mg/kg/day perfluorooctanesulfonamide: C8F1 7SO2NH2 PFOSA, FOSAmide, T-7132). The study was designed to evaluate t (FOSA, certain toxicological effects, such as hepatic peroxisome proliferatiohne,omftethaibsocliosmmpaonudnd and to compare these effects to other `compounds dosed at the same concentration... Methods Test Materials: Perfluorooctanesulfonamide (FOSA, PFOSA, FOSAmide, C8F17SO2NH2 NB# 120067-108, G. Moore `propylene glycol. 10/28/00, T-7132) was investigated. The vehicle control`was Dose Groups: `Three male rats received propylene volume of mi/kg body weight. glycol as the vehicle control by oral gavage ata Tohnreetehrmoaulgehraftosurroefctehiveedstauddyo,seaonfd 4w0ermegeuFtOhaSnAi/zeKdgobnoddayywfeiivge.ht,Avsiuasopreanlsgiaovnaogef8omngd/aymsl. ofFOSA in `This dosing propylene glycol was prepared, regiment achieved a cumulative anda dose volume of of 160 mg/k ml/kg wasadministered. ofdosing. This dose was the same as the dose of PFOS g after administered four successivedays under amendment 1 ofthis protocol `proliferation in and rats is comparable to effective dose ofPFOS found in the literature. levels for hepatic peroxisome The LD50 for FOS but the cumulative dose of PFOS 251 mg/kgfor PFOS FiOnScAomadomili,naisstaerpeodinutondferreftehrisenpcreo.tocol is A is below notknown, the LDS0 for MethodofSpecimen Collection: "iTnhetl~oiv1ergwpiaescersemaonvdefdlaasshr-afpriodzleynadsirpeocstsliybilnelaifqtueird euthanasia. nitrogen in The livers were divided taredpolypropylene n(iNtarloggeenne)hacdonetvaaipnoerras.teTdh.e Tcohnetatiinsesures wwaesrestmoorvededatto-7d0r0y Ciceanadndsh`iwpepieghdeodnafdtreyr iaclel.liquid Sera: cUlpottti0ng1,0mthleoblfoboldowoadswacsenctorlilfeucgteeddfforr1om0emaicnhutaensimaatl1i1n0t0o gxlagsastse4ruCm.ttuhbeesse,rFaowllaoswing transferred to new tubesand becentrifuged again for 10 minutes at 1100 x gat4C. 1 19/ oo TomIisO ER aTnwaloysailsi.quotsofthe serum sample (~ 0.75 ml) were saved for possible metabolite Specimen Handling: isdhein2ptpiefgidcraiatnmidoanylcihiqacuerott(b0oeftlhteohweantlaoil:vyetriscaalmpllabeosrathtaotriweesrleisftleadsahcfcroorzdeinngintloitqhueidlinvietrrsosgaenmpwleere PKreonfdeaslslorB,.DWealpltaocfeB,iPohc.hDe.miDs.tArBy.aTn.d Molecular Biology S1c0hUonoilveorfsMietydiDcriinvee UniversityofMinnesota Duluth, MN 55812-2496 iCanvotenhreoslxaaibmdoparlsaeetsoarcwyioevrfietKyaen(naPlWoyaozlesldcahbcayenpDdreeYpvatimooaufszlaByikoipcuhb1el9mi8is6hs)etadrnymdeptrhootdeisnfcoornPt4e5nt0 (cBornatednfto,rdLu1u9r7o6y)] University ofMN. and Molecular Biology a the Another 1-2 gram aliquotofthe liver sample was sent to: Dr. Michael Wempe LDaebpoarrattmoernytooffMP.hWa.rmAancdeorlsogy and Physiology 6Ro0c1hEelstmewr,ooNdeAwvYeo,rbko1x4761412 amDnera.tlaLybsioinlsiXotufelopifveterhrfesoarlmmupeoldreostchfaerraobnomanlrysastwissegroieftvhedneetsaeearmnsiagnmeepodlfelsuoarnodcparrobvoindesd.abTrhieefpasruenmtmaarngyof the previously published methods (Hansen ef a. 2001). ive samples by LC-MS/MS using Liver SampleIdentification Chartfor T-7132.1 [Sampl1 e# Animal1# [SpeciesT[TSe1x [Dos-- sGr-- ows] I17 TN [ [3 2T[IRRoO00774#5| RRaatt | W Joono --------] |W Jeanie [|= --&Ti[RooTr4Rs ||OR0Raatt7||4WW_5H[H8o0mmgoiigigddaayyFFOoSsAa--] 145 bTiaso (TRO at_|wiJoomghgraar rosa] 1 193 | hTrion Results and Discussion Average body weightsofthe FOSA control group on days four and five treatment (Table 1). gLriovueprwweerieghstisgnwifeirceanntoltysilgonwiefrictahnatnlythe bdiofdfyerwenetigbhettwweaesnstihgenitfriecaatnetdlyanindccroenatsreodl ignrtohuepsF,ObSutAtthreelaitvmeernwtgerigohutpas a percentage of `Theresults ofP4350 content liver samples indicated that (Figure 1) and Acyl CoA oxidase FOSA induced the expressio o activity (Figure 2) for the therefore a hepatic peroxisome proliferator in rats. n fthese proteins and FOSA is The concentrationsofthe (Table 2). The data show parent compound and metabolites that PFOS was the major metabo in livers were`measured AgipvpernoxFiOmaStAelaynd0.3FpOeSrcAenNt-ogfltuhcuercounmiudleawtaivseiddeonsteifwiaeds apsreaslmeinittenoifnroutmhneetdalibinovletirhteaec.lievietrhsoefr rthaets parent compound, FOSA, 3). The low percentage of or as the metabolite PFOS, one day FOSA in theliverand the appare after the last dose (Table PFOS has also been noted in in-vitro microsomal and liver nt low conversionof FOSA to slice metabolism studies (Xu et al. 2003) which alow rate, but not observed that FOSA. was converted by microsomes orcytosol to PFOS in-vitro `by liver slices at Samples It is noteworthy that the control animals (1R00742, 1R00743, significant concentrations of FOSA. Furthermore, no PFO and 1R00744) contained were found in of livers from the control Fische samples with a high background SofoFrOFSOA.SAAN-gpalruaclulerlonaniadleysis snoatmpslheoswfodrettehcistasbtluedcyrow3nce4er4netrarctaso.tnimtoaanimsniotnfaaitFneOeddSaAit.nstoThmheeeUsnpeiovirenertssutilittsmyesouefRgxog-ceyshitveosthtbaeytrtthVreaiccveoanqrtuiarunotmldiitliidveesr ofFOSA, as no metabolism to PFOS or FOSA. N-glucuhradooncciurdreed in the control wr 194 | TboIri0s7LIR Signatures: Report prepared by, AStnuddryeDwirMe.ctSoeracat, PhD, DABT Date Reviewed by, SDpaonniselorCr.eHparekseesntative Date a| 2s TonsTF References: q5B4ur.aandtfiotride,soMf.prMo.te(i1n97u6t)i.liAzinrgaptihde parnidncsiepnlseitoifpveromteetihno-ddyfeor`btihnediQnuga.ntAintaaltiBoinoocfmhiecmr7o2g,r2a4m8- Hansen, specific, Kq.uaJn,, tCiltaetmiveenc,haLr.aAct.e,rEilzlaetfisoonon,fMo.rgEa.,niacndflJuoohrnoscohne,miHc.al0s. i(n2b0i0o1l)o.giCcoamlpsoautrnidc-s. Environmental Science and Technology 35, 766-710. Lazarow, P. B. (1981). Enzymol 72, 315.9. Assay ofperoxisomal beta-oxidatioon ffatty acids. Methods `PCooosAcohx,iMd.ase.a,catinvditYyaumsaiznagkai,laRu.rKo.yl(-1C9o8A6-).baDseetderfmliuonraotmieotnorifc paessraoyx.iBsolmaol cfatBtyioapchyly-s Acta 884, 585-93. Wallace, K. B., Luebker, D. J. Butenhoff, J. L., and Seacat, A. M.(2001). Perfluorooctane sulfonate and 2-(N-ethylperfluorooctanesulfonamido)-ethyl p1e6r57o.xisome proliferators in rats, but not guinea pigs. Toxicologist 60, 348 AbsatlrcaochtolIDa:re Xu, L., Seacat, A. M., Butenhoff, J. L., and Anders, M. W. (2003). N-Ethyl-N-(2-hydroxyethyl)perfluorooctanesulfonamide Biotransformation of| microsomes, eytosol and slices. In Toxicological Sciences(,NS-uEptpFlO.SVEo)l.by72r,atp.l3iv1e4r. 19 || 196 | 7 Tables Table 1 Biological Parameters re E arr er Bots Amendneniiz____-------- ------------ ohn IDT15BodyWeight (BW),Liver Weight(LV), Th old Weight (TW) | TT = AR Sar Araos 2b 1ST2Ga 2 2e 2201ror oi ] ] day1 |day2 |day3 |day4 (days W(%) (ig) rr :| [ ;: w = mw m w onsweoam AB e iT s m we s aril 7s aians Sean So E= rr orm em 1a canoe]a ey014% ay Ci ar || Eo ewwn woassoe a Jm T wC wm s1ew LP Test [oil on ae 0.34% 0.09] mAtt t ve tr oral 008 002 oot asi oiag /97 oTrsam `Table 2 Liver FluorocarbonConcentration Hepa Cocetions oftoons sd Farcsbon Metabolic i Lives off Gives FOSA (fo F songs r Tsaex ooo m |oSiatn ed| ooom d|rroosmsya |a"pm]|e2Sdaneit] [[maomomoonosnssTTT wose roassTo [oowoeam srn|[|a oasidadasaam i] ag eomedllonoo)ee] mg/kg/day) Sons. 5 79% kTi roma `-- Teapblie c3PPcertcenotf DFolusorFocsarcboneDnosendinFLioverr en thls inLisof ts Gen OSA E-- S;EEAnima P(-- Gorsosp[TDorsae [Pmros>-- - [veroos obrros[-- ade Paeas rFoone-- -- [Fere [-- emot ES SS i (mg) |dose* (ppm) [(mg) [uber (ppm) (mg) oar ) | ree; ls Te l; d HO cH B|B gr a ee a Tn m C r--r =f | C [1 I Sidmmooonres3s[[rr2oos5saalSssTuss0e2l]T esTse0sEasll2a21] 174r0] 3e oz1]51 11130e5a7l]5r2aa5]oo0yu1 12 169 Aor0s71 FR Figures Figure 1 Cytochrome P450 contentinLiver Total CYTP450 Content in Rat Liver Following Treatment With 02000 FOSA Evrrgrectrosos t Joi | mercer i ome coun EE iR : E fe: flik ooo PERE contol bs ur a Fosa 15 ypiam Figure 2: Lauroyl CoA oxidase activity in liver LCoA-Oxidase Activity in Rat Liver Homogenate Following Treatment With FOSA foi5Fa%l)| Lm Amematnes; Segetaren * Fro] 3m] El Zeno posl in- pe wo 50 20/ EEa EEEEELINE EECCE EECEE Inn. | sega] TTT pC pe SEECEEEEEIN[EE 3i EET Fr 207 | ETE LT HTT TTT Ii EE. i nh EEE N AE ] iBehl =m=e sa"| [TF Helebik Zo3 wh Ce exeaEER[11] | [zee[1] ei T casein ET ell] Fi [Jee [TTT] 204 TTT pee M[siTeaTine ivsis]| f{Z|[| eereeeFeddNT4|TTH]T pe 11 pl]Te h[11177 208 EET ER [MSaO e Frere|] [Bees hit HEE (LILI 206 |NiE fn| | le| (SEER Ti e i13 f A| fin 27 SHEE LTI T] T Es T TTTTTTee sssemaag II| TTIII EeBezTEeteeeseeege H [feE emle bigsr He [Hl AF [T[EH& TH F E 2p iTM5Ef gf g3 I 24 = 5 :g: gz < 213z 5253 i B2E85Y82 a gi- gk BZp2%is =FI a 2 258 3%2 ae g23 Bsfg og$i5:8 $s58p BiE5BYiyd i2z $3S5Y 2 S354 fiSr 58 B g3Ia 3= F2Eg f95f2.g oPg EBfuiREfEEaEE: 3 Flizszig: 98322 o]d = 3s = Fz S. gHgEcyd aEipEEy ss:iifsd 838 o 85 Z i 2 5 :3 a E3] s : fx f : e 23 ES GsUFiSl H31E35E34 5828 s53i8g2s% 207 oo gz E is = 5 3 Piskf i25i3%s: 353L25845s3 E53ii2i2c5=2si7ddg Hr SB5E8E1:F P8i3s8s 300 BFEeSfEEify EHHE5C8E5E5S5 pins SESEELSsREE: 2 i2 Eh 235 Biz S333358358 3gEgsgt 5: 50E2SiE fiHsE; g dl BZE28RD airy nd oo : o ih Eg LE2 Of LFoO +sEFiETsOs : HEI] 2EOE E E5E2Z2T E2:2F89 R)28%82S5 b23 Tfoszi2dgniiyc Eggi:l if23 5 Sissi 52I $iS5igesgssii2ssys fire sB 2F$3E2%E3 i2F f ggiiise Rip of 3: Fissiiizsnied:d 5 diii8 is: fg3oS:: 5f. d$PaECSsER giS5nif I33 zp i=ss3l2i2f2e:g d oii32d5y g2l3 & Bifr fb3gspfiicgils gIo pfEpiiEf oiFi g 2% 2322353 E83 2 OF 27 BTEeEsZiE:887 3 275% Fizz 38 pi: 2/0 J 2 ig : qu if i 33 5 ig 32 2s 25 2:2 228 H i 35% ig i I] 55: 3 1FE:3 i i! #02 dd 55 i 232 io: HF3EIs ER i7F [1OE2EM1EL5sINEiaEH f3r5is bo5f g3bl E tiH3hiEil A PEE HT iEiH , 5i2i3: a8i5f, 1%32 % HiHEHfi: iifI5 3ij855F i8g%s:niiszf pl isPitz $: usH ii I 1 Eis HPS EELE ER OB <5% 853F 23If 4: sL3H 8E | | 2/ | iE al Hi i LgHio P is laps 3Tu5g4e3l5: aii ni 3$:453: ji Hp i 5 er HS5B1lii0Fp fipei I31L pH en HEEii finn EagitlREEliEiiic LeainnEiel aEiaigiakniiishniei A 1 DoiTaBHnRolfmL glhig aa g Hpeys, HB1i8 L fiBciR fazy sffilz : 3 i2 Z 88 z |E:s F][& FFFRAEFEEEER] :s [2 FFREEERHTIREIEE' EREE: Ii.lFi EE ad EER = ILL a5t.s [EFEFRERTIFEFRFATNi Bess [FEF IFFFATIE S5.a5ti<e [PITT&[FFFFRLg sl] slr le 2 SELLE [BER esis? : 5 = fz le 2 EPELAE N TTTT R: TT fid|T6 EE if PEER] ab HHTEEEES HE i FE ] [EFF oee; i | BfIad|F|r|E[m[[Ee[hErEEiRReiET 3(A 1p|MleITFT TEEEIEEEse ERT, ir Eh T n EREn iE I iz 2 As Eh : | | CT] | rE | Ii fe] 5 [3 i Ea |e [ee 3 | | | [2 | 5 if i|i 2 a| HE ih i f3 |e&g HHHiHi ||| SIE | 2bLeL sdo Sraraiw dosaonos n| a7 1{ ETE 2 HE 1iyz i iiEEiHf & $8 2 8 8 = (BuruzozH sofoun) Ayagoy ssepxoryony 29 oT0r7s11 ER n | UT | THER E {11 R 1 ( E L IIE[L T reEeeE ar T ]lgay A[re Ea1 1 | |saEuems 2 220 ilaiigl bi EE 74 mm || G EET L 8 i Hr E. 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