Document 500ROm8OEK29KXmg4GYjjoyze

TT pt 7 NO-repore CONFIDENTIAL Ae 2 2% 3 756 T`HSEULEPFHFOENCATTOEFD 3ONPECRHFOLLUEOSRTIENRAOTLE/DBIALLEKAYCLID METABOLISM IN 15 %-FAT FED E3-LEIDEN "CTORNAVNESRGSENIIOCNMIINCTEO ICNHVOILVEOSATNEDROOLNIFNARTATTY ACID HEPATOCYTES IN VITRO. Project study number: 031.10074 /3M-02 TTCoNNmOOnQBtuliaomlsioctniyeyonfceLsite SHTeoorrvecao PD EE iO etS prEEhvoeSestesnennn naeesaparen SSSftoromysmoamraptaneeoSenrcio.aron,int)nd. Ci AST frSSiaCanemretsoan 11242k0013266002 BeEotnvae ste ao strim sm toecesr SSutEincoCroven SFiinaslsvearasiton:04 December2006 |||| . [ rfni-- Stmo208 BR Beare | Neca 35 i ATeucthhnoorliozgaytmiboaynnOra. gPoHefArV.aseQuulaaxr,and Metabolic Diseases Signature and date: Ei i Eolrle |a L i s : 3 vm I Fon 52 L Toss TNOmotofwutMO Contents PAFSampAoEtMsdenvics p.2 Ge12z06 Testing faciity page 3 Contributors page 3 1. Introduction page 3 2. Study design 3. Resultsofinvivoexperiment (ApoE3L eiden mice) 3:1. Bodyweightandfood intakemeasurements 32. Plasma measurements 33. Measurements in metabolic cages 331 Foodand drink intake 332 333 OMrocvoinnsgu(mXp)taicotinv,itCyOzproduction and respiratory exchange rato (RER) 334 Energy expenditure 34 Fecalbile acd,sterol, andfatyacidmeasurements 35 Intestinal cholesterol absorption 36 Perigonadal ft tissue weight 37 Liver weight and hepatic pd contents 38 CLhiovleersetenrzoylm7ea-ahcydtrosxy:laHsMeGa-nCdosAterroeldu2c7t-ahsyed,roxylase page 4 page 6 pag6e page 7 page 13 page 13 page 15 page 15 page 17 page 18 page 22 page 23 page 23 page 25 4. Results of i vitro experiment (rat hepatocytes) 5. Summary and conclusions. 5.1. EffectsofPFBS in ApoE3Leiden mice 52. EffectsofPFHS in ApoE3Leiden mice 53. EffectsofPFOS in Apo3Leiden mice page 30 page 32 page 32 page 32 page 33 20135 BOwcdNMNG eSoixlseniceeo 3 Testing facility TGBuaNsuOibniQeuussaslLiUatnbyiotorfaBtiLioofrseyciences PZe.m0i.kBeodxre2e2!15 P2h3o0n1e:CE+L3e1id7e1n5181200 fax: 31715181901 Contributors `Study director: EPDr-h.moaniLelo::uilc+so3uoH1i.hs7Ce1onh5@e1n8t1n4o69n! Experimental work: IIInnnggg... ABChenrpiasHtvoaavengaeNnieeTwhiwekloop. IInngg.. WAnigmelvaanKoDuucyivsenvoorde IInngg.. RKooon vnavnandaedrelrMSarudkis IOrn.g.PEeltsebretVoPsehtoelrman Aavisor POrr.ot.HaDrn.sLPorviinsceM.n Havekes 1.INTRODUCTION oa3fnMitmhhaelascsotoumdbpiseoesur.nvdeIndsotorhndaetrchtoohleeOsbcttoearmiopnlomauonnrddosbiinolsfeigaihncttiednrtessoytntthhheaevspeiossssitinrbolpnergimmceahcrohylaecsnutiletsruomrloefloofawcerarotinnhgetpheafetfoeefccftyeslcetissn hpaeenprdaftooinrcmbbeiidlleesaatcuiidddy.suypFnttrahokemesisihn,ihhsaomwsstetuvdeeyrrlint edwiuaescpticeeofcnfaceslctunodnevditsrttoehraholalvtsehyenbheceeosnmipsiomnivuegnshdttsibgaeptoesdisniivbnollayvepidrneacvsriweoeaulsslel.dy amInehttuahbmeoalnpirisekmseehnlativpeosptbruoedtyeenitnhipenavteitsnetrivngiavtaoendde,fifneucnAtlpiokoeEf3wtLhieedidtteeysnptetcrmoainmcsepg,oeunisincdssemincsoein.ticTvhhoiflsoersatinepirmdoallloamwnoeddriebnllgesdhroauwcgsds. dliikoe.statins. Furthermore, the plasma cholesterol levels can be elevated by adjustment of the acAdhndodilteissothneoarrlotlly,cshyiannitnthheefsaiptsryehsaaevcnietdsbpreoinjnteoctobcilhnaoflioenrsemtdaetrvioiolanaionnntithheveiperofeessctetunsdcyoe,f iotnfhewththieecsthccotohmemppocouounnvndedsrssihooannvehoefpblaeoteningc Investigated. 30135 Ons peas nso pa csnzses 2.5TUDY DESIGN T0A8ph.pe0r5od.ve2as0li0g6fn,ofrDotorsvtihasettipouenrsfbfoyrohmmaenhcTieNOopfoAtlhniosasslthuaEdxvypehebaresiambneonmetenCnotdiemosmnciertidbeebeedlhoianw:sthboosfnincabsluidnyedpornof0o6ca06o-f 2T0h0e6fiunnaldesttunduymdbeesrigDnEaCnHd1t2h5e.concomitant measurement points have been depicted in schema : ST SE TTI TETT E orBurso12(o(0r 1To10oo0y) 11e 5++00000010%. PPEdrBooSscoed. ee mEsnEr--InmI-- eIr" ---- rO oouwn?L (1t0O1H1E+S+ 0000t01%% PFeHOrSSs mere e---- ----} [a -Fienasstrs ptote 2. -_ ow = . 12-]x aorromicroosowmees namedetamnairs = 4 ma 1.0.4.6) -.x Scheme 1 Experimental dean of study {p= pooled per group) Acrhafontldeeosramtie3zrewo)deeoiknes,brans7ii5snmopfaelrboiooddAypoownEe3iagLshee,imciPenasysnmtaihcetecihco(li1eg1sht-e-2raotlwe(aoHnkFd;st1oi4fg%hagclaorrdd,aot1ss%(omrcteoamosfuorfuednanmad)e0w2e45r%en fhPaeFstBifnSgo)l.(ogwTrihnoegun.p12s0thawenedgerk3os)u. p0sG.r0roe0uc6ep%isvPe5Ed aHrnSedsp(e7gcrtroieuvcpeeli4yv)HeadFnrdios0ep.e0c(0ic9vo%enltPyolF0Og.rS0o2u(3pgrPo1)uFpaS6ndai0nn 0dt1h0e%0,F1i0%0diP3eF%tOooSrf iFcinogc.thid3e.id1a.ts1,)tuaotnpdawffitohoe2drthwIenestceakkreseochfamsernastitsmoaeadnlnty iodheecrireceaesetldohst(osoenen.iagFvhigo.aag1oC.o23n),tgeraFimostotufhinbsta0dhyaewsooainngdhot1f(wtsehoeee sCAtoufdunryteh(seirpsdoevdnieadvtiiaosoinfno5ccn.oe6nocfaenhrden,s16pthouelyopca2l0s06ha)a.skebteoenne discussed wilh 10 study montor by Eat bodies determination. These compounds can oTftnarelseayiasntobgmne)eniatdcne(tdoselectedtheeEndro-etfnmoblro,pcamoesnaweasaausprioneGndaadceipdnnroc(odop1e11rs3awapman,ydrpa3rlftaesJrtmuhaaninseSda2oo0mvt0pe6alm)meiisgonhfatwfoaensdiknnsgwbpoaeknd-1(a(1n6adk1o)w.noFaeotrer8tiosnf Am7teawsSeuMreekcds.om0,pIna1nw,ye4efokarsntdh1e8(7cpeGltaaeysmsmianofactehooalnmesoetfentrthoel,ta4ontdaCntodrmi8gp,loycu5en,rdi1delcoalnseevmoenlatsastaaiftomenprsl.4ehs-fhaasvtiengseheanvesebntee1n0 sorss Tmt 5 PFTinalSapAnAXENGMTG 0032200 Aavseroraiggeinavlallyueplsaonfnetdh,eaet swuebepkosol0s, w4eraendde8t,erlimpionperdo,teiant pwreoefikle4s waenrde8,anoanleyzpeodol(eatd wseaemkpl0e, tpheer gdEir-fmoeauripelnwtcaofsmreoanmsapeloxynpzdeedce)tn.ecdeS,iwneicxttehratshlteiupwdoyepermokotne4ii-ntloiprprooofpifrloe2tse4ihnJaupvlaeyttb2e0re0ne6sn),.easnTpahelceyizaelladltytienroftprgleraaostummpaesns5ta.amwnpedleke7s,6 ww(eesrreeee o`ebxtpaeirnieednceafdteoresonvoetmiggeht diffafsetrienngt In weeks 0, 4, 6 (extra measure rp(oenisenuctlset;ssscsaoermeypaEfr-omreadiklteotcolonirepropebrsooptdoeynindedpnertocfeeirlmoeifsnoa2ft4i4ohJn-u)fl,aystw2ih0ni0gc6hs)amaipnnldeosu8.r, pInlawsemeakA5L.A6Tfevcaelsuehsaswebreeendectoelrlmecitneeddfirnopmoo3lseudbsgarmopulpesspeprertrgeroautpm.ent.group during 3 periods of 2de:t3erdmaiynse,d. aInnditiafteecdalbybtilhee Ea-cmidasi,l cfoartrtyesapcoinddsenacnedofs2te5roJlulyqu2a0n06i,iefseceasndhascobmepeonsicoilolnectweedrien wIneewkee4ka7s,wtelhle. mHeotwaebvoelric, aucptitvoitnyoow,f4weansimeaelnsopreeragsroonuypetfrtoomingvreosutipgsat1e, ft3,fu4rthaern.d analyzed in metabolic cages. The folowing parameters have been measured: food 6inhtaakvee, bienekn ipnatraakme,etfeeresditnhgeboruetssp,iratottoarly pehyxscihcaanlgaeciirvaittiyo, (ORcEoRn)suamnpdtiOoy-ncoannsdumCpOlxiporno-druecltaitoend. boFdryomentehresgey ethxepemnedtiatbuorleihcacvaegebse,enducrailncuglattheids. peBreicoadu(sweetehke f7oo--d7c.5a)n, otnhley obtehesrupapnliimeadlisn ipnowthdiesrsetdudfyoamlsion rInecweeievekd8p-o9widnetersetidnafloocdh.olHeesrteearoflteurpatlakaenihmaaslsbereencedievteedrmfionoedd.in chunks again. afInrnodwzeepneekrini1gl0oinqtuahidedalmniitfcaretogwweeenrrfeeorstadacerktieefnricmoeiudn,tatpailnoandsmowfaeliwigpahisedsdc,aolntldewcoetvesedmnatbluylalmpeRiaeNncAessoanfoafhleytashriest,lpiuavnencrdtiwmoeinr,certohqseuoiicmvkeelsry ``saatnneddromckhi-ot2lo7ec-shhtoyendrdrorolix-ay7laha-sahevyedacrtboiexvyeiltnyasihesaosalcabttieevdeitnfyrmhoeamavstehuerbeerdeenmianditenhtienegrmmipitaonrcethdoofnitdnhrtiehaelivmferirac.crtoiHosMnosGm.-alCoprAepraerdautcitoanss,e 50135 ps IHOmporsy tz EoFAoSnnpAomnENgte es osrpzuns 3.RESULTS OF IN VIVO EXPERIMENT (APOESLEIDEN MICE) "baTephlpeoewnr.edsiuSxlitgtinnoigftiihcnidasinvcrieedpouoraftl.dpirTfihemeraremyncedeaastnawvaaanslducesasulcbpuseleraqtgueredonutupscianalgrceutlhpaerteendsoednna-ttpeaadraairmneettphrreiocvfiiMdgeaudrneisnnWahnisdtepntaearybalteUes iItnefsttteharenendficgteuhre(e5sr<,e0s.au0l5ci)onlgboerpet-dwveaaelsunteetrsihaserkveaagtliuaveesnoefpintahrtehaetienadpmipceeaatnsedudirxgeropuopin(tofintdhiecastaemaesctaotliosrt)icaanlldy tshiagtnioffictahnet control group. I the tables, tis difference has been indicated by a yellow background. 3.1. Body weight and food intake measurements. APSFOsSho(wgnroiunpFsig5s.an3.d1.71) caanuds3e.d1.2a,lrtehaedytrferaotmmetnhtewsittarhttaherahmigahrkcaobnlceendtercarteiaonsse oifn PboFdHySwaeingdht wanenoidrgmhfoatoadfgoaionidtnaikunet.paktAeof,etrchoetmhcpoeantrcrahoballnvegaeltuobeaasl,lckthhotweoertvhgeerroc,uotpnhst.olbSuodbidesytewqieuniewgnahtctlokyf,t2,Ghrteohuegpsreo5uaapnni7immamalilscser,geataiutrnhneodtuogh aTinnhcdere6PaFsliBonSwg ctargoeanaictnee,ndtsrmtaiatycieeodnssbheoohfwiPenFddHdtSuhreiasnngadmtPhoFebOowShdo)ylwleeosittrgweheatitgmiehnntctrseipagensriiefoiadc.santthley icnontthroellsa.stTwheeeGkrsoouf pth4se sbteluodwy.when compared to control animals. This i in line withthe loss of fat tissue as shown VFailguurees3a.r1e.1means +BSoDd.y weight o[rroGemrosvw4TiHFbc++o0100m10e45PFOSS GrGroowEHHE +C0S.O02TRPPFSS. GGT rroo E1 +R2003 SPFOS = 1 ddl meee e t1 f s =e trI iadTE EiiTl3 8a 1s 2 0 HneR n at r5r etEE time (weoks) aEnxicmeapltsfaortethaeboduevtitahteiosnsamineftohordouignthaokuetbethheavtiroearotmfengtropueprisod5 (aFnidg 73.1(.s2e)e.above), all ther Gor3s 7 ssa A ize iFgrried3e12 Foodintake E EE TE re Si . To. me i T i: oy7 gE33,2 ?: . eTRT 3.2. Plasma measurements o Te lovinn g psthhevenbesan stains n pas the crs sy. cholesterol (Fig. 3.2.1), triglycerides (Fig. 3.2.2), lipoprotein profiles (Fig 3.2.3 - 3.2.5), ALAT TFgE fJTorm eEszs aresP20ae sE saai.AT A 3, i. --1 Pi q == = -- iil : : sn0o. t ani3adios,ePrREe:aNntsFOsongderTstand 2 0%sor ers oon lst Tass pe PT A Be, seam TihheePPFROBSSgvreouapm(o7)n, hdoeweovterdetcirsoarestsurgnnwiacsadnellyayhedeuchtewsetaekrfvorahse PFHS group (5) Mare ores th same resus avebeen abained forthe plasma tighcerd (G) ves VFiageuser3e22means 6Pl5a.sma triglyceride o R SeA mr 1aI osootro s4m7o4s5 =Gm HF 012 5 a-d +r1r 2) c20 f2is . 2ETNNY ----t---------- 20s EE --- 00d. -- : ms 4 rp : z. Time weeks) iT3Pr)oFomaHmafSeicrnoaanndtdwovelPeeRkTrOsGShoefdavavcesnrceeomsscseeonedntatTgnrGaeiTnelGvosew0v,eeslbacu1tornuawclpesoe7onkTtaha.ttoGhnirrgoohufoepsFut8BcoTo.GnsecgerecolnuoipnartnhwdoefebtFtnBpaooS0wrcec(vogoernrokttup! Iovls att sh 10 compl det 3iIThoh2eve.ld5dsifi)lefroarenennndt.ibcaTytohomelnepaaisanurttoraoicpnsiagnphetra,oevtepenhibesoassshphnhoaaowseltadderociarnoednpdbnofypewmrheeeiacnsshucrsT&imantogchjhakoersobsC(screForoeevsnee3dcr3owtl5iit.cstoo3feo5npn4ed0of FOL Tacions (Fis 32.09.02 55.50 TWVTahhIceeDilLPoonsFwsScemonteercteemoarctbcsmoeaentnintorcnlrroeepsfauaslPtteeEdednSant(o3ernouumpit4n)oDsrLhcLohwOaenLdgeatsnhdinaHhpOpeLeaprroeapnsrcteoeomfnoafvrpatoctpliroonosrennhmfnea.cstioen wlShieinsccheiophnlossapHihDodLtpbidefsioargreenthaaemraHijsDoLfrocfaomcntaisonineianmtnp,anihlrslocfvlaeecrrliaopnnpciehnagokfwmbioaholntevdesDHiLgLnRabOtyeLdhraeescvaorsne,This aT2fIrphDgaLcecatb-ai)-4rocenvrooeasnotsparkepoimennpmagnIlragyeetdddpeevtacoeoprrfrieynoacMshlreeouednag,hsserwhbhuueipcpyohGhenramolneiuoigrpnmhg4.atelrispntad(riielcgaalhttemcewaonintdnce.wcitrFmheHeaPjSsFoe;rHiSrna.eprCnfoodcn2ucecweotmeiiieotrkna/snstealcynr,regctVoinLotnHDoOLolf.aintd SHogwenvemro,rsafLeDrL.anTdoL&awnkgss,mahlaHaDpron prfies wont sack oe conel pass ith sors i Figure 3.2.3a Pooled plasma lipoprotein (cholesterol) profiles ; PFBS treatment Week 0 ==] Wee4k i jo A i i i np AL \ . Week 6 a. Week8 -- Is In I AA EL | he ALN]Syl " AA ' > Sag Fire558 Potod asa pprti Ghosphold pis ABS sit Week 0 I EEE |L In Wee4k I: A ju \ fu ot \ Week6 -- Eea e ra) Week8 LI A | EIF -- AA F322 REI i | /3 es an - | INQreportof siugy M02 rg 2 ADOF Laden mice. 04:12:2008 quesmts =Pooks ses popesErseterote AHSsr Week0 Em Week4 L. I t: IhaAN i ON Week6 -- Week 8 ne iI ii Co | : Pad SA ol i 2.40 ok sapri sot etnFS esr Week0 Week4 =) | IToa i | I io a rE I04 Ei = 3 oa for a ohne diles el i I I fo ~\ Loe A by 3 14 | Te |{ Moons eSmAa dEnedn0ope Figure 3252 Pooled plasma lipoprotein (cholesterol) profiles ; PFOS treatment Weeko == Week 2 on Bul How + ib BAR | Iii I:oe oy Week6 Weeks ii ii Figure 3.25b Pooled plasma lipoprotein (phospholipid) profiles ; PFOS treatment LS I1i 8 I w, = ar] Wee4k os I = i who a /P ATTN eoke Weeks -- Horas [ pz INO suddtt2 PEASi Agoaiten mie os122008 "(Tghreoulpip6o)prsotheoiwnepdromfialjeosrofthtehelaPrgOeS-HOtLrefartaecdtiaonniwmhailcshasresrteemaardkyabilnec.reTahsienglodwurcionngcetnhteration raenadtmIeDnLt.fraAcptoiBo-nscohnatvaeinbienegnldiepocprreoatseeidn.s have been decreased. Simiar as with PFHS, VLDL lGervoeulsps7i,mwihilcathorsthheowcoendtraolrgecroovuepraygoafinp,labustmashToCw/eTdGlloewveIlDsLataenrd2awveereykslacrognetrloalrdgiee"t,-HhOaLs pVeLaDk,L twhhiacvherremVaLiDneLdptrhoeduscatimoenohvaesrrtehceorveesrteodf(tohreitreenahtamnecnetdp)e,riboudt.iApopporsostiebilnecelxepalraannacteioins sctoiulld be tihmepagirraeddu.alIltyalinmcirgehatsebeofthteher"osluatrogfetHhDeLlopnegakhai-n imfeeofinPtFhOe Sloiwn ctohnecebnodtyr.atTihoinsPeFxpOlSaignrsoaulpso6, which in week 8 has become as large as ths peak in the group7 plasma. PSiFncSe,tthhelivveerrseenezmysmteo ableansitnreonagmliyniontvroalnvsefdeirnatshee(mAeLcAhT)anhiassmboofeanctdieotnerofmiPnFedHSa afnodw times. more than originally planned (Fig. 3.2.6). Figure326 Pooled plasma ALAT activity GGrroouupp cToHT +co0n.00E6H%S M Wareo s5TRHFE+u T0O02T n PPEPRHSS. WGouwu 6THHF +O0T0RF3PRR%.OS. [E 7: +r 01%p pros 500 || 35 400 ; < 30 2 200 533a10 & week 0 iir 4LR|| L2 week4 1 bl J week6(on fast) i A% iJ week8 aAtlatlhloumgehafsuormewepoeinkts2AgLrAouTph7arsecbeeievnedintchreeacsoentdroinl tdhieetPaFgaOiSn.-trPeFaHteSdignrcoruepasse(dgrAoLuApsT6asanwdell7), A(lgtrhouopug4h),pbluatshmearAeLtAheTraecttiuvmittyoiscoonfttreonludiseetd(agsroaupma5)rkcearusfoerdvaer(pdaratmiaalger,ectohveeriyncorfetahsies ianctthvey. tprleaastmmeanct,anbubtectahnearlessuolrteasusltwerlloomf tanheienndluacrtigoenmeonfttohfe ethxeprveessriaosn osfeethneivnethrisesntzuydmye(sbeyetFhieg P37F1B)S.had no oronly a minor effect on plasma ALAT levels. | 120135 p13 IMCs 002 PES Aon stan Spilnacsemait kweatsoneexpbeocdtieeds thhaavtethbeootenstdectoemrpmoiunneddsbemfiogrhetahnadveafaenre6ffweectekosn offatrtycaatcmiednmte.tabolism, Aocostnotsrnhoelowibnoediyn(gFliergvo.eulsp3.s.2T.57h,aenPdFtHw7So)wasetnledkr,seoseufvleteneadmtiomnreiennctsroew,aistPheFdtOhlSeeviheinlgschrieensatwseeddeosksei,6gnfioflilcoawnetd tbhey4pwleasemkas of PFBS did not significant increase the kelone bodies VFailgueusea3r2e7means P5l.asma ketone bodies [2[[WFrGororoouuTp7s.THHFEe c++oD0m00e10r8$%7P0F5S W= GorroulT5p:FHF+00002THPPFPHES.WW or rSWHFo E+T0.E0v03 %PyFFOSS _s 3E e $s 25g 2 5 4 i : z0 bi Fe i week 1 week & Sanoa,lyPsFisHSinamnedtaPboFlOiSc ciangdeuscepdefrfaoymnaecddinoxwidoatkion7,ofwhtirecahtmiesnst,upported by the results ofthe 3.3. Measurements in metabolic cages ASa1)Tc,colfutihp7meaw(at0gie.rze0aortku3ip%soon6Pf)iFnhrBeatSavhteegmrasboraeutamp,ne(4agcnrmaoaigulcepy)e.oz3ef),dTthhhienedidf0voii.lfdle0uor0awel6inln%tygfrPpoaFraiHr3maSemcenrattyoesurgpsrinouh(mpgaesrvt,oeaubtpbohele4)ieHcnaFncmdaceogantehstsruor(l0ead.gf:0rt0oru3fp%1oo(ddEgaFryaoOnuoSdpf dalaictneykr :pianrtFaaikgm.eet3(e.Fri3gs2s).h, e3O3rr.ec1so.p1nisaautonmrpdyti3e3ox.nc1h.a3an)n,dgeCfefOeidiopnrgo(dRsuiEcztReioFbniog.u(tFi3sg.s3.(.F3i.g3.33),.33.w.3h1.i1c.a2hn),dgi3mv.eo3sv3i.in2n)gf.oramFcarttuoitmoyntoh(nXe tehnoersguybselxrpaetneditthuatreis(Fugs.ed3.3f.o4r)ehnoevregybegoennercaatlciuolna,teadn.d additonal, theOr consumption eiaed 3Nsi3oz.e1siagFnnodifoiidcnanaktnddinitdfarkinen,ckaeilsntthahokauegvhetbheePnFaHbSsertrveeadtebdemtiwceeenmitghhetghraovuepseaiiefnoosdoimnetawkhe,atfemeodrieng bout 130135 Domo erAShteleto00n02s0 p14 Era Figue3311 Foor[d=-oirntoauukT4epHHFc+o0.m0e06%PES GGrroouupp3: HWEF +SO0T035FPEFSOS n" - . . 3. 33: . 2 N oi u iuw ] rfl d ow a profriet ow on G Figue 3312 Feeding bout size Time (h) [GGrrououT4HHFF c+ome0r%P0IS0GGrr8oouupp&FHFE+O0003F%F3TPEFSOS oon pe --- -- EE sos : ] 2 god 2on i | 3fon )3 eli i] 1 ondiou lead ow we Ty ow 7 Figwe3313 Drink[i<Gn-rtGaorkuoepu4pT:HHFF +Go0m.ia0l6% PTimFe HS GGrroouupp 6SWH F+T0O.I003%FFPFEOS| : ee " " oe?: iTi .n i2 . 2fe i. rl oom au ntIi i3 xety ow FT Lod ow 5 on taoras DOME eSoseCleros pts reno 5N:o5s2igMnoifviicanngt (fe)raecnticveitsyhave boon bserved between fe groups in moving acy Figure 332 XActivity [=GGrroowunp &THHFE +co0mr0o0l%6 PHS GGrroouupp3: FHFF+TO0.F00E3%EPFSOS 2 a 55 1000 JENN 2 3= uw 3H:E J <<, Fray) om uw Rl fT % oe Time (h) lis rey] w7 1r3H.iao3st.g3erred0omu,aapcrnsoki,anmhbaslluiesmetwpoatsnisionosngq,cnaiCttiOhch,aate-rpftsrrymoaadilunclcrcgtoreiaoossunepsdia.,nhhOdeirsrceoeosnppnisaorrufaamtempoerriteyoenrencsxoecfrhatbahveenetgwPbeeePeerHnanStciooabons(deRnrEPvReFa)dnOdSbePtFwEeSe-n oated mic have been ound Figure 3331 Orconsumption [[GGrroouupp 4T:- HHFE c+o0ntr.ol0%P0EH6S +GrGorouupp 36:-HHF + D0.000033%FPEFSOS S000 pe . o 3 "0 | |] | 5i 300 hii ss hSagi sRecslic a0 1000 0 ou #0 oe wm on Time (h) sors p16 MOsencs_ erFoasisaganc _ournas CiRnOEhRpircfoohdtultchotesisoemngeroofuptophisentPasrFecHolSmopwaeanrrde(dPsiFtnoOiStchaegnritonyucprfseoarwsPeaFsiOnfSOo,cunobdnutstuonmbopelyieitnocnsr.teaatFsioecrdaaUlslsywerslie,gansaiolfntihctoahunegth fiasogrrpPorEeemNfeSrn)ottnhtwaiantlhtuahseneidRnEfcorRreaeosnfeetrhiegyfcagotenynreoarlcaitadinoodxniPdoFavBteiSro-ntrheesautrseeefdloegfcrtcoeaudrp.bboyhTyhacinrsaitsnecasr.neaiTsnhedidiscpafltsianosnmtaat ft KaestfoonuendboadtyerleivnelthiinsPstFudHy.S and PFOS.treated mice and aio for te decrease inal issue Figure 33.32 Gro__CuOrTpprHoFducctoinotnrel Group3:F + GIS PFS |o-Group 6: HE +0.006%PFHS Group 6: HF + 000%3 PFOS. 500 _-- 3 4000 3000 Prionti pi 200 ieWi $$ 100 : .| um % @ @ on Time () Figure 3333 Respiratory exchange ratio (RER) [[=~GGrroouupp 74:: FHFEc+on0.t0o06l%PHS G==rGroouup6p: FHFF ++003.74%0PP0FFEO3SS 12 -- at El iSo 25%os RRNYii n $2 os & 04 1% pprrotiod %4 Time () faArlh l & on tooi35 p17 BE raa sp osm 3Ba :34eEnergaynexbapoesnpdeoiotiuvrnseataetoonss ibtohoans nbeoensscaaluaastewedllthbaytrhee OPrEcTSoannsdtFrO:S veo Fgwesse Energy expenditure GrGroou&TpHFF S+ 0o.0o06% PFHS =GGrroaup 6W:HFT+ 000003F%EPFSOS Lon igg 53 g E10 Mimnnt |P| g Hi ei, vie 35 a os |wu au 2% @ ---- @ on Time () hToengferregncgersouopbssenrveedvinathermeataSbuoecsagpereanfaeltyasbisei=gh)t be mors pronourced by ors p18 -- PES 0 sco Suson mice ou 3.4w.eFeekc6aolfbvieleatamceindt,,sfteecreoslhaanvedbfeatetnycaocliledctmeedaasnudrtesmpeindtasnd bile acid contents dFoectaelmibnield.acids have been decreased by PFHS (0.006% group 4) and even more by cPoFmOpSou(n0.d0s03h%a)v.eAslilhlokuegphhgerbioeua5acpndsdex7crweetrieonbabcelkoownhceonctoonlirdolievaolrue4s.weeks, both hese. sPiFgBnSi-ircaenattymdeinfterseenetmfsomothheacvoenitnoclregarsoeudpthoerfgecraloubi2pl.e acid excretion, which was even VFailguuersea3r4e1means BSiDl.e acid excretion CrrosTHEc+o 0P0SM0War0ea,EFHF +S04O3RPEFSS. MWarreasSTT+ OOBSRPREGS ro 1 +001 0PF0S 6 58s z5eg ; : 23g83s:?2 >; y 2 g8s4 b3y fe i Table 3.41 Fecal bile acid composition EE -- Tr ---- [Feeera2rsrTaarciurlsvo a+aois]roesssasaisllimeaeeswseroialisiooonssasaalsTreeirua)s)) E o Nfeoeesrs2osoorls iis3o 0s2sasss5aaoTa enesshsaalatteesssSssae71o 3sg86ss+vioolarzyae+r1a4d]) "woTibhtseheltrahtvoteeddr aiesncwcreelrla,sbeewyhwaiPiclFhsHPceSFanBraSensdduilPdtFnfOortSo,mchaaacnahgnaengtgheefiinenccbaiollmebplaoecsiiadtciisodynnctoohfmepesoxisscirtpitaoetndh.wbaCioylnscaiocmidtishteahnvateslryb,een ibvuetralasfotefreaaboscohrpamtnigoneainndntrsettiurnnaltobtilhaIcviedrconversion. Th later might aso happen in the 180135 INO spoofs M2 _ eriSpiEdsmene p19 Ges OlSnosleoylrspi,nigwohnrioacuhn4pdari(evPepFrrHpiSrm)oadwurcaytsicoofne.mcpaSloisnneecudetrotafhlecsiheeortloussetexcderrecatoorine,otnadisenitegednri0mf.iin2cea5dn%tbciyhnofcloersoetadesrieondlt.,akFef,eocoiadnltiennsettuaitknera:all PSibnlecaroyeraaspsiemodanjfoorcairloslseo.tteThrheiessxafcsmreemotoirrosninioncrrgleerassoseutd4ph(es(sePaeFmHFeiSg):iin3sa5l.tl1hg)e,rormueopssusl.tt Sporifonabcnaebiilnnyctertsehtaeisnsaelidnchhieocplaaetnsittceyrol eOAblssheotruvgaehdrn(osseyfeneiFhrieges.ins3c.5e.2Ts)h.ihsavf eibn oagnreseemeenntntwhitehqtuhaeniinfcyroefaseexdcrHetMeGd-pCaonAt rsteedruoclts,aswehiaccthivity Smriigghntabtee ftrhomretshueltdoieft, tchheacnogmepoinstinitoesntiwnaalsasbisgorhptlocnhiarnsgeecdreitnotnh.e PFHS and PFOS grou. It FVailgueusea3r4e2means SStDe.rol excretion GGrroouopTs4.HHFF c+o0.r0a0l6% PFHS W GrouapT5WHrFET+O0e O02T%Hs FPFFHESS. WGorroouupp: FHFF 0+0.S003%PFPEFSOS 5Group 7:HE + 001 4 PFOS 100 i5%f 5 8% 60 Ts5 a0 58 2 33g 20 3 0 5 4 3 i| 2 : pF Er3 b =0 neutral sterols i] ry 3 i phytoste=rols Table 34.2a_Fecal neutral sterol composition CC omwmwem [00 :00 00:o0[as 0a]27 202 20 03) Ii CT TRRT RAGI8N 0c0o::o0o0l[0000::00009o5aa5 +=0154]]2341 +=0i3s]]2216=soo02]) (SEIS MRORIEEON00 =00]00 +00[ 037 +07]46 30s] 17 +01] 190135 Cs 2 p20 erssinsnse ours Table 3.420 Fecal phyosterol coG mpositionomme Sp ES 276:22. 765 +1 +0087]| 857.720 ++111966l55e86r:=+ 151]] 17] 0000:+0000] 00 +00] [I PPATRT APTN A eR Re 2276c56: :+v1o11|1]4665: 5 ++022507[][66e07s75x== 210155]]] 0000 00 =0000) +00 i REMMI 5: rol 50: oso: 20] 00 =00) FVailgiuorsoa3re43means +Fat5.t0y acd oxcreion SrrosprToH++e 0 1 4771 05 Br0yG T61T4 7+T00O3S TPPS Wnayn1S+8T08 1EPFCS 1000 < 35 a0 325882 a 22s:g5 w i: i5%t aw 0 \ od Jos Table343 Fecalfatty acid composition I =emm =--e--t ----= ----| . F s ra nosa ooaors m s oreoasleoosoreotmteostoun)) E: e SooeihrieseH aoeO ustaotiSsaTsoles erooefoemttoSomntoes ttvooomn) F(GoPrcLoauMf4poatst(tyPoaPfcHtiSd)eahgarodoudspegrsii(visecdoamfneotymhaitvnhecorafearatsgfedoSmeD)dfoasohdfoanwyeadakcnas(,TsiGgw)nhiaefnircdeaanitsdfgsrroapurnpoc1due(c2rewdoembeykcstohnPerFlv,OeSr aCnodmp4oswtoikone Conv) showed a 0m decrease, wih a concomitant smal change i ay acid wats S From the ee frre S weLiatce opyrico324p(tJ1Ge)rCtr aakmeaper.styuhicrreioodueserasaWd4btogtgrrmyteyseaied, Te Fguosss Fatysatauncs i ---- wo goo if HT) * FE5 200 Donets sz p22 ersnttnsmnee oz 3.5. Intestinal cholesterol absorption Talhemiinoteswteirnael icnhdoilviedsutarlolyl ahbosuosrepdtiaonndwraesceaisvseedsasnedorbly tGohsofeocfal20du0ak1isofolIopee moefthcodo.rIinwnegek1 8 Coanriom[r"oeCulds-,cehaoonlvedesrtfeterhcoeels4a-wnddraye1pCceoiallde[,cHt}ae-dsnifdtoodrsittsahsneool{lv,leodpweirinnmlgoa4usned.aoytsB.opdFoyetwcaeesissgihwutemrae(nIdhloflmooo,gd6ein0nit%zaekeedt.hwaepnreoe)l.ed Radioactivity was determined in the fecal samples to assess the ratio of ["C}-abeled cishoklnesotwenrol abnadp[o*Hr}y-laabbesloerdbseidtos(t<an3ol%.)Siintomsitsa.nolThweasfousremdaasustehde froefceraenlceccocmhpeosuntdersince it aFbsCoTrptiodnoswaisn:g %mocdhuorlees-te1r0ol0 absorption = ([**C)*H}-dosingmixture -- ["CJ*H]-feces/) VFailguuerse 3a5r.e1_meaInntses+ti$na0l cholestaro absorption CFarma.1ro+0o00arParPosS Bn 1 +0034PY WTrTen +o 7108 5 E) _e 3H%E 5 En i E0 : : oe pro `week 8-9 aBloltghroPuFpOsS-htaveeataebdougtrohuepssa6maondch7olheasvteeroilnctraeakseedviianttehsteinoaoldchaonledstheerolsaabmseoroptrison. Sine ``(AccslhhotooelloeeFussigttgee.hrroo3nll.o8tos.yu2sntt0iph)guentssiiuf(sgie.gcxaecnIsetnpt,dtsengeadren,oucntperhsee4t)aii,nsntcaehrlidesacihsheneopcdareteHsiaMtcsGreco-hmoCulasoebstsAtoebrrrpeeotdlciuaoscnuytsaonefshdegebarscyostiauvnip4tyi(niPcnrFteHhaSes)seedsghrmeopsuatpistco bboeexsipglhaiiyneidnc2reawseeldl 3by5awnelincwrheiahs,edbneapsatcacchoolnesatelrolansyinnhcersesas,ed fecal excretion, might 2s p28 Ono suMZ_ erFAoSsmomnENssenmes __oatpzom 3.6. Perigonadal fat issue weight FViagluurees 3ar.e61means + 5.P0e.rigonadal fat weight GGrroouwTpHHFF c+o0n.r00e6l --%PEHS WGorroauup3m5:THFF S+000012R%FPFPHSS @WGarS6:WroHFFT+uw0S0p0w3H%PFpFSOSS [Porous7:1 +0.01 % pros 12 2 00 S8: os0 S5 os 38? 04 5 02 [1 * week 10 PAsulFptHphSoorutg(eghdrnoobutyp mt4)ehaaensdudercerPdeF,aOstSeisi(nmgbirgoohdutyprw6seeifaglhnted.7ac)Pdleracosrnmeaagslerepeitdniuntcolteoadvlepbleosrdmiyigfgoahntadibaselsufmeaet.atsTiuhsirssueedn,otwihonicihs sTghrhooeuulrpdesdbuectfioounnidn tpoerbiegodneacdraelafsaedtiassasuerweaflsecntoiotnosfedenecirnetahesePdFBfSat-ttiessautee.d groups and the PFHS 3.7. Liver weight and hepatic lipid contents csLioignvnteirrfosilcaadrnieteltsyioerfnotnlhgaelryggeridoncuvrpeer5ass.PeFdWHbhSye-nPtrFeeOaxtSperd(egsarsnoeiudmpaaslss6%caanuodfse7to)dtaaalndrbeotPduFyrnHwSteoi(goghnrtloy(uFspilgi.4g)h.3t.lTy7h.be1u)tr,esttthulem 0 cPoFnBtSr-or.eated group 3 also had slighty but significant enlarged livers when compared to AltihsvaetsrsohfooftwhnPeFicOnoSnF-titrgro.le3am.t7i.ec2ed,.gtrhoeulpispid6s,antrdig7l.ycIenrtihdeesoatnhedrcihvoelressttehreysleestceornst,enhtasvewbeereencoimnpcarreaabsleed wiintthhe Free cholesterol contents havenotbean changedbyany ofthe compounds. 20135 TNOepesru of M02 PAFSaineAgpcoEltdeniice p.2s ost22008 VFailguuersea3r7e1maeans + LSiDv.er weight [[[WBSGrerooou7wwTp4.:FHHee +c+o0n00t10ol6p%rPoFs.S WB roosE5:r FHFE+O0F0T2u %PFFHSS. WWGarroueS6pHHFO+00T03 RPFS 6 5 2z , 5gs 53 2 1 week 10 VFailguuerse 3ar7o.m1eans +L5i.v0.er weight as% ofbody weight [[GBWr&rG7:oFHHFoFercu++o0wr00orp.1o0l6pu%rPoFpsHS WF orm r 5E:HFF+u T00O2O4m PTPFFHHSS. W WGroou6pSHHmFT+ 0Ci00H34p FFPOFSOS gC2 Zs E3 i ny 50 Ls fH o i Bh = week 10 200135 Ose te 002 PEAS inApoEUsten mie p25 osm VFailugeusraer3e7m2eans SILi.pid contents iver [RGrroooupTHHFec+ o00o0l64 rts WMrraayT5EHe O+00012P4PPRHS. W Mroua s6:HHFE r O+0R0p F4P0SO3S crop7h + 011pr0S Zan 3 250 S 20 A 510 100 2 50 235 0 Free cholesterol : Triglycerides Cholesterolester 3s.t8e.roLliv2e7r-heyndzroyxmyelaasceti.vities: HMG-CoA reductase, cholesterol 7a-hydroxylase and byidemremonesdiyislaoatlsalteyedaacffttrivorimtsyatchheraivicevee,breiteivnsesrudsee.htaeTrvhemeriebnaeeledtenirn,rtehHmeMovmGie-cdCraoonsAdomrmaeildcuprcrtoeaspsoaemraeatsnidoanncsdhoamlniedtsotsceehrrooonhkdTr2ai-7a- have hTyhderoyxieylldasofemacitcirviotsyoimnetshepmeirtgocvheorndsriaabout the same in al groups (Fig. 3.8.1) Fig. 3.8.1 Microsomal preparations [H [GoromG me7T.HHEr Fc++o00n30vo01e67P7w0P8 W Bara o5:WHEeSr +O0F42Tr FPFFRSS WWraraspT.TS+0T0R03AEPSOS ED 5 1s 2 Z E310 3E E5 3 30 EAD Bor3s 2 INOsgetat suey iz e6F25oirn Aogos ite mice srzam0 AfrlaottmhheorcuolgnahlrrgHoel,MfGPo-rFCHPoSFAHaSnredgdrPuocFutOpasS4e. saSecieinvcmieoythwheaasyvieerlaidtnohcefrrtehoaeswemdiitcairlslospaorcetmpietasyr.aptieTohrgnissiaivsnerdsgitnhimeforcrfaoenroteeySsdDis'fsortaohrneot sblaearemgneerdaeincfdaelrlceaunrlcgaeetseddiinfaentrodtoanelcxhepesrpeabstesitecwdceheaonslettoshttearlIovalcotrsiyvwinettiyhgephsteissvhbeaervtew(Febiege.nen3t.h8oe.b2gs5re)o,ruvTephdsi.sextmihsie.gahTtcovtinaidlyicHhaMateGs-that aCnodA7re(dPuFcSt)a.se activity per iver has been increased significantly n roups 4 (PFHS), 6 (PFOS) VFailgueusea3r8e2maeans + SDH.MG-CoA reductase activity in liver microsomes F [GrSouo p7.rTwHFeE+o a0o00r.1d08p%rPoEsHS WCrooreETHFS+S00T2 4PPPHRS. WMaarroe. +TF T0O0R40TFSO3S 2 ZHz R 3515 28 $fo20 3: Sg< sEs s 22 0 ; .k i poss FVialguuerse a3r.e8m2eans S.T0o.tal HMG-CoA reductase activity per iver PGaGrrrooounwpTesHHFE+oco0otv0ep0rPoH8sS W2Cerorp5E. H+SO0F02R4 PPHSS. MWraSMTrF +O0eR00nP3FO4S 2 1000 2 E 3gE m 2s 3$25 a0 22%% 0 3eg eo LfZool Er p27 FiVgiutreess3e8m3ers + 5C5h.alestorol7 hydronyeR se activity in vermicrosomes T ni . ZHE 22g2E it: 0 Figruee 3a35mes5Totalcholestrol Tahydraoca ntvyylpoarsveer sT es L E aT sm fw iizz, SE I S 2%T 60 33 5 23% a = i3 o , A TrOoinuernoosnfcitstehoysrm,a1athhsibooitnnge teenfzoeuypme edasessfeiton lsntaedlacoeiydPsEyNfSnc(soohnFe6rmiacroosSyoomParEloanrneecholmens To tof einnddPssFOcSoarouse3ssreedmiond.ooappcoyseproorrashiriefcmantadsecrseas by PHSS gOr 4) 2am TNOsp01t syM02 p28 EiSinAvENesennss __or1z2s Tm3i8ht.eo4co)htoshneodrerimraeelgduplr0aetpobareryaiteinnocznrsye.amsetedwofainisterheeamcPaiFrdkHsaSyb-nlteehaetsthiaestd,thsatnedryoilPelF2dO7o-Sf-hmryiedtarootcxehydolnagdsrreoiuawpsa(,sgismugegagIsevuesrrt,iendFgigi.tnhtahte hobessesrvceodmipnocurnedasseionfcfraetatsyeadcitdheoxniudamtbieorn of mitochondria. This might be elated with the VFailguuerse a3.r8e.4means S.Li.ver mitochondrial preparations [[ERaFGrorpor.u HHFecs+0ao0rl085.0PF8S WBareisEmF+0Sa02.X4:PSHS WMprTT+T0T00R35, 4 53 SE32 8Eon 3 p 30 a ne AbysPexFpHrSes(sgersoupper4)magndofPmiFtOoScho(ngdrrioaulp6psroatnedin7s)tearsolwe2l7.-hydroxylass was found fo be decreasad VFailguuerse 3ar.e85m2eans SDst.erol 27-hydroxylase activity in iver mitochondria [GCSrrcoorspin7HtFc+0r001s85PrEoHsS WuarnyEV+R O0U04FT3HS. WWranSHTE+T8O00SP098 2 zS.Es 8 in gL $5 o 2ot35 n2 TNOaoot of styM02 PErienses _cetzaom eHbxoepwrieenvscesrrei,ansgaekidiheinngatcihtnetvosiyaacmpceeorugnvrtoeuropfs(.tFihge. f3e.8r.e50n)c,etsheintomtialtoscehornodr2i7a-lhyyiderlodrsyalanseinavcerywewiagshtfsouanndd VFailguuerse a3r8o.5m5eans S.To.tal sterol 27-hychroxylase activity per iver GGrroousTpTHFE c+0r1e0%P0E6S WGrooupE5:rFHESe+0T.0aH24PPyTFSR. GWrooreuETHF T+00E03 EFSOS Group 7: HF +001 % PROS 800 2 Si: :600 zE a 32R52E2&0 20 wi i EX 20135 .30 pesos 2 EErSkonseuamene osname 4c.FormoRpmoEtuShnUdLrsTsmSiigInhNtvVinIofTeuxRepOnecEreiXmtPheEenRtiIsniMpoEmrNaflToc(moReAdtTaiHppEroPoj,AeTwcthOiC-cYh1T,ESwt)hae ssubssuigagteestfeodr htehpaathitec PFAS CCbchheoooellmneepssittoneevrrueoosnlltdssiagyynsarrtthhehedessisini.n,tcrhoSeorisppnourcrveeasatfesinaoatnytponarfochjideadicsftce.abreteAnaetbnao2tlp6aioks3setnsiakv&deoinsicogaoubmr1ecole0,e2fdcoarhfs3aeooyptneaagrtacaelsnohdpotseoperefichmiooietlcnotsh.ttbrfeGdoaenrhstoatefsal Cxpenment have bean paiomod 3-4 ines (as moniancd ath ures) VFailguuerse a4.r1e meats $1. (1-4) Incorporationof ["CJ-acetate into cholesterol inrat hepatocytes in vitro [14C]-Acetate incorporation = : i 2i5s 3 2 w o ow e ot 01 1 tw compound concentration (molr) tm 1000 FViagluurees 4a.r2o Imnecaonrspo+ra$t5i.on(ovf4(")CHhutyrate into cholesterol i rat hepatocytes in vitro [14C]-Butyrate incorporation 20 2 20 = 5f wo 38 5 100 ig2 w o 0 00 01 1 1 10 `compound concentration (moll) 100 000 wars Pesan soy to trsaeatsoeos,toSehmtischaittecy srcooerbtn be hardly an effect of these compounds on cholesterol synthesis from acetate and butyrate. E ai ie SHS. Anrae `The incorporation of the longer fatty acid [**ClHaurate into cholesterol is also comparable to much stronger inhibitorifthe cholesterol synthesis from ['ClHaurate and also from ["C}palmitate (Fig. 4.4). Figure 4.3 Incorporation of ["C]-laurate into cholesterol in rat hepatocytes in vitro Tse[14C}-Laurate incorporation .fiw- Ta mrs 5 5gs8100 2 a.wo oment1%time we wm 22 : Figure 4.4 Incorporationof["C]-palmitate into cholesterol inrat hepatocytes in vitro Ts [14C]-Palmitate incorporation .; - heorer 5100 PEOS 5g58 e.wooom ,compe1ns%ate ermoe we wwmas F3D) .32 TOselssNR erFaSoitmeofileenne oom "fcieTonshnftecleucineontncmcropeardtopiouiornnnadatdisseoonopmneoenfadwpveheanarlctaaygrteieenma,ltalirhvksaealdbeullceoersnewgaaearssyeet..ofIawSteottysrhaettchpisadrnedusicsefoefndniccreiunoolofttviatslhnuedsstesdu.ridfsyBft,uraetnnndttfheccrohsntocleashntotearootbloovnwisoausosf the `siecnvodemerppaeolnufdnaedcnsttoarwrsaeayni.ndfSlitunheecnecritanhdgeiposalolalmbuiebtliaeittdecy ouopfmtptahkoiesufntardnagwneasrpsoarttpirydeesagecrniatdtdaiinturitaohnceeimnretadhmieouusmntathimase,daeclolpnietcngredhenittnrbgattohinoant he test compounds have influenced fs solubility and nanos s avalabik for uptake 1 the cls 5suh0so,erdta.llantodgeltohnegerrthreadtieosatbceolmlepdofuantdtysahcaivdeienttohecrhonloesotrroolnynlriagththeepfaftecotcsytoestuhnedionrcotrhpeorcaotnidointioofns A5b.ilotlhSoogUuicgMahlMtbAheehRatYveisotArNcoFDmorpCotOuhNnadCrsLeaUhsaSovnIetOshNeiymSiaarrecdhiesmciucsasledsihreurceurseesp,aratthoely.show differences in their ta5Bh.no1etd.hckoPneFttrEBoofSnlf-eetgbcrroteodsauiotpeesf1d,.PgmNrFeootBuadSbposil2iinfc(Ap0epa.or0ra1Ewm%e3ee)Ltrnaaeenricdsodo33uesnn(sd0mm.ei0ian3cs%beu)ordseyhdownewiemgdhe,nto1a0ob0rdosmicanaltglaekdsei,fpffeearceasnlcatesrAowLilAthT aw`eSnimdgahflta,tdtiypfiaecdeindcocenextscernehttaisvoean,nbdieneetennsztsiyonmaelenciahnoclpeltsatsesmroallaibdseovrpetsi:aonsiighstsudeecwreeiagshet ainndcheolersterol fasrneadectmtieroindsgyticonebtrehdeeiscn,ocnwrtheoialcshpeawadtattsemesr,e.flbeuctteIdDiLnILmDiLnoarncdhHanDgLesweorfethseompeowphraottilnowpeartttheamns,thVoLseD:L wImnhitcieecr.hemsTtihigenhgstaissmutiphcepeasrmstahltolhwbeupdtreaslcigsenodiifaningcainnntoctirinoecna.rseebadustehebipeacftaoiucaslcehoboifleetsateceirdloalerxg7ce-rhsettyiaconrndoaixrnydtahdseeevgiaarctoitouuni2py Tdiehscervseaualgsugeeedsitptwleaadsssmnmaoaltilsitidantcilsroteviaceasslelyinsibginliefaiccaindtsynthesis mightbrelated fo the slighty 5Thsth2eaer.tsgtdruoodEvuyfapfpte4eorcinatosndsi.imonTafflhoPseoFhdhaHiivgnSehteaibkrneed,eAonwpshoetirdEceh3agtLrleoeedduiwp1dit5eah0n0.lm.r00ig20ec%6el%oosfosfPoFPfHFbSHo)Sdysthwhoerwiogueghdhtoaiunthteha e71o0wmwdoetoekeksofof ttrrheeemasateimneaenntdi.moaTlnhset,rheawtfhoitrchehr,,oufagofhmotuarwtetshshkorr2t,eersheoorsfleothnemgieecraetpwemererinoodt,ppusethrbiooawdcs.kdTohsniimsciorlnaetrsruovllatledudieeitsnaan8drmeecoavseurryeodf TAilhnttehhoePuFcgHohnSt-frrooleoagtirnomtueapnk;tehiiondwgenvroeotru,chp4saonhgmaeed,tdthihefefrboaolncocwyeiswnegrieegmhfatfinesetcde.adily decreased in he last 3 w`pbwheoeradeitykgscof.anatanT)d.habiTlseefccaaotlnnamctableseursdee;exfdfpitelfcmatriiowngmeahdisnbbcpeyrreobtaebhtsaetebpedlrrypotmlboiaamnseeemnanatscakuleurotsesoesnpdielnbafbysaomtdIaitniecessrspeuataensnde(daamsefaapatrsteumyfraaererckdedenarcsfoexoifrodawthifoaotnl.e oc(xotinednsaduteminpocntyiootvnoewraancrdadrscboodnhecycodrmreiaattsaeendotxRiedEnaeRtr:igongy,reoaxusppemnsediaiztseuuwrraeeswdarisnattehnhetismameantlca)eb,doAlindctdchiaetsgoeenaaanlnia,mlayOlseri.s Anotherindication for increased oxidation s the increased auantty of mitochon, sors 33 TNOeoort of sty W802 PTFoSalinrAeporpt Enmics 0122006 `iosxoildaatteidonf,rhoempatthiecsaicgtniivfiitcyanatnldyleonrlmarRgNeAd'lsiovefrs.enFzoyrmfeursthienrvsoulpvepdoritn offatitnycarceiadsoexdidfaatttiyona,ci0d. AtPrcleyaalst-mmCaeonActhoopxleiersidtoades,reo,wlhsaihncodhultwdraibsgelypmcaeerraailsdlueersleewdde.rbey astcrohnagnlgyedeincrliepaosperdotebiynPprFoHfSi:throughout the AtbpoyowaBa-rdcdeoscnrltaearaigseneri,nignlefhsresapcdatteiinocnsseV(LpVDaLLrDtLpi,reoIsdDuwLcataisnodnobLasnDedLr)vdewedec.rrTeehadesesecedreupaphsteeandko;elmwcehonenarveecraassni,obanesohfeixfHptDliaLniHnbeDydL tiInhnelsiuivvemermr.sairzye,weiivgehrtfuinvceiriolnispihdacvoentbeentesndcihdannogtedc,hawnhgiec)h, iisnfcurretahseerdsupplpaosrmtaedALbAyTanvailnucerse,ase iaMinrtceorrceahatosenedldriimHiatMliGnsgt-eeCrnoolzAy2m7r-eehsdyudicnrtoacxhsyoellaaecsstteievraiocttylivaainntddy wdbaielsceracechaiasdnesgdyencdthohalesessewtseel,rlorles7pae-chtyidverloyx.ylase, which ``eCxhcarnetgieosn iannhdepinatteiscticnhaollcehsotleersotlebriolleaabcsiodrpstyinotnh.esPiFsHmSigihntchraevaseeidnffelcuaelnccehdoltehsetefreoclalapndidfatty ainctiedsteixncarleutpitonakaenldcodnevcerrseiaosnedanfdecsalmabillecahcaindgeexscreintifoon.odOinntatkhee coatnhenrothabned,ruclehdanogutesinitnhis rPeFsHpSec.t. There is a rend observed towards increased intestinal cholesterol absorption by fSgbaervnaeetresaslw.hoFifoctrhhteahriveserrreegaeufslfoeanctteistdmobifygPhPtFPHbAeSRoGaf.ruesseitmoifluatrorthterhoisneveosftiagaPtePtAheReaxgporneisssti,oen.ogf. oltihkeer Aefsfekcntoownn,intseuscthinaclofmupnoctuinodnss ilsonwoetr kpnloawsnm.a lipids and decrease fat issue as well, but their `o4AfsdPimFseHanpStp,ieoafnroeelddl,oawbaeondvdbeyt,h8einwpteahreeakgmsreootuenprcs5onmatenraoilsmaudlriseetdwiagtraheitanu,2mtewhdeeemekofsfreecttroserabltyemsPesnFrtHapaSitdtalhsyetsoheiecgonhnetisrntogldroosuep. veranellmuaaerisgn.eeTddhaasntigdwniapflsiacnasnomttalytAhiLenAccrTaesasesteaidyneuadlplsitoonmswteeawenhckaes6t,thahiengdhceafrseect:ahlaTnbhicleoenlatircvoielrd.seKrxeecmtreaotinineoenbdowdaaiesist.steill rt`ehcdeoummcpieacdne.y.,STwohielit gvmiaivlgeuhetisnbsoiefdtephlianatstdhmuiarsiPmnagFttHtehSri.scpoenrcieondtrPatFiHonSs,wawshincohtwyielt btoetadlelyteerlmiiminneadtebdy fSroNm 53. Effects of PFOS in ApoE3Leiden mice "mITnihsceteaenfincfeetcshtismsoosrfteuPdFpyrOwoSenor(uegnrvcoeeurdpy. 6s;i0m.i0l0a3or%)thoonsethoef PpaFrHaSm,etheorwsevmeera,suthreedeffinecAtpsowEe3rLeeiindesenveral `LdToehscasrtemoafes(aendpsRetErhRai,t ggwrhooiuncpahat6sdthimaosiswlcueee)d,shasotpwrreoendgf:leyreinnccerfeoarsfeadtpolxaidsamtaioknetoovneer bcaordbioehsyadnrdatae oxidation, ``aacniddaoxciodantcioomni.taAnddtiltyioinnaclrleya,siendcreenaesregdyqeuxpaenntdiietusroe;f mailtloicnhdoincdartiioan,sfhoarvaenbeiencnriesaosleadtefdatftryom tTPihlpeaopslrimvoaetrescihnoofplretoshfteielsreeoalasnaionmbdasltser,rivgelydcewriitdhesPFwHeSr.e sAtproonBg-lcyodnetcarienaisnegdf,rawcittihonasswiemirleardecchraenagseedi;n ttwhrheeear"tetmaaesrn,gteap"e-srHhiDifoLtdpinneaHtkhDiiLsngctrrooewuaaps.reddsilnartgiemre,, lseusgsgdesetnisnegptahratticPiFesOwSasisoabcsceurmvuelda.tiInngtedreusrtiinnggltyh,e ors p34 THGmpora sug M02 EAinSliepnorAt oENnGcdeen 0s122008 FpinulcrartsehamesaremdAo.LrAeH,Teptlahetevilecilve.mriwLciervioegrsohpmtdaisln,cHriMegGals-yeCcdeoraAildmreoessdtaucn3tdtaiscmheoaelcstewisvititethryycwloanescstoemirinstcawrneetarlseyedsiingacnnridfeiacsaendtly cdheoclreesatseerdo.l 7a-hydroxylase and mitochondrial sterol 27-hycroxylase activits were. PacFiOd Sexcdirdetniootncahnadncgoempfoesciatlicohno.leOstnertohleaontdhefrathtaynadc,idthexcrraettoioonf,dainfderdenetcrneuasaeld fsetcearlolbsiiln tabhseofrpetcieosn.haTshbeeleantecrhmaingghetdb,eanadrefPlFecOtSioinnocfreianscerdeassiegdnihfaipcaatnitcncfhoslteisntaelroclhoslyensttheersoils, but other changes In intestinal cholesterol metabolism cannot be ruled out. ATiskewaitPh PPFAHRS,agtohneisPt.FOTShsmencothiaonninsemedofsafcutritohnermiingvhetstbigeat(ipoanr.ty) understood as working "coTnhterooblsdeirevta)tairoenailnmgorsotuipde7nt(i2cawleteokhsoosfea ihnigghreorupdo6.seAnofdPaFltOhSoufgohllcohwoeledsbteyr1ol0lweeveelksssoenem 1lraermrgaeeci"onHveeDdrLaattaforrasci4tmiiwolnea.srkTlshe,veesltehteahlrIpoaourpgerhooilunetdiintchapetaiwlothneosrlntehacptoenPrtiFionOduSeadncdtoontccheoantnttPraaiFtnOitoShnesissinnaotmtheeellmiaimrgciene:ated rmofoetnahkdeeyytsfr,reaoatmnmtedhnehtubpmoedaryin.osd,,PasFhnOoduSdleditscekornnmfioinwranmtittoohnihosafpvtreheseauplmoplniagosnme.laimPiFnOaSt-iocnonhceanlirfatiinornass,at the end Tdsahitega.hotnMludycirfheemrleaenrstskacfboalmtepyodasciiiftdeisroenhnoacfveefbabtetetyewnaeceifndosu.gnrdSouoipnti6h.nantfdeec7essitsfoaftfitoygnuranoacduilpidn7at,bhwseohfriepccthailoanelxs(coarnpedot/isoosrness a conversion) seemed to be increased when PFOS is withdrawn from the foo. Tdiafbelreen5.t1mgeiavseusraendopvaerravmieteeworsf.the efects of PFBS, PFHS and PFOS treatment on the 40135 Tralo--5.1: Overview offectof PBS, PFHSand PFOS restartonmessed 1i 0 A bots | I| Bi i 5 a |) FESR] i LLEE I iiE HF iiedaI i | H. ia i Sep Leb H } .it3 iH aiii .. - = 3 i 1 AoengMi02KloFinTalveN rsioO n eport0ao 122l 006 CONFIDENTIAL | | AOPFPSETNUDDIYX T3MO-T02NO-REPORT | i Project study number: 031.1074/ 34-02 tH | -- [il I{ TTNNOOBQiuoaslictiyoonfceLisfe |{ SF23o0emCEencrsosn" ASseo:n i PfF riotne T TSSa1Iva0n0 HoFsaptemeadntotel ii1 neeseslaDeotparent 3 SJusoymoSne uPhe.D.CnH D,AST i: `V0CeordCpaeonrtatteDereSpBoauernkmtiesnntgt220022 435 SPsLrPohanule, Foo Minnesota 55144-1000 +es1i517 1062 uif . y sEumaaltatcyegronofifocnomn or ou Conen &i1 i Sts ng On E Final version: 04 December 2006 i ||1 pDDrao2rcs506vGCeuretser:r220080 ` ! ------ | ATuetchhonroilzaotgiyonmbaynaOgrerPoHfAV.ascQuulaaxr,and Metabolic Diseases: 0 nf | Page AT of A172 iLe ' # MopendicloTNO.qepoof M02 Finslversion p.37 04122006 Contents Contents. Ipa)ge A. Code ESL mice before and after matching 03 B. BB1o.dyIwndeiivgidhutaalnbdodfyoowdeiignhttaske measurements B2. Food intake measurements. naooas no? C. PC1l.asPmlaasmmeaascuhorleemsetenrtosl measurements AA1133 CCC23a... PPPoolooallseemddapltilrpiaogsphrmcoaetreAiiLdneAprTmoefmialeseaumsreuearmseeumnretensmtesnts. AS nA274 C5. Plasma ketone bodies measurements 2s D. MD1e.asCuordeememnitcse ianndmemteatbaobloilciccacgaegses a228 D0D234... AFXvoaeocrdtiavaginetydbmoderdaiynswkueriiengtmhaetknetmsim.ceeasinurmeetmaebnotlsic cages 2A289 38 D5. e0x;cchoannsguemrpattiioonm,eCaOs;urperomdeuncttsion and respiratory Ast 06. Energy expenditure 50 E. FEAe.cMaFleeabcsialluerwaeeciimdge,hntsttmesreoalsaunrdemfeanttysa,cibdodmyewaesiugrhetmaenndtsfood intake. As3 As E23.. E4. FFFeeecccaaalll fbsaittleteryoalaccimidedammseeuaarsseuumrreeenmmteesnnttss, 8803 At07 F. Intestinal cholesterol absorption A122 G. LiG1v.erLwieveirghwte,igphetriagnodnapdearlifgaotnawdeailghftatawnediglhptd contents iver AA1t2z77 G2. Lipid contents iver Az H. HinMiGv-erCmoiAcrreodsuocmteasseanadndstcehroolle-s2t7e-rhoyld-rToax-yhlyadsreoxacytliavsietyaicntiivviteyr mHiit.ocHhMonGd-rCioaA reductase and in iver mic-r2o7s-ohmyedsr.oxylase cholesterol-7a-hydroxylase dria activity A136 A136 H2. Sterol activity in iver mitochon Alas 1. Measurements in viro experiment (rat hepatocytes) A156 Page A2 of A172 Appendicto TNO.reort of 3.02 Final version _AppendiAx. Gode ESL mice before and after matching E e E er e e e s e etee a i ect rte eerr ] ] [w e m eoe r rd ] r toto] a e sa rr ] i m e e ese e tor r ere] ]] eaet a e me e e wrr h te5 o]rh] e E e ett ee h re o e e ee ao r e e] ] e = r =ate r er : a e ae ee rr r] H] [C o o sea w s r ams r hges aes a enat Re saee]| CT E Caoe e rrr fe e herira etoh s aa Ae gBtas ien| ee [ E rrea er ] ] A | e A et er e teor eto] ] t+] m a r ere se to t e] a [oe [ -- oe aeo -- w o-- Tfot otho-- olH a ed a =a= i a m Tao se a TTooT H afaeee ng ee E[ [Et eEee eo Ss ah ta ses e-------- ----t ----1------ ----------| 5.38 0122008 PaA3g ofAe 172 AppendixtoTNO-reportof M02 Finalversion `Appendix B. Bodyweight and food intake measurements. 81. Individual bodyweights [F E r =[e a=a rH Et mm e rauetr mr arera te tae)r] EEPP E eeeere e e a ertae t er r rt e eerrre a a eeere r ee ra] rae] E E EE P e e es ee e ee ttt eeeer tttha r re rebea erat bbbbeae eaarrtiae E EE E PBeee ee etae e eat--ttteteteeet rttrttteharrarrae raaattrnabbaarerr riteaaann) E E F F E e e e a e e a fe at bt bh teeete e e aa aeearee eeetr arr]]]] E E E E F eee e te e e r abea a e ee e e e a ar eaenaeo)]d] E E F E E e e e e e e e eee e rr e aaeee eeeaeaae]rr]])] EE E E E e s st tt e t er a a reat e t e aees arrere e tae aiaa]) EEE E E ess tt e a e e ee t ttebe e em be r baer e rra]e EE EE E rstre s e aetr eet the es hre aat b b baae e erre taae aar E E EE E sm ste t s amteree e ieaaar amnamebmaaearmrrmoara) E E E Es e ee t e ae ae t r bae aere a] a] E E EE e t e e e e m e e e e a m aa e e raa a err aa T]r]m) EFE E E eEtee e m te ee o r aeeaeee] e rar aaaoa]r EEE E E eerie b ee t E e el t r rt re a eE Hbe a ei] m e]] [oni Tecesran]o713161 308 |391 310] p39 04-12-2006 Pa Adog fAe 172 fomto THO aptof We 2 Enatvasion .40 oerpzn E TEE a TERETE Eee Ee ae ae ge ad de e ee Page AS of A172 `Aopen10 TNOrepoof r0t2 Final version p41 06.12.2006 SE ---- re -------- [J JoJET7 oooc:STaNrNSooycJ-eoy= oooL |i ToBro|JlEerrcocsos:JJ etm oivc:aa tal Rol J CJ oooorcrsSoB=oBBYEoovcrJ ScfoooccooISNREJ<oor<o:J ooJesSSEUE o-o-cccr:|| Ceof ofosrI BY oc| Foor:SErovcx:J oaRRE o<c<| [JE oo SoJYcsSE vcJY ooJ ocBE | Fac} foe] oxiSRoceoJ o>SNv0 CTR eT le 5 Ler ee le > er 5 m= id bE oy on co oa ol s=RRE cDccso:e BBEE occoooossEREEYoo+RBRE oofooooooSSYR= YoocJJ oJ0oo:SRBB05oc7:|1 [5sRREEEEo=.oNE BcosoSBYY> osBBiR oocoSSTR)Yooovoo:JJ ~ oscSoSo| [[o=RSE oorwoofSiil oovseRSEE oooonrSEE ooSYcoJ ooc>eSR Boooo:| [[[2foo3l8atlooouotrrs:8 RRife ipfeoo2 zomfocr]] Ro Io{feocooo0r I GFoalRlIlooorctc:sff0Caaissfii2nmtoooecrs Bipes1geoatl oooirrr:s] E [E 5m Jools Jpg osJERe 75ROY<1}ERY 05 { e ors7s 1 O1 pes1 o7r|| [{v5stSat [so oooSeoootr:oBBBmiiingttcacor':oosRRRRpOOEmRREYYooor>ooscEBBBElEoeBltoooooorsrr[FiEelstonnlnooalis:knia[ka minoooo0oor.osrt0Bep itoRBaiaisotitaolros|| [o [=Fosoo foo:fRiE 5 {orocsRRBUUORYooBvsi:Be Bie o000c1sFuKernldil)c oofaie na oorucssBREcoocl: oft ov rooboll ovcs ttosi oc m(cComo aotv= Roa FocsosoRRERYRRYc10o0BRifit ooor Fboull]co Jouno o0s7ss Reo t1os.] Page AG of A172 `Aopendit toTNO report of 3M.02 Final version p.42 04122006 82. Food intake measurements Lo TeTFerar[eFeaete[moon E sForcdorica mFausre[ios||Foind t]e E 2 E CS T E2S0 J S . 2J 002 Ye ||| [FC oesweo eeTo mosers mo0os0TT[ooo 0sSS iwa mwmsssS maassr [[[77711135aa%m]]]| C C S S TS J . BBCC E 2B J T C 1 2 2222 2 |||| C C 2 E S S 0 J J 2001 YJJ 2 ||| ES 0 CS J 2| C r SE S 200 S CE Crr T pJ pi 2 || C 1 1T ww JT--ou--] Page AT of A172 p Append toTNO.reportof M02 Finalversin -- --_-- 0 --HEE rr ait EL e HHHa em Git Ha LsL oiln y hi Hi I i HH F= eili fl I] ll il Ll di re HH afHE H e EnHH ET Ee Le usee | mi i i HLH L i Li HH inti = ! 1 FEEiLb Ee ilies i ilCl Attia st EeI FH eLe i dy Fii [E im i Sl a BL - i] = FEL IH pedeheof A172 "7 x nixto TNO: 2 : IlE E dr lras i ae e ted Ht a re ee lfeil e edfaem a m ie)| F E P oso ee ee s e ae e ee aa areree] ] E Eee eee eeere a] E P P F e e se e e re eae r r+ pe e r e ee_+e +e tae +e_rP+e a_ee --+e +&| F E E e EEee rr e rae a eta etrhemem F E Ee e E ee F ere E r ee re B E ee e e e r t e eeeee ee ete oe or e e e eeesp Sea te e + t F Ee treetba ee ae e ttw ee a er e|| e e terete eate ee taerre ae e te ar tan ter| w F B Pe o e e s reeerrr eae e r o eee p e e ae s a e rts re] eee]] Bee ete rebee teen oEeE be ee a F EeEo ee E a etee3S eot n--+ S+ttar+ PaA9g ofAe 172 Aopen toTHO copon r 002 Ertan pas oa122008 Lo fT et Ti aalmeesR sEe ie n geeA e mre|E weei| e ] | sgg ense] fPeE steatn eR eneo mne a e n et] e ] n ]|| E F F E-- e r+s e Ea w s -- a FabH n se ia -- m e a e+ e sr ae| FE P ita ee e T ee ee fJeH E m cifons] | mn e a s --cageo|a m n n m n t e e 1 n nein E] e H E m En saa E a e m e ee R mm ffE E Eresoetmi----man te w e e a T nw e ee eee e e PI e --o t--rs er Ee Ee h e e o eee aeee] s i]] B Er a t or a t eem r r arr e 7e EL r CX e Ca 7e t m ee aNOo 1 e e J e r Nr Ca Ae |3| fh P ee aR ea e ee s ss e e e Fat Fa ae1ar] eea eee] ] E Bm E EC P F e F-- E+ mae rte m r m e e + e a ar t pHeeeeee] ] EErEe reeReere eeer a eea e a e e erreie ee]] BEEer a a --T----T rs 1 + + 2 Page MOOAT72 ~Agpencisto TNO-reofp4o.0r2 Fins version ln e P Mal =e r=e e e dP F E Er ee e E F P e se a e r r ra e F E r ra s r r e a e e F r F Pr er s i a e te e s FP E e t r e ea e E E E e rt r e e ao e a E E E r e ta e at t r a ee a h e m EF E E --t eC+e 1 -- ma+ + aa . = t a| b E E E r s e e ar t e e s p E et e e re| EEE Pe Err me e e t| e ee a r -: | EE E e e ra ae ar tee r ts rE a E B Ee e + re o e em e aa = E FE r F ia rr e ea s e r e F E Er e E eE o osa err e e Fa etH ] a mr eeee C aa a E E E me ae eH r e e E ]| aeese em em a pots 04:122006 Page Al1of A172 p47 `Aopendix toTNO repoof r4:t02 Final version 06.12.2005 ses e ) m J = r reoF irs mao pRsepal se aortfasee:otfaeiprtSrLiearpslTfearprof]y --aroem fF C GR n Gd om med Ga ol a r rE c ECR R c R RER Rs RRco BB CR R ea EE oFioa L liRro=r al EB ofr eeof of Bi bE Dw ml ma ce onl Ft Bc Ro Bo ScJ [m E e ere [iperel LertRRalcCCRpNei= ontuCcosiRnlcSoEnaiFcnotoc|JrueiJ<not<o{Jrater|| e rfp tEc E oti5n3 o etr n13o AofJO ca3 to0caa ooSrSE3a2 J 1333t1of| E E t E oRoCEee EoEEoTvE hE o E SC to E ES r C RR l p RoRREc FOR 5 op oc ftv| Page A12 ofA172 pas rs -- 2: AC1o.poPnldaiscmCa.cPhloalsstmralmmeesassuurraammaerntss E E i E Eh E Ep r m Rh E m e e E e r e E ee e E hcyeTmc)=) EE L E omdR eR E H e E eE eeeeee EE L oE maEo mne n e mle ea e a es e EPF H EbEemtitamEnE teseh e e E a rE E e a i e EaEe te HHE F H EEHE EEE EEe e E eE e eE EeE EeeeEe eeeaeH)E)] H E F E Er E e ee e e ee a ee E HH bH bbE et e E t C e Sh e eeeeeaeEe EE eBW ER ETR CEE He EEE R E T e e e e e e e e E E EE E ReE aee e e e EeH r B E H E H iren i e rahe hs D at rm e ra aS tr e ae r teHaaeFaeeHe) FE H erhiRa aE sr he HRm RE e ee esH ee e EE B E E EEE D EEi E i e e H n meeEE eE e ee uRmE e Ee wt Eam eBEEEo E Tw boErh Ee E Re w Sel Ee e E e t e t) ] E EH EEE h aee Page A13 of A172 pt coool 2 Fialversion or122008 C mosmm------------ E CoJrwr rJ 3ot3 Ie oe0o r LayecR3eee1rxa3 y| = F Foc nJooen:|LepaarrI)Drpoisa Ree0t0?0I on fordsa [oss|leJoss =EE C =rIC C3 i R I 3foto CYtR RoeHBCYEEC0 pts I I ICer a crfomto!| [EE E rriapet acrtoH cpiaeppH E erpai N redBntoefisapaiE =ie|rE h uB ni i{nI agpa R apnlge patearn) PaAt4gofAe172 AopoeTnNO ep f 2 Fa vrsion .50 sume C C2. PlJ asma vigyoeride mear suroment ts e e e E F b c o re mE ET Te E e e rEe eErI e E e Ne F BP F L E ors R e r S H E He e eee E HE E Fe F F FE oorr er r eee o r i s ereb rrer E s t Hrn e i eee eeaeee e E e HHn ie E F E E E E r o er E Ee e Ee e E Hn R ee ere eeH EreeeE Eo EEon RhnEm mn Ime inabR eernrRE b ee eRHSESE HEeEEae n2 ECE E oee r e e e es e e e ee e Heee ae F EE En rp rbe n i e r r s i aE br Tr h a ne rE d e eHe e eEe Hr EeEeee een) EE E e P ErhE eEr E e e E EEEe Ee E E erEeeE T E EEEEECER oHhs mEsE E E = E E e E H EeE Eee E E EEeEe C C P T e H he P r E E Eobmtmte p e e E e e e rees e eeere eaeeeaoiiyee E E PR ee eE e rm e Enreee eee e aeee teea ee EE E E E E E h T E E a e e m E h E m e ee eee reE e es e e E ErE e E Err r e e e HE EE EE EEpE e e e e rae EEE r Pa A15g ofAe 172 p51 Aopen to TNO port of 02 Final version 04122006 ssE es e EEESI3 E 3 E T3e1areee 3re3mto|sr3 ri T5r3as| = F E C eeC rI) forIi [eI =z) fC e) C oo =a fos) oor] [oFrioElR a pipap=R esLipaoptcR JapapoarFIfouarb at p+ apalLaas)| CER = ToI JoJ [CE EroRobaE pGahl bmE pepb Gssn o apao apnc{apapoDfoc gaspE oaers|apagt oadlofgiparoalt] CE Eei Ro atFt oofa o> Pa A16g of Ae 172 Aopento TNO portof 02 Fiaiversion G3. Pood lpoprotein prof measurements .52 05122008 Week0 me EE E rrr ee ec a tr bree e eaem r Preese ate a Ee o ae 1 ssa ae 1 ee He E E Eor ee e e ee H H e u e ee b ea te EE E E r re re rte reb a e a e e H e tb rae ee--te t aaee e e r r aeh e e a eea he hbbeeeeee E EE Err bar bb e ta er PHhbeae essea e as F HHr oha r ae e oe emeebfba 5 [E E ZEZr e elm nob nr [H EEe e Jee] ns r oe[Es SE Jt oe]e ne er e[Re 5e 1e7e|] EE E re er e ee r e e r e H ee e eehbee] EE E E E E oe F e e e e r t e e e b e r e H m e a e e H Ha e e e e ee her e e e e ae e e e ) EEE EE E oEoe e ee e e e a ta a ee aaaaeemee e E FFHe ErhhE ea e eeeeh e eeeeta h ahFFeee EE E eee t e ee eHe eee e eee eee]e Page A17 of A172 Sopa to TNO reportat M02 Eats p53 outpz008 EfE wrerekmd a ee e L2 m Deoee=se eeDD p E p r pmEE edyd p e err o ee e er a pEF Li P E hEhrper eI toI rm iTE e e aer er L re ereb ee r I e p al m i iC ee E e ee e er eE rr E e e RmE n H prpEEp r HEm E E Ba Ms eE e E bo e ee mrH lee b lla .) n D R h H eE rma mE el re E B ml eE re e me em rs e eeE r n pos e e mi e ea ra E rE EE E er eeeE E E pE r T EE e eE r r rE eEeeE e mreEr e e Ee e re ee m e e ee e eeeer eee| ES Semee bee EE E EE ea E ay E rere E E EEEEE t E EE E Ee EEE EEr EEeE e r e E ree e ee rte er e e R e r e e e e S nersee Page A18 of A172 saves Tota tase pst -- [EB ErEe e er [r r 2a fe e]5e . [e 2e 2t Pe]55 EE E E E EEEEe E e e ee e e e e e e e e t E E E B E B E E e o e e e r E e e e e r a at a B B B E E EEEo e o e e r a s ee ee e e ao H B Ee aHE EEEEeEEEeEEEe E eE eE E e re e E e e eeee e e e e e ree ee EESEeSEESee EE.=E === ===Se ES SSE e SSsSSSe Ssie EEE EEE rE e EEEETEEe eEE e e TEe Page A19 of A172 Aopend M02ixtFoinalT versN ion Ooport04o 122f 006 CEE EEEEe CEE ET 59 Page A20 of A172 gendi0x2 toTFinNalvO erso.n 1oport012o20f06 v.55 A M FEEEEE EE] = E E E E re e r e e e b =r e m e T e e Te eee r r re e e em F E E E E E re e e r e e e e e m e e e e e e m e e e t oe e r e e oe m EE E E re e e e r ere e ee e o e e ee e e e e r rm e ee e B E E E B s r e e e e e e ee rr a e ee e a ee e r e e e B B E E ee ee a ere ee e e err r ere] EZE-E EHE sE anEt F] Em eei ar e Em =F = SESSESSSSESSS = E SE eree e r eeE eere e rleersee e ee a ee eee] PEER EEE EE EEE Page A21 Of A172 to THO sport on p57 " Week Comet - E E Eee se ee S e e e e e S e e a a EE E E E B E ree s e e m e e ee + e m e ae r r a r Er E a i a e ee ECB B B BEEEe e Ee a e Ee eh e e r r r r e e e e r r ee ea r r e er r hei e B B E B EE e e m e ee r re a i Cite amt+eaa EE E EEeeEEE ErE s E E e aaEEPmeEee r e e ee re EEe E ee e e ee EE E EeESeESe Ee SEIStERe E Se Se Ss Sss e Ss EEE EEE EEE EEEE ree Page A22 of A172 Aopensixto TNO reportof 04.2 ina version p58 0122008 = [EE E =eeEe e Zmmee lSeEee e [ee 5E 2 om e e] 5e 5 e e T [2 emm e [oe e]7eee= e|] E EE Erb b e e ere r r r + e ee e Tee e e eTe e ase ee r e ee m E B E E E E r r rb e e e e eerr e ra e e ee e e e e e e Te e e s e ee e r ee B E EeEr e er ree e e er a eer = E E ee eer rew e oe Pena EE ee ete ee orere e rre retrea Beee ee EE mer e ee e e e t Eee E EEE 1a 1 e e e1 e EEE EEe S ESES ECE ERIEBE SeE ssSsSesssf| PageA23 of A172 endictoTNO-reportof p50 App M02 Finalversion 04122006 4. Pooled plasma ALAT measurements [[Gm epi= freoms [a [we peo rs om[271[3[ss| 1 5] CGrooup up3 HHFF+00003T%%PpErBBSs|||| w7es|| 33[oests| fiese [Jwiiszi]] [[[GGCrooouwwprseHrFEsr0oo0cno2oisswsppprrroossI | 1 T1 0 avi6o2[|0 N 55511TX 13a s71[ | |A1. s0ori|||sa0 oe0i0|]| Pa A24g ofAe 172 Aopen o INO potof 14.2 Fetes 5. Plasma ketone bodios measurements Wesk1 dw Ei Em " | E BEER a NE Ee Eee E =E = ee E = ere EE SI E iS=cE cs== p60 out220m == ere Ee EEEeElre SH ERE Pa A25g ofAe 172 weske ApendMi 02xtoFiT nalvN ersioO n -epot04o .12.f 2008 mmm Ie M | eee r--- = - ee -- Eeee -- Page A26 of A172. cs ntco TNeO report of 4.02 Fina version e h E r Tr E rE h -- e eR r -- E E E E hb Re E a e E Fee F E be ee n e e| E E E E Eb b e b br r e a E Er br a e E E Er r e b E e e E e E b eh re r re R e e t E EF EE Ee EEb E F F E E re r eb e e e a e| R E E e Eee e e e e ee e e tt a e FF F E e e e E te m E Ee e me e ae e ae e r e e ] EBE EEE E E e re e e e a P FE E e r e r e e e E E EbE er Fe e e e ee ea e a FE o E n r E e E E n e i e ]| E EEEBaEEee e e cr e e e e e ] ] ] EEEe EERE EE[ f e eram E a e a| ERIS R= p.62 oat Page A27 ofA172 Appendix io TNO-rport of 1402 Final version Appendix D. Measurements in metabolic cages D1.Code mice and metabolic cages i i [z iw d| T ET o] CII -- FEzT | I| + IE TA | i| I-- I -- I p63 04122008 02. Average bodyweight mice in metabolic cages [[rZoeomearl seb B [ogi= mwo tabsobg cag e u |endmwee abrol] ie [ cage]vw ainsmeetw asboloiccaa [[ w esTs e % T 1 w x mmr sT : | a mn es | [(r 2220 5o0s mmssT Te wr i 0m Twmmnem | o [o er[om To s T e n wrT [ma emm mw [ [o oo Ta r T T Tw ee r aT Te w r s a mawa w e | [oo e e 7T Tes e Ts srT Tmw o | ss 20 zs | as | mw | Page A28 of A172 Bt ans RS Footnae messremers EB E FEE Wo ----m ------a------C --a----d=E r =Ie ------a E --a H EEBHEE rEEobEpE eEoprE sarreeE r re ee reeaslpeb b E = ier re se E Ha e e BHB EB BEEErbEo o dietsr r irna aAre eE t rrr r rEeiaE rs s esle eE b eerrveeab e er ree e e ee een e aEe e n BE BB EErEEr E bo ee r rr s aE re rab arE ee ma --iaies Erbin rei e B BD = ete ra= ne eee r e e= e r eeeree BB E B E EoErr a eE rir E rs eE rraamEBms= rroseler= e eere e== r =e= a a = e EB B BB EEpoEsr o o oriti r r eeinred i e esrrbee e re m m Pe m P e oeee ee e e er seee H H eaea r a e a B B E E B Ehr e r r E ab r o rMn R a ee e rte E r a a imE Ha e e p r oEtmS ee s n ea Et i e sy E= S BEBE BrooEbE eemrraerrae rbrE iseriregisEo e e p E a e e S te e B E B E B Br r r r o r i E eb e e r rr err E ra r eea ea e e e e e ee e er e r e e e ee e e a a a e BBE raermenreE n --i l Bere EH SHB eree neeraeh he--Ee y PageA29of A172 4x0 TN " Ee -- Final version p65 122 -- FiE t m mmm1 n E Mi iT rm Mme mt? ER s e E H FrHs ie ie Pe ae] EB rber r nraame e e se e e Pebam ee rae er] ersbE eenee er Ferre rae He PeeHa es baTs ee) E E hreae e e E e eP Fe ea eHHee art ee ee aee)) E oamR 2em am mt ns Erb reee ee r rHerbal Lae Hey Page A30 of A172 `Acpendixto TNO.report of 34.02 Fina version p.66 04:12.2006 rr rsse [eero ][nal p LolFol 007[Iala]c5o[Jnyolopcc"lBalyalopeLIsnlolp| [ee] Bflo0e2s00]JYoso 030IRo[cosces BYoi| Joes [urass]] JJoocecs]| [ieJ 0cos RY 05 cooSRYbo+oooosJNEYRYoccw| BY coc] BocaBY Pocee fi[erss]J [[zzxzJJ [zz6J [1] [[zzse]] [24] c0ocIBYJMo0eRsE] 0ccJBBYY+ooooos| oo0cccJIR os00 coSMRRY<scoccJSRRY oo x oJsocee::| 0ocIY o0 sMYccJR ooc Josec| coo Yosoc NERYFoscscecBYRYoces| ofzozoo0]SREv0ocMREB3ofcooceccsSMJRREE++cooooocBSYBRYE0o5i651 BY0555BRc0 oBRo0 cRY coc JERRYooo [ooJ ccooooSSRYY 05051:sBREocMBBYooio:o|JR 0BRE0Joes] [ss] [Yoo] Bi oscNEYccoBYs0es a[ass]] [cooMY E0Emcm BBBYYYo0c.:ccoocccJBIBYYY ++oococoooBYPR0oYo| [ie] Pfoeo| BYo-ooBBY cco B0Ycos] fee] BE05+ IY0 IR cocJPY ooRORYo | [[r=l] BPE0o0o57}IY 0BBYPi<oceocoSSRYY ocsoscRBARRRoo0 cc| [ses] oo] BoisBoccc Boco [[5s2o9sJJ coooooISYY 00005s7]IYc0os coo Yovic RBYiBYocc<occsccJBYRYcococoxcJcBBOYIRR oocs| [[5e1sSJ roocceeooJJRREEB110o0vvrSSBRRRYYooo++ oooooBMEYYRYec0<so6SRARRRYEYoo++ ocoooJR oc | [[eoatf] co eoJRFp0a1o1SJRRYco+ooooBRMc0 ocYicRMAARRY 5 00cocRR coo [eas] <oYoriBR oooBPJ coo osoHSRRYY ccoocxB0R cas focooJY ovcSRY oooNRE iccNERY ososPRocY| [rer] oc RYBboooooss7]NJERYY ocBYPi 0 oc MARY c0o0c0ERARRYE 30 5>MARY oos2c05cR BB)OY oR 0ccos Page A31 of A172 AppencisloTNO-reofp4o.0r2 Finalversion p67 04.12.2006 Fooding bout size measurements. --r err n --mm)J 1fl a mn dmTm m E E [r i sor n s soom ot --sf wis --fes [tT oem m m 5 t sor{eo n onmem nf1 om a rt io ot on|[soo omu[----su sumer----fs feeoomm [[osrir ooioasu{oomm {{soiua{|svoart|--eeoou --ffuomds EE Bs ss e os nee T-soo0a[mn --stwme--t --ttefeoomm[[osroioon {oomm {osuuaee{{svtmsTT--seoon----ffuoumn T E EE s3 oE tsom nE mT~sa on so m m u n--ts ) esm f [ari om u {o1 m {m c m 5otu n ass 7n feun] Ese todmo1on oom0----sisn --tefom [oT rion {sm 3 Ts1o mTm s3um s+on n enn1o51 ESE oso Tor--vam ms--tt1o [sm isson {oi1 voun voT--s3mm--nfoe h E R at omnm tmo o|--soi--n r teon [oT ri{som tvme om m m sorTn n ahoe EF E so o r oSmob booeomas E t eoaoe wt wso masr----e fts ooftoomm [[FT toa ireecmm [ i {ve n{s o sum t {evsme n aseoos o u--fne omds oe one e0aeToormo [m soo Tomi oonn {om ma|--omr--1oms| Es 0 m 7 7 r m 1m m n n ERm fl m PaA32gofAe 172 `Aogendis toTNOreport of 34:02 Fina version p.68 ox12.2008 ErietoeFotetomtom|[tito osteo tS +t en mmmfm m Bre m aa ten a f E m e He Hee e se) Eee tesa Fare ne ee ae a 1m nf Pm E om om a amesame] aio|ooa oor om tarsi] EaEsEt m om oe me aor tm on ieem|[Faeer ooatoon oor eee oo ea sensu] ee PageA33 ofA172 AppentodTiNxO aportof 4.02 Final vesion p69 04122008 seio][ofl 5Tlffos5ssT]olof[ossJ[oyfBlooussc[MBaElootc55c0[EcPipIiRn oo5ooo|)) [sFeo]o oscMRoc] fozo0 BBEoooocss.:SREREsoocococccoRARE51 [zl ocBY i] Boos] Joo] [ro [rss] oesBRoooIREococ RY ocos RY oo ORYoos Joo] osc] Jose) 0.555 RY 056:IB Jro-200 Joes] 0-200BR 0.545ER 0.200| Jr{+o..0e00|SEYoccSRRJososssc)SRY c0 cc RY|Lso.s50:5| ERY0.200] [265Joos" RY50 :5 Bo200SRYo ocBURYooo| En [EX GE Jro.4e6) com oom ozo) J0.465[EY 0057| fore focSIRJoosos]) EQ BlEccR ccMRT|o0v5i5sEFRRYY0osssi6]| [Ex JoosJYo+iRo ceoJR05: RY 0200][1peo0s| os) |0.057JEN 2000 EY 0.02[5ERR 0.114] [e[rso]olooo BocoxSRYcfooicsSBRBYY coccoooMSRYYoossBBOAYY5 0 || [f2ai4t]) Jo200) J0o.sc0o EYY5o0ccoocS[RERY 6 oo 5 i5 s[ERY465 [a[riec] JosSRBYY vc0ososlMnccroocSBPBYYB5soo:oo5oHBMARBRYYE0 occcoo>BBPIRUARYo0o55c0) pEesn] [orJ Gc+oEoSRRcfooE0scYs}RoOssEcSRYBcYo5oosmoBIBcRYoo+cccoocBIRR E1oRE61 21] BYcoor cocJY ooJYcM [522] f[5es]] Jo.02) Joss] JBooeo]) BY Jo.025SEY |0.265JY 0o20.c00o20c5I| YJJ0o02cso0cs03BBY oc| [[sssco]J ooc RYi foi0oo.zo6co}lMiARYJ0co.io45ss6MIRYEY oo4co6co5[|RRYYJoo ccsocscBEBYURROYo66o6| bfocc:IREococBBY ococMARY cocPOYocc| f[ee[soesso]}J J0oc4e3cSRYo0ocf5ooo0coacslIREJ0oo+.o4o5oBBMABERYYYBo1 0v2i655sMB[NEERRYYoR6o00cc5 oocc5 cBPP3UIIRRRYoos1 cos [cosJ eFoocccoJRYYBYooo+vco:]]JEcosRBYBlE cococooocSRBRYYYoo+osissoPPIRMYoocc|| FFoooczsocso[[RRYYMooooiioss]]] BBllBccccooccNMBYYY ooosocosoc PPRYY PR oscec|| oc| <BBEoovoR]JRE055 RP0YY3 oc00:1s11c5sSBRRYY o0 00s-:7E[PRIRYoossass)| Page A34 of A172 `AopendtisoTNG.report of 34.02 Drink intake measurements Final version 5.70 os12.2008 Bk [ie on amaTs3 ao] vo o2mToo rue{contefon1n 03) 7 10 Cr E T are 3 re rier E r re Tin fo1u3l]Fr E oier owTrr inOE mn rHi oom) 1111133 T 1 7 37m 110 C1 1 E ems ea safes)Fave i11 wrne fo17n13] C1 EoRss e ee0e r1 ei Ter a fem n)Foeeeeto E a on T nee n es]13 e He ha Er earb He a EshE abeees m arehe be te ha) br ahe ecb Tmhy r Irr s + 13] Er I C C rr r r 1 1 CE Tr 7 wn 1 11 r 2 2 rn 1 311) 1 1 1 0 3 E d 0 1 111 T C 0 100r C 1 35 7 1 11 F 0 eEr erm e 1 1 e a Fm ee t BE+ a ee pese te2)21111 0 m 1 om 1+ 571| E E ee E E Be C 0E ee 1 ew E es aE ea] 71 Page A35 of A172 Appencic lo TNOeportof4.02 Fina version p71 0122008 Erbe e r tb i re r, as r Ht a) a Serena ena) En re e ee) imebie a] EE E E m rtm s imeorm t mt irae s nm|fhm tmmmmm b n nm e e teonm ERspeerenee hire be ieteen EEEre EEbE r e niepeeA aE tssea=erEllPPseeRrrEmlaeREmEe EAhEe Aeea e ceea) mm] EE EE r e re es set errrre ieetoeenoee rs ete e eu ] )FFPPEeRhEeESsSet PE rRn ese b bR e ee e s eE r n eee)snr Err er a a ebe ie es F Ea s rioe r ooae e Te ae e pae] Fa ertier bieaa s bet eee a n Eat to Tee te PealFatesbeabaa en E Er e eim reb a a --e am p re --a Fo] a] FP ertb are aba en e | E CE Eoamrm ss e eeae e TTiiina e ree e e a r re] ] ] ]mfe F ara bo e rn er e a eb aa t aee a erat e|| E EFoErCer s ah s e ea) d] F DF are ebH ere br aa t an e a EE Eirp rbee e s bebea a --ao --a H] a PPF ierrper b h eeeb rbbe eeaee e e ae ce a]] Coreen arte] (pitreToatie] PaA36gofAe 172 pendix 0 TNO repofM0012 Fini version p72 012.2006 tw [ole p Jalalp 1ololp lolol plolol Iolo 5| [s[roe]cJoFososcJRE0Mfo2oi0sJMJRREEsooo+ooooco]JERRY=oc<cJNR FcosocucJR:ERRE o2c0o)s foesM0i | Joocecs] H+frooomo)l rs} ooooooNMERREYBf0ooscsoosccIYo coc PRcYoc [1.000] [srsos]} (cHoEoDJ o+ oo SRY. Bcc) [zz cooJRoooJYc+ooIREcoc RY cc fz} FososJ [o1oc]] B: EoD osc I [[2ozr9.l]J cooMEYJooosoe] oooSRYPHoc.ococcSBRYY|+1+.o00o0o00PERR o0543]| foesSRYoooRY cccJR cco| [eFoes]sSofodsessc] Bcc foFsoecs M0os Bi ocoPRY | Boc.coocoJBYY+oooooo BPRYRoRcoc| I 0cocBBY +cco PAY osc PiBcocooscBEYYoo+ oooPRRoc| [ss] PByooseoso oB.oooosBBBEYY ooooooPRPYRoRcoc|| [EsOz oEcEcSRYEcsms] SfcEcEccD JEY 0ERR Eomo [[ssoosoJofccooosccMRREY oci0 oosos] Pb4o0cc0c0BJRYE+ccocooPAo Yoo EE Ec | [s[[soosrz]JJooccooccJRYY soosoeJRco ocMRBiScEc:EocDccIMAYRY+oEo0EPRY os [[osssss]] BoYoeoso: PiBooccooscBBEY+cocoooBY [eas] :: mEmm PBY0o5s5IBBBYYo+ ooooBRYAY cs FooBIRY 0cocJREio osHRY so cSRY ooBY 0{.010055JJYY cv00ooccSIRoYo>MRREE scooccNMRYE+ccoooo PPARRY o 4i | RococIRN 05MP socIRD cco RI P0 age|AST OT Tea ition one CES B La p P -- i e o o e -- m -- e e e me Rtr roe anleg|lLS F E eE -- rme ea s iT-- EES t r tee e e a eEe El | er em BBEE E eEofeermere teal rreaerraaennas Ebr Pp C C E a tae rrdep H aee he a e a hSn Pee PB C E E E oh rtae erre E E me i oREEieeeneae pppH =h=eb =c eeas rc sB aeE Ei a aac se r e Er EEE B DE rE rr Coes E E E E PH h s H r T ie Sr R rs aiei es snd e P [ p er e ae e h ape se e s e e a oaeSan eed ba B HSE Bier dr berr eEe me smere eeee ene ara eiteit teSesad e m E be ee es ee e e hhe n n e e ea s EEE Roa cE ee N aRl EEEE EEEEEREEm EDErTRERERi a EErTE e E ETCn E NEEE E PP o P Re n E Er e e a EeRa s eere EEF EEeieLrabEbprerE mrerirHsibrsrebE airtereterrsrS earin)rey HE e ebEieee ns e ao e ee e ee e aee BE EBEmE rEeRE aT eSbE eE rES rey PPEe eeErre e e e Hea es BEE FeeTas nsToaTeer seo [sas] E EEb nabre ErRr rebeiErEteE Fa aE m serTa wn Tos] [Tess[seo | wea| 519 | asi| vas Tus) BPD eeree e i ee a es e [Ta[ove| eas| ass|wa| imsTad) E E E E EE E EE B Se ee s a Fa Tana amn o omaTe as Teer) [Tas | ares|aor|200|vois| ews mors) Pa A38g of Ae 172 Agpendieto TNO teg01of M02 Final version p74 0122008 Em rbt er t r em e ema | eme ser stieeeas arrm Em E T rSr m m em ar e m es am m errt a m a r m e a i E f ee s erh r r e ae a a ] ae f m E m r tm m r me rrm m m m ae em m te f S f f f eeme e m eae a EEE r E r re m r E m r m Tmrea mam rreemm r m a amn e e tf E PEt ere reeebbeee e ae H ea e e aaa ]| B E E E Err bera m E eerHroEr R erHbbeea ra a me E sm e S H SEeeEe eeee be beee ee e ees b e e e e aeso e e a a] m E m E op r Em mm m E m a e e arta2)JS f n E ee ee m e e e ae ] EFm F aerrra aarrar aa r m r s ae a a ae r ) aJSS eere Eee f EEraer m e ae a a aee a] ] oEm r m ramtm erm imsmmm mHm m E e m m my m S r e e ee e+ e a] EEFE EeaErrtasErmbrEaE etrs reEetmea m ReSHrHam a Ee) E]e e H m mS e ee rr ereset h e r e ae ee a es aHe H] ] ea] I E fEm E mr e mm em emJ o f ebee h m m ea | m m m T TRE mm m m m m immm omm m mm eJ J ]J cJ T | EFm E eer sCmm a i m emsim eef e mmr af J beeree she ere m ea ee ] E E m ir e m a emer e me m Ea ma rJSSf eerreeHeeae r Ha ee e ] ae] Pa A39g of Ae 172 `Appendix to TNO-report of 4.02 Fina version 5.75 04122008 XS ps ore Wey Ue [FoJoCoorsetsSCiO oceoO roRgW Soc7iRSCEYYoiriossNJCYYDEo+crvcc CBKCeoi oe r[[eooroo -JJooosvsse|sJREoScEesoJRREEoccoccSMRRKYoJscoecocJMRYYYoocccscccBBBiRYoooosol)) [[[riezcfe loJocosso:|JEcYooe| RYcocMYJoococMJYYoocscecc|BIlRose ef] focooJ cocBBYcocRRocBRR occ [iss Jose SEs RYos Yocs RYooo [221] [zz Jose| oocezcSJEiYRfco0oocsecrJBBYYRYococoso):cNNYYooocoBIR fcoeos] (=oo sc oJSoolfciIEoooeoBYBBocoYcBRYYooacccosPRPYRYo1cc Fooflloo Joe| JossJYocBYos [[ouzsl] oes RY foeosoIYooccNBPBYYcooJBAYYoocccPBRRYoccic i [ozfoozo0[YoSocoo7SEYoccoooc[BBYYocoo oocMYRYo0o::|Ro ozs)[peo| [E20n0 BJmCoyEcSREEccoooEsm RECceomEcMY(cEcmcMAfRYoCccc:EcplPRY0Gsce)) [os JossSRY coo EososBBY coo Yo2:s ee] Bi ccocBo ocBBY coocSRY o-oo PRY occ | [f4r] JoSREHcooosJJREcoocsMIYB occocoBYRY o5s1c5 BPURYY +ioc) [oof oi YovoJREoc:NPY ocvsNAYo ic PY 01 [ez Joie JRE RYoccMY 0IREoccc BY co [ees]of+ooo BYoocs REoBocssRRYY ococcYRY occc P PRY Y<cio| | [ose {SooiccoJYJooeosJREcoosoBHERYY ococrSRRYYoocsBPURY R11Ys =] oriosc MbJooocs)osBNYoooosBYBBoo0lcIo:BYBYY ooooosoBPPlRRYY oococic| r[irsor]J osseSRBYl co2eos0BRRYEcosocoMYBY ccooccBMBRRY 1 v0c BPBR0ccYYo | {osSRY oooMREocBMYocBY occRRR ocee| PaAd0gofAe 172 5.76 `Acpendixto TNO report of 34.02 Final version 04:12:2006 5. 0;consumptioCnO,; production and respiratory exchange ratio measurements 0,consumption --rr-- ---- E EE er aeTaa e ee Hfaal] e am ie eee seean e e saH h EC E Ef o e hai eTe ae m3 eeTe]) oeereaeeee aeaaess ntha11 oE b iasw amee oee a t aea fPaae3] f ee eee a-- Seo om a ae-- essettaar]] EB F irer e tbe aestrat e m e aoh ) a) S CF eeeea btieer Paaeb e HHaeaee e a8st n aan aa E EFBEoCr e reete e eebre eeirrars a a h1ha]a PP P D ere se o E b r sas a teesss t e a saee me HhHaaael E E CRe ss EEe erE a a asTr se ao he a]) en Forbes aan PereHa e at a a t a H EE C S E o oaei s a aashsi oeHolanlFF F oraa een eette aaa e e 8 a 52 h ea) C CCC C Eorsaeo o ew rs oremwoars Ta oeoeoa w a shwei h es heh rha he)l FF F F oraseee Ha aeo He r eeas a ta seaaem sse aase e eaa e--he ae| Cees ia eeTon me Di] C eates oareme FC o C eieso ses aa m war reeeeeTo s s e ahnae h) ])FPFaR aieEa Ea E eEeE m a aat h a E C F Ce oB o trr s oemaettaaaai e aeaets a wea r tn--hstaea)l)FF D F arbe erHe C me brbaab aerrr a tboE na e 5 --r ai a e) a fa blal Esereser For e ranf e eaa r ser s ) eae aah Faria tr as ie a ou a l C e h Re ei maha1) C E oE AEeeAtRtmeA mam smE]| eo rT1 3 mm PageA41 ofA172 0 TNO tpt of G2 3 nr 012: Loa E n Iprmmmap E sietietinirse ee]slts E an Enn t e||l etm ommega ob aTisesim ten raes-- 1)ca -- ognTe]e]| EB Eerbebteertaetaa ta or+ btes DD b obea beb bm e oooerw s ttHuao) BEferreteramtm=mr l C T or mi btm aeet s Ht a | E Bre erras set taee te PF eeeterterbae re--r eta a o BEE rere He Ferberbeet Te BBE Behfr ssrte eet ttrr eeErrr a aae s taa oahn tt se SBC b eee rbbat eb a re se tbeta b teetrt = mwae u a ittTe= a a ae EEErrrEbteerttaetsea taathee FFeebbeerreee ata e EBreet re e er tt =a a tn berstee= r Merershb tr ae b eeb r aT e Ta ee e o FEE BEEerrt reer rrt b aeeeerr e rrm eoar tr= or aa=t+ +htt anfnFFP FF e eertreee brt aer nbreeet t t e be e ee a a ar TTa Tr Ta a oae BE EE arteer r rb eertb teetaa re aae t tr ta aaaar rttHtmens BF ereotbbe eatrrr aeetb t et ee t te e n t= b a at a m Ta Te Ts aaer) Brea e tat Ferbeeta saa Ta Page A42 of A172 ~Aspendix to TNO-rport of 34.02 inalversion p78 04122008 [amet omaToe fonssm susVu Tonated [[[erogieos]][aTeTJ5 oseL|oloJTopssL]alajifposr)i[eloBcloczsl|JolYI ool+LolPRo+Eospo )|| [se] fossa] fozooliRoYosscSJRYYo0oss BRIRoocozs| jr24] JFooosesess|) jioo]odpors oocBioYic:}MYJoo2u00s])BY oases] oz) fire] [oz00} jJoorud]] joey B0o2o0s0|BBYoom s BY [res} [Fes} yo} Jose] yo: Joss) Joe] 02040 J0-20S0E 0.656| zi} Joz} Jo.ii<] Jose] Jro.35 E0200] [225] 0200) oad [uaz ons 0114} fazRY co:| JJo0..543Ev E060606] [EXE] [Foo 0.225| oz] J00:7IE 0.020 Joass| Jo.1sc 053 ps] joa jJoos.s)] mE jrfooozssER owas [z3} JJooises) MfRoEcozosrBSRYY o0o-soso|JREo + Bi o-oo f[[2ss2ae4]]] ofsoscssBYJ0oE.o2o0o0IMIEYRYE o06co5oo6o}|BY|002.20000PERRYY0J0o4s0ss]|| [pci] [er] [ossJoBioco]sI ovcJY BY cicccsMYRYoo02c0PBYRY 0cos [ssc oo| BY 5o :SRY0200RAY 0200 [3a9} |oscJY 114] + BEER JFo200Ea 0.543 [24] Boo RYco =BBY +oooSREY055BAREooo] fas] ros Mg: Yoomoumn)| Jro0zoSYBEoocomccMBERYE2 00 00004 fiom) [ss] oe:Mocfoees| RYcocHRYBcoooIYoc [es] Jo.11IE 0200} J 0.566 ERY0465] |0.05 Ego + HE [oo | By0057 0.029} Joss] [[53s22o95.]] ose) BEY cocJovic] JoosREBYYBYo1iio01oo:s}JMYYoJ0o-2ooz0so0])ROPRRYY o0 vsi | [se5] }0casJYPY 0c0.o20s0|IRRo.c20JREBR6o.6 cosSIERY 00.200025RREYY00.250405| [535] [ez JoseJRBYEfGoo coRcoRoS}RRcoovooSNERRYYBYo<cc0o 5coNJKEARRRYY0c022o000BBYPRY0700 [[[eeeses3]J] s0ooJRfEBYzoccs}JRE1o0v1oJNPYYPico<o<csccMREUBYY1coooio1:c:PRBiR} Ycoess i1.000SEY 0666| EEE EEE [ees] {[oososoS[YoMocoo1oo7l) RRorovro RPYY0ooc50o4or3I RERYY 0 o1so14cERABRRYG40505ss6||| [24] |1.000SyB06o%)| BY1 0c00osoBEYY 05c5o6cPRRYY 6vsic|| Fox IY 28 0l00o2v20e00]4MIR co oH YPYBY0ic03o4oc3JBBRNNEP o2o00 o0svoR0:RRORRRRYY o01s1ec4e| Page A43 ofA172 7 Appendix " Final version 122 CO;production LF Est r I o e a ao e t s e s e m me t] m e ae tai ae t) s| E t D|pB r r -- ere eb a r e-- rae e rreeee t ca a me geea na| E E EmE r irmm r mi Aa neEtmr m e eeieatHaee mne Ta mesa o er m tte) Hee)]E DDD iEiEeom aEtEe aeee mTEh eEe tEm oea a e ae a = ee ) 1aa)] BEEE rRbbeEaars ttaaeesresr tHH t eorS e e rSa esa --e etHa)ee S DMe ee ire ee e a e eHr E eea a eee s e ase e e eea a)) r ea E E E EE r i re s eE E a s e ai rreees e a E waae e ] r)d Eaee a C BH aP eR re eEe Ee reeeeee eea e e s a heel Er E B ran e r Rb E e EEe ae beee ma as ar at mallB Ho eTEbEe eEH r e e Ee e EEs tEa eEhasaatm a) FF EeEtEeE s te b an e ee ra e e eoee eree eToaes raasea t]a E FFE iar trtea ssT rl seb a erem Tm roeaata a ssa seae n ee) [H Faeaoe treaeaee ahree reote a ooaeeo | ee d ] l [EE ES EEE r EEse Ee E s Am mms memea mer) Fe ee Ae wmme me mmme a --a--l -- EerotanaFoto Cie ee EE E E rrbe es o teoenerereoa a n a ssue H ealo[FPoiecleeer t|t oee w{Haser eTtoa eaee sa aea me ra] FF F F oo e one sese r a mt ce hsss e--er pllel e eeenee eeFr e eeen{Haoaeneeo Tooma nea o ns nn aeal ]l F EE E EEe Ea i E e Aa bEb eAn r t Em oe ne em aa a me --m isal D B aAiEee oAe mem mtr mome nmema r eao-- a s a] E E rE t Eetse [Boe f fe aetse e taeta as Hal PageAd ofA172 Aopen 0 TNO te01of 4-02 Finalversion .80 ot122008 I---- marae] BEEree Ere rete ota Cre iseaeteea --r e atm a Eames ea eee eae a EEF rerrsae ttea r rrr aee rtree am so re H te PCPeeereres eae sH b ee ea a a e Fa ee EEreteset rhe ea r EE mreEm Ee aE tem a eem ) r ffllP m me n ar naeab m nAemam=un Erberteteta mt Perera a Eerie e Ferreremma FF Feooerrbm earaae r Peaete taambe emte) a) FFPe eeere erreEee erra e a e e e a Sa a Ha a Ferber teteraTa] Sertabarata PaAdSgofAe172 Appendicto TNO.report of 4.02 Fina version p81 0x12.2006 [f[i7o9e%t]g1opoTrtratsfa3m3a Monods]Nr3a p3 v1a8v[1r3oBonrsCBraEnTdoerrEaEEgyo2s0] Fas cJooeso:JSocovere:JREocsJRE cfooieceSRYYooocBBOiREocco co [ss froze lilos|e Joo| [rFoossessSeBRREYJooocs-oso:ooMNIocooco BBY Rf0ous! F[iciolcl FireJ oJoossvsJEoRcYoovcooiosJMMRYEEEocosvooc|MSRREYcc0 soccJRcoooooBIORYEfoorocn) im[szosr fEooseEoosMRRYEcDoooomss7JMRYEoE1vE+o| RYccoomsSRYfooocexosBRoEBc0o)oos) [[(oa1scSeofoSoooconenJJRoEooo1oosBfREooovocc|| oocemeBloc)| E[[znooirmfooyc:| Eociocm] om roa] Cfoomc oooomBoieEenEs|| [[seszs lf-oooses|sMocooMER cforco REcJoosceSRiBccsBIR oBloo)c {oooJY ococ| Joses SovBR ozo0)[pos| [[[oasrssclsSNfoosoooeroSMRRRYY cscioososNBRYEJcc+ooocco:|SRRYocooocSRRYEFccooocooBPUARREYPooooso))l fa[ass]eSO oseeSE coooooJMRYEoo:cssc RYYcoocJRYY+o55o: oBRYORY+o| [anc SoSREoor BEccesMY cocPRY oc rl JoMo vc BoosBREoroRR oe| [[scceotJJ FooossssoeeSJRMYEo02i0o0cBBRo0v0isBSBRYYBoooovocosMMMEYYRYooiccocccBBBIIlRRoocossrss [ss JooJREc0ooBcooSRY c0oo RYocBURYis [or JoeJo cooBYcocSURE ooo RoPUR occ [[[ecosotorJ JoososseceJSJRERY+coo ooeoosoBoMocoicsMMRYYBococoscccSBRRRcY coscsBPRYIORoY+o0! csccJY ococJYoccSRY cocMY cs PoYc | [oo JoJREoooMocco MY ocooRY Bosc roourJSTY oroseeeJJRYEooo0 osJSRYY ccoocoMMYYooicss PPRRY0053ss Fer{ooosooMYc ovoMl BNoovBREEB o2o0MARY ro B icRO Toco] PaAd6gofAe 172 p82 nlio TNcOtenorof Final 2: Respiratory exchange rato --r-- ET. EE Erm7 r rr r r b m eeaar ooae me l mmaamr r m mef os 3 m m eooart owe o ouo m Toowmte ooo oommnr------fmm fuuraun)) E E m m rmR m emmm m meaeem ss f hem meaf f le H e m mm cmm r m ooe Haeen a m eee E E F arr r a re eR err e mE et see ea E o n as e ih eion)FFFF oaote ne i oe m owe o eeoe newe a aaer n einneom EE R E EaRe Es eemr e bbeeemR s e a ee se ae h e he 1 aealFT o aaartTe H eM Hh eM eee seoeenea r a ae ames r e en EE E rb RE eE e ame E m a f F T eo a ee tr teeet eeae aaaaesase| m E imm mE R rm er re emE E me m l Seefl e m eo e rmeo1 r n o es eaa| E E F F ee a ar m e ee s Pr Heem ea t e e t hhe el F o e asehe ao e eeeeeew e e e oonue se ee e ee n E n EEmR m e Ee e en m a t h e f aTERTR orro ee om mooo 1) ee mee ee Hee m m m m mf f 1fmm m m m1 FF F orreo e srmer o ob omb m o mm e e e en s f r s ) fom FFFm oaEirCooae -oeomnHoootmnem m +He oeum+a e is--e ffam| mr s m nmmm m 1mffl n r rnr mn mn) ERE see re m ae1 e0 ee ben ea eae f1ee) PageA47of A172 Appendix 0 TNO-rofeMp02o Final version p83 os122008 fo rr -- : E he r e te e He ae te s en ) m T o r o e hhm oee oe oem m ooi mm ere) Er ee Fea Fie eehe on oor E ateS eTe em) e s F om o foomm o oonm n F oomm--e eeena| ESe eaa eee oe eee] EE re eE Re ereHe eee aaa] EE E R R E rteeeeHH baa a s E af R R e eeeeeeee ee Te eenee eeeee EE F arsr te ee trermo e t --ee ee e a--s e Rhaa Fe fee E E o o r a e eoommeHoe e oeeemees e eae ao ssoeeae]]]| E Ee r EeHHeea e e Re ee eehee eeoene e ee e] ] EE R RERE EEE E R m H HE a E e fa ae m f re e e eeeae mr eeee Hee m am E r E ssm a E e bm eebe t mm ne mea--e ao E Fre re e eee e e RRee ee en es eee] eee oneee] E RE r BeRmbm esm e F m o elm memi m eim m ea PageA4Bof A172 `Aopendic to TNO regort of M02. Final version p84 04122006 [6enGlef1 Jo5ocsoTssNJobyMtco5ocsSTElfoo5es|tSolYd coooMlpYoi fvoiesTsEoalloBloo|)c| [[[erooosi JoocsossSSREoJo+ooenc sJBREoocvsioo REYoSoc+ooooocJRR oceoscRBRRYJ1o0occss)c) r[[zrsooo}ff ooseessSSRYfYr0oo5~0c]sJEooviecJJ Yofco oooosoHRoEl0oiis0BBiYo0o5) ros [rss Jrloooseeessss}SRYooococSREofrossfMRoEJoocsoeo:BJURREYoo+cocoBRRoJocosces)|| [[[rzzoiolsfJ oossoooosoSSRoYcwMoovseosJJJRRoEE2so01oc0sJREoocvoocuooosBSBBREYEsc0oocossoBPBRYR01000 )1| [[ozzssSfoofosccioss:SYS0=oss|JREoooSooJroiossoo:BBRdoYoco-ocoscBBoBRllE o+0oeo)) oooSocecJEooo RE ooo cooBocc) [zoofl [zcJ oooisssosSSSRREEocvcsooeooJJSEoowcooce:s|JBYEvo0oo}oJNfcooccBR B0. l [[ssoiszfo} oloccsossSSRRYYoofosscescsJSERNoEooooricooJSRRYYYcocoooosoBAREoE+ci0ossBBBRYo00scs))[T5055] [[[5a7soas scoloosooe| SSvYcSRREsorooomss RSYYoooooosocRRR cEos:MBYEfosss]| o[feesnoilofil-mooocivaicscBYoceJi ieooncSoESiomsSBcSJooooscso:SRYYoc:|BAREfof+oisoosms)]! fst S05 EococSI ocxJY os:NYooPORE | fa[seseol] [sir oJsoesse RRYYBcoocvccee|JRoSooreB|EYBoocococRfooococc:ocPBIOPREovcl) [s[sscsoosr li-lSooossoeeoss|RY YcoooJEfrSooovccJJYYcoc0oeoesoBBRYcoBRcsE|:DO oo s[ososl oloxoosos}SRRYYoo<ccJRErJovocseSoRYYocsGessRBBRloocoosooBPBORYYE+o) fe[osizsssJ oSScooeoosooSSSRRYYE+cooscss[RoSocorce RBEYcoooococcNSRYRoo+ccvc< BlBYBoooocs [feossotl oJooioscosoMSERRYYYococoeeoesSJNESccocEocSMRcRYYoc0cooosoNBRARYRERovcooocc|BoBBOYYoc|| foeososfoJoico RNYY 0oc::|(NofrviocJYfoRoovceeo}|RE occ BY[ofoosoos)] Cel or co oc BB Bo [ror Jcooess|SRY coxJEvrooemsIJRoioooBBRREcococcPORRORY0o20s0]| Page A49of A172 pos fot THD ptt y 06.Enorgyexpands EP B B Rm E R E eo e e )e a e e em E |P[ e E P e Eo fee e e s e e mee meae| EBE BB E eeroBe g eene = e E e e n EE E E ee E ee E eE e = f E EE BB B B EeeE eb de ee E 0 sE e r em e p ce e EE BE H B H B BB oEomCanr se e e i emm =--e e e n ee n obE t r er e e e e e e a e e E I H m em + e E [ e 1 BHE EH E H eeEer E b i r e e e e --m e osr r n E P Eerm E eeHre =Tia] BB B B BB iea eo e Er e o a r e re --e h r n e b Hae bee re n u2 B H B E EeHrEr E E a E E--= E E e EEe eHEB eeE eE e E EEr i = E BEER BREE Pa A50g ofAe 172 THO-report of3 inal vesi p.8s 122 EEEmereE e| am y t |=||Po a ahsm aea ty iat R H e e | | SFE C EEertb e ei arm t ae r tr a eet r e ain l b n ) r a FB B P C erhe ea te e nr be op b ab e e e ar = e s rHs =aa B FEFE B eoo brrer s b o bee bt rb e s te-- a e r r e ---- ah t --s t a --s a )PPE bB AereeEbb bpeEr eorerrhm eeerrna = aa e ea e eH ee--r a a-- eo--H)ae E EE BE rebred er tme aeiertt it n eei --i s --e e ae ) --s-- Fb P F p erbe e e er re e r t re r b r o e e e e e c n ) e e r BE B B EB robEr o s a e e mb e ertb r aearer ----a ed----a ----nis DB B P F er re e ri eeber r ner teeu e b E e ar e eH a r a EE E E EsEpEe s er na tb a Ee --d u--r n us)FED B EerrEbbh eae r e r bb eb aae e rr is e a EEE E Fri bi ee s r snee r re i erb ts n i orhe a e --ee 6hs --r s) )8 PB P b Pee ia e e se eR r t b e b b e e e i a e i n s or| F EBHEogh e bE reer ttbe e d ea es l e --smr --h88]) E Bonse ea at ) bPFH B EaeeeErrbre reebasaenr h brHiaeb e b e e e ar a r = a H) oinn) FP E E F E rE er e e A b bbem r ib e es E se m --epr e eREB E BF Fe E ee e rEmbar r eAb = er b ae ee e ee ee r H) --a EFE E E Heerreesom eg bb tee errb-- e bieer eeremni --ea -- --t --e ie ti ee) a a ----a -- ----)a) SFF B LB eeeEprrSsbhe eiee asretee e b t b h e ea e b e sr e a ae a a a Ha EB br E be b E he bBeh PageAS1 Of A172 Appendix toTNO report of 34-02 Fina version p87 00122008 Er 3co FFeoos] rJoosses|JREvrooscesJorJo2cov0si0rJJJYYYooc0ooooos]IRYYoovo=oRRJ B0o102o40s0]!) rroosse| r[oossa o|Si Jo] Fori JYBooocooo]JY[oco:sPcBiooeoo) neeoli-ousJY ooJiRoo BEYosc) io FSooono} Jon [aFroze0s0SRBYY 0coscJRREccccc| Bocoo) ros] ozs: JRO B00 [ars foes| foes: fosee] [zsJ oe| fonsSY0<] Joss [sii [osc ofooessesscJJPRYEJococccco:|JREEoocvcc|JREoozoooMR occRYJo0ooss)) o[co:oo| Joes| Bozo] ro.JPYAPcooooccooBMJRYYE2os00oc0|cEIRRRY0s0e20e0)[e0051 [essssl] 1oooJYoozooo} PoocCo:cBR0E2E0E0BRocc| en ma Eoonl ma: m fooocBdY oE [zcJocooSBR ocerJYoooSRY ocooMEY0200 BY c0 o fies] BococJYiocJY cccMYoc: I oo BMoooJ oc PiScoccoJYo00o5o7 BPRYo0ec [sss Joe| roi: Yowc Josss) [ese] roses PPJYooocooocosJRRE oo-roorroSRRARYRooocooooMBRBYYY0000<c70cRBiYBR s00 5cs| [=] Bi cocJYooisBYoc foie [eof oess] oooJY o0oo RY0-0BIR 020) Page AS2 of A172 sDCt a2 in -- pec. oli ci, st and tyacid esos ---- ottand dina argcoer ces re rrm ae E e e eT E E E o Ee e a Te eE e ee ee a Ee . ee =e Heee eLLLLE ELL BEE e E E E E F EeeeL e E es e EE ee eeea EBe eeee E E F Ee r e e e ePe el e e Fee Ee E E Ee ECNee eee ea E eei ee Ee ET e ene F E E e ECT E e T SSe e IMTE=EE EESTIeMEe Ee=ee E EE E e= e e e BHee . e e Pe5 Lm ee l e eePe2eEIN EF E e a D e e e Re ,ye eyae ie t eoeh ie E E EIEhEEBE e alEmEeE EeE ee w e Er e F Eornas re E Eo s s e Eesm . D B =BE a Eeyi ee oee er eg EEE ee Pogo AIT2 p69 Aspen e e to TNOs -eporte of3.0s 2. ----Foalve -- ersion--------) 0u122008 e -- rTa +n ] a +ps Tre] ma F F h ie ee e = we eFP a a e s oe we P F ar ee e l eoewee||| r Ee ee Eee L Eae e Erw t E = a E Eoe F e re aee e e F Eea e e E FaEw e a we | Ee EEe ELE a me E e ee ae ae= ey E E ee e e ae EE F F EEEr aea E F=R e= o. we FaEee eEaa ow | HF E ee e e=e= E= = E F Fe ae e a EO o = =e P a =t = PY BE = E E c e E h m Re e a e e a Le P = a e ae e r R . e es ePF a eaeee ew ewe| S e e b e P E F Faeth r eeh . F ae --8 F =--e o. F= e =Ea aow| FE F ero e a e Ee n e Bee e r e eBe Er a er E Ere e r es e a Ee e] e B EST r E] Pa wo Per a ow baae]sa]|| Page A54of A172 reo TOspot 02 es wa E eF E eeEr Ee C S SE--E NNm -- e e --E eA--=a ----ra e-- E EHe Ee RE EeaEee eaa Ee] E E ie LLEELee EEEe E E E E Ee i ea e ee e LLLeF EEEr eLLe ewe 3 = E F E E re e t e Hee e e ErFEEE E E b E e E b e e l e e he e E ee eEi ae elre ae eRe = Ee E E E F FEe ee Ee e Se CHe INPBEeSElEee e geFBe =EE e |e E F EEe Esue e nS=Le EeME=e EE SH= SeeEe EE BSEe== HT E EB i E eE e e e ee e eE sSe E=Se Ee E Se S=Sa s EE E E E EEsE se E snE e C ic =e DE E BeE Ea e ee = e e = e BT e =e SE r E ESE E i e r E E o repe r E e rea=be E E e E e teee eeeteE e e EEeee EEE i Hl Page AS5of A172 Append 10TNO rop0 of 3402 Final version pot os122008 Fecalweight 1 l g in d 0 idIenl s0 [ S| 7 C | 2T|e665e6s0T|TGseoYaws|] msoosmsNO OGvid5oem || |E[= CTSTC eeee0nooEwaCs A oemwww--T--aaw nn|| [1 T=Teeeerefromaal oemwe aowwwn || EoCT eee 0r 7 s SO a ome | F So ETTCeee0 eeosr vrTssaNsa| vre5eneNO [1 omoye | || [0 |T 5eeT5e eeen mrsfT eomr s!sees1oe nmSO s| ooomo7owmm |||| [= eenTowel ore sum | |C 20 >C Tess0 as [Le2eE7e]O O0 sE e.erSO | e GL.:a ,: || (= 0 5 A--7-- SO _L _| ESR homo7os| | | 0 || eeeeeeessss [|[ovrzsoeermni|| geveouowes | | | o20vomamm0 ||| | [ [E = o eTT C =TTeoCsewsea0 nuTTTooreeoman4ls]E ]Z svoaossemes TN osoee mw ||| C CL i1 0 5700 1 -- || 5 81 2 [7EMSS se|7 |"e1eeo00[sIsars| m1ossO| Wa=es | PaASGgofAe 172 AppendixtoTNO portof 3002 Final version p92 0122008 l r.iMl d0 Mi 0 =acrne0 [ F | T7 T =TTeeeeseaerwnmTT[TegaeommmsiessI TowoEweeiesrsds 1aii2iGo0eoiesm2 |||| f [ ETT 1 c =TeCseeTeesneoT[ Teo7 esssIO s aoemre S 1Wo seioes2 oemm || CO CC50 30CC7N8 TT eee eee reeSSOOTaBw X 2 s f = eeTes eoemeoe TTTTeei ommsie soesrmaire wTsT--e e osi3m mi7zmeer|||| [ [ T e T[oeoz [rsm ose | ttoaeos]] Bl E oe| 1E :T[ee Ce eee s0 e eT[TveeoseossshsT T z| eT vs io as sS ss m | B aase e smaos T our || Bo Eo 2 I| [ | +T Teoce oow0 s[T[rgooroveees0[| vseme7issmAM e T|iI saoesw ]| | 7 ZT [Toeoo7 m TTS eCeTmmne] Ttoowu Toton ur | [E[ 5 oOT TJ Ts TeCsTeonmT[[Core mom0em lwse oo wses w |r E s2] o0F me] BEES EEE TTeeee4n72T0 sreeen| rm ow]| mn | rw | B[EeomIm ss[2 2C Tesee0 rwse[[8 oootE no|| ssmewesA . ) ||rmusw|]| E C20 C0 12730 6 Page A57 of A172 Aopendixto TNOreportof 2 Enatverson pss ot1200 Ta a eoar| g 9 g faces 2 T 7 77 2TTeee beeeasnm [TTseroermamrT T[ m B s sa e e veer e |ottsoooemmorso|||| =T=Tem[on sar |2s | |[6T|=5TT[eeeemeses T[rfoooe0nn6sT| ro el osuar Ta| m2mo0orsre ||| E E|E o T ET 2 C 1eerE 0[sAases|C A g5 AieO s | Y CCaYEve7s20|| | 1a 1Teesss[7eaas | asa | 08613 | |o5 T | 5TT1eeemsesesezT[|rr96eo20an5[[| vvaeosiseear || |oo2ws62ne6s5 | || P70 |o 6|T 7 {["eeeoeees[Toooseneree|] eboraom srs | |o2sn0a4e0m||] [SEs BST I [TseCeeLwr [[roEooEmmC|7 Torsmes [|dNoeEmemn || BEBSMMS| o | 7 |66635|7802| 76ses | 09928 | BBEoSS 25| 2 |6o6o6o730[|vA9e4Ze3r|| 7oosar | 1 o23w538s4 ||| | 28 [[sot T TTeeeesmassCiOTT[ooT7oOsae[[ ]T se ssd osis o0 tool Toostasms ||| | [so Tee [ras [vem ow | | 00 00 +A-- <1 | E5 c 7 570020 2300 1SO 2 1 Pa AS8g of Ae 172. 27 7 `AppendixtoTNO port of M02. inal version Feces weight, bodyweight and food intake GC samples oiai]] FEForeen] Eid Ca Fe mereeree r Ff oe a essme e ]| EF RS eeee||| [mao dow we a: | Fi snee] 1 1 re) rreearl] FFereier)d EraJ FS eor e meee a eie a ey ) 1| Fm einesmea Tr] Fo esmi e x w neee J1 F (I mso n se w e we a or Smsos seve-- re-- e s | F Fo r r i a t ee o e e Co e FFrreoeSR ve| ce |oee e |aie CCoorSe = s ro a aesue e ECOoIeNJR we|ine e| si ef es a we 5 re [re Es as CEE ar 8 p94 2008 Page AoSfA1972 nix toTNO reporot f 34.02 2. Fecal bile acid measurements Fina version p.35 12: A TM suns o[oe mmee] Tee E1T"wopi[]Treniaee) Ciaownpooossrs|a|| 2168548720 m0n wi wtoe|| otmane|| mmeos| [oomsee 6 | ase | wa| om fCmeiicenoonnrd| 35% |_a11 womsre pi | aSZose ||| 2uoems || da[wwsre ||| oovsoommw wor erT7 o [wlT 5] rortvoseonyT (BIE TI] ams ] E enE -- ouccis [E] e |B e] [ ~I|OI [eEbm] [oigsres| psricosomiuietvnsn|| J21=23 2wTmor 17a52 ||| ooosssess||| eoSnosa||| ooomimes Cemnvreccanhoorsnee||_ S276117 pmeaarre | wwVaroz||| ooaotsre|| s[Wseiealrz|| oovueers wtwonCT T[CEEE TT Ce] rT 209 o TTwl T omieces 2T5ma o er m PrSEe ] wraed sCaupyrceionhnnae|I 047 oar wTo |ooeem |s2s|| oomm es a | aon | mo| om wC[rrroaevmven]]| I720-3A1 hwepiosle | Iw14lo5|| oorsscse || tsdaelssse| [oeouadsr terse |_2 or | om |B oe somopusleon[TIC Tr]Tw sn ec Tommy wr ooym ||E3o2n8s wenn || 0io8n6s PageABO of A172 Aopen THO sponof 402 Eoalversion EE -- pjeie Iil ra e crFH II iI S d r= FA I RE cl sora oen oEwneIe e ||IBmmnr]|lS=mme mA[[a|] oemR reolnCELCE=]Tw 7Tw]7] ---- seoourss ICl o[7 5| mmeal | E 2 L| EPmor eEa|]I EBee | oan||i [Hr oaPEn|eTm|=e|| EieS oSmE |u| Em | mom saeomeemll) gmm ||eEL|mEmre | |0[hW waNEl| e|E ]B] S0iE ettolryCT ER TCB EmT ] wr o 35 ToeT ] 33 1] a swcoo ne C mT E |R7R|T eEmmT r ]| ge | T [ om |a S ee|mEoem tcmomemlee me m| Emm e| | w2m || 1P)| se ehweere | [|RSWa|e |eEaamSESEE]]||W|] am EGmE rtm CHECTm]TewoereTero aT on|| mmmee oktTioene||] mS2e2 ps6 oe122008 0 Page AB1 of A172 cto TNOoer of 1452 Enaverson -- TE osom=oeeen) EPFm ES roOa oHEEEl lNl a =B Be ] EI Ed Ias CS o HE wo [wwE e]H eonyC [FBE ] T CTF ET s]s T Tw] F 30] EE -- S -- ra ||H | 22== EHawwwFeeoRB o|uE e|[c]rwaalH amm0t]me|]E mil]l H =oomm eswao[[GBEE]TTCBEEE]TwT o 5Two y n) --_---- | I r= Pee GE i rs ad ocS l oeonalmlee |nH 22t8 Hawm weH meTe o||oumB I l ox|lme a[2meE ||l] mw2a]2] H eaommm l m= a meee ==|| =2 wna | me ps7 oz PageA62of A172 /01 `Aopen to TNO-report of 4.02 Finalversion A sips ITT r=]Pa eoeo TI ili ForresSEar) [OE me ome Gwomosooetne|| 7054013 oTaonor ntoss || 52 | ooosesmss||| waass || so | ooosxaser wsioeoanooan]ls 17|_57158%3 sEruOys | BwZeEr] | oeosrte|| rwwesls| [ose aoe i onsnJ -- EE omnis[CreaEsSS e Pomeaei] owoancoenoe||| 11t1o87e5 Cmcian| 71 mTwo so | | ooonoo||| ooaomsmeo||| wimmmeas| [o[oommmee simes | 06| oie| ma | osm wiinsncrosna)|_ 2to6r Katss | 2os || oieos|| s5e || oooom x CoT7o TeT o] i am -- EE sruonr I 2055 rlPS eT I E llESforiel os | eo| dom [mr [ome cPeoomwiyeme|l Tien 771 wrinome| 751 mhseiidas | f27o75x||| o5dmo0ne|| md[eeess|| [ooamumers we | an | ame | mes| Sn nos|_ 363 i Vs | ot | WY aoe or CE Tw Tem ow rococo BEETTT wm] wes icvovouran ||| mai1me0s6 ewntoetro|] a35n2e6 p.58 12: Page AG3 ofA172 a ---- ire i Ed SmEEEEEET] E82 E|EBEElS@E|]E os ws [CEE] i CETw 5Two) EE Cea]iaa] mmE= ELEEsBs|Ee|S]] i8E oa [TET] CETw Tw] o wm] EE ---- roctystnry[TTT] uo eSeElrIiHUEiRrENeIlEIFTiERBT REe ] == EL OR (BEE E ol CE Tw rermeieon T CHR mo] ase aE== =ZE|H[8 wo Tee]5] 7 `Aogendixto TNO report of W.02 inal version 5.100 04122006 wes SATS scrim [TT F Coe|W e e E ] P T a Ta|= bees)" oooonnrnesn||| 1554797 9 wGnsro | a6os|| oi | ooonamarn ||| soeio|| mr | ooammwee rSpoiinnceoaormnnaa)| 02|_5210 uCmo 0 | mZrr ||I vem oie | [ais | Wr | om oom ow I i: on 2 orton [TTTTT TET] eer EE smucnona[ ~TE C SI E T=P T |r a] | ee SweoI oonioinaea| 1S5007 ooToeor iso| | oo[oasm|| w[mose7|| oo[owmuees w[triiiokronoesn|| 24508 sGo2s a || 3o30a]|| 5oYe6ws2|| aa[mB]ra| oo[ammms Pe CerTo +TwT 7] omenTET] ws -- [ AE | wel m ToSeo ] eee]] Ssaenoosmcerreaosme|| 72[07TT00 omarss | wsTss |o[mvoomse | [mh iaess|ao[okse Wsweinnuoconoinosae]]| 62[123989 edeokrso|_ 7 saToorrms || Sw9|5o|| do12Ee%|| Ed[nee]se|| oo0siee = 20 | om [2] oi reenopltnoy[FETTT5 w] J are 0 CL | ccooymle || a2e6s foomnes|| irceess oie] 3326 Page AGS of A172 Aopen to TNO capo of10.2 Eeatvarson p.101 pe -- A wmimslnl] l m r |v|] PfsoI =wm eL[EiaiE]seEe]wes] =] reaarsts] ccwwooernele]| dSarem || a| viE |0v]o | sEwieeu e |[[uw|ae|lRabiemmem|eiem]]ee| eomwewmer oo oto [T CCTET] T Oy ]o zm C 2 | EE oe[=a PS mESe]~P a T [A -] 20ree] | oo] om |e] a Sowemn]| =2 2 | wwom |]n nlulmm imele]]]a|e am om w eto C OFH ] E [CTE]T1ETTwo [=iEE FEwl p =ie =eP ar T m o -- a rre] )] o--wm_m)|| 2= aaTwoem|a]|oeus||momum|em]w|ee] | oeeoommam wno ee] 3 wve |lu| umae]me aom ost oto T CTE W CE ]Twws 5Tm]=] = e = |= y ooen |i[2E=2f Page AB6 of A172 oped 0 THO ot 312. osverin aenoe I =|| = =|:= = mown]| em 2w E lemEle] E oa wa [EE] reson [FETTT] am A -------- Cremea] ee] EmAe)E E wEoEmA |e em =S=HHH we HEH EEE ] H cE wo [EE] rerio[TTTTm] wes a NS EEE eee ee| = RE|: E ao E|u|E AE|W om 2w vew EfE ] ]E] coee = psy[ITT] se EE eEe]E52 PaAGTgofAe1T2 106 AppendixtoTNO-reportof M-02____Finalversion A. pred| ICA Tot loEzA]E = eSpomceie]o]I2421 awtsOl| EmlA2E|atiame| EaimwYe)| ome om oso [ CmTTB] wo [Eee]P= ee] vcaemnn]]|| i=2o =|: E aw w m ne |o E om |Gl ] |me mSE ] m] e]]|] ] =aoommm= = C= PC e mn S tS Sensn,]H ||| 5325m HHFwm wwI wAooo uuH ]Bwn E lH s e aRlm] em E]I ] H u=aoesm = CED etn CELT Em] =e oa=m||| 8dae p.103 04-12-2006 Page A88 of A172 Aopen THO porof 3402 Foalversion p.108 ox1z.2008 A spnionys Ir= PE e SST oe aI] iom|SLsae|domomsd SfJraoineecn]sd wwwERwe o||lIeaeslm|I a]oel|Ie|a82|[ wuoneme etwtnerC[EEE]RTTCm=]Tw 5T&T ww -- n1 eron[s = TOgemmos a rrl e | e aA |] b Fo x om | om] om pfesoroee F2E:) owm o| wa|eW || 0ooem LR eT en voterCELTTw] w= EE ---- ro | Ei |2 T ST Soow al]r u| ]om|| AAawm ee||= SLeomm rosm oell == wSvero | |2l0wu|l|asemn|lem]e|| om w [EM [Ce TeTrTmTw ortonTTT mr] on [ a=| | =|am=m re 2 PaABSgofAeTT2 Aopen THO ep of 402 Fina version p.105 012205 -- EE pelrie[CEE T sE wE r ]Ta |eP | omr | aT e |Ee om cnFneonaen we m www w w|||e ] aofomm||| amaweeE]|| ooonmm pee vl las ae | oe ==] CoIw fs Tm wr) tyCTTTB] 0 EE -- Jso.| = TWiEsiEE e [Pe Ie TT FEES| arSrLee]] w | wf om rweoe]||| =Z2o awwSwee o||onmf[s e|fslm]0a||s]omwmme|| =ooaemm = -- EE ---- aemolr [ lET E | V mm a] | Per a|eiaes |eopoio] m Ccwmonaool]l| dBimmee | vi||v]e| | eel mEobesme ow | W|| [BD])maimaEm]m] | wowrem] || ow] ao2iemmm w BE] [CTIT&T 0 a ao|||| mm=e= tn | Pa A70g ofAe 172 AppendixtoTNO-eportof M02 Finalversion EEE J peaen],r|IE1 e Bwoole unr so |] | ] aa] =oomm ase]n)| Te | Twr ]|luw| loomeaml]fmm] oeooem - Cr EIT w eT 5] ects [HRT] emo EE swwmooeoen|elrmw L|]Tw TEwEm ||e] | eoma||]ee] op oe twmeelH mr|E ]3 |HowwB r E |wu| aH mmE |u| H aoee S - [CE] C=T +Tm w] ow] metystoory[T TI TR] smo p.108 04122006 PRET er sonnel --|| == BowE oan]| rr es] a am eS wnc])|H 3=2% HH wTw ha a H l|a| H mammm|aE ]w]e H aacmE orw to m [T TE] T TT O ] wan0SO S 30 | on]|$e20 1/0 Page A71of A172 p.107 ixto 2 oi220 CIE e Sere mS aT] | ] ee]| J p--wea]]e 2I=E) E aTme o||r lEweel|sm]or]mr| owT aer||Shoem a] = rreeree]l 218 wSwi|on | u mla |emolom||a]mwre] | ooemm w FC IeTTTT ot [ETT[BE] wr aE-- awnutetase I TT =)rSeWE e |Iwz|iom|rrwTs=wR | raemal fr--aonno--e]l||_ awwAweeo e||f5 one]AlIamomowele|||aI eaw]se]|| uoms P otto T CTTT] 0 1a ET EV ER FE 7 a| a Sam[ m e|v m serR ewm e |a oe] m SmsE | ana]p=a scweooeml]| Ef| wrm Te]||]] [WaEdRmn || aE [wbBEm eEa]m|aaas||| oaommm i2 To ! > lel we] wm orm TETTT] a =w|| m=a oTaan || amee pire S3 Page A72of A172 a -------- Foavrsion ---- sro mel EE m || T e | | oE e eB a ]] E|e ]T om] sSoaime e | ooL] omwl cceorraeleneel]m| ]mSn e||]vie | wuoenw || |Ba[xBW||i|tmeel|wam|ee]| | aaoa om wa [CEE] Cw E T TT T 75 1] ote CEETTT sw] a SR AET owienl [eSm erER ]||Poa e|Er mAe|eo] e oconnn|]| B52e3 we a Twmweo mo|aan]]B(s awmseeml ]|)mmwees]]|| aoemmm pte CHE= T] en wz J [el Sra em T] AS =a=e |=Olo]s cprmnels]i| IY 320 aamSfrwo||u]woS|r|]eome||| momwesem]s||| 0eoomum mT ene]e wo |eel|imm)asa]] - CETerrToT= orton TTTTS] som oon| | me oc |F=H p.108 2: Page A73of A172 Appendixto TNO-reportofM02 Finalversion p.109 04:12:2006 nel[= lenRaEE] ~LP L SRT R re pma = Bgllz z E x2l E -=zrlE z( mzlilz mmE lee = == wo octatone [ [[EETE]TW T E] ams EE PaH[E CIF EEIE TesB ms]E-VEP oRilT eAR rs =e] S EmH ilEE glH a E n\H BlH = oa [TE] cette [[ TTTo T w] wnt oo = Sa =eT r I n= r= Per Z 2E 2 E E I rIE ZB iIlBml|ElArE2d] 13E=el a S ieH 3 U ZrT R (s 22E z 32 = etw terT [T S] T TS] om swan ---- i-- EE Aee]ESa20 3 Page A74 of A172 Aopencixto TNO por of 34.2 einaiverson I -- aoFroosoyn =wr| ormorm|| aaf| 2oomm SCae nlmal]s a e-tn CELT[WT] I CE ffFrhoieyand U aw S Sn oeE wa [m R] pm ] anooRw ||omsouemm H fe w [EO p.110 ot122008 CS3EFeoomnenEEAgmw wlem oa E nB[(ARaiule alE ]si]|I E aelwoesm]]]| a um ess-enCETTE] oes =8 Tin|| Ge Page ATS OfA172 Vd ptt LE -- -- mE | Cem ETaDn ] |os| ]P ow|E eawlre| e ooem] S fwraevrneed|IH 2 EE aooH eleH me]m] E oonme|| H ooem t=nC(ETDETTT] 2 I -------- Crem Toe] oe etweeln| | =2 RW = Ts 2w ww STo oo o oE sw w m wl] s e E llm|E awmowme|||]]eessaRm - CT ETw TT no i mmmm EREE Caemme] e ee] t2 aTwToneeenl)|| wm mw w omolE ]ea m lol) lE as E] ooa=aioms ewyC CETT]wen 75 Page A76 of A172 AopantioTNO spon of 42 Fnalversion ET -- swmooonlee mlml|raE|l|PEc EwEee ||I swreme|aomlmi || er hese|| rsd oemn acmS reeleeelHwvmmoe |E |fV o|m |H waomeeI|SH mS |l]svH m| E aE Ww]] RH aamR - es me mp E EEETE T ] i c--c= moem--eem ]enlS l leimm HL Df|vm] ||oE mmoom%eSH||H w B wB|wlElamS dnE ]S]l|wAeH wen ||f s E Eoe e-yCET m ] en n TE asi I rll Ps I li ErdaSUl ftyroonid Te awwre | || wom [ne]mee lo]ua|||w]aEaes] || om oooeao eoeanCEC To]Ter ToT= =[E room| me oe | me p.112 cerzz008 6 Page A77 of A172 Aopendixto TNO eporof 84.2 Final version EE praeossne [elim Tom|E ]aE ] T |eE |o wsrr]|eeemm Crrooeen wwo we lE|| eam] | maaee]| aom -- eo lw las] wr] a e Ww [E] e [EITToT=] SS w J |e I =Pc Fw O uIEEEi ] IRE I PELm ow | TenneAn!|| 5 Ewo EEal|wuml|a]]mIlaErmwA|ryR] os e= etoenT [CCTEE]TC TE ]Tnw er TwTo) pr--i_--ses[EaB E ]e ET r Eoe e-pes ] fpo rete EawA|o Ea|e]] ei=m o==inm He = (wl HH eown C[ 0 EETCmE]I= we srw |a oe | ae tee] $22 p113 our2208 Page A78 of A172 Appendix o TNO-rportof 3.02 EE Final version SL p14 04122008 -- HEHEHE ie I -- ovo CEE] on CEE EEE. et co Com] ono Com li ,, EE EEE pea cso Cm coo Comm YZ Page A114of A172 I p15 -- I------ a coo CEE) lN-------------- uo CoE I -------- ------------ EEE EEE oso CEE] LT -------------- EEE oiso mCommm)mmmm mm Co =a] ------------ EERE Om oso CEE 7/9 Page A1o1fA1572 Aopengixto TNO reportof34.02 Final version p15 0122008 HEHE | HE| P= oso CEE) EES | SS SS TE EEE EERE P w (= EETwT IE er] s=o[CEomlm ewee we] HiHHHH[BEHEHE | aE CECE Te Tm cso Com) ~ CeCereer so CEE) [20 Page A116of A172 I FE | EEE 1 cc CE HEHEHE wo [EET Ww]wee omer]owewn fC El HEBEL wo [ESSE TsTw) 1/2) Page A117 of A172 fogendixtoNOepotof M02 Finalversion 06122006 EE EERE Ee beEpi EE Re eR bla EE EEEEE ee Ee e ER R EE ae Feist ceeise ssa eb Ereah kb L mEeH en Llee] eCe Ippeaeen| secitn| ee |eens |ies mest |Rann| feFe EEE EEE EE EE ee LE ae apie]| ERERRRERES) | EREEaRRR GrEac| | FRERACESR| | EaRbae BERRRrERE FFeeEs E al FFere mentTEBoana eer |Ekma an boohfeeennle | Bneeeeoaaol | Raocnaen | EE ETETT ese]| Ea el| Eomsane]| Posehn]| aon]| Pore]| Pacem | soE E fciesmeeE | Eos E oh][neE nT i] |pT erE e T|MEelon ni [n manae na[| de eel pall ed I ---- [22 Page A118of A172 p19 Aopen TNO pot 002 Etvasin re. EE] [mE Fa eErEP C . EcIFRR8CaCaiRy marEil ee paErB smCB amrR aanE y] = E F= oe[=s= EB eBeH 1 BE oBcB5H ooof [o F Fa oos m meeR SeLlilpoE[eepRsTaetaptelea]rEeep erecBLearledeiasoecTBaplaeapleRcocoo;ebl=cfJIegetaBeolsocoodepfaaor]rBle==1fJe0are2p=td]y F FeefosB Rc Bce Bo=c BY =<J E = =o E--= --r n e -- i e leeeenC8pc=eiBEe oBofHmEE <a BoEgiElOEBa rE o5e eeB]igd Real a F a e aE lo E aBei /23 Page A119of A172 [EC ------ Foalversion p.120 oxt22008 Fatty acid balance E l R eHr Re e HC e s R i e , e e e a n oe e n>. fL FF re reo p tR eeE eRn ee OHt EEe ei a e tee E e epee R e e e ee e ei a eea lTEe F h b oo E nie m e e r e e l e n e e e ee ee a oe Ee E cReA e a r Eee ee e = g , r ess e s e e e e e = FP o L C egbHee l r B o n SeHR e Ed eH e 8 g me rHae a e il erae e aeoo s a o Bmn eee P F F C Bob o A ieeP .nr Hr eErre ae eF r r e r e e e em m r a er eaBee eee a o -- e O E r -- e dg -- rr E s+eE e e e e me eT ae e mheeen +eERoe--R E EmBB Re s eEe o o a EAee B e EB E me EEee leloa rieiErrn n E adE a ea e a ner hbn reep n re rs r eee E ra aea a e sr a e es e uen aaneee Ee Ep EF a C p eE rR ri E e e r r e h SREpS r poee ere pE Re Ee Eer r Bae-- E e t e e oe e se nae e ae e r ebe a e ee e rhaeeee ESaaeeee [CerremSF pE m Ef p E a e ef atE p et a a e TE E E EEEs EEEEor e a E e Hr ep e a a a e e e e a e a e B a S EHl E a e aBE e E -- on Page A120 of A172 pz -- Enatvrson 1 [om =} oe[eeraevrtfa amepfpanosF sLemEaipec|_guaise|gnoeroer [fama) re] Ihe 5 or a Elle olal o Lolol po 1olaf o Talelool i 7 Talato papeearrfWai teres iparaasLasers | arnr | foepel Liipeter 12. 5 Page A121 of A172 pndixtoTNalvGesi.n qeport41o220f08 p12 Ao 02 Fin 0 Appendix . nesinal chester absorption i BS ES CE = Sn 1eeor) EE Cree Cr ET EE =8 BEE EE me mo _E C2 = 22 EE -- E22 i. -- = oF =5 EE 2 2g == 26 Page A122of A172 p12 CE WNWE SOUR) Cr) BE EE i. =o -- gE -- Bf bm -- Big iE -- Br wm ue B25 2 - Cri EE = Ey ee EE = z= Bogs EE = HSL CE = 127 Page A123 of A172 p12 = 22 5 ZI Bt EE = = Sp gm = i - =8 = Cr) EES = Ea E = EE = om a Bx S Bg = 2= a Ci r= EE om os = Eom z STE mow 12g Page A124 of A172 `AppetnodTiNxO-reopf or0t2 [a F e =Tr-- ae ---- F | F F ----r a r--| | F = ---- F a --r-- i r --| = F F--r -- = =| H F F F iT-- -- r t r -- -- F = r re -- e e -- = a --. -- F F a i r r -- = a. F a s -- a. F Fr r| P a Es --|| BBE eea5sror e----waa ae+ ---- || E E a-- n -- 1 || E E 5 F o E t| F a a-- --a -- -- FF F o--r a --| E a + a-- e -- E E Br e s ee a s| e 5 i ii r --3-- Ee Final version p.125 05:12:2008 Page A125 of A172 opto NOt sti -- Er pnlaL-- n LEE LE -- LnNoh p-- e| ah Glo]5 LololpLala Rd p Lalof p ole][lo][nla]| [Tom CRATER a Al , 130 Page A126 of A172 Aopen to TNO upon of 8.02 Fina version p12 out22008 AppendiGx. Liverweightand perigonadal fat weight and pid contents ver G1. Liver weight and perigonadalfatweight a E E EE r e r E e E E Fa h E E E p E pb he n re e e a a a ee r t r E E EE ph a f p pe e a E F ErrEo ea E ee e e EE p E E Epe f r a a e e h F r e e E h p e e e ee E o e e | p p E Er E e e E eE e e a E E Ea f re e e Ea E eo E F r To EE e ae a EB ERe E s ee| EE e EE e A e EE T E Erer E h a a a E E E E E E Ae R e a ne iE m E EI EE aE at ra e s ae e B E E H e rr e m ae nr e m S E ope e psE -- a m e e s O a n Ea e B Page A127 of A172 /3/ paz ndicio TN u Fins version 12: ee [ e rCi areer= ia LnRooLeR=Le [ ese r REe EC ori <= | [m= rer =LiEplalpieecTit omlearlsEseo TRpaEaleaplEstToplaosTspe)=To[oluoel2 ins Iiopafpfs aetTo aolpael! reaot] [(m [ rmse rVooer LMaReErIaSrefSistommerst3 s>ie3 RbeRerR oesRL3 a ierol nJooqRlee2r<roermosJRpkae2ga+or=mfEeS eEArJao!] eeEE RE Rl Page A128of A172 an Fina vers G2. Lipid contents ver Liverhomogenates; protein concentration [TF E] ET E He aeea] ha H E m m e ss m n s n sma e m| E n E Fa i e s ae e e t 1 ahnem| H E ee e athe m E m eem m aaa E E E F e a e a es aae e te te E E me s e ma a m e e| nm 1 | I Es e . se e 7m|| E m e em n n i | E He ee eta are a E E Ee e r isa a te e e e ae E ee aea s E EEe s me e a a e F E Fe e e t Tae ae taaeear Ese aa E FoE on E ] p.129 os122 Page A120 of ATT2 5.130 HPTLC measurements, iver lipids: EF C cm7 e t12s37 0)CLio2sCC7oe ErT) EwEw)| oF Een 7 3 Jr A 32 C1 || C7 = CB JX20 1 | r n BCJ= T 0a CE || T En 7 oe [oie2a s n Xo | eC 1 013 EE m nT w 7 T7 = o0 ma--iJ5[site0[e e 0 es|| rn777 n | 7 1 3 FIT 7 T7 reomesmlc ss e 7a0fam200 sn] [+ I Cuieme[sao~ te0i[eaand|--s1m1s Gam~e~]is[3 [2 rm 0| FEwwTCC ra=eaesoTni seTa esn ec [oaarTae wmee r1|| meBC me0Tim | T I T 1 --so [eoioaC e|]| CTo=welC56A6 ws30Tone0 3m 70 02 | = J1 cA m massa --a15 m | 273 ve[e3e % enpage A130 of A' `Appendix to TNO.repor of M02. Final version [Em]PesTewarsi oe) OC 2 = A1 I C 7 E7C A co 2: rc | [sm cI eTa 1 on Tol 2mTn s -- | [ls frmsasTors am|| p.131 04122006 I 7 7 r 0 +o A || w 7 e 1 rs mc1 || I a w esmoese] ae[oc es [esr[or ||| c= rw 35 1 || c [ ws 7 m --33e aToa n 2r 0 || 2A | Frc 1 | [oT{F Freon--ao 5o 5[ser 55]| 7 Fr0 70 3 ||| 7 0 | CF0 r 0 c 0 1 | et2 co | [7 Co oseemelsss [Soorn[a1 vr 30]| 7 Fr 2 313 7 || co F 0 o 1 7 11 ||| 7 7 || S B T c 2 || Cm 2 1 | 7 SC7 2 ||| rs 1A | Page A131 of A172 `Append to TNO report of 4.02 Finalversion p.132 04122006 7 Ai | LF ro Ee 7 ws. 1 | | LF | C = I I A 1 > || I 0 072 22> || 1AA 7 || 2 c 2r I || 2 Lc 0 o 7 31 S||| cr| S r c] 0 || 7 ce r =emc 2r 5 +7 3 |||| 0Cl 0 1| I CC 7001C|| 3 | 2 LTmoF aoemiiI enI!e sr Tn osT[0s amE mm[sa510r----]| w Ic 0 7 172 x ||| 7 2 |o Tec esTAso soa mu2r T wes]||| EI E C e57 Ce 1 7 || | E S T Jcrm=osmenl coL sTa7 mTn]|| o 7 F c0C Ccer 11 15 7w+|||| c L C7F r <n|| 2 2 0 1 || Page A132of A172 AootoaTOnoptt of 12 -- 7 FeEF o e7 se rt t 7immmie e n es||| ---- I Choesterlester] 57113213[23s| vier | t re [2 Toghoendes|"550 |i215[iessro| 2517 | Freee I 3TiCromesatarsctlaonsdiaorrs|] 34891 5|31202843||o2r0i6s9e5| | 200 ess | | E F a F 7 e Ee at e ie a m e e Bt naoe aan s t e| I Croesterdlester| 8333172|vss0z|__ sor | F = ees here fTCrolestoroloster| 495 | 35273 | sases| sos | f---- 5[imemaisandra] 406 | 274|oss 200 | E E= n 7 re i mi s ----[Chole| s54t 6|a5r 34 |cral orse | sii 170er| F F 3= Jimee e maistnsars] 35 68ae e P"2e2t0t8|ee26725|s t 2+00 | 7 P 7 ees t aa ara| Fe Ferte =r F F ee s intm ert m te eea n--rr ICrolesterolester| 485 |3578 |iseie1| i115 | 7 7 3mt Fm | fTCholesterdlester| 485 | 3505| tsaeer| 233 | 7 C14[Cromocleste] wm | 445] 5i8|aerss|7] p13 oerz208 137 Page A133of A172 -- Hr-H0 ---- Append to TNO-repor of 3.02 inl version I Em mmem meree e map )es] Fe a r ae e 1 Fes a +e e me m| Ym p34 oe122008 Ett Ee ee er F |e ee ee ee e Fr s ee e e e e eseee e ea ae a etaeu = 1a e| F e Fe e ore e a er e ea Fe t--e+ee +m 50 Fr h=e ta e e ee ere sa Ee ae a aifrsa e | Feta ea ite1ama1m| | e1e | Ero Bae r a Fa aHeos Es aee Ees eaaa r a -- -- o e ----r--er mhme1e eee eae] | Eee area | Fo ese a | E E r e ee e --e a] e | F E B me e t raaenm oaree e aas a a an | Ba 1 1 e mnae | | E Ea f r ertee] | Page A134of A172 apt mt2 rosin I. e -- FF E [ elE ee =R E RE RCRE m El ECeEES-- R E E Re REE RNEr eEr [[E oamreoE se]eR leoE Rm ehaS heme lel ep tela pdalel8telal p Tol-T 5 Lolof Tolat 2| [E E oomemE N isl E pe Rt eE slE oh iE Lel {gmedwt{gowts|gowtus| gown'| gos ros =] Pago MESA AIT2 5.13 [RT ates serezes ZH.raHyMtGo-eCooAsrsedcucotasienatndrcmhiolcehsotnersoile-a-hydroxylase activity in iver microsomes and sterol1 HMG.Coeisardchest Toyotas cy in hrmicsors PE CErasEtitT EctomcaE asnraE omtomnris--L ttT rsseirnsnesE [Ne=nemE iE atmm miE E nmnmne e BBB B B B EeEEEEH E E EeE tEE E e ee EhE S E BE B B EE e ESeSEE mEE EESE R e e oRbb n R eenEe e e SR aEbEt eaeE ESS S S SE EESRSSeS S E ESSSSSS Ea SSESSS E R eSSSSSEr EHiccS SSifiH. . E SSsSSteSEEESSSSESeRRgEggree gEEgbrSSeSSSsESSessssbn SSSseSSesSeSSRnE oEecsEiiiEe iiEEissne sSessSsffsStfC ESSSES B E BSeESSSESSnS I.SSE Se S SSSSSSES Sn SIiSHEsSSit S SiSEeI s SSSiSsSS SSBSEB SSBeggbggERggegsScE SSSSSrSSS=ShocsSSSEh.SSeSSsSSsSSSaooSonE sEEiIiiiiEirEeStnnsSsSasStEssfsoS SE E SB ESSSSSSSSEES iISSSe E E SSS-SSSE SSSISoSeEh E E iEi. cSSSssSgSSBSB SsSsEiEgsgREEseSsSR 22SEeS2eScS2S2snSES2eeSi2S2RSN2oE SESsEsi2iEuSiES0nSESseSt2sfC EEE B SE B SSSSEESSSSS E e SSSs/.SSS=siEcS E e ioS:sSSsSESE SSEBEgEgeEe SgSsbhaSSS22r S=tScS2ESSSseS22i S2SSSEiaffIiSEe 2SsSsySe EE S BEB B SSSeES e SSSSS eE SESSSS me SScSeES aE Eoc. : t SSsns SSSESB EeERggEREerSgSESESEoSSSSssecSR nnSsSSSSSsEEnosoiSEieiiEeinnncSSSeSseesfssl E E S E S EBSSESE S SSSSSS E S S SSSSSE R S SStSSSEEEocS E S S Se SSoE fSScSSsSEESSSE BE SSSSgEEEEiggESsSE e oSSSSSSSE ScSSSsSoE S=SSoeaE osSsE e cSeiiiEE tsSSnnSfffst EE ES S S E E SSEiE SSSSE S S e SSSS SSSr SEEeeE S S e SSiie E SSffSggE B SSSEE RSEggEenNgESne E Sa2esSSSSnsscE sSSemSoessnsSe C icfItiEEbtnSsEsSeSffltf BE EEREEE R BBEoEEr REDeEaE s Page A136 of A172 AmendixtoTNO-report of M02 Finalverson 04422006 E1 eEeESe SESESa Se SC E| Ee ==Ce st+s|| E E| H E ee+ e S+ e ee e e e+ eee eeee+ | Fr bE = b b e e = =reree bee lebe e ee = EE e EE1E Eme mSE Se Ee Se E Se Se Sne Sa t oSs ts F E =e e e E = F b=S es-- Ss sss =e--=--neseS= 8s e2--8s e-2 EEEere e E B EE e e ee e ee e E + ee EE mretE re Fo Tr eame aa ge e ne + | P E e e ae e eye EE eae LE Bea pe pee Ere Et ee | = PageA137of A172 5.138 Gholesterol-7-a-hydroxylase activity measurements in microsomes. temastuasneddarp2do02rm0aiOnFsu0452sb0gpspgaperrotydeeilnamination [CF-- oy[rgomn]e W mo o == |b o Eaa i Shim ngr O O-- E EI 1.2 re XC m J1 J -- Sn Nooerimme[[svooiso [2oso [[oo s omo || --m -- | [woN m [sssowmTo2maeTToomer ---- | [ ----woewhw omrem[[8vvomeo 1[|o2omv%eeTo r oor mw T[ [ z m e | | ] -- [wwo oormm|[osm sasio T[1o2osoae i m Toor ooomm rm | || e m --------e | x| [ -- --r-- vorimo o w e[[oraseir zT[|ooosor w ssnTT|oomi oooonT | e --a--m ] s ]| -- [ [ Z_-- w [owweo[ n[[rsssmeomw rmc[T[e onooeo owm [TT m oo ooe oroo | | i [ m m m z] ] e m| e x s [ [-- N-- o [rsioso uTToomom sTTooi oo T | w = TTA =NP oemc[7e91a0m--[vo [oe ooom t7] 5m] 5 7YO CY -- I =wnooprrmmaToroos [|omnseT [oooo| [| --Jr =neoT erma I eNorompmea eee7 asem T[|ooomCYwa e [ [Too1 mw eo J [T---- m --o-- m] ] worm oosTneToo|| Joba worm eams Tod mo e n om m o=s ----1| --2 wormTomEeC 1 na ooo [ --| J woprme 7st [as om =J wormma| rFmso|rzooe] ons -- oem[som [om [oop| CTI -ewwooprnmmea 7vasme|onmoe 1oowmee | [] 1 Page A138 of A172 p13 Append to TNG rego of 34:02 Fina version os122006 [rm ee ee [ -- a --Twm ioowAaeoemhmaeo I|Trasiomsw |e oasoman[|i oooowo i m momg3s 7sgory | 0C | [|WoPHmx| 68 | ose | oom [1] 1 womimxTei Tad T0000 11 [F --&T|TwDoPsHmmex[| eBEmTS[oOSmS ooome | oe || [|NADPHmxT8480 1 nd ooo 1 [IWDPHmx| Bas2 | gwar | ois| aes | T1woNAsOPHmix18e09s1rT122n8o6 | 1oomier [ [| | SE -- I|C7w.ohosPmHTmmxS[Er2o87wJ100t7 2R m8|X ooomJ l | 72 | C S| wpm[eas | | NAoPHmx| 808 Goes | ns | or ooo | a DPmx| Tia [I~woPHmx[ea odes| oo | | 036 | oom | oas | | SS 1 NabRhma JS C7 eed 1 na Too S 1 5J X EX | FT"mwoaosPHimisx| eeis06 || nese 1 | oooims |[| | FS 7 E TC L NADPHmx| ears | oase | oo |-- -- os --| F OT C~. wADPWmx| NN 7 CB e027 | oess | oo | | | FF1 worMmxT6222 | na I 000 NADPHmx| ease| veer |oo || val | [ Jw C noa. 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A AI.S| | weorumk| ewe0 1onseTow [1 ----JTr 8480u Taseo r10e 260 1 rms -- 1 tvoRme| 7ses 1oad oom7 -- 1 eee | waPrimx1945s Toss Too | -- | weokHme1 vanToonToo 1 --1 eee... | NAoPWmiTes1og7aToole | | f-- | twoPumx esas 1 Too 1ooe0 1 | --TNA ET0rO 1 saP es R ogee wm deme| [|woewmi eres aseTo |0 f-- 1 veoPamx"808s Toes|ooee [SE WADPHm [Tee |ees |oar| ssw | SS C7. CK J 5J = -- f----Tr estow Tne o T00 R 00m TT me 1 See I T noPWma[esse|oYeFw | r o1wJ [|| I woPimxT7935Tos | oo [| I 1 SY:STI | SNoPHmxT8607|assy| oom [| ----J twommk19261 ooss1ooos TT 1 | SnoFHmxToe | os | oo | NabPHmk |" 8a67 1 ooo | ooo [| 11 J snaoPumxT"oz52 Toga Joos0 TT -- 1 -NaoRmk| 87e1Too | ooo 1 1 FF roeame aseTor ome Taz -- swore[ae1206 | oarI 1 ee em te [oo | oes 1 f-- FZ Este NS 1 owaopumaTate T3700 T0208 [1 J neoermeTeeseT Dos 1 oo0s oem [BayTas og dee 1 naoeumaIo0261ases| oar| | CL. TSNC | fF -- -ooPHmxT36601 0s6 | oo0s6 [ J noRHmeTests Ths1000 | 1 SS CY. 130 7I | J twoeRma[sor To0uToot 1] fTonaoprmaTavsToss | ooss =e SS YS XL A: J fT neoprma ee J toPHmb woos 1oses | ows eres 00s ooo || TNeoPHmaTaser| 23% | ose re J waoeHmb Tez Toes | ooo || fTowaoeHmu aor T2620 | ome1 Seer -- [waoPMmaTas TsToo 1] -- | naoPumaTse T4028 Too| | 1 -WaDPHmk |a42s 1 00% | oon | 1] Page A140 of A172 po141 `Aopendix to TNC:report of 34.02 Final version 04.12.2006 [ wmoi w w [ow o oJ ar m om m Toitng ec ore) || --TwI T osT[aapm2O ToT s m [e 1 oo m A z-- m [| I wtwooimmm[eaamesT[onmse | |oowwes |[|| ---- [1owpormm|[a3s0T|noeSzaios[ [owo s |T= ]| 1 Twos aEEmToms[oA w I | mwwooomrems eooo| oamse ob soe oowwe [| omT 7m --I11 wvooeTnmmiae[E T voiiosTTooomns ooooT m -- ----o--r--m|]1 I I wmoommmme CoYe.asS rTTonC sT esToooo 3 ||| [I | mmNoaoemmnemmsea vooasme| TT o ose oo e T e 7m]| f = T J =emmvaooormmimmmme[[iooosoworT [ss ews o | ooo wme [s| m] | -- o I twvoorrimm[oesEmsST[inesF s[oooA opo || CT-wormms[ean| naToo nt notdette Page A141of A172 Aspen TNOrepoof r0t2. Finalversion pz os122008 I F F ale fl e eeWYe a e E 5 eme e e e e e e e= a a e -- e a n a-- ar F a Far n e e ee ee F E e Fa e e e e e eameer f + a e = a ee | E E E te e te a F s e EEEF a ee e |a = E e F e e a e e + i. F a a e eer Fee e+e EE E_ET rr = atE-- eor ea-- a ne ae1er E Ee r a a e e s a ae EB EE e e aaee | Bs E E + e a + = = i Ee e ee e = n ----F--Ft 1 &+a =| EE EErEEe a e ee a a e e ar EE ae Ea ee Page A142of A172 Acpendix o TNO portof M02 Final version HMG CoA reductase activity measurements p43 04122006 E E ene eew ar A 2 e e aa 1 | E e e Aae ree e1edn 1| E Eee eba se e eT ee en pe a a a Er a tatetae e t t E Eer rsr a eta E Et e en e e e e+n e ey| Ereae] Eee ea f ae ry Ere a a E ar oe] Em et 1 | Eeees mntes iiet1,i a s n 11t e e = e n t| E 1 mes SL we cemmcoam ses ene on [ E8 e psu -------- r= Page A143 of A172 n.18t p CSe S e E eOeEtE ee e e E a Ee e e E e E Ee SE e e eES sEhRTe e e e--a--e..e e e -esp0 eo e o EE ee eee m e e ens e rs. es 2 e = e E EEe E e eeeeee tenses 72 omacams wor had = = snd-- EE EE E = = = E = E e e = = a a =e e ee e e e E= - ee ----- e==eee seniEP e e--Ee -- s = e Tw pT ee --w -- n se i ee EE as ews SE we -- O80 Page A144of A172 do woz Fina version p45 JYRPY we te m TwTsm ew wews o FnTseorT Tww re w w wwme |s _2 _|| me sn w Tw w F F ee Tm a 0 7w wmw ee"s2we oi] F ae Tw wm Twa e w To aa ew FeT ww sae Fm eToweeTs w e e T we e e w f Tee e w ww Tw] [T Cee n r sw w aTm ae ow on 3 roomssanias my SS5i20nr50 ange -- ---- omoooams aesisisi wage i ------p--h- Spec siityCWoCmHiIcG 45012236|msop4iGnnCon cmon: 03% m Page A145 of A172 Somtcsnn4c2 estan C r Fr r e --e ---- F S S EE e o o E e EF E t e EEESE Ee Se e E ==e EL = E E E Ee n e E e e EE C E Ee SEEiESe E SE=t e E==e E ==E=S Fa E E E E = -- e e e EC E E E e eES=a E a e ==5= . EE E ESsssi- s = =iSS= E=E - E =S=i S==-- S== = = = E E E Ea-.e r e T e r e E i EESS a EE EESS E E E Ee r e e|| g e Ess =s==s==e Si S5=8=S == Ee Feeeee EE == SE S Ee EE ee = sz Page A146 of A172 `AopteoTnNOdtiegso of 4.2 Fins version p17 os12.2006 8i 3A= 3 3 = 3 3Reoa11 3 3 ot 3 ES | Pca [o[roeeDERrRo eCSTErR e SaR e sa paasS elpc pso I pet pia| PCEe [cocR R Rc CMe co JI JJ ieCJERT=hL=E RssCRAReteRok Reot eeeo eaets CormmeteHso HBece BoBefooos alc: atcc c JBoo. Page A147 of A172 pendixtoTNOvepotofW0? Fnalveson H2. Cholesterol27-hydroxylase actiy in mitochondria Protein concentration isolated mitochondria nts [o =pe re ] eem e s Es 1v a r eem n eoee]] m 1: 7| E E n 1 a m e n nr m ee | m m mmm m m i E E 3 sa es tt 1e s me e ma n r am a | a 1 t1t m ar a tome on 1| eE E n 1 e me m mam a taaae E 1 ee| E en 5 ra|| m a 7 e1 an| a e n r ema e oe t n tee s to] o e] m na 7 1m|| e E E s eo e n=e| s e om 7 a s m nr e m p48 os1z008 Page A148 of A172 cpanel TNOceptof M2 Foot version Amount scated mitochondria Crh r ere = pEo eoo e eeee e E Eee e he wr rree e e Ee ES =S==S==SS== Eee E ber ee g e n E Ee re tr = E Ee e EF o = Er e -- r FE eee EB HE ET E ee E Ee ee EEE a p14 ot22008 Page A149 of A172 Appencitto TNO report of 4.02 Final version 5.150 04122006 Cholesterol-27-hycroxylase activity measurements intemalstaunsddapr2dr200amsiOsnHa:ycu0 4b2at0igop.noprrodtee5 tinermin3 ation i] Eaoi]] i er270] 3i27 lOFlarma A | Z-rronfsce33h E [ 7 [IEnaoo prma[[r0a3p es[[CaotsE7eei 0[[C ooCmsT.euS n J |---- --e [Jor [riz[osToor 1] I [INaopmia [2 ia| ess[-- osCO s------ C [JE 7 woeriFFmrrx[sar |ome0[oCoZ so [ E E7 7r2n Forr | oso es[iAoeEAr [oCCsCYCYem -- a|| I C F 7 C -- [C vor 87s oes[0 oom7 |]| [towrorims|[089%5[tnoa [oososm |] [I2I0C voorF pmmms|[roio see0{Ctve2iso0sr 0[Aoosmete --|| [I [ JE rvom bF ima||Io6z2 o eoosm[ ToCor E.rAo2 m T I[[ CI=CrRoriw na |[osszo ws[|soet7ss2r 0[ooCo0rerm m a ----|| [ [ [J o E [T0oo733s%5eTp ooeseseirso [TToooei smmT 1 on r ] ]1 [I 7 Fo 7 opFmr o Eot2 He[eCrTi0r [Oo7 sCsTm w]ny| I [[ J7 rvobrima||T82e2e290||CvnoEraesT Too m3eS I IEr CeJ 1 een]| [[[FEov]p=otrivmomrmaa[[[o0ae0zs|[eansiaa|[[ooorsossm e wwo----] [ opl m| 7o22e%3|[v wansap| [oomo oem | | CTJ nhvoobprim [a7r3e6ss[|eoszams[ [oas n or] Page A150 of A172 p151 [Ee AppendixtoTNO-report of M02 Finalversion |[ S Socarw semse[oT0so 176{|aamsaer | [ogo esm mie taas sriyno] e) CS A HE | SWAoPHmx| Tere |aera| oss| | NETm iJ EY A JA | [[81 sNwAoDpPHrmmhx|"8ir0o0r8T|etTeeerr| | j----1TePHmax T7705 T0066 | om ost I 0006 1 dass | | fT waoprma1votsz|Taces os I ETFT TJ NYNO A JI | | NADPHmaTTA | ees ome | soa | I oT ETi CK o CJ r 7J I m| [w "eee THisig| osor 1 | EfCfCtu. orkF mx16C7777801 0E7n7a0T0000iJ 1---- 1] | [= [J1oSws aoh PHma T"7o3s521 Tsnoa T|00o0se0r| | | |Z [I wNoApOHHmmas oss [iar| ose| sore "vos| os21 joem | IEr 71T Fr a 20 Cs | SE. C7220 AX: A | [oom mc ozs10a Tomo| [1 ff w tvaooPrHmmx1"6es4i5s 1167n6asT |ooose 1[|| jE [1 NADPHmx|B| 133 naoPHm "aes T9816 | | oes | oss 1 700m | || 0C --te. oPaF mxT82 775 1o0er| ZB A.A 1| || hADeHmiT9086[trot | oo | | 3J0C=n7 aoPiomr a A720 I<T ose2 |7307 | ose | .J | fe huADPHmx178864T0051| oo00 | morn tossTas [022i1 J RoRmb 8083[naT0000 | 1 [8 SC ADT R mF x 078 OK 222210600 10am----1 75 | fej hvADP maT7768 TT na Too 1 [oommxTsovt | aos | ome |------------] J =mAoPrma1 8is[ease| ff vipamxT8251 100% | [[FEI haDDPeHmmaT| ooaers|7e2e1e7|| osor | oom | oodseer -- --| 7m----| F[ SJNAaDDPPAHmmbxT[69955669 |77061282| | ome | 041 | 7am] 1 [NJADhaPDPHHm mxo 17e 544t2 T|004084s| | 0000 [| oazz | Tse 1 feJJhvaNoADPPHHmmxx17511005888179n5a4T|00o0s0e| | | EEC.F 0 C7 7 jf] NADPHma"6460 | 260| oewr| 7S| -- CTwaoPHmb18926 1 na | 0000 J 1 04:12:06 Page A151 of A172 `Aspendix toTNOteport of W.02 Final version p.152 04122006 J[ [owsa ohmm|[[oode ewo [[am] ms2e7He ][Tvoao2m7mOsF Tome| m 7mFoeomstygowsgoSaary [[ [ Jeosnm o o [[io7e5os%m[[oe8oa%mmo [[ooowoaenn Tm e] m]| [ [I [JJr tCNEoer soHpmmesTT|w oo8se3e0s4T||oso 0measeT [r c on oom7r7o --| m [----| e|| CJtC IC +wNv7 oopmtmmss[[0o2r03[[2 C ovosC sle [[ooCsEeg| |m -- 722 ||| I [ T o [9060 |73 [Y ooJ r|--| [ IJCwEwoopmtims|[0 eorsis[[sntae|o omo1---- | | [[ owwoerm mo TT[tsoiaoso orrr[To eeoae aosmmeT[t ooom soserim o zm e]| [[ 0JCwr EomurmwT[Tsoe:ss0[nooa01{oomoe0 25 | [[wJoomshms[[osiees[[o6am[r oooorm | [oE EeT m oarTse [01 oor ------ -- 7 || [ [J oshimeJ [Teasms1{0o0ssse T[ooses m| [ [J ooshw m T[oosrsI|o nvazToor soC o TT m [ [J twostoinreo TT sroaiw1snsasm 1[--ooa0 e||| [J[ osho T[ rona To oa s [T--oop oss r i s m| [ [CJO C oosr r sasseT1o anmaeT0Toop oo e = [ [ [Jmrooomermeo o T[oessaooatmeoT [[aodoo aeetre o T[TTooososooesrn --T --m m --m| moron Toa naT0001] IJT room mme[T owe[o eseT e C os T|| [J Isoos [omer os ------ | [[oJeworpmeTToaa0e0[[n7a5 Toomooo CI Roooepimms orseesrTo7osrT Tooom | s] ] Page A152 of A172 Aopento TNO spor of 4:02 Final version p.183 08122008 Ls oe en| mixaded moi protein) [7[[38| I| +WnNaaAooDsPPHHHmmmibsx[||8082867302 [||na4s2a0e3|| | o0002z0308 || [438] | 0 | [8[[8JN[+aWNoAspOPkHmmsx[| [ri9es63re3 |[|76n4aa13|| | oo0o3s303e | | Bow | [ [5 [[88T=S SoNNoAAeDpCPT PiHHmmmmsbxx[| 1[-- 7es7060o87836r%76t {| ||a4Grn5ea ear1a5|| | |o o _oo0s2aop s3m0| | i [m a1 az] o ]|| [[1-- womms[[ozsse [|snoar[| o_owss[ || 1s]| [[[O -- T IswNNoAADmoPPsHHmmmsxx[[[76F550940808[| |n802a%29| || oa31 oooaot | | 8208 | [C20 NAGPHmx| 9543 | 7346| oes [[|ISwNoAsDsPHmmsx[[s6m0%e [|7o4wa [|owoates | 7351 | |[|| [C[[40II-+wnNaaADooPePHHHmmmxsx[[|7a76o68s966[[|oo20os6ts7 ||| ooowtsesr |[ [71 3| s ]| [8[0 |J+e NAoDpPHto mnbxs|| 8a6s3s C [|s340 m65|| o02m01 | | sem | [3[0 1I+wNoAorDPmHmmbsx [|7872671 [|n54a62[|oo03m30 | -- ores | 1] [S[[0TI1w5noNaAoDspPrHmmmsxx[[|o79a93s6%s3 [| 1vBe5nt3a7[||oooosdeos| | [71 o] a ]| [T T[OoJ]snNAahDvPPHmmmsxx|1[5a57m866e8t| | 44n76a1266T|1oo0o2ma || [1 de] s]| Page A153 of A172 sen Os sn C r E C : e r E e STe K F p H H e Ee--e e s --e e e _e r r _e aar Fe F E EBEoE oe e r e eeE E eee e e e | Fe e g F e b eo re e e r s e r s e e s e e e ee ee Fe e r oee e e e e e e e ee ee F e F E EE e rre E e ee e e E e e e n FE = N E r e e e e E e e E E E E E E E nr io e e o S es e e es oe EE L E P F . Ee ee -- er E E = - r ee-- e e e Fp = S FFeoo e ee e E EE B E e. eee E L re -- N e E e i| F E EEE er s t BE n.18e - Page A154 of A172 Aopendixto TNO eporof 42 ---- p.155 ot122008 E e I rs eTo c eorms3 eaE ali iEc e2Cmaeaxr3Ras a3miomFaEoeehaErEe]| [rE c i orms eLiCpleerfEt eeTsELporlt aapleR es pIomlleE ap]ceot Liople-pTep ILfoleo]eot11 otlearlestLporlaoioa) [rr E e e owe rsl e eILpo mpelire arS= HepR ioei:tBpmNerFellafeiooerF re -eeocpeotlr ipatBfe=lo| PageA155of A172 Anta Osotz J osr22008 I. Measurements in vitro experiment (rat hepatocytes) protei-- nconcentWrA ation determination FrlT psaerT a an Te] cf a P aea e es 11 --1-- E e P e Po r r e m e e e e e y bebs e e b e e be e e ber e e e n mee r y a be e P e p E rooe e m re tm e e mtpe ee eennpbereehlreS e be et e bee ea e ee e ] in baey E L p E mebo e b eeh m e rp e eb nss e e a eennbm e e e H me e e bbre t l e mms e e ee e bpbe m aaenn l m e L ome e e ee n re i ms n ee b ee bne)nbp e e r ele r ee pi n e ne pee e me eee bpbd e eeeee pe ebeC ee fteree r meresbp E ep BEe bee bA eee f e rame e psen pe. niel en been fF E i rmE mhT et eT lrywo ep Eq r p Tr h w beea me r a] e -sAe Freeaw Rare pe EP E E E R i r e 1 e re r -- e 1 pr er 1 aaser a er e --1t] or| ss Ete Ee eel Pe 1 e --1 Jf 1 a 1 11 Freer) pre ny Error) (Smee Tome Tome Fpt-- tt A a WO WC SS 7x JO WN.SO SSSN SO 0| Ep E me Ta r me-- TooT y][ e owel Towea r Tae | P E Eo e r re e e teee fe eeepe eee ew ET s -- --1 A[es11 --T1 Page A156 of A172 dito TN Final FlerTe e] Ee F F F E e re t ar t t t e ] ] er pF meatelwbe Eeee Et rmmt m r EE E E --i----e --e t e twr| oF E E P m r r r r e t e r e e E P E E a e r p e w --ae r--tr|| F E P E Pe r emr e t r --) ---- Ea E E E ee r e t m--rs t e t --twr|| E E F E E e t e e t te a t e ) t ] E Pe pe E F P F F ee t e e t r e r e e e ] r E E P Frt e ea a e t E E EE temt e a a E E me pe ) y F E P et te e t t t] e F E F erebt er t] E E E raE t E o] BR I I A P1857 12: Page A1of5A1772 fraERRm I a F AE s LLBi Fi EE Time) Aeeynd M2RromT Fienasl vN esversieon OWpN orpA.160 Experimentx2 Proteinconcent determiination : er Tg = i : rl Ew Iz - en] = w : we] nS RS-- -- EE===-- 2 a| P F 7 eett mec a ] 1 r a : t i =r m = | = E bebeo e >| -- = = =o E = er rerF]r] P E E i w e pe [ p r] e i Fs i : --p ae-- ea re f ---- sFoe rr]| F E mr] re P= mr] = 7>2 esm= SrsmL: r e ]: Rl Ef(=r-- s-- nr aEm---- wFFarrr] eE e e n Pe e r= e pe = er == E E [2 a B s : rFra] E P : r a c o or] F= ea r= t| = wttr r] e cm [ = EEiE P C o =r E PE E e E r : ae r e t F ] e r t F-- or] E ES di ar E =] nt e P1= ee e s] s FF[a=ree]]J]pE r ratrSr i t e: r ] a e = r ] m Fa-- --r!] =me eE w 71 p= e er i4 J wa aS miR ---- Fe --f1in--r P P= t re t Ea || Page A160ofA172 Mopendx to TNO-eportof M02 Fimalversn E E E E a e E se E E Etae te FE e E r= = P P rr p ee E E Ee e e ee e ] E EEEne a PE E o e a E Ee e E = Ee E e e E EE a--=e --=tm E e e re t b =e m --t= E E re ety E E E Et a t e e t a = E-- --=--rtw E e E e e e e e E eee E E Ei r E ] r 6S p.161 oa122006 Page A161 ofA172 Fin Ee -- i LA RR a rr I re ies i eHsE Hr i | fr La lini Loe.J Bopencitlo TNO-repor of 02 Experiments Fina version Protein concentration determination potn= con rg rongim)n con|vgrrowr] I I [1r 283%" ~Tr 7c 7To| 83s85a| [SC rcawrom Trr wwo w3 r] [S[ C 3c0 m C a300o 14 r t r 17 see rmros |s 1me2 0e7]| [s Cs moter an1a] [[3I eEraT2Y r 220500 TT&aT r Tmsss20[|e29o40]| [II 2E0 2 sosw7e 16{amsL3| [[ eZc a= ne Trwomowss{|same3 a|] Fe e amc memTr omeYo | (Cor ie mr ano] [E I 2817 2ee72 |v r 4| 1tor7as3| 363 83 | E vom rrww = 7LT1 O2 E E m me e s Tr o o 0[s s zemy E Ta s 1 1r m TTooroapse {smurn] s o EC Br r 1 2 || [7 [ 2 C4 om1 o 2 TTTa rr 0 2 aooT|7 o0 wsmaamse6 | || s236 70m00s8i||] C | [FSIoaIr s1s9e o5ee8 |Ta 4r Te |782r 319551|2o6on1e2ss|| CEIT1 2er0csomT T rw1 + --tmima ms7esso3[[] assmos]]|| [ee os122008 Page A164 of A172 AspenTdNOirtepor of 02 Finalverson ae] proteiWncoowncie(my) gi) |apevrwaoh] [[[[o88 ErI--lssttwseseseosssttaTT[s&ao4--TT|J vvoaarssroso[||aamesrooss3]]] ris [amo] [CCCeoeero|T Is T zm s2 sora T T10 a a r||o0 swdeosroTT|6 aaiamaas]]] [[CCeeeToszioos [es ome TTa&a[||wriitoen2naes[||sssmimao]]] [ICCor IC zest 4[time7s20| 022] 2 CS co 7 [[Er[ 7Es0 2 s2asa7oa T T2 1&4 roTT& s owrwnsoso[T t ||73a20m63o]|] [[rroe0[ 2010s 6I TTaam || si1omiXszo[s |i2 saro]] [[r7e zsosr20 Ta4[foeosst[|e5rmers| [CEI oOt 7 seercso I [vaXsoooo[amr] [CC Ce20 T00wT&3 2 T sioa[= sms] [[CC5 ooer ssie--TTr&&o[|7 sfiussz[[oarwzao|] [SCCee0|O stoocsomTa 1 [r ss2| mo] [CC oor saomsoeIr T T&o7 T ow52 e5[{ras0]| [[oE Cso Tos2mtooe 1T n S &[o 7 vrwse || asmr morr|] [o[ [ T o 9 C st2 s01 0 4 2 4 T |wv0 i<uo 71 6e26T[|2a1 305 i00r232s|]] II E207 XS B2 EY0B0 X [C I Cs 7 75050 TI&0 Tw2 izB [amT r] (C Cros0TIr TaB a | sm7Y Ts mo] p.165 os122008 Page A165of A172 PT Hitti Pte I BR JT ASA ADA AE U INA | [lr | Hi---- ee iE HE EE Er = e rh n -- m-- E WA `Appendix fo TNO.report of 4.02 Experimen4t Fina version p.168 04-12.2006 Protein concentration determination [v[ oi[wp iz]=e eor] )e | porwet| ns mm7oJT5 om14sTT 1 worTeor]|| A7 3 72S 0 5732000N3 1T 1 = = 1 | 7 meTe720 I1 7 B 5TaeY wei]| 2 | [iZ s TTweoweesstTT asme uesr[T5sT Tioeoessr [||wsseosss]]] 77 0 3 1 7 OA T Te7rS20T3 w7 +a1 TT5 7m32 7o rw0 cor]| [ F 3 7 a TT2 a3 aweam a TT m une s|Tey F 73 Je a 5 F e e r wr10) SF S C m7 neo X CW nsA im | s i Tu 5 asT aT a a a sTao aai5 n]s] 1 [T 20T7 aosw TsTr oss2[sa7vs2]| F 172 a S ae +Fn e1 rLae)| O e C Ja J Fanee17)-- tL J77 J SS Y S c Y 22 ro| e S S eeTim Tm |m m amoeS F T Tse T TY s imwrosr --wwser--]]|| s [ 8 o moT Tw wamose em TT+ Tr e iou o uorssoao eoeor]]] Ce sTmT|awmees1 T5T| voossz| e30i07]] Page A168 of A172 `Appendix TNOreport of 34:02 Final version p.169 04122008 [ [wio i1wmeoizrz | 5 1ro o9 0[mpVGepr3owtoeln| [ [ [s ee s tT T1ouZisoseas ||1u22h0n 0o8at0s0]| [o 5 5 |o soosrro |] sre] [ [e 2 eT Zemeererne | |IT 2wtoomotos | | | 5 55T || a svioewss | o | sssis eas| ] [ eT2e 2s08 1| asiarT ss[ |5 5 || oiszess2||ssre0r6|] e e T 7TemtT oemr1 |ossvuees [ |F 5 |mier s | ass] [o 2Te azmeue |1 o msawa we | 55u 1|a e wis a |ar]] F[ m m awe awozmX [ |5 5T|iweeers|| msmas|] [S e eT T Toom2vmeimsT 17o amm e m | | |O5 55S s |vvt2iewers0 ai||s30r7v 5s | o 7 m2 1T 7 pm[5 | CvLei |BsoT0| |sor] Fe eTo221s a78m5 TT sma2T s1r5m7 [ | [555| |mtfiousmses1||s3sr60o]| [e eT 2aoJ 8si 1 1 zOeirKsst |N 5 5T[orwraroS o || saawrsr || [ [A e eT TZiteernsTisI7 zse |[ 5Ss OTvoosr25 ai3rs0||E333s0Y 9]| e oT T Te2n o0semT T1uiaeose ses [ [5 5 5 ||o vjooaz ierr || 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A172 oieto TNO oerof 14.2 Foal ES puss-- ies[CH eSEereR] EPTET r oITRT | fSpaa oene ueVe |oosswOnR [Rw an|ae aw|e]me | | Be | oame -- ; AII ete T TETEE] we eS emsmtamlan [|Te |E[7e Re EBe]|| E o | S vmm |e mL e |] om mce soilmnem | R S[ | |v| 1] | Wewtwwe || ( ww Ee|mmmm l||]a2we | oo0ioeE ton[TET[Tm] ans r a= E [T E S FA a CN IA s reIE] ] ee] ndnaoene]l| i2 mmWFAoooaIw]|u|I-A| amwA]e | ue oe | wo lum] | F= E e =| = ennn |e p13 12: PaAT gofA1e72 SERRTC Foal version p17 oe1z2008 I r= Pe I iEr rsd ooS m--no|el| H|&_22 EHwam =oE ooam]H w luwlmam]mH | m]|E wwee|] | H oaaaae=enm t=yCT CT=E TT] oe CA R ScS rcmanslmlH mBEBr|GwU ]iVm H 2B BI B w[R l] ] lE 8EamE H ]mawm] oH soo=emes = FO CTE wrsrwr= etn-- CEEgm--] amon] [er aE ma]r o e e a Sei o--nr)t|E E 22a Eaw= =weo|w]8u 8zlE ] LLEoo==f] oud - CT o er Tow e = etn[ETTTE] wes REA oy|| araee Pa ABOg ofAe 172 Appendixto TNO-eportofM02 Finalversion 04:12:06 : ps jaa a : = E L|E = oE e E r = = = Ei = i } = I0eIe e = r onI|J ee wniii= oo e= sH= = ia = | E EEee S H : a E t ji = E fe E i a r oe siit a fas pE e SEE E Ee ! = ee on EEEes Ei: ki ne 77 i Pag [3 nixto TNO: Fina version 12: [ss ews CDEETme EpTriremea[bearers asetfseasaitaTTarmmereaas TLi eareear]] F r F F i eo a E a c C tea oE R B a OE E co BBecsa ilof Jc Bc Rc coJ 5 [rn F E eree aL C mcoxJLpetaoB E tlLpneR R iiJnt Lue ocB BotoJ|E E RgagsB R =J| soai0nd|JsoNes<at1| =E F m=m e e =Ella EsEETall RoE ecRToEleE deE oeBToElC ae oIololBE o2cCLaffl B2ao1oloTav| EF F Eroo aeme eSCo SEceB BE BcoCR lRRRE <C BMR BE = R E B fI I oeI [si ews LEmrhea feaareis par3 oEle3be3ri e maai<nti s3eBpoare ffaa3earta) F F F F Fa a e or e e z e c a R EeEREiE]H CBEHCRi CBBEEE [EA S i SEo E Ea F e h i ai Ce TC BSC ES Page ABZ of A172 Appendixto TNOweportof 302 Faas moss Finalversion EE ----_-- m Z|iT l]5 Pri [TIlSElEF]d 53 8 E mE =l=I ERwS AEa 2Om E | E R|EBEE |Ed)EA EE8 " EE eon[TIETTT ---- THE] EFSEelmm[aal |)|T |aPrawceEeS M||HBl] |E HB|Ea]H 55 5 sEmmmaEE0) OEE E(8H8E8E% - -- EE -- cesar [EETTETTT] CE aE =z: ER EAE EA = = mne | FBoRE| EE|REse w Om oneCEETI]wr om = = = = &= 04122008 PaAB3gofAe 172 SEE sae -- -- -------- TEE EEE = S TEEEi lRwIe: )Efon| EulRe | E8|BIE)8EEA)REAEk:8E EEE BE gre ovoerrs CETTETPE] S rss I: ir ProFni|EI ESTKi]lEF2E]] TE | = = =8 mEBRoHEeInHP|I|gELEeEs swreCCHEETTB) EE -- =aH === 2H w" BmOE]oaEEu ee] 8 - wore CREDIT) p17 serps 182 PageAB4 of A172 `Appendix to TN reportof 4.02 EE Final version p.179 04122008 == = + v sala] T] os T fd m _ m oT m , = c deat ca == onan Page ABS of A172 133 Aopen 0 NG toofp4o2 Eravasn EEEE = moI l r=|Te| Swn=I [mlSel|P] ]v].8 m sComontm [am a | Taf m=Wwm | |[uHf ak|8lEWsS]ES2 vom |v] ow ale lv]e o-rCT EE B] EE Emls I ee ={oTl e | rs I le |S i| TdFire Fet mm oolts | etie] l| e7 od=ZeTwm ||fzoBn f|(83]2Es](]lS|Fz]2sSE Ea E- = roesane CT HEETaw w ] z poo[lFeCy]EITgeEeEemL a] =CBRP eiT A lET A ] =Eel=EEwEmle| oOBmEO|E|(u|E|YE] 2|Ee| E28 cwe CCEEE] TTET] = sTomEo F32o owmzme 5 p30 cera Page ABB of A172 AppendixtoTNO-report ofM02 IEEE -- Finalversion =) oe Ep me ||| Zola En Lo dm --- I -------- [= Esm ~FTe ea] ee = =u Vs| in Boel EEEE dam || on | ow | ww [CIEE] cura[T IRE E TR ] [Ea] eee) Ta 5 w| | o | oo E EGsoa=l]E lTEaomEn|E| EoE | ||EE WA ||E EAE R E 88 mm] ow Golo] [EE] eenCHROOT] J wr = & Ts 367 F ooo a7 Sa S= ees 422 wpda 04-12:2006 /35 Pa ABTg ofAe 172 nda TOs 052 oss -- E FEosirlctE SEET | T O=OBoeE EE EEE T[|(TB SFT R EYT ]FeE2= SEE EE c-meCT EEmE HT] A EE -- Ce Teme eee] E E=lEH mELomW OE |EEEEEE E EEE -------- Lx:| weave CE THL ] 0L 0} CE F= ol F[| wOeE EE EEE a [TEE80E] RHEE = Cn Gone CT ER IT] mw = EES 2 p.182 see 136 Page A88 of A172 xfoTNO-reportofp.183 Apendi3-02 Finalversion 04122006 A ---- 2: cop e Ce ERTTF) EE --, sE ri E = oun mm (BETE --] A ----. N 2 e s] osCTI HT) 137 Page AB9 of A172 cea cris i =lz= == [ElEdEF]5455 ESmaHmEaE]EAEoEa n(EiEREsE E]RE ere CEETBE] EE ---- Z Tenlae]foE n| 2we L [|G] EE8 S]|| ns 02 ElEeElE] O=E [a8E8E5E8 w [Em ereCETTm) EE, Cele Pe = 7 a | BEL] E = ==|| B =FolE EalE EilE|E|88 wsrsCEEHTE)T TC r) l 2 ci Eo = old & p18 ownms 188 Page A90 of A172 p.185 A ---- ee lee] con-- ren)HH o=e= e2e dBs ieoo l|0E| nI8Es)IEE2Eow EEL = = lglglsle woenC [CEEE] LTS Ram i E = F=Ek r |ee == aBToo|lzE2O|EE: | =zg% = |g]8 3 omerCL [EHETE ] TT] Il Ze PEeE MlRiIro E == =|m8 BZwzo olaE|lalalaellE]gE||]|EsE w Cm) rte[TTR OC]Em] m) ZZ E =a = 197 Page A91of A172. AntaTHO potr02 F-- i-- Il= ld = 4 T = S= mlllema | e]an| l2=oee 2 |e|(|alEla|iElss|F)) 8E gE E mm] we JaldlF] a [CE] ---- wee E SEE LE = = E TfrH aE ]|lHE |i POo Eoi Ll El [|(| E|YEF|l EEdEE8aE mEmmel]Els oBwE | JEHEE F E8 EEsweoEeE l CT COET)ETT]--l c l sd = E===5ea7 dE 2woo|E lE8oER l8e|l5EE]|EE|EE B= = w= lElale] i w CET ase(TESTO --TEE--] =H @ 5.188 ostzz00e Pa A92g ofAe 172 sens H|Tceo-- ---- | p.187 ae w [mm union CETTETE] A --" JMal 5 2-8 ll 4 5 TE we| m| wm fom] | 8 EEE OR [HEE osoue CETTE] A -- = = = =i= i: w= B=e |ou eaEERr]E]AEEEEE =| = 2 EEF] 8 o-sCETTETCovr)TwTemE = == ==z8= ==z2 Ei = Page A93 of A172 sone o ppt2 Eraverion oe2208 A, sen[T TERTT] I es (TREET--5--] A" =: onus[T I Tr or ] PaASHgofAeTT2 172 p.189 SOO RU ROR] Com Pmce m RA Tem ---- B= =CHHIEE cesCH THE T S5T 7] 17% Page A95of A172 soniaMOspt 02 TW EE leE m an| owmm sm Eel ow E w ||E]e E | 5 S Ee alr KA c IE EA EA EAE [EEE] mn EEmEi eseCERTTET2801 0 x| AS TE = m= r E83 EL 8 w CE = = =z z =5 E=oeoR|lEE8lE5|Ed]EEEE85 ER EIEAR AR: avo= nsCT = EETT] = EB p.190 ostz2008 194 PageA96of A172 Appendix TNOveportof 3-02 Finalversion I ir V0 S| [Elsa] Fae E EEEE E ElEEEeE L]R EE s E EO2E gE( (Efl] EiYeHeEE]zE8 CEE] emeenC[ FE TE 0--) |E =Sll FL = EElli wlea=s]E ]PwwR e e= a [E 222a252488= | am]E mEeE|] [2Te | dEgL]H RE8 we CEE) Ey ais = ld ee = EESSmiEmEEnlEE ]e]ERE2oFw |(R2igElE58EH|EeE88]HEE888E vo [EET] con CH TRTr T ] EEEE oomm pei = priced pd soe er &" 04:122006 YELS Page AS7 of A172 Aopen THO spon or M02 Einalverson = HEERHEEE EE [CEl=eTm eal] e] Sole [| em |B] E |e Vial wen | [vee [| S| we| 0 co=reC CEE] I SC ---- -- [CE] eae) E= E ae|| ioew ||S]||5]EE aEmmwelIa E E [i E S[oem ( E3 NH[EEEEE o-reC CEECE rEs)rrrersn ET -- [C=] ~ PTs] = To H mo| om| ab:n | 0ah] 0 Soe|| 3e%w SSm oom |Evl| eFf owWE [ n| n||E|Eem Ce| we |e [i [8G | = C= = ==mc] = cn E7 E-r =3 p.192 aet2208 PaASHgofA1e72 rico TNGcoprof 02 Enaversion |eI ---- I 5Po IlFd) prcerd o oo a i GaCooo]]tt|| oaes S =mm=oel |B|n[nE aeBlullEaaagEEll]]ee|]lsseae = I" Teoer l ese oon[TELE] E EE E [CEE] Trae prscCimosaomlllaww ||[|]e||WwooSn Tom mlm || [ (H w Go BE wS [oEo [S wa Bo |naume = | ||lo l[aE laR lv] le orsCOSTET_T] EeEr -- Zcoam[e = e RSEl E Ns E~ Ee Te] as scTCoonomrmlsl| cvEvmemm|ac|i[|Wa | ool we fm |uwWewmm om ||| 3[|mBu6| |GEw0E ey[@|e | 9v%e =HEHHIEH coewsoen E CEETTE[TCErE]TeTwTEE comms 347 Po oCSfemnea m7 F==nr VEesia1 weny p.193 122005 PaA99gofAeT72 nto Tot tz sin ------ ee[CF ITEEaEs ~ Pm eEna TnEe] ] Tol om fon| wie | |S || SE Sa 0 | | asus oeousl CETTETg] I -- EEEEE = EElE uenR]] 5wEm h[vE8REEe|e TE cmmllnEw RomE |E|EEE |8E -ence [TTH OC]R ve] m) = =H5H - fl cc 0 mn =EE algEE]@ om om p19 surname 179 Page A100 of A172 Aopento TNO pod of 402 Final vesion C -- Es == al rr PCvoIme]il.rd| i em fTar= ae]n HRHEwo wwEe El l|E |H ala nE aE |lEen |H ls [zm omeCHET TEE] [CEE Tee osf= Torn=ne])]| H 2w4m EH m =A BceooH E lauluH ElnleasE l]a enlH E eE seeC CEETTE] al 5Pr Il spromce) PY campez r l] Vom| he ame | 5o|lewe [euem]e | ow fo SE] ea EI RE owu-nCCHEET] gm] praeiit 4 =r orate 4127 &r Se " p.195 rz Page A101of A172 199 AppendixtoTNOreportofM02 Finalversion EE 04122006 =Aal r= iIelile[ s | E = =| m w w= oouuen]aE sE|e =)| fos www lal alGalEelEe -= oreCERT 5] SS Te= es [EchE o | E me]| 2Pl[07e| oaim|le8% f= E e H EwwlE omoB [|[|sR|aEw]]EEa Sh wl Ela lvlE - oreCELT Tg) CE aSE Ili or amel Aal I alEo | TEle] el: =SEe=mEEwEC]LET oLBoemOLE|JuE| aii - Cm cos r CEE m] n ES E =B 2 5 & ne a " Page A102of A172 RR sn sem Cre e BRe EEN) == = Eels e wu [EET] C m =T e w e] uw oesCTBT TT_585--] F EE SnS TOI ee lee] E=|a.E aFR|E| E8 R[| =z EREAREAEAR [Er oc. i enCT TE Tr O ] EE CS a Il a a) Fo td Zo EEA RAE) = =e H zEZ||i#|lazl]az] = [m EE= ]mm] ow = = = =& RE 249] Page A103 of A172 fopendix to NO-epoof 02 Finalversion oe122006 EE ET a ee IH Ee A H iAReR er eARiGe re eieii EE i a eeT eneee Rea EE aey iil olec a tli eeelee ee ae ts es ie | Iline eR | hs Eee EE ---- rig [EEL da r Se If he e rr HR0 iida Pm ee e leee l e re e eel ee M b EEe Es t eea ee e E s a El E 203 `Aopendie to TNC.teport of 4.02 inal version 5.200 0x122006 Cer Ta E Si sBseD e RE [ e CC oe mI mo e asJ Se BT alCc R oRceSBli ooe-BBRNBcoc BE BcJ J BSEccRI RR=s| ae Cotmeesso c= BBBccoc Boorreeo.seolS=oo:B J =BcCc < ficoSU = [m [en vespCaIeEfEeRtSfa omervaitLo{aRanR anodiear+l |gerRL dv|grin+dsL |poApnotc| oes Ro RoR oc > ScoBI Cm [aoermree cCc oS o EnRN5R5= oE< JE B b<=aJ R oaJ ooao [www gSeeerse Lbaeet,atL |geese|gnarlL|gEusL int | cRarst oatr] [[ EoaC mreloo SoBE BEcooRl RH BB Cl Eo rlcol oFieCo < o | [ --m-- s SET w:foR n o RR en oo.aS llcool co | CoRI ooRI co Rl co A +c Fol +oIl | PageA1o0fA1672 `Agpendin to TNO repo of M02 4. Fecal faty acid measurements Ea E -- Final version -- p.201 0122006 EEE] e-no CE] HEHEHE c-o CET] EE-- E -- w (E TET TWSTE EE] os CE] I ow cw C-- om ---- - owe [TEER = Ew ETere ws oso [CERT] d05 Page A107 of A172 Asso HeOnotnof 42 Foaesion p.202 senzmn ETE ETT, a - Ereerrae - I-- I w ETTTw) ww CESTW I --A Tew) -- - OETETEES CETree 204 Page A108of A172 Appendix TNO.rep0 of 34.02 Final version I .203 012.2008 o-we CE EDTTTETes] owoo (CEEEETE I --I -- - ErT er E LT EE 201 Page A109 of A172 `Aopen to TNOreport of 1.02 Final version p.204 04:12:2006 EEEIEEEERLET pe = EEEEERIEEREEE] et oso [mE] Eeono CIEE] EEE EERE cso [CTE] ono [HE] Page A110of A172 294 oTNO-reportofp.205 AppendMi 02xt Finalversien 04122006 HEHHEHEENHEE A I Tere, Crerereren ono CE) on Cm) HEHEHE ------ |T-- - CETETSTET] one CE =~ CElem on Cm) EE. W-- E I ere Leer pr oso CI Page A111 of A172 J07 AopendintoTNO-eportof M02 Finalversen 04122006 TT --------I -------- EERE EER ererermT = re Cere re FEET EERE CErETrETeTES - Gere A I-- FEEEEEEELEE CE CEC Tm E rm eres Page A1T2GFATT2 2p A JO J cera LLL]LEE aa EEETI EERE w CE TEE oesyS m) ww CEST]Teese) EA SSS a HEHHEH[HEREEE ee EERE | EE SS SRTE ] EER TR AST w E (EET T TEEE Ter] ow [EEmE Tw E we] | oso [CE] ose CIE] | Page A113 of A172 | 21/