Document vmK9nMwjjNJ6zBky1dq83Lzq

LDDa.iVrre.rcMyto.R,rP. hJo.Dh.n,sDonABT September 19, 2001 C3MorpMoerdaticeaTloDxeicpoalrotgmy ent received OPPT CBIC - o te 3P66SlrM55Otj.11oPhBC77nao33eusx3n3ol,tn3e9M1@r327,21N7mB2830um5OFi5lmadf1xf.i3icnc3oge-m30222200-02-E-02 U4CDAOW1ESfto0tafcnRisEcu:MhTenmiInSvoFSegiefnrIttcorEoTtteninDPoeom,trxn,oDMiecSc8nCeAW(tSesaI2us)lLi0bCPn4sgrot6oao0Ctnrecedcenitnsitoeanrto(Ar74g0e7n)cy 2001SEP 26 AM: 55 SeHQ'09o(-msx3 RPeer:flTuoSrCoAbut8a(nee)sSuUlfBonSyTlAFNluToIrAidLe:RCIASKS#N3O75T-I7C2E-4(SUPPLEMENTAL) ON: Dear Sir: fr3drksinnelecaMneusshryuoooueasrwrehlllo/tivanwandstetos.eie(dwoxTce(PniebokPScrByn)siBCe.ttdSytuSAwhuFAdFaeciy8ntt)athneesiai(midesnlTnSloxaaSurctlnlaeCbi2sdotavss8aArteetmey-aaxldd.snt8pa.atecolnyTioasCelShierlnumlueedciRrpbnoeiftitseinutossctoa.nktaarxtPcnlaNiTdBtttcsiioihooiaSiotsgetlnyFneinacRsmswleienio,xewosahfckhdsbeah4iasrNlrba7teaeeein,rotptdivoeitoo1siabdc3m6nrtste0i2ceeos,lordtaJindvunfunaiaedilandlctdyyceab4atdlewa1i7oamt9sS9itrni9telogehypr9pufny)popstisemnreeaacmrnnttiofhhdd(nlbni6uehseihshonarienirrslsx2soeaatt/ep8tuobdrin,orpouaaotstanty2uatwt,ao0hnrtco5ixe0oetuxih0slactoiu)enic.gltdifyitocyhinasalydnl ot fic3nolModrtuPhhsBiatnrsSgiaFe,ls.ahtn3aydMbglliioesuhncsaeeeldspePraxaBwhcSatoiuFcrsektsspa:vlfaesecnuleytpipleaalxsitiepaodonssaiudtierruerrleiilmnismgpitiicertahd(ta8oirn-rghytiernprgTmr.oWetdeAcia)titoeonfw,0ic.t1hhemtmhgei/cmfaol3llpo(r0wo.t0ien0cg8tivpepm) =r*o <^> Please contact me at 651.733-9218, for further information. Sincerely -, V H /V X ^ LDairreryctRor. -JoChnorspoonr,aDte.VT.oMxi.c, oPlho.gDy., DABT IENnHclAosLuAreT: IOCNomApDleMte IRNeIpSoTrtR: A"TTI-O74N99TOTOCXDICRIATTYSSFTOURDY4 WBYEERKESP"EAT DOSE rR\En nCEt IuVnEoD 8EHQ-99-14523 BEHQ-99-14523 89010000317 89010000317 CONFIDENTIAL & ro3 MIN 252/004312 T-7499 TOXICITY STUDY BY REPEAT DOSE INHALATION ADMINISTRATION TO CD RATS FOR 4 WEEKS Sponsor MUTP3S3.MMtoSO.ixnAP.iMCnac.Beuoeesolnldo,oxtitegca3rya35BlS25Du2e1ire03lvdp,3iiac-nr3egt2sm,22e02n,0t-,2E-02, Research Laboratory HACPERWHENaeluuocmp2nnGooo8ttbnliiLrlnnrb4teAigguHydisddNrgSsRyooeuD,,nnose,.hadLdi:ri,efe2, 2SAciuengcuesst Ltd., 2001 Page 1 of 251 MIN 252/004312 COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS PTrhaectisctuedsytanddeasrcdrisbaenddiIncothnissidreerptohret dwaatas gceonnedrautcetdedtoibnecvoamlidp:liance with the following Good Laboratory The UK Good Laboratory Practice Regulations 1999 (Statutory Instrument No 3106). OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM (98) 17. EC Commission Directive, 1999/11/EC of 8 March 1999 (Official Journal No L 77/8). UP17anrAitte7ud9g2uS,sttaF1tee9sd8e9Er.anlviRroengmisetenrt,al2P9roNteocvtieomnbAerge1n9c8y3, (aTnSdCAsu)bTseitqlueen4t0 aCmoednedmofenFtedFeerdalerRalegRuleagtiisotnesr Iaanbvfoaoivlream.baletioton HreugnatridnigndgonteLstifseubSsctiaenncceeschfoarraccotmerpislaiatinocne, wnaitmhetloye tGheoobdatLcahbnouramtobreyrParnadctiecxeprireygudlaatteio, nwsagsivneont DHStueurndetykinDWgdir.oeCnctoLoorim,febSs,ciBe.nScce.s, M.Sc., Ltd. Date 3SMponTsooxr,icology Services. Date Submitter. Date :2 : QUALITY ASSURANCE STATEMENT MIN 252/004312 The following have been inspected or audited in relation to this study: Study Phases Inspected Date of Inspection Date of Reporting Protocol audit 31 October 2000 31 October 2000 Study based inspections TCRESSaxteeluimscpndtooipycsrslduauipnslbrregsseatipuagadnnricsateticoonntrol )))))) 13 November 2000 13 November 2000 Blood sampling 8 December 2000 8 December 2000 PRoecstomrdosrateumdist )) 11 December 2000 11 December 2000 Report audit 6 April 2001 9 April 2001 PCroomtopcaonly: MAnanaaugdeimt oenftthase ipnrdoitcoactoedl faobrotvhei.s study was conducted and reported to die Study Director and tShteudSytudbyasDedireicntsopreacntidonCso:mInpsapneycMtioannsaagnemd eanutdaitss ionfdpichaatseedsaobof vthei.s study were conducted and reported to rrPoerpuootcirneteessdabtnoadsarepedppreiontipstprivieaectetpiorConocsme:dpAuartneoysrMeambaonpualotgytehemedetonimnt. ethtihsistysptuedoyfwstausdiynwpreoregrceasrsriiendspoeuctt.ionTsheasned wauedreitsproofmopthtleyr RcoenpdourctteAduadnidt:reTphoirsterdeptoortht ehSastubdeyeDniraeucdtoitredanbdyCtohmepQanuyalMityanAagsseumraennct easDinedpiacratmtedenatb. ovTeh. is audit was Ttohreefmleectht tohdes,rapwrodceadtau.res and observations were found to be accurately described and the reported results HTDGrueraopncuatyiprntSmgMcdeaaonrnnftaeoLg, feifrQe, uSacliietyncAesssLurtdan. ce, :3: Date CONTRIBUTING SCIENTISTS STUDY MANAGEMENT DSteurdeykDWir.eCctooorm. bs, B.Sc., M.Sc., AStnutdhyonSyupMer.vBisoowr.den, B.Sc. (Hons.), C.I.A.T., TOXICOLOGY PDCroiipnli.cRnip.JC.a.lHPTaaortdhxy.ic,(oTBloo.Sxgicics.,ot.Plohg.yD).,, M.I.Biol., C.Biol., AEROSOL TECHNOLOGY AND ANALYSIS ISaenctSio. nGHilkeiasdo,nA, Mero.Aso.,lPThe.cDh.n, ology and Analysis. CLINICAL SCIENCES CDHeaenvatdirdaolCfLrDoaaobtkoa,raQBtou.Sarycli.t(SyHe,rovnisc)e,sP. h.D., STATISTICS GHeraahda,mDeFp.aHrtemaleenyt,oBf.SStca.t,isMti.cSsc. ., A.R.C.S., PATHOLOGY DSairmecmtoureloMf PcaCthoormlogicyk. :4 : MIN 252/004312 MIN 252/004312 CONTENTS Page SUMMARY................................................................................................................................... 7 INTRODUCTION......................................................................................................................... 8 RELEVANT STUDY DATES.................................................................................................... 9 TEST SUBSTANCE.................................................................................................................... 10 EXPERIMENTAL PROCEDURE.............................................................................................. 11 RESULTS...................................................................................................................................... 25 DISCUSSION................................................................................................................................ 29 REFERENCES............................................. 30 FIGURES 1. Bodyweights - group mean values (g).............................................................................. 31 TABLES 2431.... FCBCololiiodnndyiiccwcaaoellnissgsiigghunntmsssp-pdtouigorsrnitoneu-gxpgpemrxoopseuuoaprsneumvr-eaelia-unnecgsivrdao(elgunup)ce.es.d..i(s.sg.u.t./rm.ai.b.nm.u.i.ma.t.ir.oa.y.nl..).....o......f.......o......b........s....e......r.....v......a......t...i....o......n........s....................................................................................................................... 11165781209........ FFFFFFFFuuuuuuuunnnnnnnncccccccctttitittttiiiioiiooooooonnnnnnnnaaaaaaaalllllllloooooooobbbbbbbbsssssssseeeeeeeerrrrrrrrvvvvvvvvaaaaaaaattttittittiiooiiiioooooonnnnnnnnbabbbbbblaaaaaaattbtttttttttteetateeeeerrtrrrrrtyyyeyyyy::r:::::ByGALRTCo-raeeocidpnmatguyirdrlvspwiobintienrtouegerygpuiangccmtfgshuoooutrtuouhasmentncs---ttmtpsisvgglg-a-airrrrtyoooygyguuu-rrompoppogfuuommmroppnobeeeimumtaasaopennneerramaivvvvnnnaaaaegtlvllvaiuuu-aonaeeelnlgvsussus..r.aee....o....lss....uu............pe............s......m.......................e.............a.............n.....................v.................a............l..........u................e..............s.................. :5: 33333256 4344444435742687 MIN 252/004312 Page 1111134567..... MHBMOlraiaogcecoarmrodnoasswcctcohoeolepiopmgigichciystppst-ara-yttghhg-rooorlgloouorupggopyyumm-p-eemeaiaxnnenpcvaiavndanaeldvulnueaecedlseus.i.sen(.u.gsc...m)i..d......me......n.....a.c.....i.e...y.......s.......u........m.............m...............a......r......y.................................................................................................................................................................................................................................................................. 4656639317 APPENDICES 12345679801.......... BITCnACFBHAoduollldbadiiointenndsvyaocomiiiilwdtcctdliituaaauooeictllmtiannhoegsslaaeleiihoollpggmtrsgaconnsgpiytobss-sahemst-np(noreipyrltnmrwiorevndi-egneeadai-nivietniilcginvtioxdoasdhdicplndutfpioosafvruaomiosl-ilansudmblvotdruiavatneeiaoftlna)xdtluureglp-inevuvaisosycaceii.sd.ntlt(s.-iuiu.udgo.v.r..eaiin.)e.iv.nl.st..a..yi-.d..v.l.d....i..-a..ou.iv.....nl..ba..iu..i..dd..ls..n.e...iu.e..w.ds.v...r.a...i.i.v.e(.vl..d...g.ae..i.o.u...d.tk)....iba....u.l.o....l.ys...a..n....ed....l..oa...r..a...v..l.vb...i....a...lbas.....y.l...e.ta...u...i...r.to...oe..tv.......ebs.n....a...r...s.s.(..t.y..e...s.i......o..r.e.-........v..nc..........a.i.so......n....t.....n....i.d.......o....d...i.......n..v....s..........si).........d....................u..................a.................l...................o....................b...................s..............e.................r.............v...................a.................t..........i...........o..................n....................s 11111177622678678080226809 ADMINISTRATION OF T-7499 BY INHALATION TO RATS............................................ 219 :6: SUMMARY MIN 252/004312 6gTrhhoroueuper,sgarcaotiudnpagsyaofocforrna5ttrscoo(l,enawsceahcsuoetixfvp5eodmseaadylsetosaaawinredoenk5lyff.eomr 4alewse)eokfstuhseinCgrla:CwDholBe-Rbosdtryaienxwpoesruereexspyossteemd .toAT-f7o4u9rt9h, Tanhde Hstiugdhydmoseeangraonuaplsysreedspceocnticveenltyr.ations of T-7499 were 47, 162 and 459 ppm for the Low, Intermediate The following comments are made in summary: Clinical signs observed during exposure included circling movement and lethargy. weCflafielnkciictnaaglnsdoinggnetosneeosrba(slaelybrvnreoedrsmoilmavlemdgeapditri)aiotaernltydopheoyxsppteoersaxucprtoeivstiuhtryee.fionTlclholeuwsdeiendsgivgdonacsya.wliseirnegcaonndsiasgteitnattiwonithwhaenneuhraontdolxedic, Tathtaeinoivnegraaldl emgreeaenobfosdtaytwisteiicgahltsigganiinfsicafonrceafllorteasllt treasttsm(aWleese.ks 0 to 4) were lower than controls, A reduction in food consumption was evident for Groups 4 male rats. bTlhoeorde cwheasmnisotryefpfaercatmoeftetrrse.atment on the functional observational battery or haematological and Nineocrrgoapnsywreeivgehatsle. d no treatment-related macroscopic findings and no treatment-related differences Histopathological examination of the respiratory tract revealed no treatment-related findings. Conclusion Acfoolnlnosoiwstioenbngstedwravyiethdanaedfttfrheaecnrtseileewnvateslenf(ofNeecOvtEiodLne)ntcwheeaosnfensruovstotaueissnteasdbylsnitseehumerdohtodaxduircigintegyneitnrhaitlshlyestfruuednsyoc.tlivoenHdaolpworibeovsreerrtv,oacteiloxinnpiocbsaaultrteesrigythn. es :7 : INTRODUCTION MIN 252/004312 Tcahshsaeemspsbuetrrhpseo, soseyfstohtefemttheicissttossutxubidcsytapnpoceteerfnTotir-am7l4ei9dn9,raafttosHr ut5onctroiennpgsedeaoctunatidLvmiefeidnaiSysctsriaeatniwcoenesebkLyfiomirnih4teadwla,etHeiokunsn,.tiinngwdohno,leE-bnogdlaynedxwpoassutroe TanhdeDsteuvdeylowpamsepnet:rfToermsteindginofcComhepmliiacnacles (w4i1t2h)t.he guidelines of the Organisation for Economic Co-operation Twalavhbaaesoilrttaaehbtseoitlrisisteupysbe.scotifaensccooemfwpcarhseohaiecdnemsidivnueiestbetaorcekrdegbgryuoluianntohdaryldaatriteoaq,nu,riearelapmtoiesnnsgitbstloeanrcodluinttheiceafolsrtraaancidncidpweaantshtaoslleoelgxeicpctoaeslduproeanrianamcmectaoenur.sn,tTaohtfeotrhuaert Ea xppreolsiumrienlaervyeilnshwaelarteiosnelteocxteicditoynstthuedyba(HsisLSofsctuodnysunlutamtiobnerwMitIhNth2e51S/p0o0n3s3o4r1a)n. d on the results obtained from The in-life experimental phase of the study was undertaken between 13 November and 30 January 2001. :8: RELEVANT STUDY DATES Approved by: HSSttRuuddCyyMDSpairnoeancgstoeormr ent 223606 OOOccctttooobbbeeerrr 222000000000 Animals arrived at HRC: 1November 2000 Allocation to groups: 1November 2000 Exposures commenced: 13 November 2000 Functional observation battery (FOB) and Motor activity testing (MAT): WWWWWeeeeeeeeeekkkkk-42311((((FSSS(FuhhhulooollrrrlFtttFOFFFOOOOBBBBB/M/)))MAATT)) 2299158DaannNNeddcooevv11m0ee0mmbDNebbeoreecv2rree022mm000b0b00ee00rr22000000 Haematology/blood chemistry: Week 4 8 December 2000 Terminal kill: Week 5 11 December 2000 Histopathology completed: 30 January 2001 MIN 252/004312 :9: Tradename: Chemical name: Other name: Intended use: Appearance: Storage conditions: Amount received: Batch number: Assay: Expiry date: Date received: Supplier: TEST SUBSTANCE MIN 252/004312 T-7499 Perfluorobutyl sulfonyl fluoride PBSF None stated Clear colourless liquid (presented in a steel pressure vessel) AbymthbeienStptoenmsoprerature or in a refrigerator, unless otherwise stated Ca. 35 kg None stated PPeerrfflluuoorroobsuultfyollasunlefonyl fluoride 962-9-48%% None stated 26 April 2000 Sponsor : 10 : EXPERIMENTAL PROCEDURE MIN 252/004312 ANIMALS KFstirefantiytn, (oo2nf5 1SmNparolaevgeuamne-dbDe2ar5w2fl0ee0ym0o.arleig)inra,tws,eargeeodbtaapinperodxfirmomateClyha6rlweseeRkisv,eor f(UthKe)CLrilm:CiDted,BMRa,nastocaneRsaoraedanMdaerrgivateed, tOnarluelnmaotcmabaretererinsdvta.ttalo,ttoa1olloefda4noinmgrtaohluesptswa,iele.raecThehxoeafma5niinmmeadallesfsowarenardebn5aocfrcemlmimaalaliettiises.esdTahfnoedraasnitgimlnesaaslsot f7woedrvaeeyrstthbeienllfouhrneeiaqclutohemlyamnieddnecnreatminfdieeondmt bolyyf The identification of individual rats in the 4 groups were as follows: Group 4R231e((((sILCHneooitrgewvnhret,rddodoolo)ssseee))) 11AM6116-----a11E2leR0505at numbe2323FrFse1661m-----2334aJl5050e tIfrrneoamvtimethweentoangfritmohueaplnsn,euetmhdebfaeonrrsi.mthaelFnuunmcbtieorninalg OsybssteermvawtioasnasluBchattthearyt ittowbaes npoetrfeoarsmyetdo widiethnotiufyt kantorewaltemdegnet ogfrotuhpe FTbyohlealorlweemtitneagrintwhinreigtcteaonnmimomnaeltnshc,ee5mtameilnaatlenosdfaewnxdeproe5surfeertemasiantlheeeds,raewsseeprroevteeasnswtsiiagelnrereedkpiltlaolecdteh.meernetssedrvueringrgotuhpe. aTcchleimseawtisearteioindepnetriifoiedd. ACCOMMODATION phETbaETbooaxhahscslcpeedoikhotirrrinsaaabocugtntenassrdngeerwwtdoeftolpeoehoomrraoaoeenpkdrekihpawnreaonlpwaiuocttcshieroeneidrlnsddeoeitnirauav5pirttinlsehodoaildenuecfagseastlvsdhllaaeobemscibetrdaseeoelgaelotriomelhdomweseoebhnmaapeestntoie.eatfdexstcy,srhsiytinaiobd.cdgiealdaistgiEt.hctyeiaeaoTcgtonoghehfarelociiblnnouycahlpa,tslatgeauelaecfasrntstpyteideaworwnnnttedhhiarmoeeesedfasnsilttsnuueatesmsarxpattibcneomrsenlfeeerdaststpetahseadoerrasiafotneatetnxdelheplfertvroahcoeacseamnunkgtrispeime.ltasahppatPefeeleislrtdrartiaescowtctdodsiaacn,wsbitetnairicaniathnchoeyaehttsmhndwgegleiwrreinsotdheihgutdirhpdnfoirwaunwoitpilnhatiysthtes..., MIN 252/004312 TT2hafu1hhfmeeec2itstdeetmiudCtydpt.yheaernahDsdtoucelirdv5eeii5nnaattginifodircno1rsieo0nlm%ftaertogivtmrreeiemtsyhptphueoeecmfrstatieihvtdueeirrtlayseynt.uogadefnysRtd.hewerceeohlrroaedtlidevodienfgrhraerunolmagotemiidsviewtwlyyeerwrseeherore1erc9to.s5dredutterotdaotiu2obs1nie.n0agmnCaadiKnacnteoadninnts3eiCd9dleertwaeordisthp5n8ianon%trlietmroceoiltahrsdtaievvoree.f Lighting was controlled to give 12 hours light (0600 - 1800 hours) and 12 hours dark per 24 hours. DIET Wfoohdile(SiDnSthReiartcaangdesM, oalulsreaNtsoh.a1dSaQccCesmsotodifaiewdemigahiendteqnuanancetitdyieot,f SstpaencdiaalrdDqieutaslSiteyr-vciocnetsr,oWlleidthlaambo, rEastsoerxy).rat TolofhdteghreeedtweinastsHcnuoonmtiipnnofgoudrnmodnactLiooiufneldaSvrceaiaielsanobcnleeasbtAolyricbnhediviecexastp.eecthteadt atonybenopnr-ensuentrtieinntthsuebdsiteatn.ceThliekealnyatloytiincaflludeantcaehtahveeebffeeecnt Tcstaaugpdeysw.. atTerhewwasataevraibloabttlleesfrwomeremeomulpdteieddpaonlydprreofpilylleednedawilaytearnbdotthtloersoautgahlllyticmleeasnwedhialet itnhteerrvaatslswdeurreinign tthhee Tsyhsetreemwcaosulndoreinafsoornmabaltyiobneaevxapileacbtleedttoo ibnedpicraetseenthtaint athneydsruinbkstianngcwe altiekre.ly to influence the effect of the test RWLfoierfasethueuSlrtm)csiaaeosnnfcccteohosnen.dsruAuocmnutegtpidlntiieaobnnyp,Whtvhyieazs.tisecCurapoltapuaklnnieecdsril,ictDAhseingrmgeulciiidctaieanvlleiWna8ena0sat/le7oyr7ns8Sews/eEraovEtefiCcrwe.qsautLaetlrdit,aythfasrvoomeurbcteehee(nsEamEmaCpdlediniargvecaptioilvainbetl,reeGtloartaiHnfhugantmtoinwFgdaintoeanrl The analytical data have been lodged in Huntingdon Life Sciences Archives. ADMINISTRATION The vaporous test substance was administered for 6 hours a day, for 5 consecutive days a week, for 4 weeks. tvTshtahaepeinotelrueasrsttssaltiswmqtueeoeirdslepaihennextdropegosplstaaeasidssns,ltiewonsgistthihsnettaeocneoltihnncetotreoierlxln/vptaealospvstouoaurleturmmcgoheesanpomehrfbae0etr.oer7.r5sinamnw3d.hwoTlhaese-bdtoeidslutytaeetdmxpwoosistphuhrecerleecahwnaamasibrperprorsdioucrcotenodsttbrhuyecmtreeedstueflrrtioanmngt T45h0e ptparmge(tHcigohncdeonster)a.tioCnosntfroorl reaxtpsorseucreeivwederaeir 5o0nlyp.pm (Low dose), 150 ppm (Intermediate dose) and : 12 : MIN 252/004312 pDreetsaeinlsteodfinaAdmDiMnisItNraItSioTnRAanTdIOanNalOysFisTo-7f4t9h9eBtYestINaHtmAoLspAheTrIeOs NtoTgeOthRerAwTSithaptpheendreesdutlotsthoisbtraeipnoedrt are CLINICAL INVESTIGATIONS rDwDeeaaclyotlebrddaoesoadnkt.od. tshIingenagedrdodruietpicooonrb,dsosebrovsfeaartvilolanaticsotinavnsidtrieeelsxarateimnlagitnitanotgioitnnodstihvoeiudtdulaianyletadoniidnmatyahlrsisuwnpneroirnecgemdaunarddeemwtaheirrnoetuergnehacnoocurdet etodhfeitnhsettuhsdetyudSwytu,edraeys Mortality Tdahyrofuogrhdoeuatdtohremstourdibyu, nadllacnaigmeaslsw. ere checked in the morning and again at the end of the normal working Clinical signs DInadtievdidaunadl asniigmnaeldrerceocrodrsdsweorfe ampapienatraainnceed,ocnhtahnegbeasainsdofd.isappearance of clinical signs were maintained. any observation, considered to be of possible importance, made at any time during the study; a(pnryioorbtsoerevxaptoiosnu,rec)o,nosnidreertuedrntotobheooldfipnogscsiabgleesi(mapltoerrtaenxcpeo,smuraed)eanddurdiunrgintrgandsafielyr tcoheecxkpso;sure cages aspceacriealfualtteexnatimoninwataisongimveandteowtheeekdelyteccotimonmoefnacvinagila1blwe reeeskpipraritoorrytosotuhnedss.tart of exposures when rDteersumprioinnngadlienkxgipllo.sismurielarsliygntos dwieeretesrtecsourbdsetadncaes. aLgirtohuep crleinspicoanlsseigwnsheirnediavlilduvaislibdlaeta,andiemaathls caopdpeea7rerdefetors btoe BODYWEIGHT cbEoeamfcohmrereaentxcwpinoagssuw1reewiogenheetkhdpeordinoayar r.troiBvdaoildefyoswtlaleoritwgohinft gewxrapasonasdluosrome.reaclDloourcrdaientidgopnthr. ieoTrthrteeoawntmeeciergnohtptpsoyefr. ieoadc,hbroadtywwaesigrehctowrdaesdrewceoerkdleyd, FOOD CONSUMPTION Tsthaertqoufaenxtpitoysuorfefsouondticlotnhseuemneddobfytheeascthudcya.ge of rats was recorded weekly, commencing 1 week prior to the : 13 : MIN 252/004312 FUNCTIONAL OBSERVATION BATTERY Neurobehavioural screening tDdsiaamumyresienwotgifhmtteheenesotnsifntougdtdaeywyx,.paasoNfsbueoandtlc)aatnalilocnrnedadatplsarowicobrersoretseorsvtstaehttsuietodengydariiolnnubitpoaiasntt.teeiordOyna.bayns(edorvbmasoetirtovonarstaiowctniesvriemtymawdaeedreperdep-uetrrriefnoagrtmmtheeendttaratenaadtpmpinreonWxtiepmeeakrtieo4ld)y, (btohunet AAdetfsauhillolerdftuebnneceltdoiowbn:aatlteorbysewrvaastipoenraflobrmatetedrydwurainsgpeWrfoeermksed1,du2rianngdth3e. prTeh-terefautmncetniotnpaelrioobds,earnvdatidounrainl gbaWtteeeryk 4is. tT(ohpbherseee-arbtnvraeiatmattietoamrlny.esncwotTmeahrnepedrimsseWeacddoeen4edkwsei4sttehstootnhoflefyoo)obbbscessoerevmrrvvapteairtroiisobnenlsid.sndhwTathaonsedthhfpineergsrttfr/osesparemettmcewiefdaincstinpcteoetsnrhtfideonirgttmieosoentdfoatwfhretehhneeaannaaiinnmniimdtailaat.llh.leyAhflialnnatdlhliesnseget Observations in the hand: FERPVOPESiuaaaxeoclllsroaoccpiecvepuaaetraplhrobieirtvptsrfeichaaoiernttatlaienyciolrmco/menaltlnaoonoouccsvohsfrueiniacnmrnoehgdanattlvnhiionudengllsiani(noeginamss,eatlorfefrmohmaonrsdth,lietnwcgia)tgchees Observation in the arena: AOALRGPRieeescrclovoscactieeooruevislrmrrnisedrotgmeycifnpntcceiagrcoooenrenuosuteonunonstftfsaccgleoanoitvf/pufaolsesictouanrlesb, otrleumseosr,su,rtiwneitches : 14 : Manipulations^: MIN 252/004312 PGLBBATARTuaaoooirupgipniudddplphdicyyirlpttishowionrittnaneerrrgegceemfycilhnsfhgerspopgehxrtreootefatehrltsnrs(esatppgs(pxltfoeeuo)loanrnrreyseseese(*apnCodn)sheindlimb) Aaptparonpyrpiaotien.t during the observations, additional comments were made as flee text where considered J The manipulations were only made during die pre-treatment period and Week 4. pArletsheonutgehd binodthyiws seeigcthitonwaass trheicsohradsedb,eenno cdoivsceuressdioenlseowfhaenrye pinosthsiebrleepeofrfte. cts of treatment on bodyweight is hFaosr btheeenobosmeirtvteadtiofrnosm, ifpraellseanntiamtiaolns iinntahlel rgerpoourptsalftahioleudghtoitshhaoswbeaegnivreecnorsdigend souncthheasralwacrdiamtaatsihoneetthis sign wMaostomroancittiovrietyd uwsainsgpearCfoorumlbeodubmefIonrferai-nRiteiadtiAonctiovfittyreMatomneintotrainngd Sdyusrtienmg 1t.he fourth week of treatment and roTloechcccioousrmrdrseoeydntso:ctreemtashce(teiuvvstiieetmysnteisas)nspoprieefnsnfeetranacti-enhredd.cnaodteemgtoeocrvytoermisteoanlts,mo olronecicotoomrrdoeatdco.trivaiFtnyod.r rneTopnho-erltoifncooglmlothwoitsionrdgaatccaat,itveoigntoylry.ietshHeoitefimanceutimsvpibteyenrtaorinef pDFloaartcaetedwsatiinsntcogo,tlhldeeecsctieagdgneeasvt,eetdrhyea2nteimmstiansleussteswisoeanrnepdprwolagrcriteatmdenmsoienngtwolyaasifnsltotaoprtpoeydb.sdeiTsrkvh.aetitoenstcsaegsesiso. n fOonrceeacahllaannimimaal lws ahsad1 hboeuern. TpehrefofrumncetdioinnaRl ooobmse0rv0a7t.ional battery was performed in Room 011 and the motor activity monitoring was Analysis and presentation of the behavioural screening data PTsatnrhreaee-lntyrgfseotihasltl,moohweifninntdvgdalardiitmaaantbwaceeswrpeelfaroayenl,laorlbwoyousedtediydnwebbleyyyigashnWutabliyajlelnsiciadstmeotdsef'mtvotpaeressirattaanttuc(iWsreetif.icolallillaTomwahneseasdely1bsd9yi7as:tS1a/t2ruew)daeerfnrionet'rgsa'nfaaantleddysosta.escdet-irvueisltiayntegcdoaurnoetsnspe,o-wngsraiepy. 1 System supplied by Coulboum Instruments, Lehigh Valley, PA, U.S.A. : 15 : MIN 252/004312 Tttmfpehruheraermeqnprunsoereesesnorpep.cfsooynrtoThtsoifeenhfreghedopaeocosogcbfrrbusteteierhenrreeevgnona,crcrtaeaiesottpyenftopgohareorelterreeoideacnfcawadhlrspeebodrsgeieparniotonnatunsogpsfseo.reuerictm(Ahith.healeterr.ahksposoartubbeaegalsrsetheselerendvd:tioaufonftrfierioonhranegarbnaenslrcadeeecnlbcsittnaoi.obtgrtnde,e,tirnwayagreno,heuansessoaatmrlhb,eteeweegtrnicert.osc,hpuhahop,nansadv,sleeffeosldibrnweacieenhngr,etihtveaceetbclnau.f)soel,sanlitlffdoeoipdewrdoitfinhonignert As there were no remarkable differences in the incidence of observations, no statistical analyses were performed on the categorical data. tTveahsretia(CnWcoeuillflbioaolmluomswe1ad9c7tbi1vy/i2tSy)tudfdoaertnaat'wsde'forseteeas-rtn.eallayteseddruesspinognsae.oneP-rwe-atyreaatnmaleynstisdoatfavawreiarnecaenfaollylsoewdedbybyanWaliyllsiiasmosf' tWdaaihtlaeednwtteahsset ahcnaataselbygesoeerdincaruelspdionargttaeJdsou,ngucgnkelhessetesedrteha-eTpreoepsssspitobranlesteedssiftwfe(eHrreeonldcleiarnebdceettiwrone&ealnWicnoonnlfatertuo, rle1a.9n7i3m;aSlstaatnXdatcrte,at1e9d92g)r.oupAs,twthoe Key to the functional observational battery Ease o23f remoesvlaaisglyhfrt(ollyimttalwceakrgewesiasrtdance) Ease o23f handlesilanisggyht(llyittalwe krewsiasrtdance) SalivatYion (osniglynsocbosreerdveifdpresent) T(1h,e2n,3uminbcerresasaisnsgocdieagterdeewoitfhefsfaelcivt)ation indicate the degree of effect N sign not observed Arousa345l raseoledmrutec(wendohraamlteharitlgn)hes(sslight excitement, tense, over reaction to external stimuli) : 16 : I Gait UATHu auhwnbunoannalrobkcmrhlienmaegltdaogloaanisttsoeesss - see additional comments T(1h,e2n,3uminbcerresasaisnsgocdieagterdeewoitfhefgfaeictti)ndicate the degree of effect Mobility impaired: mobility ofthe rat impaired due to gait abnormalities Approa4260351ch wofbasnrnptaoeahicpelfrekkrfzeross/eatrascoupectnmaaiholcseysttnsiapoawrnnoadb-yesseneifafsdditionalcomments Touch4260351 towwtfnuurtoeaarhrennllrekkzessrsseatrscobeatwaaiooccapktnyipwoonasri-dtesseseidaedditional comments Startle4O2351 onnenetxaooohratrerimgtrecwgearaeeaciltarbtcfaicllohteitennioordcennhrslepy-sospenoesneasdeditional comments MIN 252/004312 : 17 : MIN 252/004312 Tail pin21ch 346O5 jtnwr23wofAuuuroteamarhnne1llreskkepzsrwssaaesrwcsiaatvefttuwowahiairoocrynaaawlntyeyisoraniewnrtmsdtip-utmhosrenneadesetiauadttrhendlaiyttioisnnalotcoamtumrnenitnsdicates that the response included a turn such as Rightin1g refleimx mediate reaction VocaliYNsing ssiiggnn onbosteorbvseedrved T(1h,e2n,3uminbcerresaasisnsgoclioauteddnewssithofvvooccaalilsisininggi)ndicate the degree of effect Pupil rBeflex reflex observed both eyes Urine NMLS snmlmoanroagedleleoarabammsteeooruauvnmnettdoooubbnssteerorvvbeesdderved Rearing and activity counts rTCmehoaaeurdinenfltgwsoohwwreaeonsrefevcthmoeeurantadhtreeeednaoanefivwrmeearaasyrlimmntigmaorvaekenetdddhaeaolclaftffnioviinmuittryoafle6wleihetftqeiennudtaotlbhoaoernteheaanosfiomf(r"teahslqefssueeweatrseeqcrsuel"ea)ai.rnresAot.hfceaoausruneptnpafo.orrtAaincgtcivosiuutynrftawcfaoesr. : 18 : MIN 252/004312 LABORATORY INVESTIGATIONS Sample collection Bdeloporidvastaiomnpolefsfofoodr ,hpareimorattooleoxgpyosaunrdebolnooDdaych2e6m. istry were collected from all animals following overnight rSaatms wpleerseohfelvdenuonudserbilsooofdluwraenreeawnaitehsdthraewsina.from the retro-orbital sinus using sterile glass pipettes while the The blood samples collected were put into tubes containing the following anticoagulants: ECHDietprTaaAtrein --- fffooorrr hbclalooetomtidnacgtohtleeomsgtsiicc(aa0ll.5iinnmvveelsswttiigghaaottliieoonnbsslo((o00d..57) ml ml whole whole blood) blood) aTbhberehvaiaetmedattoitlloeg(ifcoarl uasnedinbalpopoedndcihceems aicnadl tainbvleess)t,igmaetitohnosdspaenrfdotrhmeeudniatsreoflmisteeadsubreemloewn,t. together with an Haematology The following estimations were performed using a Bayer-Technicon H*1E haematology analyser: Units Haematocrit (Hct) L/L Haemoglobin (Hb) g/dL Red cell count (RBC) x1012/L Mean corpuscular haemoglobin (MCH) Mean corpuscular haemoglobin concentration (MCHC) Pg g/dL Mean corpuscular volume (MCV) fL Total white cell count (WBC) x107L Differential WBC count NLyemutprohpohciylstes EBLMoaaorssgnoineopochupyinlhtsseitlsasined cells ((NLyemutprohpohciylt)e) ((((LMBEUoaossCnoino)pochpyilht)ei)l) }} }}}} x109/L MIN 252/004312 Units hCyepllo/hmypoerprchhorloomgya:siat)hweerme roesctordceodmamsofonllomwso:rphological changes (anisocytosis, micro/macrocytosis, ++++++ ==== mmsnlioaogrdhaketberndatoermalities detected Platelet count (Pit) x 109/L The following estimations were performed using the appropriate methodology, as described below: Reticulocyte count (Retie) - Sysmex R3000 Reticulocyte Counter %cells)(of red The following were performed using an ACL 3000 Plus analyser using the appropriate IL reagents: Prothrombin time (PT) - method of Quick, H.A., (1942) sec ARacptiavpaotertd, PSa.Ir.t,ia(1l9T6h1r)omboplastin Time (APTT) - method ofProctor, R.R and sec Blood chemistry Tinhdeicfaotelldo,wsitnagndpaarrdamHiettaecrhwi 9as17anmaelythsoeddowloitghieasn: Hitachi 917 clinical chemistry analyser using, except where Triglycerides (Trig) Total protein (Total Prot) mmol/L g/L Protein electrophoretogram (Beckman Paragon system) Albumin (Alb) g/L and % Alpha 1 globulin (al Glob) g/L and % Alpha 2 globulin (a2 Glob) g/L and % Beta globulin (Beta Glob) Gamma globulin (Gamma Glob) g/L and % g/L and % Albumin/globulin ratio (A/G Ratio) : 20 : I Urea (Urea) Alkaline phosphatase (Aik. Phos) - reaction temperature 37C Bilirubin - total (Bili. Total) Creatinine (Creat) Sodium (Na) Potassium (K) Calcium (Ca Total) Phosphorus (Phos) Chloride (Cl) Cholesterol - total (Choi Total) Glucose (Glue) - hexokinase mediated Alanine aminotransferase (ALT) - reaction temperature 37C Aspartate aminotransferase (AST) - reaction temperature 37C MIN 252/004312 Units mmol/L U/L umol/L umol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L U/L U/L SACRIFICE Apelrlfgorromuepds owneDreakyil2le9doffotlhloewstiundgy5. consecutive days of exposure a week, for 4 weeks. The terminal kill was Afronmimtahles bwrearcehikailllaerdterbieys.an intraperitoneal injection of pentobarbitone sodium followed by exsanguination : 21 : MIN 252/004312 MACROSCOPIC EXAMINATION AND ORGAN WEIGHTS All rats were subjected to a detailed macroscopic examination. wTheeighfoedll:owing organs from all animals killed at the scheduled sacrifice were dissected free of fat and kehapdiediardnerintedyaylssmides tlleuivsntegerss Bilateral organs were weighed together. FIXATION OF TISSUES wtSeasemtreepsplaernse,dseoerrpvtiehddeidiwnymhbouidlfeef,esrowefdhthi1ce0h%fwolfeloorrewmfiianxlgiendo, riegnxaBcneo/ptuitsitsnhu'eseses,oytelousgt,ieowtnhheaircnhwdwitthheernaentpyrraenmsseafrecvrreroedsdciontopDi7ca0avl%liydsaaolcbnon'shoofrimlx. aatliveen,tiatineds abfaaabeeaeyodnpblrrjodccrsiimoaeeinirrlemtaouedmeiejtnsoconeuualsouincemtorcdaornumdha(nmnulyptumaelithmsim*haacnmdorlahu(iieyrcgtdmaniouetectrrssisiatfc**eci)cxtaatinodnmmeadruklla)* nphhhkmoolllllauaiyaapivaeevcvmmtrdnamnsraataryehmeigndaadrcrecrenprntlsieecr*sdymehx(etyr(nhtea(rumsuiiaseean(ainalnlrscl2rolralntoc(blvuaeylbilldilonlerglunreagoaeaibscoldvcanlbsrraelnaiteedunesran(enclaesmd)dnssgpdh*)g)si(d*asia(n*ciaa3opnagulnidu)rllcesildicoba)va*uruelrtl)leleasagx)rri,*,aonnds)* tttttuuvppssssssssohrehekkacppttariraesnyyetiomlieiligcutniarnenrgmrlsevinhuiteeioaatnustinauaeuntrsalic*aearaydtma*srcyewlnlyso(bmevgiiwrltrnedlahuvaicsdtsenilchdccusdeelldpsreerviasn(irtxgahtibhgiyhfru)oricdastion)* * Including nasal cavity, paranasal sinuses and nasopharynx : 22 : I MIN 252/004312 HISTOPATHOLOGY ptHahnrieodscttoe4esp.stsaeetTshdhowaelenosredgeitcssitastaalsiiunneeeexsddawmwweiiintrthehathPieoamAnemSbse-awhdtedoeaexrmedylapiitnneorxapfnyoadlrriamnef.ofeidsninownfaoxralealxnsadcmhsieendcauttilioeondnsbtaiyspspluirgeoshxti(mmmaiactrerkolyesdc4o-wp5ei.tphmS*e)tchtfiioocnkr sGwcreuortuefprcosumt1, Fstoarintiensgtems ettihssoudeosloognylyto(raenmimovael nnounm-sbpeerscif1ic-5haaenmd a1to6x-2y0li)n, s0t.a5i%nin(gv./v) hydrochloric acid was used in the Histopathological examination was only performed on any abnormal tissues arising from Groups 2 and 3. STATISTICAL ANALYSIS All statistical analyses were performed separately for males and females. FBbooodrdyyawwlleeiipggahhrtta, mdoaregttaaenrwswetreheieganhaatnlaaynlsydesdcelsuinswiicneagrlewpaeptiehgrohfoltorggmayienddsa.tuaTs.ihneg fothlleowinidnigvisdeuqaulenacneimoaflstaastistthicealetxepsetsriwmaesntuasledunfoitr. Im7f5et%thhe)o,ddtsha.etaOpctrohoneprsowirstitisoepn:reodof maninimanatllsywoifthonvealpuaerstidcuiflfaerrevnatlufreom(reltahteivme ofrdeequweanscyanoaflytsheedmboydaepepxroceperidaetde B(sataatrbttlhleeetvt'a1sr%tiaensltecvewesalt)sruhacepttpuelrrieoegdceotnuoelditteybstewfooabrstahfioentueendrod. g, eaneloitgyaroitfhvmariicantrcaensbfeotrwmeaetniotnrewatamsetnrtise;dwtohesreeesiigfnaifmicoarnet mrIdSefotsousnprdeoeo-enrtnsehstilea'gas,nntetitDdftwieucrsoanetnnsgwpteroaothtsun'espsuteesstreewo(dsWget.erien(lDlebiiauetmyninnswge, tact1s,o9m7d11ep9ta5ear5cnetddea,dn1gd9(roo7ru21p)9i,f6mo4a)er.asinaftsitFshwofeaerrcreetsoewcrpyoaamsrtrapeataevnrisedftdwoenruomcs-egiantrfigooounrWpawilclnaoioasmmnf-posma'urtoniesndsoo)tn,tfosoa,nrnidcaa tIr(wrDefaasnusspisnogufnnnos,esirfemd1i.c9a(a6tSn4ioht)i.nrh)le,eFtyeog,rrroo1sgue9epp7nsa7er)iaw,ttyeeorortewfiofvc-aotgrhmireaoprnuaecpreewdcwaosmausspeinvparigrdiesesoSennhncsite,rl(aeafyonW'rds iaclncoonounoxl-dnop-nanmroraoatmnnbokeettorsirnucemimcteotrsevetsestpdfoo(Wbnrysieala,codDlxooousgenna-,nrri'est1hl9amt4tee5isdc)t bWFoohdreyorwregeaaipgnphwrtowepihgriihactthed,maatiang,ahldytiseinisffilonuafelnccboeovdathryiewanoecrigegahwntawwsaeuisgsuehdstse.idnapslaccoevaorfiaatneailnysaisn oafttveamrpiatntcoeailnlotwhefoabrodvifefesreeqnuceenscien. For microscopic findings Fisher's exact test was employed to detect treatment-related differences. : 23 : MIN 252/004312 LOCATION OF STUDY RECORDS AofllthreawSpdoantaso, rs.amples and specimens arising from the performance of this study will remain the property PTarrycophceievsdiunorgfessm.amayplbeeadnidspsopseecdimoefnaftthearttahreepuenrsiouditsabsltea,tebdyinreaHsuonntionfgdinosntaLbiilfietyS, cfioerncloesn,gSttearnmdarredteOntpioernatainndg Aasoreurflaccltshhhooievtntiheammebfarelote,rsetarfhaeimaeeplpseSbl.reepisiooIndnfagsrnooeadqfrgufrwsieepvieseledtlceiybdwmee,iaetHcrhnosustnnfhtrateaoincnmSdtgepdadtoholnlaensnrodLadrawi.htfeeisdoSaanctdaiwvewnihcciieleclshsbowteuhigelrlheStctaotouinnndteiytndhuDebeiryreteotcHutrruoennrt,atsdiiinnigsgntpdhsooestnhamelLaofitirfenerfaiualSrlcrtsheiepsenurocbrretje.estecAnitnttfiotoietnasr HthuenftiinnagldroenpoLrtifien Sitcsieanrccheisvwe iilnldreeftianiintetlhye. Quality Assurance records relevant to this study and a copy of : 24 I RESULTS MIN 252/004312 CHAMBER ATMOSPHERE CONDITIONS Chamber analysed concentration of T-7499 The data are presented in ADMINISTRATION OF T-7499 BY INHALATION TO RATS The data are summarised below: Group 423 (((IHLnoitgewrh,dddooossseee))) CTahragmetber 41555000 coMn41c5e46e9a72nAntrnaatiloysne(dp22sp1d29m) Tcohneceancthraietivoends. chamber concentrations for Groups 2 to 4 were in good agreement with the target CLINICAL OBSERVATIONS Mortality There were no unscheduled deaths. Clinical signs The data are presented as follows: ATTAaappbbpplleeeenn21ddiixx 21 ---- pppdrroueesr-itaenenxgxdpepopxosopsusuortreseeu-xr-epinio-ndscugiivrrdeioedu-nupcaienlddwissiutevmreiidbkmuulyaatilrooydbnasoielrfyvooabtbsisoeenrrvsvaattioionnss DClaiynic1aolfstihgensexopbosseurrveesd. aLsetahagrrgoyupwaressepvoindseentdfuorrinGgroeuxppsos3uarendin4cdluudreindgctihrecleinxgpomsuorveesm. ent for Group 4 on TaCGnhlrdioinsui4cps.aisglHn2s,ywig3panesasrnaaocdlsbtio4sve.iertvyvWiedwdaealniksmtinfomogbleslooednwriavittenoedgelysftohp(leaolbosenwtxopiernoxmgspuaotlrhseuegsraeefioxt)iprnwoGcsalruusordueeepsvdi2fdovreroanctGtsafrldioosuluilrnpoignswg3ianWnagdnetdaehgke4it3ead.xtuiporoinnsguwrhWesenefeohkrasGn2drolaeundpdsfo33.r rOf3faeontmflselotamphwlrreaieilonreeargrottadocptacsreyiaxpo.sprrioiotonosrusetrsoxeipgeoonxnsspuDowrseaeuyroree2np6oD.rneaAsDyetan1ayt7lp.l1ro9Vito.horeWctroaatlieilmskxiipnensogg,sauaonlrnledo.toafAgetishgtea(ittaaaibobtninooovnwremhwseaihnlgengnhsaahihnta)adndwlderaldeessdwpowrelevraseesedpnprptrerefsioseoernrnGtttofrfoooeruxrtppwao4oGsufrGeormeruoptauhlp3ee : 25 : MIN 252/004312 Asigt nosthaerretcimonessi,dderryedskininciadbernatsailoannsdtonotatirles,laitrerditatoblterebaethmaevnito.ur and pale coloured teeth were observed. These Bodyweight The data are presented as follows: AFTiapgbpulerene3d1ix 3 --- iggnrrdooiuuvppidmmuaeelaanvnavvluaalleuusee(ssg(()gg)) Tdehgereoeveorfasltlamtisetaicnalbsoidgynwifieciagnhctegfaoinr salflotersatlml taelsets.rats (Weeks 0 to 4) were lower than controls, attaining a Food consumption The data are presented as follows: Table 4 - group mean values (g/animal) A reduction in food consumption was evident for Group 4 male rats. The food consumption of other test rats was considered not to be affected by treatment. Functional observation battery The data are presented as follows: TTTTTTAAATTaaaaaaaapppbbbbbbbbppplllllllleeeeeeeeeeennn79658111ddd120iiixxx 456 ----------- tbrtaagclgieoanedcoorrmnatdiotdudapirdiuivlplyivtibnpsietwinitotiogdrygrmseaunueucitcfmanaeguomoollghrusoucmotaetpnhtnoscbs-etattmpss--nsirevlgy-tm-aggririvytorrogneygooauf-rnrtuulompiootopopgsuubcomnrp-mpsmoonseemmiuimeeratnpvoaaonedeanrntamiaiotvnvnivnvraoeigaavdlanavullauncusuaelateleusviulsvseaeoislstubyes-esrivnadtiiovnidsual values (seconds) Flseuuvmnecmlstiaournypa.ltoob4s5e0rvaptpiomn,alshboawtteerdy nfoindeivnigdsenfcoer oanfimneaulrsottroexaitceidty.witThhTe -f7o4l9lo9wfionrg fcooumr wmeeenktss,aaret emxpadoesurine : 26 : I MIN 252/004312 In the hand observations In the hand observations were unaffected by treatment. Arena observations Aberetrneaatombesenrtv-raetliaotnesd.showed some inter-group variation but the differences were not considered to aCdinbiofsfmteehnrpeecanearceroeedsnfwawsiiemdthrueirlcaiconrongnrtresWosidlpseeo,ernfeksedem2topaolabetfsetetirdrnenuasaetlmdtlouterrnneitana,tgtueardlpatrhlgeovrcouaeurgdiphaisntigsoschnooo.rrwesseudfbosrreedqmuucaeelnedtswawceteeivrkeistyuonafnatfdrfeeracettamerdien.ngt,Isncthoetrhseees M anipulations D4csoo5umn0rsiepnidwpgemhrWaetdwerteeeorkdebue4cloeawdossefboretcrtfiheoaaartteenmdctehwonomitst,hemltoearnnefacdcteiomnmngetenrnoftot.lsoo.tfspMtrleaeayatnmmevenaaltsuauenrsedmfothernetsthseesfdeoifrgferfroeeumnpcaseleswswerreeerc,eehitovhwienregevfeo1rr5,e0anlsooort Motor activity TcvbshoieheonedwwturtoeieomdltfaoeitnnhniscemaprteeluaaanlrcsstakeldidnvouaarflriocdnitcaiogotvsimioWatnygoe.ette-oimrkreel4aasct.otiiofvTntirstheyheaisfpteomardenmindftfaetwlhreehesnecorreeepcsapesofi,avsiiiinlnneggdcodt1noi5rtre0aaccspthtipo,iemnfvesemwosaaftlasectshilsoartnewiccgeaeerli,vstwhiingaegnnrie5ftih0ccaoaotnnrcssihe1do5eaw0rnednpd,pbmtioyn LABORATORY INVESTIGATIONS Haematology The data are presented as follows: ATapbpleen1d3ix 7 -- ginrdoiuvpidmuaelanvavlauleuses There were no treatment-related findings. 27 : Blood chemistry The data are presented as follows: ATapbpleen1d4ix 8 -- ginrdoiuvpidmuaelanvavlauleuses There were no treatment-related findings. MIN 252/004312 TERMINAL STUDIES Macroscopic pathology The data are presented as follows: TAapbpleen1d5ix 10 -- iinncdiidveidnucaelspuamthmolaorgyical findings There were no treatment-related findings. Organ weights The data are presented as follows: ATapbpleen1d6ix 9 -- ginrdoiuvpidmueaal nvavlauleuse(sg()g) There were no treatment-related findings. Microscopic pathology The data are presented as follows: TAapbpleen1d7ix 10 -- einxdpiavniddueadlipnactihdoelnocgeicsaulmfimndairnygs Treatment related findings There were no microscopic findings that were considered to be related to treatment with T-7499. Incidental findings All microscopic findings were considered to be incidental and of no toxicological importance. : 28 : I DISCUSSION MIN 252/004312 Icnontsheicsutsitvuedyd,ayrsatesacwhewreeeekx,pfoosre4dwbeyekisn.hTalhaetiostnudtoy mtheeanvaepxpoousruoref lTev-7e4ls9w9 efroer467, h1o6u2rsanedac4h59dpapym, f.or 5 Tcohnesuomveprtaiolln mwaesanalsbooedvyiwdeenigthftorgmaianlefaonrimalallsteinstGgrroouupp4s. was lower than controls. A reduced food Aneinxencupilromuosdtaueolrdsxeievcpxorepcofoacfleseiecdsdtautariaenotsdn.cgoaeCnnncdleeinrnaaigtclrilaatylatitrosieoingsnsonlsowvfwehde1en6rp2erhipaoalpnrsmdtoolen(edGoxtapreonodudsupfhorye3rp)tahealrlenatdfceots4iltvl5oig9twyrop.inupgpTmshdea(oGsyne.roosucigcpna4ssi)wownesererpeocosletntsehixasrptgeoinsctudrweuirtaihnndga tTcrrehehleaaemttrmeeidsewtndrayti-fsrfpeenalraoreatnemecdfeeffsteiencirntdsi.oonfrgNgstar.eencartwompeesignyhtrtoes.vneaHthleiesdtfounpnoactthtriooelanotagmliceoanblts-eerexrvlaaamtteiidonnamatilaocbnraootstfecrtoyhpeiocrrefshipnaiderimantgoatsroylaotnrgdaiccntaolretavrneedaatlmebdleonontdoAfcoolnlnosoiwstioenbngstedwravyiethdanaedfttfrheaecnrtseilewenvateslenf(ofNeecOvtEiodLne)ntcwheeaosnf ensruovstotaueissnteasdbylsnitseehumerdohtodaxduircigintegyneitnrhaitlshlyestfruuednsyoc.tlivoenHdaolpworbieovsreerrtv,oacteilxoinpniocbsaaultrteesrigythn. es : 29 : REFERENCES MIN 252/004312 BARTLETT, M.S. (1937) Properties of sufficiency and statistical test. Proc.Roy. Soc., A 160,268. DACIE, J.V. and LEWIS, S.M. (1966) in: (EDS). Practiced Haematology. 3rded. p. 28 and/or p. 140. DUNN, O.J. (1964) Multiple comparisons using rank sums. Technometrics, 6,241. JD.UANmN. SEtaTtT. ,ACss.Woc..,(5109,5510)9A6.multiple comparison procedure for comparing several treatments with a control. DUNNETT, C.W. (1964) New tables for multiple comparisons with a control. Biometrics, 20,482. FISHER, R.A. (1950) in: (Eds). StatisticalMethodsfor Research Workers. Oliver and Boyd, Edinburgh. YHoOrLk.LANDER M, WOLFE DA (1973) Nonparametric Statistical Methods. John Wiley & Sons, New PROCTOR, R.R. AND RAPAPORT, S.I. (1961) The partial thromboplastin time with kaolin. Am. J. Clin. Path., 36,212. 2Q4U. ICK, A.J. (1942) in: (EDS). The Haemorrhagic Diseases and the Physiology o f Haemostasis, chill p. SHIRLEY, E. (1977) A non-parametric equivalent of Williams' test for contrasting increasing dose levels of a treatment. Biometrics, 33, 386. StatXact-Turbo User Manual (1992), Cytel Software Corporation, Cambridge MA. WcoImLpLaIrAedMwSi,thDa.Aze. r(o19d7o1se) cAontterostl.fBoirodmifefterircesn,c2e7s, b1e0t3w-e1e1n7.treatment means when several dose levels are W51I9L-5L3I1A.MS, D.A. (1972) The comparison of several dose levels with a zero dose control. Biometrics, 28, WRIGHT, B.M. (1950) A new dust-feed mechanism. J. Sc. Instr. 27, 12. WILCOXON, F. (1945) Individual comparisons by ranking methods. Biometrics Bulletin, 1, 80-83. : 30 : FIGURE 1 Bodyweights - group mean values (g) Bodyweight (g) MIN 252/004312 TABLE 1 Clinical signs during exposure - group distribution of observations Group Sign (Con1trol) (Low dose) (Inter, dose) (High dose) No abnormalities detected No abnormalities detected NLeothaabrngoicrmalities detected CLeirtchlairnggicm_o_v_e_m__e_n_t _____ Exposure number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 V V 'i V V V ^ v v VVVVVVVVVV V V V V V V V V vv V vvvvvvvv TABLE 2 Clinical signs post exposure - incidence summary Group Sign (Co1nMtrol) (Low2Mdose) (Inte3r,Mdose) (Hig4hMdose) No abnormalities detected No abnormalities detected No abnormalities detected NAVgooictaaabltinesodinrwmghawelihntieehnsahndadenlteeddcletedd 51 25 35 45 55 65 75 85 95 1N50uEmx1pb51oersusrh1e5o2nwuimn15g3bseirgn154 155 156 157 158 159 250 5 55555555 5 5 5 5 5 5 5 5 5 5 5 5 55555555 5 5 5 5 5 5 5 5 5 5 5 411 5 5 5 5 5 5 5 5 5 411 5 5 5 5 5 5 5 5 5 MIN 252/004312 TABLE 2 (Clinical signs post exposure - continued) Group Sign (CoInFtrol) No abnormalities detected 51 25 35 45 55 65 75 85 95 15N0 uEmx15pb1oersus1rh5e2onwuim1n53gbesirg51n4 155 516 517 518 519 250 (Low2Fdose) NAVHoogycipataeablrtinaesocdintriwmgvehawelihntieehnsahndadenlteeddcletedd 5 5 411 411 411 411 411 411 233 5 2331 5 14* 411 2l3bb 411 233 411 223 411 (Inter3,Fdose) NAVWHgoyoaipcltakaeablritinneasocdgintriowmgvnheawetloihnteieehsnsa(hnadadbenlnteedodclretmeddal gait) 232 3111 411 3 2 5 2211 5 51 4 5 322 1 3 2 32 2 31 4 1 411 2 3 3 2 3 2 4 1 (Hig4hFdose) AWVHgyoaipcltkaealirtineascgdintiowgvnehwetohneehsna(hnaadbnlnedodlremdal gait) 55 555 555 555 555 511 25 51 1 5 5 2251 5 25 5 5 511 5 5 5 5c AAAlllsssooo ppprrreeessseeennnttt aaattt ttthhheee ppprrreee---dddooossseee ooobbbssseeerrrvvvaaatttiiiooonnnpppoooiiinnnttt fffooorrr aaanlnliimmGraaollunnpuumm4 bbfeeemrrs3a36le6FFraatsnd 40F MIN 252/004312 TABLE 3 Bodyweight - group mean values (g) Print Mo: 0008 Group Compound : Exposure level(ppm): 1 Control 0 234 -- T-7499 - 50 150 450 Printed: 26-FEB-01 Xybion protocol number: MIN 252 U} SEX: ------------- -- Ma l e -------- --J>L M A L E -- GROUP: 1 2 3 4 1 2 3 4 Ul WEEK -1 220 217 0 277 269 1 317 303 2 353 324 3 380 347 4 387 352 Bodyweight gain (g/animal) * Week 0-4 110 83 ad 10.02 22.0 % of control 75 216 275 302 326 348 355 80 18.8 73 216 268 279 299 320 332 ** 64 21.6 58 155 158 180 181 190 191 200 198 211 202 212 204 38 23 7.9 12.2 - 61 158 160 183 184 181 184 187 186 198 200 204 203 21 19 7.4 12.5 55 50 p<sd Standard deviation Williams' test: * 0.05, **p< 0.01 MIN 252/004312 TABLE 4 Food consumption - group mean values (g/animal) Print Ho: 0009 Group Conpound Exposure level (ppm) : 1 Control 0 23 ----- 1-7499 50 150 4 450 u> On Printed: 26-FEB-01 Xybion protocol number: HIM 252 GROUP: i 2 3 4 i 2 3 4 WEEK -1 204 197 195 203 144 140 136 143 1 211 196 199 180 131 140 119 123 2 218 192 200 180 139 134 121 125 3 226 192 198 181 137 131 128 136 4 194 185 186 174 139 128 125 130 Cumulative (g/animal) Mask 1-4 849 765 783 715 546 533 493 514 % of control - 90 92 84 - 98 90 94 MIN 252/004312 I MIN 252/004312 TABLE 5 Functional observational battery - group summary of observations !'re-dose Sex Group Number OBSERVATIONS: IN THE HAND removing from cage, easy handling, easy salivation vocalising IN THE ARENA grooming arousal, alert defecation urine GAIT Walking on Toes Hunched Unable to Assess MANIPULATIONS approach, a reaction touch, a reaction startle (present) righting, immediately tail pinch, a reaction pupil reflex 1 5 5 5 0 0 0 5 2 1 0 0 0 5 5 4 5 5 5 -Male------23 55 55 54 00 00 00 55 23 42 11 00 00 55 55 55 55 55 55 Numbers reflect the number ofanimals showing the response 4 5 5 3 0 0 0 5 1 1 3 0 0 5 4 5 5 5 5 ----- Female 123 555 555 553 000 011 000 555 000 200 2 13 000 000 555 545 555 555 555 555 4 5 5 4 0 0 0 4 0 1 4 0 0 5 5 5 5 5 5 : 37 : TABLE 5 (Functional observational battery - continued) MIN 252/004312 Week 1 Sex -Male------- Female----- Group 1 2 3 4 1 2 3 4 Number 5 5 5 5 5 5 5 5 OBSERVATIONS: IN THE HAND removing from cage, easy handling, easy salivation vocalising 54 5 54 5 5 5 554 3 54 4 3 0000002 1 0000 12 10 IN THE ARENA grooming arousal, alert defecation urine 00000000 55554555 23300000 23 110 100 GAIT Walking on Toes Hunched Unable to Assess 02223323 00000000 00000000 Numbers reflect the number of animals showing the response : 38 : I TABLES (Functional observational battery - continued) MIN 252/004312 Week 2 Sex Male ----- Female Group 1 2 3 4 1 2 3 4 Number 5 5 5 5 5 5 5 5 OBSERVATIONS: IN THE HAND removing from cage, easy handling, easy salivation vocalising 55555555 5 552 5 3 53 0 000 0 0 00 00002200 IN THE ARENA grooming arousal, alert defecation urine 00000000 55555555 23 100 000 22 130 0 00 GAIT Walking on Toes Hunched Unable to Assess 0 124 5 3 55 000200 11 00000 100 Numbers reflect the number ofanimals showing the response : 39 : TABLES (Functional observational battery - continued) MIN 252/004312 Week 3 Sex ----- Male Group 1 2 3 Number 5 5 5 OBSERVATIONS: IN THE HAND removing from cage, easy handling, easy salivation vocalising 4 55 5 45 1 01 0 00 IN THE ARENA grooming arousal, alert defecation urine 1 00 5 55 0 00 2 32 GAIT Walking on Toes Unable to Asses Hunched 1 23 0 00 0 01 4 5 5 4 0 0 0 5 0 1 3 0 1 Numbers reflect the number ofanimals showing the response ----- Female----1 23 5 55 4 5 5 5 55 5 4 33 0 2 11 3 5 00 0 0 00 5 545 0 0 00 0 0 00 4 5 55 0 0 00 0 121 : 40 : I TABLES (Functional observational battery - continued) MIN 252/004312 Week 4 Sex Group Number OBSERVATIONS: IN THE HAND removing from cage, easy handling, easy salivation vocalising IN THE ARENA grooming arousal, alert defecation urine GAIT Walking on Toes Hunched Unable to Assess MANIPULATIONS approach, a reaction touch, a reaction startle (present) righting, immediately tail pinch, a reaction pupil reflex I 5 5 5 0 0 0 5 0 1 2 0 0 5 3 5 5 5 5 -Male------23 55 55 45 00 00 00 55 10 22 20 00 01 55 53 55 55 55 55 Numbers reflect the number ofanimals showing the response 4 5 5 5 1 0 0 5 0 3 2 2 0 5 5 5 5 5 5 ----- Femah 123 555 555 54 5 030 12 0 000 555 000 000 555 112 000 555 455 555 555 555 555 4 5 5 5 0 0 0 5 0 0 5 2 0 5 5 5 5 5 4 : 41 : MIN 252/004312 TABLE 6 Functional observation battery: Activity counts - group mean values Pre-dose Group 1 2 3 4 Mean activity counts Males Females 10 12 14 13 11 13 14 12 Week 2 Group 1 2 3 4 Mean activity counts Males Females 12 17 11 9* 11 13* 11 9** Week 4 Group 1 2 3 4 *p><0.05 Williams Test **/7<0.01 Williams Test Week 1 Group 1 2 3 4 Week 3 Group 1 2 3 4 Mean activity counts Males Females 11 13 13 11 9 11 13 10 Mean activity counts Males Females 11 15 12 14 9 14 10 10 Mean activity counts Males Females 10 16 13 14 11 14 11 12 : 42 : MIN 252/004312 TABLE7 Functional observation battery: Rearing counts - group mean values Pre-dose Group 1 2 3 4 Mean rearing counts Males Females 65 75 54 66 Week 2 Group 1 2 3 4 Mean rearing counts Males Females 68 64 75 6 5** Week 4 Group 1 2 3 4 **/7<0.0I Williams Test Week 1 Group 1 2 3 4 Week 3 Group 1 2 3 4 Mean rearing counts Males Females 66 65 45 55 Mean rearing counts Males Females 57 75 47 45 Mean rearing counts Males Females 67 65 56 56 MIN 252/004312 TABLE 8 Functional observation battery: Grip strength - group mean values Pre-dose Group Forelimbs 1 2 3 4 Hindlimbs 1 2 3 4 Grip strength (kg) Males Females 0.78 0.77 0.73 0.71 0.67 0.75 0.68 0.66 0.69 0.77 0.65 0.62 0.65 0.69 0.69 0.65 No differences of statistical significance (p> 0.05) Week 4 Group Forelimbs 1 2 3 4 Hindlimbs 1 2 3 4 Grip strength (kg) Males Females 1.31 1.10 1.17 1.12 1.23 1.10 1.28 1.08 1.06 0.97 1.09 1.02 1.03 0.96 1.14 1.00 I TABLE 9 MIN 252/004312 Functional observation battery: Landing footsplay - group mean values Pre-dose Group 1 2 3 4 Mean splay values (cm) Males Females 9.6 10.2 10.3 10.5 11.9 8.7 9.1 8.8 *p<0.05 Williams test **/><0.01 Williams test Week 4 Group 1 2 3 4 Mean splay values (cm) Males Females 11.2 10.0 11.0 9.0 11.8 7.1* 9.0 6.8** : 45 : MIN 252/004312 TABLE 10 Functional observation battery: Temperature - group mean values Pre-dose Group 1 2 3 4 Mean temperature (C) Males Females 38.0 37.8 38.0 38.4+ 38.0 37.9 37.9 38.4+ +p<0.05 Student's `t' -test Week 4 Group 1 2 3 4 Mean temperature (C) Males Females 37.8 38.4 37.8 38.5 37.3 38.1 37.7 38.1 : 46 : MIN 252/004312 TABLE 11 Functional observation battery: Bodyweights - group mean values Pre-dose Group 1 2 3 4 Mean bodyweights (g) Males Females 245 166 242 165 243 166 238 166 **p<0.01 Williams Test Week 4 Group 1 2 3 4 Mean bodyweights (g) Males Females 384 208 349 205 351 199 323** 198 47 : MIN 252/004312 TABLE 12 Functional observation battery: Coulbourn activity monitoring - group mean values Pre-dose Group 1 2 3 4 Ladrguerimngov1ehmpoeeurnriotosdb(sienrsveactioonnds) Males Females 605 514 446 686 589 737 771 465 No differences of statistical significance (p> 0.05) Week 4 Group 1 2 3 4 Ladrguerinmgov1ehmpoeeurnriotosdb(sienrvseactioonnds) Males Females 772 484 601 722 338 679 610 557 TABLE 13 Haematology - group mean values Group Hot Hb RBC Retie MCH MCHC L/L 3/dL XlO- 12 / b % pg g/di* MCV fL NBC X10-9/L Neutr ophil X10-9/L H CD H 1M Mean 0.471 16.4 8.26 3.02 19.9 34.9 57.1 16.84 SD 0.0117 0.53 0.235 0.395 0.73 0.52 1.60 3.968 0.633 o-p- n 5 5 5 5 5 5 5 5 5 2M Mean 0.461 16.3 8.22 2.30 19.8 35.4 56.1 14.87 2.13 SD 0.0206 0.68 0.287 0.211 0.33 0.33 0.75 2.805 1.259 n 5555 555 55 3M Mean 0.460 SD 0.0246 n5 4M Mean 0.450 SD 0.0120 n4 Williams' test: *p< 0.05 16.2 0.84 5 16.1 0.53 4 8.04 0.402 5 8.15 0.423 4 2.34 0.487 5 2.55 0.651 4 20.1 0.94 5 19.7 0.54 4 35.2 0.51 5 * 35.7 0.54 4 57.2 2.42 5 55.3 1.44 4 14.88 3.347 5 1.989 4 1.32 0.362 5 2.22 0.547 4 MIN 252/004312 * H <0 H TABLE 13 (Haematology - continued) Group Lymph ocyte X10-9/L Bos in ophil X10-9/L Baso phil X10-9/L Mono cyte X10-9/L LUC X10-9/L Pit X10-9/L PT sec 1M Mean 14.46 0.16 0.07 0.18 0.16 959 13.0 SD 3.702 0.059 0.038 0.073 0.076 295.6 1.03 n 5 55 555 5 2M Mean 12.20 SD 1.741 n5 3H Mean 13. IS SD 3.111 n5 4M Mean 13.26 SD 2.032 n4 Williams test: *p< 0.05 0.13 0.039 5 * 0.09 0.013 5 * 0.10 0.028 4 0.05 0.017 5 0.06 0.026 5 0.07 0.017 4 0.17 0.045 5 0.12 0.046 5 0.26 0.050 4 0.18 0.041 5 0.11 0.062 5 0.24 0.042 4 1116 83.1 5 1124 122.3 5 1023 245.8 4 12.9 0.26 5 13.5 0.61 5 13.3 0.80 5 APTT sec 18.7 1.87 5 18.1 2.86 5 19.5 1.05 5 16.8 2.82 5 MIN 252/004312 Group HCt L/L iF Mean SD n 2F Mean SD n 3F Mean SD n 4F Mean SD n Williamsi' test: 0.433 0.0160 5 * 0.409 0.0077 5 * 0.401 0.0133 5* 0.411 0.0209 5 * p < 0.05 Hb g/dL 15.3 0.40 5 * 14.5 0.40 5 * 14.4 0.49 5* 14.9 0.63 5 TABLE 13 (Haematology - continued) RBC Retie XlO- 12/L % MCH P3 7.71 1.300 5 2.52 0.321 5 19.9 0.49 5 7.39 1.161 5 2.03 0.182 5 19.7 0.66 5 7.24 1.204 5 2.17 0.507 5 19.9 0.54 5 7.47 1.332 5 2.22 0.712 5 19.9 0.61 5 MCHC g/dL 35.4 0.66 5 35.6 0.55 5 35.9 0.13 5 36.2 0.83 5 MCV NBC Neutr th xlO-9/L ophil X10-9/L 56.1 0.89 5 10.12 2.122 5 0.95 0.358 5 55.4 1.18 5 55.4 1.26 5 55.0 2.25 5 7.75 3.126 5 * 5.70 1.223 5 * 7.76 1.170 5 0.84 0.235 5 0.88 0.296 5 1.25 0.536 5 MIN 252/004312 TABLE 13 (Haematology - continued) Group Lymph ocyte X10-9/L Boain ophil X10-9/L Baso phil X10-9/L Mono cyte X10-9/L LUC X10-9/L Pit X10-9/L PT sec APTT sec IF Mean 8.68 0.18 0.03 0.14 0.14 1071 13.7 17.1 SO 1.983 0.077 0.012 0.052 0.050 171.9 0.48 1.40 n 55S 55555 KJi ts> 2F ** Mean 6.62 0.10 0.01 0.10 0.08 1049 13.8 16.8 SO 2.810 0.021 0.009 0.053 0.036 94.5 0.49 2.38 n 555 55555 3F * * * * M Mean 4.62 0.09 0.01 0.07 0.04 965 13.5 14.3 SO 1.105 0.027 0.007 0.025 0.005 155.8 1.47 2.41 n 5 55 5555 5 t4F * * * * * # Mean 6.27 0.09 0.02 0.07 0.07 1075 14.4 16.3 SD 1.089 0.036 0.008 0.021 0.016 83.7 0.11 1.46 Williams'ntest: Dunnett's test: Shirley's test: J#*ppp<<< 50.05, 00..0055, ** 5 p< M p< 0.01 0.01 5 5 5 5 5 5 MIN 252/004312 TABLE 14 Blood chemistry - group mean values Group Aik. PbOB U/L 1M Mean 436 SD 109.0 n5 2H Mean SD n 447 56.8 5 3M Mean SD n 446 62.4 5 4M Mean SD n Williams' test: 381 33.5 5 *p< 0.05 ALT tl/L AST U/L 44 86 4.3 16.1 55 43 75 3.8 5.2 55 38 72 5.4 5.5 55 42 94 5.6 6.1 55 Bili. Total umol/L Urea mmol/L Crt umol/L Glue mmol/L Choi Total mmol/L Trig mmol/L 2 5.75 0.5 0.728 55 43 7.40 1.50 0.71 1.8 0.622 0.181 0.243 5555 2 5.57 0.5 0.589 55 41 7.22 1.69 0.81 2.2 0.622 0.250 0.185 5555 1 5.22 0.5 0.557 55 2 5.77 0.7 0.615 55 41 6.81 1.38 0.50 1.6 0.873 0.191 0.140 5 55 5 * 44 7.07 1.16 0.61 4.3 0.924 0.166 0.245 5555 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Ha K mmol/L mmol/L 1M Mean SD n 140 4.2 1.5 0.65 55 2M Mean SD n 1.0140 3.8 0.20 55 3M Mean SD n 139 3.9 0.8 0.17 55 4M Mean SD n Williams' test: Shirley's test: 140 0.9 5 * p < 0 .05, p<i i 0.01 3.7 0.27 5 **jp< 0.01 CI mmol/L Ca Total mnol/L Fhos mmol/L 1.0100 2.74 2.37 0.089 0.080 555 * 101 2.63 2.16 1.1 0.086 0.114 555 * 100 2.63 2.43 1.1 0.071 0.329 555 * ** 101 2.58 2.24 0.5 0.078 0.069 55 5 Total Prot g/L 64 0.8 5 62 2.1 5 ** 61 1.1 5 ** 61 2.2 5 Alb g/L 31 1.3 5 32 0.5 5 30 1.5 5 32 1.7 5 al Glob g/L 11.04 5 ** 11 0.9 5 ** 12 0.8 5 ** 12 0.8 5 a2 Glob g/L 0.04 5 4 0.8 5 0.04 5 tt 0.03 5 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Beta Glgo/Lb 1M Mean SD n 12 0.4 5 2M Mean SD n 12 1.3 5 3M Mean SD n 11 1.1 5 4M Mean SD n Williams' test: 11 0.8 5 **/K 0.01 Gamma Ggl/bob A/G Ratio 3 0.96 0.5 0.083 55 3 0.9 5 1.06 0.098 S 3 0.5 5 3 0.4 5 1.01 0.076 5 ** 1.13 0.074 5 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Alb t 1M Mean 49.1 SD 2.50 n5 2M Mean 51.1 SD 2.40 n5 3M Mean SD n 50.1 2.07 5 4M Mean SD n DWuinllniaemtt'ss' tteesstt:: * 52.9 1.58 5 #*#pp<<00.0.051 al Slob % 21.6 1.25 5 * 18.6 1.32 5 * 20.0 1.63 5 * 19.2 1.47 5 a2 Slob % 6.3 0.32 5 6.2 0.78 5 7.0 0.17 5 ## 5.0 0.28 5 Beta Slob t 18.9 0.65 5 19.9 1.46 5 18.5 1.60 5 18.2 1.52 5 Qamraa Slob % 4.1 1.03 5 4.3 0.98 5 4.4 0.95 5 4.6 0.97 5 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Qroup Aik. Phoa U/L ALT U/L AST U/L IF Mean 237 38 81 SD 36.7 3.8 3.6 n 555 2F Mean SD n 215 51.0 5 41 81 7.8 13.0 55 3F Mean SD n 266 36.4 5 33 83 5.3 10.9 55 4F Mew SD n Williams' test: 240 39.6 5 33 5.7 5 *^<0.05, **/K0.01 76 8.0 5 Bili. Total umol/L Urea mmol/L Crt umol/L Glue mmol/L Choi Total mmol/L Trig mmol/L 3 7.15 0.9 0.981 55 * 2 6.52 0.5 0.812 55 * 2 5.74 0.4 0.458 55 * 2 6.19 0.4 1.069 55 44 6.79 2.09 0.53 4.1 0.738 0.433 0.194 5555 47 7.45 1.91 0.50 3.1 0.521 0.154 0.253 5 5 55 ** 44 6.59 1.52 0.46 2.4 1.043 0.085 0.129 5555 ** 43 7.03 1.23 0.51 1.1 0.659 0.345 0.347 5 5 55 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Ha mmol/L K mmol/L Cl mmol/L Ca Total mmol/L Phoa mmol/L IF Mean SD n 140 3.7 0.8 0.11 55 101 2.66 1.91 1.1 0.082 0.174 555 2F Mean SD n 140 3.5 0.9 0.21 55 101 1.5 5 3F Mean SD n 139 3.5 0.8 0.30 55 101 1.4 5 4F Mean SD n Williams' test: ** 137 1.1 5 * p < 0.05, 3.4 0.25 5 * V < o .o i 100 1.8 5 2.59 0.135 5 * 2.50 0.084 5 * 2.51 0.083 5 1.73 0.227 5 1.72 0.200 5 1.80 0.133 5 Total Prot g/L 64 1.8 S 64 2.6 5 59 5.3 5 60 3.3 5 Alb g/L 34 0.9 5 35 2.9 5 33 3.0 5 33 2.2 5 al Glob g/L 11 0.5 5 11 0.4 5 10 1.0 5 10 1.5 5 a2 Glob g/L 4 0.4 5 4 0.4 5 4 0.5 5 * 3 0.5 5 MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Beta Olob g/L Gamma Glob g/L A/G Ratio IF Mean SD n 12 3 1.15 0.9 0.5 0.059 555 2F Mean SD n 12 3 1.19 1.1 0.4 0.135 555 3F Mean SD n 10 3 1.22 1.1 0.8 0.025 s55 4F Mean 11 3 1.20 SD 1.3 0.8 0.078 n 555 No differences of statistical significance (p> 0.05) MIN 252/004312 TABLE 14 (Blood chemistry - continued) Group Alb % IP Mean SD n 53.5 1.47 5 2F Mean SD n 54.3 2.94 5 3F Mean SD n 54.5 0.70 5 4F Mean SD n Williams'1test: 54.4 1.10 5 * p < 0.05 al Glob % 2 Glob % Beta Glob % Gamma tGlob 16.7 0.74 5 6.3 0.45 5 18.1 1.14 5 5.3 0.97 5 17.3 0.73 5 6.2 0.49 5 17.9 1.83 5 4.3 0.69 5 16.9 0.54 5 16.7 1.76 5 6.2 0.13 5 * 5.7 0.41 5 17.7 1.07 5 17.8 1.83 5 4.8 0.59 5 5.3 1.45 5 TABLE 15 Macroscopic pathology - group distribution of findings P rin t Ho: 0013 Group Compound Exposure level(ppm): i Control 0 234 -- T-7499 - SO 150 450 Printad: 26-FEB-01 Xybion protocol number: HIM 252 -- N U M B E R -O F -A N IM A L S - ORGAN AND KEYWORD (S) OR PHRASE SEX: --- MALE-- -- -- -- FEMALE-- GROUP: -l- -2- -3- -4- -1- -2- -3- -4- NUMBER: 5 5 5 5 5 5 5 5 ** TOP OF LIST ** ADRENALS ........................................ EPIDIDYMIDES .................................... 5 55 55 555 5 5 5 500 00 HEART ........................................... KIDNEYS ......................................... PELVIC DILATION LARYNX .......................................... L I V E R ........................................... MEDIAN CLEFT PALE AREA(S) LOBE(S) NECROTIC iLONGS BRONCHI ................................. DARK AREA(S) 55555555 55555555 0 0 1 0 10 00 5555555s 5 5 5 55 5 55 00001000 00001000 555s555s 0 0 0 100 10 NASAL TURBINATES ................................ SPLEEN .......................................... TESTES .......................................... TRACHEA ......................................... TRACHEAL BIFURC.................................. LN MANDIBULAR................................... CYSTIC ENLARGEMENT ENLARGED 5 5 5 5 5 5 55 5 5 5 5 5 5 55 555s0000 55555555 5 5 5 55 5 55 5 5 5 55 5 55 10000000 000 10010 MIN 252/004312 TABLE 15 (Macroscopic pathology - continued) P rin t No: 0013 Group : Compound Exposure level(ppm): 1 Control 0 234 -- T-7499 - 50 150 450 Printed: 26-FEB-01 Xybion protocol nuatoor: MIN 252 -- N UM B E R -O F -AN IM A L S - SEX; --- MALE------ ___ -- FEMALE-- GROUP: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN AND KEYWORD (S) OR PHRASE Os ** FROM PREVIOUS PAGE ** LN MANDIBULAR................................ CONGESTED NUMBER: 5 5 5 5 5 5 5 5 555 55555 000 10000 LN PANCREATIC....... ........................ ENLARGED 555 55555 000 10000 S K I N ........................................ SCAB(S) 5s5 55555 000 10000 STOMACH ..................................... ANTRUM WHITE NODULE (S) 555 55555 0 0 0 10 110 UTERUS ................... .................. . FLUID DISTENTION 00 0 0 5555 00 0 00 122 V A G I N A ...................................... PALE ARA(S) ** END OF LIST ** 00005555 6 00 0 00 10 MIN 252/004312 TABLE 16 Organ weights - group mean values (g) Group : Compound Exposure level (ppm) : 1 Control 0 234 -- T-7499 - 50 150 450 P rin t No: 0014 Printed: 26-FEB-01 Xybion protocol number: MIN 252 SEX; GROUP; NUMBER: t1I 111 riii-4 iii i 1 ABSOLUTE ORGAN WEIGHTS ------- MALE---------------- -- 2--- -- 3-- -- 4-- 555 IO N MEAN sd TERMINAL BODY WEIGHT 9) ** 5 386.0 5 357.7 5 358.2 5 331.7 : 11.2 37.0 23.1 34.0 ADRENALS N MEAN sd 5 0.054 0.007 5 0.059 0.005 5 0.058 0.004 5 0.054 0.011 EPIDIDYMIDES N MEAN sd 5 0.978 0.032 5 0.972 0.092 5 0.920 0.083 5 0.904 0.061 HEART N MEAN sd 5 1.305 0.174 5 1.549 0.234 5 1.277 0.205 5 1.106 0.167 KIDNEYS N MEAN sd 5 2.44 0.05 5 2.39 0.30 5 2.35 0.19 5 2.22 0.31 Williams' test: ** p< 0.01 Dunnett's test: <# p< 0.01 BODYWEIGHTS ADJUSTED VALUES 1-- -- 2-- -- 3-- -- 4-- N MEAN N MEAN 5 1.168 HEART * 55 1.552 1.278 5 1.239 5 2.27 KIDNEYS 5 2.39 5 2.35 5 2.38 MIN 252/004312 TABLE 16 (Organ weights - continued) Group : Coopound Exposure level(ppm) ts 1 Control 0 234 -- T-7499 - 50 150 450 Print Not 0014 Printed! 26-FSB-01 Xybion protocol nuaber: M m 252 SKI: GROUP: NUMBER: ABSOLUTE ORGAN WEIGHTS -...... MALE..... .......... ---2........ 3........ 4--5555 N MEAN sd : : 5 14.77 1.10 LIVER * 5 13.07 2.06 5 12.43 0.87 * 5 12.69 1.79 N MEAN sd : 5 1.259 0.072 LUNGS 5 1.306 0.158 5 1.248 0.095 5 1.180 0.054 TESTES N MEAN sd : * 5 3.31 0.22 5 3.48 0.58 5 3.29 0.10 5 3.31 0.26 Milliuu' teat: * p< 0.05 BODYWEIOHTS ADJUSTED VALUES -- 1-- -- 2........ 3........ 4--- N: MEAN : 5 13.83 LIVER 5 13.09 5 12.44 5 13.61 Kt MEAN : 5 1.203 LUNGS 5 1.308 5 1.249 5 1.234 MIN 252/004312 TABLE 16 (Organ weights - continued) Group Compound Exposure level(ppe): 1 Control 0 2 34 -- T-7499 - 50 150 450 P rin t No: 0015 Printed: 26-FEB-01 Xybion protocol number: MIN 252 SEX: GROUP: NUMBER: ABSOLUTE ORGAN HEIGHTS ------- FEMALE--------------- -- 2-- -- 3-- -- 4-- 5555 N MEAN Sd TERMINAL BODY HEIGHT 9) 5 214.8 12.9 5 210.8 13.9 5 206.7 11.1 5 205.0 18.8 ADRENALS N MEAN sd 5 0.057 0.010 5 0.069 0.008 5 0.053 0.004 5 0.069 0.010 HEART N MEAN sd 5 0.924 0.163 5 0.898 0.100 5 0.891 0.124 5 0.840 0.121 KIDNEYS N MEAN sd 5 1.37 0.14 5 1.48 0.10 5 1.59 0.37 5 1.66 0.36 LIVER N MEAN sd : 5 7.81 1.00 5 7.68 0.52 Dunnett' s test: # p < 0 .0 5 5 7.34 0.50 5 7.56 0.54 N MEAN N MEAN N MEAN BODYHEIGHTS ADJUSTED VALUES -- 1-- -- -2-- -- 3-- -- 4-- 5 0.055 5 0.903 ADRENALS * 55 0.069 0.053 HEART 5 0.893 5 0.901 * 5 0.070 5 0.856 5 7.63 LIVER 5 7.63 5 7.41 5 7.69 MIN 252/004312 TABLE 16 (Organ weights - continued) Group 1234 Compound Control -- T-7499 - Exposure level(ppm): 0 so 150 450 00\\ SEX: GROUP : NUMBER: ABSOLUTE ORGAN WEIGHTS --------------- FEMALE---------------- 1-- -- 2-- -- 3-- -- 4-- 5555 LUNGS N MEAN sd 5 0.998 0.090 5 0.938 0.060 5 0.946 0.067 No d if f e r e n c e s o f s t a t i s t i c a l s ig n if ic a n c e 5 0.949 0.064 Print No: 00X5 Printed: 26-FEB-01 Xybion protocol number; MIN 252 TABLE 17 Microscopic pathology - expanded incidence summary P rin t Ho: 0016 Oroup Compound Exposure level(ppm): 1 Control 0 2 34 -- T-7499 - 50 150 450 Printed: 26-FEB-01 Xybion protocol number: MIN 252 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- S E X : ------ MALE------ ----- FEMALE----GROUP: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN AND FINDING DESCRIPTION NUMBER: 5 5 5 5 5 5 5 5 ** TOP OF LIST ** HEART .................... -- VALVULAR ENDOCARDITIS 0 0 0 0 0 0 0NUMBER EXAMINED: 5 0 0 5 5 0 0 5 1 KIDNEYS ............................................. NUMBER EXAMINED 5 0 1 5 5 0 0 5 OS 0-- CORTICAL TUBULAR BASOPHILIA 30110 00 0-- MEDULLARY TUBULAR BASOPHILIA 10000 00 0-- DILATED MEDULLARY TUBULES CONTAINING EOSINOPHILIC 0 0 100 00 COLLOID -- TUBULAR HYPERPLASIA, SIMPLE -- INTERSTITIAL INFLAMMATION -- UNILATERAL PELVIC DILATATION 0 0 0 01 0 0 0 0 0 0 00 0 0 1 11 0 1 0 000 LARYNX .............................................. NUMBER EXAMINED: -- EPITHELIAL HYPERPLASIA - ARYTENOIDS -- EPITHELIAL HYPERPLASIA - VENTRAL POUCH -- INFLAMMATORY CELLS IN LAMINA PROPRIA (VENTRAL AND VENTROLATERAL) -- INFLAMMATORY CELLS IN LAMINA PROPRIA (VENTRAL) 50 0 55005 10 0 1 10 00 10 0 0 0 0 0 0 10 00 000 0 00 0 0 0 0 0 1 LIVER ............................................... NUMBER EXAMINED 5 0 0 5 5 0 0 5 1 0 0-- SIDEROPHAGES, BILE DUCT HYPERPLASIA, FIBROSIS AND 0000 0 MINERALISATION - RIGHT ANTERIOR LOBE MIN 252/004312 LUNGS 6 BRONCHI ................. -- ALVEOLITIS -- FOAMY ALVEOLAR MACROPHAGES -- ALVEOLAR OSSEOUS METAPLASIA -- EXTRAVASATION OF EOSINOPHILS -- SUBPLEURAL INFLAMMATION NASAL TURBINATES ....................... -- TRANSITIONAL EPITHELIAL HYPERPLASIA NUMBER EXAMINED: 25 00 0 0 5 0 5 2 0 0 1 1 5 0 000 000 002 0 00 0 0 00 00 00 0 10 10 0 1 0 0 0 0 0 0 00 00 0NUMBER EXAMINED: 5 2 00 00 5 0 5 5 TABLE 17 (Microscopic pathology - continued) P rin t No: 0016 Group Compound Exposure laval(ppm); 1 Control 0 2 34 - T-7499 - 50 150 450 Printad: 26-FEB-01 Xybion protocol nunber: MIN 252 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- j\ii tit i i ii i ii i i Ii Ii i Ii Ii i i ii iii Ii i i ii Iii i i Ii ii iii iii i i S E X : ------ MALE------ ----- FEMALE GROUP: -i- -2- -3- -4- -1- -2- -3- -4- ORGAN AND FINDING DESCRIPTION NUMBER: 5 5 5 5 5 5 5 5 ------------------------------------------------------ --a -- -a - --Mi-- --Si-- -a - -a - -a - --a-- ** FROM PREVIOUS PAGE ** NASAL TURBINATES .................................... NUMBER EXAMINED: 5 0 0 5 5 0 0 5 -- RESPIRATORY EPITHELIAL HYPERPLASIA 1 0 00 00 0 0 -- OEDEMA IN STENO'S GLAND 00 00000 1 TRACHEA .............................................. NUMBER EXAMINED: 5 0 0 5 5 0 0 5 -- SUBEPITHELIAL INFLAMMATION 300 i2000 IN MANDIBULAR........................................ NUMBER EXAMINED: 1 0 0 2 0 0 1 0 -- CYSTIC SINUSES 10000000 -- INCREASED CELLULARITY - GENERALISED 0 00 100 10 -- BINDS ERYTHROCYTOSIS/ERYTHROPHAGOCYTOSIS 0 00 100 00 12 PANCREATIC ........................................ NUMBER EXAMINED: 0 0 0 1 0 0 0 0 -- SINUS ERYTHROCYTOSIS/ERYTHROPHAGOCYTOSI8 0 0 0 10 0 0 0 S K I N ................................................ NUMBER EXAMINED: 0 0 0 1 0 0 0 0 -- SCAB 0 00 10000 -- EPIDERMAL HYPERPLASIA 00010000 -- EPIDERMAL ULCERATION 0 0010000 --DERMAL INFLAMMATION 000 10 0 0 0 STOMACH .............................................. NUMBER EXAMINED: 0 0 0 1 0 1 1 0 -- ECTOPIC NONGLANDULAR EPITHELIUM IN GLANDULAR MUCOSA, 0 0 0 10 110 FOCAL U T E R U S .................. ............ ............... NUMBER EXAMINED: 0 0 0 0 0 1 2 2 -- LUMINAL DILATATION 0 0 00 0 122 VAGINA ............................................... NUMBER EXAMINED: 0 0 0 0 0 0 1 0 -- PROMINENT PROSTATIC TISSUE CM SEROSAL ASPECT 0 0 0 0 0 0 10 ** END OF LIST ** MIN 252/004312 APPENDIX 1 Clinical signs pre and post exposure - individual daily observations Group (Co1nMtrol) (Low2Mdose) (Inte3r,Mdose) (Hig4hMdose) (CoInFtrol) (Low2Fdose) (Inter3,Fdose) (Hig4hFdose) An1un6mi-m2bae0lr No abnormalities detected Observations 6-10 No abnormalities detected 11 -15 No abnormalities detected 42351 NNNAAooggoiiaaattaabbbttnnneeoooddrrraammmnnaaaddllliiivvtttiiiooeeeccsssaadddlliieeesstttiieeenncccggttteeewwdddhheennhhaannddlleedd,, EExxppoossuurree 111 31-35 No abnormalities detected 36 43337890 AAwAAH15gghggy.iiiipettttneVaaaartttthoaeeeeacdddcdntaiaawdwlvinnleshheddi,edenvvEn,ngooxEhwcchpxaaaaohnpllnsiiedossudnlsiirlennueehdggrda,e1,nwws1EdE9hhxlxeee,p1dpnno,5oshhsEuaaaurxnnnerpddedsosllee31sdd9u7t,,,roaeEEn1s9xx1d,3pp,poo1tr1o1sse5uu,-9rre1,eeax4sn1p11da9o9n,asd1u1n71rd1e6aa11nn4t5ddot.op12H7r02y.e0p-aHeneayxrndappdpcoeptrrsieravue-cere-te,iexvExpe1pox,5ospE.usoxurVsepruoseorces11au531lri1esainn11gd 26 27 2298 30 AAEAAE(HWaxxggggybappiiiipnlttttookeaaoaassrittttrunueeeaemdcdddrgreeatiassowalavnnnng29ehddda,,etaiov4Entvnve),oxod,oshc6pcEcaa,(1oaalnxa0ls1ilbidpsui2ssinlorineitneosnodgsugrg,mrw19ewE5waht,xlh1oehp71egne17onanth1soiaht.ua)hnnaW1r,aden9dEn.dal1dex1ll9Hkedl1pe.d.i,oydn,EsHp,guVExeyErorpxoepanxopcsecpasotrt4ouoalisiv,ecrsuseeuts6irin,er,v(1egEae.1s1b,x2w6HnpE1.tohoyxorestppWmoun1eor4arsaeh3a,lul,ack1rg1ntei06ai7ndsvi,glteae9)1,,ndo8aEEd,n,nxxEdapt1pxno1o1odpe.s0ssouu2Wsrr(0ueea.arsbel1nk9Hs4oinayr1ngmp1deaoral1ann0cgdttaiovi1tee4)s,,. 21 22 23 24 25 HAtAAAAeEgg11ox0aa3xygggggpii.5ppiititiitott))ot,ttoeaaaaWa,,ssr1ttttt2uEEaueeeee1a0rcdxddxdrdleeatkppaiwwsn2aavionowndnn0ehhsds1ghdd,u.uee1pterrEnnoo3Hervenvx.neshshoo5yp-aacc,ht22Wpeoonnaaa7xesettdldln,aprooiiuslalssdlo8eerkiic28(le,dsdnneiat0un,i,ggad1b9vrgn1EaE,neewwndoxxo,a2EdrphhpnEn10mxpoeeod0x.tpnnsrsaoptueo1ulHoeohh-r6srsgseeeyua.a2ausxsn(pnr0iarpeWdedt1e1b)soarll,neaeatn9stoEodcdloud1kt,,rrx6ip5imevpnt.EEroeaog2Veaxx,nsl-0ppooeudE5.goocnxr,xasesa1pHiptuusl1to1oioy)rr.1s2se,eespiusssunEVetraorgr(xeone1a1apw8sccdb2ottatn09oohaislvo.ine1uaser556ndriHnm,..e.nhd1sEygaa0pxWlW12nwVe1ptgdotraaohaoolalleisec2ckktud8na)t0ii,ir,lnnvaehiaEEggsenani9xx,dnndooppEdgnnp1oolxe0rsspwtteduuooto-,rroheeeseeEessuxss2nxpr0((2e1poaahaobsbt9toaonusnnnudroo87derrrpelmmaa2ersnned0aadd1.,ll : 69 : APPENDIX 2 Clinical signs (pre-exposure) - individual weekly observations P rin t No: 0019 Group 1234 Compound Control Exposure level (ppm) 0 50 150 450 Printed: 26-FEB-01 Xybion protocol number: MIN 252 CATEGORY A N U A L DEATH NX OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 1M WEEKS 1-5 16 7 17 7 18 7 19 7 20 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group Compound Exposure level (ppm) : 1 Control 0 234 -- T-7499 - 50 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP; 2M 67 77 87 97 10 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS WEEKS 1-5 P rin t No: 0019 Printed: 26-FEB-01 Xyfoion protocol number: MIN 252 MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group Compound Exposure level(ppm) i2 34 Control T-7499 - 0 50 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP 3M ii 7 12 7 13 7 14 7 15 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS WEEKS 1-5 Print No: 0019 Printed: 26-FEB-01 Xybion protocol nuaber: MIN 252 MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group Compound Exposure level(ppa) 1 Control 0 234 ----- T-7499 50 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4M 17 27 37 47 57 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 STAINING BROWN DORSAL BODY SURFACE 5 ANIMAL HAS NO SIGNIFICANT FINDINGS H(D i 5 ANIMAL HAS NO FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS WEEKS 1-5 5 Print No: 0019 Printed: 26-FEB-01 Xybion protocol nunbar: MIN 252 MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group i2 34 Compound Control Exposure level(ppm) 0 SO 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: IF 3 i 31 7 32 7 33 7 34 7 35 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDING8 WEEKS 1-5 P rin t No: 0019 Printed: 26-FEB-01 Xybion protocol number MIN 252 i MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) P rin t Ho: 0019 Group Coapound Exposure lttvol (ppm) 1 Control 0 234 ----- T-7499 ---50 150 450 Printed: 26-FEB-01 Xybion protocol lumbar: MIN 252 ^U4\ CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODS DEATH QUALIFIER GROUP: 2F WEEKS 1-5 36 7 37 7 38 7 39 7 40 7 5 SKIN ABRASION DRY TAIL 5 BEHAVIOUR IRRITABLE 5 ANIMAL BAS NO SIGNIFICANT FINDING8 s SKIN ABRASION DRY TAIL s ANIMAL BAS NO SIGNIFICANT FINDINGS 4 2 3-5 MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group 1234 Coapound Control Exposure laval (ppa) 0 50 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 3F 26 7 27 7 28 7 29 7 30 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS MEEKS 1-5 P e in t Ho: 0019 Printed: 26-FEB-01 Xybion protocol nrabar: MIN 252 MIN 252/004312 APPENDIX 2 (Clinical signs (pre-exposure) - continued) Group i234 Compound Control Exposure level (ppm) 0 SO 150 450 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP; 4F 21 7 22 7 23 7 24 7 25 7 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 ANIMAL HAS NO SIGNIFICANT FINDINGS 5 TEETH ABNORMAL COLOUR PALE MEEKS 1-5 4-5 P r i n t Mo: 0019 Printad: 26-FEB-01 Xybion protocol nunber: MIN 252 MIN 252/004312 APPENDIX 3 Bodyweights - individual values (g) Group 1234 Coapound Control T-7499 ---- Exposure level(ppsi): 0 50 150 450 GROUAPNI MAL WEE0 K WEE1 K 2 MEEK WEE3 K WEE4 K 1M 16 292 336 374 393 401 .* 17 275 312 345 366 373 18 279 322 359 389 397 00 19 277 318 349 376 379 20 263 296 336 375 386 2M 6 274 306 332 361 371 7 293 333 357 378 389 8 241 263 272 289 289 9 275 307 329 348 348 10 265 303 331 361 364 3M 11 267 281 313 332 338 12 278 306 327 355 356 13 281 315 346 381 391 14 277 308 329 350 355 15 273 298 315 325 333 4M 1 264 277 290 311 323 2 250 255 273 285 298 3 293 306 334 363 388 4 270 286 308 334 343 5 265 271 292 308 306 P r i n t Mo: 0020 Printad: 26-FEB-01 Xybion protocol numbar: MIN 252 MIN 252/004312 APPENDIX 3 (Bodyweights - continued) Group Compound Exposure laval(ppm): 1 Control 0 23 4 ----- T-7499 ---- 50 150 450 GROUP ANIMAL WEEK 0 WEEK 1 WEEK 2 WEEK 3 WEEK 4 SO) IF 31 165 185 197 208 208 32 193 208 214 229 231 33 171 181 192 200 194 34 183 188 194 199 212 35 IBB 1B8 205 217 216 2F 36 181 186 186 181 188 37 189 204 214 224 220 38 178 195 199 202 200 39 189 190 201 200 210 40 165 179 192 204 203 3F 26 174 161 165 191 196 27 191 194 206 206 214 28 166 177 175 180 190 29 189 193 206 217 218 30 194 182 185 198 203 4F 21 186 178 177 187 186 22 163 163 170 181 184 23 190 195 199 225 222 24 202 197 203 213 219 25 179 185 180 194 207 Print No: 0021 Printad: 26-FEB-01 Xybion protocol number: MIN 252 MIN 252/004312 MIN 252/004312 APPENDIX 4 Functional observational battery - individual observations Pre-dose Group 1 Males 16 OBSERVATIONS IN HVRTHeaomncEdaollHviisniAingnNggD N22 degree - IN AARBTUHoercroiatlnEiuurveissnAiatpcgylRroeccuEsoonNeuutnnAnttt 1N4251 Gait - MAATRTSNtoaipagIiuptrlPhuctrplUtorheiinnanLscgcAhhrTefIlOexNS 33331- Pupviol creaflilseexs TBeomdypwereaitguhrte((g)C) B37.6 248 GLARfhNIoPiDnreSdIllNTiimmRGbEbFNOGOTTHSP(KLGA)Yt (cm) J 007..68.690 Values represent the mean of 2 trials 17 Anim1a8l number 19 N22 22 N -- N4601 N4990 -- 37B323321-.8 23231 2B 38.0 249 238 001..188.502 001..285.832 N22 - 1N4500 - 3243138B- .1 253 009...86009 20 N221L428-6 32311 - B38.3 238 008..76.111 : 80 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 2 Males OBSERVATIONS 6 IN THE HAND RHeamndolviningg 22 IN VTHocdEaelAgisrReineEgNA N - BAARoecroaltuiurvissniatcgylocucoonuutnntt 13482 UGraiinte present M- MANIPULATIONS ATopupcrhoach 23 PRTSutaiagpitvrlhuitolpltrecirinnneasgfclilhsereexfslex BTeomdypwereaitguhret ((g)C) 351 238B.2 244 GRIP STRENGTH (KG) J fhoinredlliimmbb 00..4587 LANDING FOOTSPLAY (cm) J 10.1 Values represent the mean of 2 trials 7 Anima8l number 9 N22 N22 -- 142 145 41 09 M- S - 22361 33301 37B2-.3 -1 38B.4 252 226 00..7750 9.7 008..65.418 N22TN140616 23331 - 38B-.1 247 001..478.782 10 N22 - M14085 - 33331 - 37B.9 241 009..86.079 : 81 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 3 Maies 11 OBSERVATIONS IN IN RVHARUTABTHHeeoocraroimatnlcdnEEiuuradveeoisslnHlAigivaitpcsgrnyliARroeingneceucENgsgoonNeDuutnnAnttt N22 - 402S5 MAAGTRTSNtoaipaagIiuiprlPhtctrplUtoheiinanLcgcAhhrTefIlOexNS - 32331 PTBueompdvtuiyoplrwcernearseafliiltgseuehxrste((g)C) 238B.1 237 GLARfhNIoPinDredSIllTNiimmRGbbEFNOGOTTHSP(KLGA)Y%(cm) % 00..8526 13.1 Values represent the mean of 2 trials Animal number 12 13 14 N22 N22 -- TN1140150 1N407-5 30 32 31 31 3 2B 3B- 37.8 37.9 242 242 001..155.152 009..45.667 N22 - 1N2451 - 3232-1 37B- .8 253 001..387.172 15 N23143S8-0 32231 B238.4 243 001..287.067 : 82 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 4 Males 1 OBSERVATIONS IN THE HAND RHeamndolviningg 22 IN VTHocdEaelAgisrRieneEgNA N - Activity count 16 Arousal 4 Rearing count 5 Bolus count 0 GUariint e present TN1 MANIPULATIONS TAopupcrhoach 23 RStiagrhtlteing reflex 31 Taitvluoprcninasclihses 3 1- PTuepmilpreeraflteuxre (C) 38B.2 GRBIPodSyTwReEigNhGt (TgH) (KG) J 228 hfoinredlliimmbb 00..6729 LANDING FOOTSPLAY (cm) $ 9.2 Values represent the mean of 2 trials 2 Anima3l number 4 N23- N22- TN140614 T2N40811 23 36 31 31 32 - B- B22 37.8 37.7 221 259 00..5654 11.7 009..65.288 N22N46-81 33311 - 38B- .0 244 008..7.6516 5 N23 - 140S41 - 22331 137B.9 240 007...87149 : 83 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 1 Females 31 OBSERVATIONS IN THE HAND IN VRHABARUTGHoeeocarraoimanltcidnEuituradveeoissllnAgiivaitpcsgnryliRireongnececuEggsoonNeuutnnAnttt N22 - N4720- MAATRTSNtaoipagIiuprlPhctrplUtoheiinaLncgcAhhrTefIlOexNS 22331 Puptvuiolrcnreasflilseexs TBeomdypwereaitguhrte((g)C) 237B.1 155 GRfhIoPinreSdllTiimmRbEbNGTH (KG) { 00..6833 L%ANDINVGaluFeOsOreTpSrePsLeAntYth(ecmm)ea$n of 2 trials10.6 32 Animal number 33 34 22 22 NN -- 46 143 20 06 NS -- 33261 32351 337B.8 - 37B- .7 176 161 00..7853 10.1 006..66.753 N22 - T1N40716 38B33362-1.6 159 001..278.909 35 N22T1470S62 32331 2B 37.8 179 001..078.707 : 84 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 2 Females 36 OBSERVATIONS IN RTHemEoHviAnNg D 2 Handling 2 Vocdaelgisreineg N2 IN AUARBTHoerrcoialtnEuiurveissnAiaptcgylRroecucEsooneNuutnnAnttt N14203 MAAGNpaIipPtrUoaLcAhTIONS - 3 PTRTSutaoiagpiutvrlhiuctolpltrhecirinnneasgfclilhsereexsflex BTeomdypwereaitguhrte((g)C) 2431 238B.9 164 GRIP STRENGTH (KG) % forelimb 0.82 hindlimb 0.68 LANDING FOOTSPLAY (cm) J 11.5 $ Values represent the mean of 2 trials 37 Anim3a8l number 39 22 N2 N2 -- 142 143 36 00 NN -- 22331 22331 -1 38B.4 - B38.4 170 168 001..175.578 001..177.334 N22N14061 - 32311 2B 37.7 169 009..6.6234 40 N22 - TN140616 23231 - 38B.6 153 008..46.939 : 85 : MIN 252/004312 APPENDIX 4 (Functional observational battery-continued) Pre-dose Group 3 Females 26 OBSERVATIONS IN RHTHeamnEdolHviniAnggND Vocdaelgisrieneg N23- IMNAAABTRUGATRTPBTSNutaoHeoeiocrpraagIoipimualtdiptvrlEnPuihtuurictyrvoplepiUlstroshAnweiiceirnatnapcLngeayrslRecgoracflAiehicluctEhgsersuoToneehNexruutfsIntelOnAnt(et(gtxN)CS) N14021 - 33331 -1 38B.1 155 LGARfhNIoPiDnreSdIllNTiimmRGbEbFNOGOTTHS(PKLGA)YJ(cm) J 00..6574 5.1 $ Values represent the mean of 2 trials 27 Anim2a8l number 29 N23 N22 -- T1N46012 140 N03- 38B23032-1.0 22451 37B3- .4 170 159 007..88.236 001..175.554 N22 - T2N46021 32231 - B37.5 171 001..088.368 30 Y221 TN144011 32231 B238.5 175 008..66.884 : 86 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Pre-dose Group 4 Females 21 OBSERVATIONS IN RHTHeamnEdolHviniAngNg D 22 Vocalising N IN THdEeAgrReeENA - AArcotiuvsiatyl count 141 UGRBeoariailntureisnpcgroecusoneutnntt TN072 MANIPULATIONS RATTStaoipagiuprlhctrpltoheiinancgchhreflex 32231 tvuorcnaslises -1 Pupil reflex B Temperature (C) 38.5 Bodyweight (g) 167 GRhfIoPinredSllTiimmRbbENGTH (KG) $ 00..5619 LANDING FOOTSPLAY (cm) t 10.1 { Values represent the mean of 2 trials 22 Animal number 23 24 N23- N22- N22- TN190412 TN146014 S9530- 36331 23331 - B- B-1 38.5 38.1 154 172 23331 - B38.4 179 008..65.155 001..187.094 005..6.8840 25 N22 - TN144013 23331 2B 38.7 160 008..56.616 Group I Males IIONNBVRURHAABGTSTSaHEHeoeroaraciloamilntcRddniEEtuiurvadveeeioVssalnlAHggiivaipttcAsgrnryiliRAroeoiengneTceecunENgsgoIonNeODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) W eekl MIN 252/004312 16 17 Anim1a8l number 19 20 NN22-- NN22-- NN22-- 1N442-1 142 M50- 1N490-0 NN22-47 N34- NN22-147 M05- : 88 : Group 2 Males IIONNBARUGRVABTHTSSaHeoHeErracoaoilamiltncddRniEEtuiurvdaveeeiosVsalnlAggiHaivipttcgAsrrnyiliRAroeeiongenTceeucnENgsogoIneNOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 1 MIN 252/004312 6 7 Anima8l number 9 10 N32 N22 N22 N- N- N--- N22 N- - 48 147 140 145 91 37 05 60 M - TS1 N - TN1 N22 N- - 464S8- Group 3 Males IOINNBVRHAGARBUTTSSaEHHeeooraacrloimaniltcRiddEnEtuiurvadveeeVoissallHnAggiiviattpAcsgnrriyliARoieroengTneceecnuENgsgoIoneNODuutnnANntttS APPENDIX 4 (Functional observational battery - continued) Week 1 MIN 252/004312 11 12 Animal number 13 14 15 N22 N22 N22 N- N- N- -- - N22 N- - T4403S1 11N4012 - 1N4053 - TN42911 NN23-N4337- : 90 : Group 4 Males OBSERVATIONS IN IN URBGHVAARTTSaHHoeeraraocilomalintcddniEEutiurvdaveeeiossalnAlHggiivaipttcsgrrniyliRAroeeoingenceeucnENgsgooneNuDutnnAnttt APPENDIX 4 (Functional observational battery - continued) Week 1 MIN 252/004312 1 2 Anima3l number 4 5 NN22-- NN22-- N23 N-- NN22-- N23 N- - 146S02 N4407 N0946 N14301 N0429 T2 - T1 - - 91 T Group 1 Females IIONNBURHVAARBGTTSSaHHeoEerraoacliomalnitcRddinEEutiurvdaveeeoiVssallnHAggiviaipttcAsgrrniyilARrooeeingneTceeuncENgsgoIoneNODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 1 MIN 252/004312 31 32 Animal number 33 34 35 22 22 32 N- N- N- Y2 N- N - N22 N- - N1407-1 N4490- TN159017 T2N140111 N22 N- - T1N46015 : 92 : Group 2 Females IIONNBUAARBGVTRHTSSaoHeHrraecEoaiolailmtncnddRiEEtuiurvadveeeisoVsalnlAiggHiavipttcgsArrnyiliRAroeieongencTeeucnENgsgoonIeNuODutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 1 MIN 252/004312 36 37 Anim3a8l number 39 40 N22 N22 N22 N22 N- Y2- Y2 N- - 146 144 48 00 SN - T1 N1420-2 TN140714 N23 N- - TN145012 : 93 : Group 3 Females OINBRHVTSSaEHeaolmncRdidEvdaeeoVallHggivitAsnrriiAeieongTneenNggIODNS IN ARUABGTHeorrcaoialitnEutiurveissnAiaptcgylRroecucEsoonNeuutnnAnttt APPENDIX 4 (Functional observational battery - continued) W eekl MIN 252/004312 26 27 Anim2a8l number 29 30 NN22-- N22 N- NY22-1 NN22-- N14607 - N1470-1 T1N47014 T2N49002 YY2231 N4950 - : 94 : Group 4 Females OINBRHVTSSaHEeaolmncddRiEvdaeeoVallggHivitAsrrniiAeeiongnTeenNggIODNS IN UAARBGTHoerrcaioaltinEuiturveissnAiatpcgylRroecucEsoonNeuutnnAnttt APPENDIX 4 (Functional observational battery - continued) W eekl MIN 252/004312 21 22 Anim2a3l number 24 25 23 22 22 N N- NY1 N N- - -- N23 N- N22 N- - T2HN4095U1 T2HN4081U1 TN146013 N1460-2 N4606- 95 : Group 1 Males IIONNBURVAABGHRTTSHEHeoerroacaoimaintlcRdnEEtiuurdaveeoiVssllnAHgiviaitpcAgsnryliRAroeingTneceucENgsgoIonNeODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 16 17 Anim1a8l number 19 20 N22- N22- N22- 12 11 9 444 101 40 07 N - M- N- N22 N22 -- 12 14 N442 - , M44-1 Group 2 Males IOINNBHVAARRBUGTTSHHEoeeraocraoimantlicRdEnEiuturdaveeioVssllnAHigivaitpcAsgnryliRAroeingneTcecuENgsgooInNeOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 6 7 Anima8l number 9 10 N22 N22 N22 N22 N22 --- - - 141 143 142 141 140 M92 N55 N50 N30 93S - T2 - - - : 97 : Group 3 Males OIINNBHVAARBGTRUTSHEHeoeraocaroiamntilcRdnEEituurdaveeioVssllnAHigivaiptcAgsnryliARroeingenTceucENgsgoIoneNOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 11 12 Anim1a3l number 14 15 N22- N22 - N22 - 4287 14080 14604 TN1 TN1 N - N22 - 1460S-2 N22 - N1407-0 : 98 : Group 4 Males IOINNBAARURHVBGTTSEHHeoerrcaoaoiamtlnicRdnEEiuturdaveeioVsslnHAlgiiavitpcAgsrnyliARroeingnTeceucENgsgoIonNeOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 1 2 Anima3l number 4 5 22 23 23 NNN -- - N22- N23 - 141 143 143 49 140 05 100 60 40 05 TM1 Ts2 T2HNU1 s - T1HNU1 : 99 : Group 1 Females IIONNBAABUGHRTVTRSHoHEerraecaooiaimltncRdnEEtuiurdaveeioVsslnlAgHiivaitpcAgsrnyliRAroeingneTcecuENgsgooIneNODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MEN 252/004312 31 32 Animal number 33 34 35 Y222 Y222 N22- N22- N22- TN147024 TN146012 TN149019 T1N49017 T2N40832 100 : Group 2 Females OIINNBAVARBURHGTTSEHHoeecroraaoiamtlnicRdnEEiuturadveeioVsslnliHAgivaitpcAsgnryliARroiengnTececuENgsgoIoneNOuDutnnAnNtttS APPENDIX4 (Functional observational battery - continued) Week 2 MIN 252/004312 36 37 Anim3a8l number 39 40 22 22 23 Y2 Y1 N - N22 - N23- N4308- TN145012 TN146024 NU4250 TN43071 101 : Group 3 Females IIONNBUVAARBTRHGTSHHEoeerorcaaioamlitncRdnEEtuiuradveeioVsslnAlHgiivaitpcAsgrnylRiArieongneTceucENgsgoIonNeOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 26 27 Anim2a8l number 29 30 222 N2 - N2 - N2- T2N40810 TN144012 TN44082 N22- N22 - TN140513 T2N1450H3U I : 102 : Group 4 Females IIONNBVAARBUHGTRTSHoeHEeorracaoiamlintcRdnEEtuiuradveeioVsslnlAgHiivaiptcAsgnrylRiAroiengneTceucENgsgooInNeOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 2 MIN 252/004312 21 22 Anim2a3l number 24 25 N32 N22 N22 N23 N22 --- - - T2HN14501U1 TN149020 TN42052 TN49502 TN140522 Group 1 Males IIONNBABURVARGTHGTSSaHoHeEerroacarloiomalintcRdidnEEutiuorvadveeeoisVsmallnAgHgiivaitptcAsgrnriyiliRAreiooenngneTceeucnEgNgsgoIoneNOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 16 17 Anim1a8l number 19 20 22 22 22 NN- NY1 N N- -- - NN1490-0 TNN94401 N144S0-1 N22 N- NN4480 - N23 N- - 1YS4902 - : 104 : Group 2 Males IIONNBRHVGAARBUGTTSSaHHeoeEraocrarlooiamitlncddRinEEtiuoruvadveeeiosVmsalnlAHgigivaittpcgsArrnyiiliARreooienngneTceeucnEgNgsgooIneNODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 6 7 Animal number 8 9 10 222 NN2-- N2 N- - NN2-- N143 N146 N141 S07 - S07- TN302 N22 N- - NN3940 - NN23-TN1S40816 : 105 : Group 3 Males IIONNBURAARBGHVGTTSSaHHeoeErracaorloiomalitncRddinEEutiurovdaveeeoisVsmalnlHAiggivaitptcAgsnrryiiliARroieeonngneTceeucnEgNgsgoIoneNODuutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 11 12 Anim1a3l number 14 15 N22 N- N22 Y22 N-- N-1 N22 N- N22 N- - TNN144013 TN1S40811 NN1407-1 N141 T0S51 T1HNN4309U1 : 106 : Group 4 Males IIONNBBAARGRHVGUTSTSaHoeHEeracrroaolioamlitncddRinEEtuiurovadveeeiosVsmalnlAgHgiivaipttcAgsrrniyiliRAroeioenngneTceeucnEgNgsgooIneNOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 1 2 Anima3l number 4 5 22 23 22 NNN N- N- N--- NN22-- N14S40-2 TNN40612 T2HNN14084U1 TNN146013 N22 N- NN4940- 107 : Group I Females OBSERVATIONS IN VRHTSaHeoalmncdidEvadeeoallgHgivitsrnriiAeieongneenNggD IN UAARBTGGHeorrcaroioalitnEutiuroveissmnAiaptcgyilRronecucEgsoonNeuutnnAnttt APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 31 32 Anim3a3l number 34 35 NY222- N22 Y-1 N22 N- - NY22-1 NN22- - TNN140711 NN1405-4 T2NN140511 TNN146022 T2NN40802 : 108 : ] Group 2 Females IIONNBUAARBRVGTHGSTSaHEoHeerrcoarailooamlitncRddniEEutiurovadveeeioVssmalnlAHggiivaipttcAgsrrniyiliRAroeeionngeTnceuecnEgNgsgoIoneNOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 36 37 Anim3a8l number 39 40 22 23 22 NNY Y2 Y2- Y21 N22 Y2- YY2221 TNN143015 TNN107524 TNN143010 T2HNN14066U1 TNN140816 109 : Group 3 Females OBSERVATIONS IN IN ARHVARBUTTSGGaHHeeoraocrraloiomanlticiddnEEuiturvodaveeeoissamllnHAggiivaittpcgsnrriyiliRAroeoeinngneceeucnEgNgsgoonNeDuutnnAnttt APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 26 27 Anim2a8l number 29 30 NN23- NN22- NY221 -- - N22 N- N23 N-- T21NN40100 TNN140827 TN1N42020 T2HNN14604U1 T2HN1N4050U1 110 : Group 4 Females IOINNBHVAARBUGTRGTSSaHEHoeearroacrolioamilntcRddinEEtuiurovadveeeioVssmallnAHggiivaipttcAsgrnriyilRiAroeioenngneTceuecEngNgsgoIonNeOuDutnnAnNtttS APPENDIX 4 (Functional observational battery - continued) Week 3 MIN 252/004312 Animal number 21 22 23 24 25 N22 N- NN32-- NN22- - TNN149027 TNN46082 TNN42081 NY32-1 N22 N-- TNN149013 T2HNN14404U1 111 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 1 Males 16 OINBTSEHREVHAATNIODNS IMNARHVAABUPRGATRTTBSTSNuatoaeHeoeioarocpraaglIopiimmuantlidcpvddtidrlnEPiuhtuiuvrctdayvroeeelpepoilUstsarohllnewggAigiceirivniattnpacnLsgnaerrriyrliseRcgoeeoriecalfngAneiichleeecnuhtEggsregsuooTneehxNeruufstIntelnOAnet(t(tgxN)CS) NN22-TN40831 337B93365---1.52 GL%ARhfNIoPiDnredSIllNTiVimmRGabbEluFNeOsGOrTeTHpSreP(KsLeGAnt)Yt%h(ecmm)ea%n o f 2 1110..11.594 trials Animal number 17 18 19 NN22-- NN22-- N4905- TN140714 337YB26032-11.29 38YB33321-1.1 398 011..493.382 11..5316 11.3 NN22-N4905337B36337---1.99 01..9077 8.9 20 NN22-M1408-7 337YB363328-1.48 01..9512 10.1 : 112 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 2 Males 6 OBSERVATIONS IN RTHemEoHviAnNg D 2 VHSaoalncddivadeeallggiitsrrnioieegneeng N2 N-- IN THE ARENA AArcotiuvsiatyl count 49 Rearing count 6 BUorilnues pcroeusnetnt 4S MAGNaIiPt ULATIONS - RTATStaoipagiutprlhutcrpltrhoeiinnanscgchhreflex 363511 Pupvdioel gcreareflileseexs - B- Temperature (C) 37.8 Bodyweight (g) 366 GRfIoPreSlTimRbENGTH (KG) % 0.99 hindlimb 0.83 LANDING FOOTSPLAY (cm) J 12.8 $ Values represent the mean of 2 trials 7 Anima8l number 9 N22 N22 N- N- -- 144 143 50 70 NN T1 - 32631 33331 --- B- B37.3 38.7 372 291 111..212.134 018..9.0695 N22 N- - 1S4077 - 32331 - - B38.0 350 111.2.32.144 10 N23 N- - TN146013 23331 - B37.0 368 11..2287 8.9 MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 3 Males 11 OBSERVATIONS IN RTHVSaHeaolmncidEvdaeoallHgviitsnriiAeoingnenNggD NN22- IN AUTARBGHeorrcaoialtidnEuiturveeissnAgiatpcgrylReroececuEsoonNeuutnnAnttt - N1408-0 MAARTTPSNutaoipagIipuptvrdlPhuitcroepllUtorheigcirnnanLerascgecflAhilehsreTeexfsIlOexNS TBeomdypwereaitguhrte((g)C) 3332--1 37B-.6 333 GRIP STRENGTH (KG) t hfoinredlliimmbb 01..9312 LANDING FOOTSPLAY (cm) J 14.3 X Values represent the mean of 2 trials 12 Anim1a3l number 14 22 NN2- NN2-- 143 4S0- 1N450-1 3331-1 -- 3323--1- BB 37.2 37.0 355 381 011..180.864 019..9.1416 NN22-429S037YB32321-1.0 352 1111..43.500 15 NN22- - NU4220 3333--1 37B-.8 332 111..212.376 : 114 : 1 MIN 252/004312 APPENDIX 4 (Functional observational battery --continued) Week 4 Group 4 Males 1 OBSERVATIONS IN TRHemEoHviAnNg D 2 Handling 2 VSaolcdivaealgitsrioeineng N N- IN AATHrcotdEiuvesAgiatrylReecEoNuAnt - 142 RBGUeoariailntureisnpcgroecusoneutnntt HU30S1 MANIPULATIONS PRTATSutaoipagpiuvptdrlhiutcorelpltrhoegicirnnanearscgecflilhehsereexsflex 32331 - B- BTeomdypwereaitguhrte((g)C) 37.6 311 GRhfIoPinredSllTiimmRbbENGTH (KG) % 11..3158 LANDING FOOTSPLAY (cm) % 10.8 J Values represent the mean of 2 trials 2 Anima3l number 4 NY221 - T1HN14051U1 33301 YB1 37.4 288 1111..01.624 N22 N- - 1440S8 - 23361 YB1 37.8 378 11..4129 8.6 N22 N- - 10S497 - 23361 - B38.8 337 117...21452 5 N22 N- - TN42091 33331 - B37.3 302 116...03317 : 115 MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 1 Females OIIMNNBAPRVARBUTHAATRTTSGSSNutaaoHEHeeoiorcrpaaaglpIiiomaultnicpRtvdddrlinEEPuithiuruvtcadvroeeelepiolVUstrshoalenlAHgggiicriinvaittnpacAnLsgearrrniysliRAcgroioeeecflngAnTeichleeeucnhENgsersgoIToneexNeODuustfInnlOAnNetttxNSS TBeomdypwereaitguhrte((g)C) 31 NY22-1 TN42071 38YB23323-1.4 208 GLARfhNIoPiDnreSdIllNTiimmRGbEbFNOGOTTHSP(KLGA)YJ(cm) J 011..091.539 J Values represent the mean of 2 trials Animal number 32 33 34 NN22-- NN22-- NN22-- TN140813 TN140728 T2HN9430U1 236-11 38YB3.5 4336-1 38B-.2 221 196 38YB332361-.6 208 118...01550 007..95.454 111.1.21.276 35 NN22-TN149026 38YB33233-1.3 207 111..120.269 : 116 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 2 Females OBSERVATIONS 36 IN THE HAND RHeamndolviningg 22 VSaolciddvaeealggitsrriieeoneeng N Y2- IN THE ARENA RAAercoatiurvisniatgyl count count 426 BUGorailinutes pcroeusnetnt TN01 MAATNopIupPcrUohaLcAhTIONS 36 RTStaiagitvrlhutopltreicinnnasgclihsreefslex 431 Y- degree 2 PTuempipl reeraflteuxre (C) 38B.8 Bodyweight (g) 188 GRIP STRENGTH (KG)} hfoinredlliimmbb 01..9135 LANDING FOOTSPLAY (cm) { 9.2 Values represent the mean of 2 trials 37 Animal number 38 39 YY2211 T1HN4609U1 23301 Y2 38B.2 220 117...00086 NY321 - TN4206l 33361 Y2 B 38.2 204 119...00488 N22 N- - T2N140222 33051 Y2 38B.6 209 119...21824 40 NY221 - TN140512 33351 Y2 B 38.6 202 019...72739 : 117 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 3 Females 26 IONBVRHTSSaHEeoalmncRddiEvadeeoVallHggivitAsrrniiAieeongnTeenNggIODNS NN22-- IMNAARBURPAATTBTGTSNuteaoHeoiocrrpaagIpiomiaultdiptvrdnlEPuihtruuitcyvroeleppilUstsrhonewAiigceirnatnpacnLgearrylseRcgroecaflAiechilecuhtEgsresuoToneehxNeruuftsItnelnOAnet(t(gtxN)CS) TN140852 318YB33338-11.99 GRhfIoPinredSllTiimmRbbENGTH (KG) J L%ANDINVGaluFeOsOreTpSrePsLeAntYth(ecmm)eatn o f 2 005..96.050 trials 27 Anim2a8l number 29 NN22-- NN22-- TN44081 T2HN4081U1 3333-137B-.8 213 318YB33322281.20 116...21617 118...11809 NN22-TN140552 37YB3330-11.7 214 119...11253 30 NN22-T2HN9405U1 38YB3366-11.3 199 016..7.0228 : 118 : MIN 252/004312 APPENDIX 4 (Functional observational battery - continued) Week 4 Group 4 Females 21 OBSERVATIONS IN THE HAND RHeamndolviningg 22 Salivation N Vocdaelgisrieneg N- degree IN THE ARENA - Activity count 9 Arousal 4 URBGoeraiailntureisnpcgroecusoneutnntt T3HN40U1 MANIPULATIONS ATRTStaoipagiutpvdrlhuctroepltrhoegciinnanrascegclihehsreesflex 33231 Y2- Pupil reflex B TBeomdypwereaitguhrte((g)C) 3188.02 GRIP STRENGTH (KG) % fhoinredlliimmbb 01..8033 LANDING FOOTSPLAY (cm) t 7.4 Values represent the mean of 2 trials 22 Anim2a3l number 24 N22 N- - T2HN4406U1 33231 Y2 B 38.2 178 006..89.519 N22 N- - TN42091 23231 YR1 37.7 223 11..1290 7.4 N22 N- - TN140513 63331 - B38.7 209 114...20639 25 N22 N- - TN140812 30351 YB2 37.9 202 01..9076 8.1 : 119 : MIN 252/004312 APPENDIX 5 Total time spent in locomotor activity - individual values (seconds) Animal Predose Group 1M (Control) 16 17 18 19 20 741 234 709 635 705 Group 2M (Low dose) 67 446922 8 545 9 550 10 180 Group 3M (Inter, dose) 11 587 12 878 1134 269 538 15 674 Group 4M (High dose) 1 522 2 426 3 1198 4 987 5 721 Week 4 486 426 1073 888 985 479258 545 1072 163 321 655 148 250 315 639 864 649 517 381 : 120 : APPENDIX 5 (Total time spent in locomotor activity - continued) Animal Predose Group IF (Control) 31 32 33 34 35 401 786 674 273 437 Group 2F (Low dose) 36 37 38 39 40 872 418 603 727 810 Group 3F (Inter, dose) 26 876 27 483 28 1000 29 30 935715 Group 4F (High dose) 2212 23 24 25 568 280 448 330 699 Week 4 681 552 550 229 410 617 915 1172 631 277 937 446 797 917 297 789 516 672 2 806 MIN 252/004312 : 121 : MIN 252/004312 APPENDIX 6 Additional comments from functional observational battery - individual observations Pre-dose Group 1 Sax M No additional comments : 122 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 2 Sex M 7 (7 ) Slight brown nasal staining In the arena: Scratching 8 {8 ) Touch response: Rears : 123 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 3 Sex M 11 (11 ) Slight brown nasal staining 12 (12 ) Touch response: Rears 14 (14 ) Slight brown nasal staining 15 (15 ) During manipulations: Soft pale faeces : 124 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 4 Sex M 4 (4 ) Slight brown nasal staining 5 (5 ) Temperature: Slight body tremors : 125 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Pre-dose Group 1 Sex F 31 (31 ) Slight brown nasal staining : 126 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 2 Sex F 36 (36 ) During manipulations: Vocalising moderately Landing footsplay: Awkward to handle 37 (37 ) Slight brown nasal staining 40 (40 ) In the arena: Scratching : 127 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 3 Sex F 27 (27 ) Touch response: Rears 30 (30 ) Slight brown nasal staining : 128 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Pre-dose Group 4 Sex F 21 (21 ) Slight brown nasal staining 22 (22 ) In the arena: Scratching : 129 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 1 Sex M 16 (16 ) In the arena: Outside digit right hindlimb curled under 19 (19 ) Slight brown nasal staining : 130 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 1 Group 2 Sex M 6 (6 ) Slight brown nasal staining 8 (8 ) Slight brown nasal staining 10 (10 ) Slight brown nasal staining Scab left forepaw : 131 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 3 Sex M 12 (12 ) Slight brown nasal staining 15 (15 ) Slight brown nasal staining : 132 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 4 Sex M 4 (4 ) Slight brown nasal staining Slight hair loss forelimbs : 133 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 1 Sex F No additional comments : 134 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 2 Sex F 37 (37 ) Slight brown nasal staining 40 (40 ) Slight brown staining head and ears In the arena: Climbed on front edge of arena : 135 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 3 Sex F 27 (27 ) Slight hair loss face 29 (29 ) Slight hair loss neck In the arena: Climbed on front edge of arena : 136 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 1 Group 4 Sex F 21 (21 ) Slight brown nasal staining 22 (22 ) In the arena: Scratching 23 (23 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 25 (25 ) Lower teeth pale In the arena: Climbed on front edge of arena : 137 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 2 Group 1 Sex M 18 (18 ) Slight brown nasal staining : 138 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 2 Group 2 Sex M 8 (8 ) Slight brown nasal staining 9 (9 ) Slight brown nasal staining 10 (10 ) In the arena: Climbed on front edge of arena : 139 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 2 Group 3 Sex M 11 (11 ) Slight brown nasal staining 12 (12 ) In the arena: Climbed on front edge of arena 14 (14 ) Slight brown nasal staining : 140 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 2 Group 4 Sex M 2 (2 ) Slight brown nasal staining 3 (3 ) Slight brown staining head and neck 4 (4 ) Slight brown nasal staining and slight brown staining head and neck In the arena: Climbed on front edge of arena 5 (5 ) Slight brown nasal staining and slight brown staining head and neck : 141 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 2 Group 1 Sex F 31 (31 ) In the arena: Climbed on front edge of arena : 142 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 2 Group 2 Sex F 36 (36 ) In the arena: Climbed on front edge of arena 37 (37 ) In the arena: Climbed on front edge of arena 38 (38 ) In the arena: Climbed on front edge of arena 39 (39 ) Tip of tail missing In the arena: Climbed and walked on front edge of arena 40 (40 ) In the arena: Climbed on front edge of arena : 143 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 2 Group 3 Sex F 27 (27 ) Slight hair loss head In the arena: Climbed on front edge of arena 28 (28 ) In the arena: Climbed on front edge of arena 29 (29 ) Moderate hair loss head and neck In the arena: Scratching and climbed on front edge of arena 30 (30 ) Scabs right ear and slight hair loss neck : 144 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 2 Group 4 Sex F 21 (21 ) Slight brown nasal staining In the arena: Scratching, eyes slight to h closed on occasions and climbed on front edge of arena 22 (22 ) In the arena: Climbed on front edge of arena 23 (23 ) In the arena: Climbed and walked on front edge of arena 24 (24 ) Slight lack of grooming rump In the arena: Climbed on front edge of arena 25 (25 ) Eyes slight to H closed on occasions and lower teeth pale In the arena: Climbed on front edge of arena and jumped out of arena : 145 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 3 Group 1 Sex M 17 (17 ) Slight lack of grooming rump 20 (20 ) Slight lack of grooming rump : 146 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 3 Group 2 Sex M 6 (6 ) In the arena: Climbed on front edge of arena 7 (7 ) In the arena: Climbed on front edge of arena 8 (8 ) Moderate brown nasal staining Slight brown staining head, neck and muzzle 9 (9 ) Slight brown nasal staining Slight brown staining head and neck 10 (10 ) Slight brown nasal staining Slight lack of grooming rump In the arena: Climbed on front edge of arena : 147 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 3 Group 3 Sex M 12 (12 ) In the arena: Climbed on front edge of arena 13 (13 ) In the arena: Climbed on front edge of arena 14 (14 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 15 (15 ) In the arena: Climbed on front edge of arena : 148 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 3 Group 4 Sex M 1 (1 ) Slight brown staining neck In the arena: Hunched on occasions 2 (2 ) Slight brown staining neck In the arena: Eyes slightly closed on occasions 3 (3 ) Slight brown staining head and neck 4 (4 ) Moderate brown nasal staining Slight brown staining neck Slight lack of grooming rump 5 (5 ) Slight brown nasal staining Slight brown staining head and neck MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 3 Group 1 Sex F 31 (31 ) In the arena: Climbed on front edge of arena 33 (33 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 34 (34 ) In the arena: Climbed on front edge of arena : 150 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 3 Group 2 Sex F 36 (36 ) Moderate brown staining head, neck and ears Slight lack of grooming rump In the arena: Climbed and walked on front edge of arena 37 (37 ) Slight brown staining head and neck Slight hair loss neck In the arena: Climbed and walked on front edge of arena 38 (38 ) Slight brown nasal staining In the arena: Climbed and walked on front edge of arena 39 (39 ) Slight brown staining head and neck Tip of tail missing In the arena: Climbed on front edge of arena 40 (40 ) In the arena: Climbed on front edge of arena : 151 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 3 Group 3 Sex F 26 (26 ) In the arena: Climbed on front edge of arena 27 (27 ) In the arena: Climbed on front edge of arena 28 (28 ) Slight hair loss neck Slight brown staining ears In the arena: Climbed and walked on front edge of arena 29 (29 ) In the arena: Climbed on front edge of arena 30 (30 ) Slight hair loss and brown staining neck Slight brown nasal staining Small scab right ear In the arena: Climbed on front edge of arena : 152 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 3 Group 4 Sex F 21 (21 ) Slight lack of grooming rump 22 (22 ) Slight brown nasal staining Slight lack of grooming rump In the arena: Climbed on front edge of arena 23 (23 ) Slight hair loss dorsal Slight brown staining head In the arena: Climbed and walked on front edge of arena 24 (24 ) Slight lack of grooming rump Small nick right ear In the arena: Climbed on front edge of arena 25 (25 ) Lower teeth pale Slight lack of grooming rump Slight brown staining head Eyes slightly closed on occasions In the arena: Climbed on front edge of arena : 153 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 1 Sex M 16 (16 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 17 (17 ) Approach response: Bit probe 18 (18 ) Slight brown staining head 19 (19 ) In the arena: Climbed on front edge of arena : 154 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 2 Sex M 6 (6 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 7 (7 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 8 (8 ) Slight brown nasal staining Slight brown staining head and neck In the arena: Climbed on front edge of arena 9 (9 ) Slight brown staining head and neck : 155 : APPENDIX 6 MIN 252/004312 (Additional comments from functional observational battery - continued) Week 4 Group 3 Sex M 11 (11 ) Slight brown nasal staining In the arena: Climbed on front edge of arena 12 (12 ) Slight brown staining head and neck In the arena: Climbed on front edge of arena 13 (13 ) In the arena: Climbed on front edge of arena 14 (14 ) In the arena: Climbed on front edge of arena 15 (15 ) Slight brown nasal staining Moderate brown staining head and neck In the arena: Climbed and sat on front edge of arena : 156 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 4 Sex M 1 (1 ) Slight brown staining neck Slight brown nasal staining In the arena: Climbed on front edge of arena 2 (2 ) Moderate brown staining head and neck In the arena: Scratching Touch response: Rears 3 (3 ) Approach response : Bit probe 4 (4 ) In the arena: Climbed on front edge of arena and scratching 5 (5 ) Moderate brown staining head and neck Slight lack of grooming rump : 157 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 1 Sex F 31 (31 ) Slight brown staining ears In the arena: Climbed on front edge of arena During manipulations: Moderately vocalising throughout 32 (32 ) Slight brown staining ears and head 33 (33 ) In the arena: Climbed on front edge of arena 34 (34 ) Slight brown staining ears and head Slight lack of grooming rump In the arena: Climbed on front edge of arena : 158 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 2 Sex F 36 (36 ) Moderate brown staining head and neck In the arena: Climbed and walked on front edge of arena During manipulations: Loud/aggressively vocalising throughout and awkward to handle 37 (37 ) Moderate brown staining head and neck In the arena: Climbed and walked on front edge of arena Approach response: Attempted to bite probe Touch response: Rears 38 (38 ) Slight brown staining head and neck In the arena: Climbed and walked on front edge of arena Temperature: Moderately vocalising 39 (39 ) Slight lack of grooming rump Tip of tail missing Slight brown staining head and neck In the arena: Climbed and walked on front edge of arena Touch response: Rears 40 (40 ) Slight lack of grooming rump In the arena: Climbed and walked on front edge of arena During manipulations: Moderately vocalising throughout : 159 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 3 Sex F 26 (26 ) In the arena: Climbed on front edge of arena 27 (27 ) Slight hair loss right hindlimb Slight brown staining head and neck In the arena: Climbed and walked on front edge of arena 28 (28 ) Slight brown nasal staining Slight hair loss head In the arena: Climbed and walked on front edge of arena Pupil reflex: Right eye pupil dilated under light at 1st attempt 29 (29 ) In the arena: Climbed on front edge of arena Touch response: Rears 30 (30 ) Slight lack of grooming rump Slight brown staining head Small nick out of Right ear In the arena: Climbed on front edge of arena : 160 : MIN 252/004312 APPENDIX 6 (Additional comments from functional observational battery - continued) Week 4 Group 4 Sex F 21 (21 ) In the arena: Eyes slight to H closed on occasions and climbed on front edge of arena 22 (22 ) Slight brown nasal staining Slight brown staining head In the arena: Climbed and walked on front edge of arena 23 (23 ) Slight brown staining head and neck Slight lack of grooming rump In the arena: Climbed and walked on front edge of arena Pupil reflex: Left pupil dilated under light on first attempt, on 2nd attempt constricted as normal 24 (24 ) Moderate lack of grooming rump Small nick out of right ear 25 (25 ) Moderate lack of grooming rump Lower teeth pale In the arena: Climbed on front edge of arena Approach response: Bit Probe Touch response: Rears : 161 : APPENDIX 7 Haematology - individual values Group Animal Bet L/L IN 16 0.488 17 0.477 18 0.459 19 0.462 20 0.471 2M 6 0.452 7 0.471 8 0.434 9 0.459 10 0.489 3M 11 0.483 12 0.453 13 0.483 14 0.455 15 0.424 4M 1 0.463 2 0.453 3 0.434 4 CTD 5 0.450 CTD Clotted sample Hb g/dD 17.2 16.8 16.1 16.0 16.1 16.1 16.4 15.4 16.4 17.3 16.8 16.3 16.9 16.0 14.8 16.3 16.5 15.3 CTD 16.1 BBC Retie X10-12/L 8.23 8.59 7.93 8.31 8.25 3.06 2.37 3.16 3.44 3.09 8.00 8.37 7.92 8.19 8.63 2.47 2.49 2.00 2.38 2.17 8.64 7.85 7.90 8.23 7.60 2.68 2.05 2.52 2.80 1.63 8.65 8.27 7.64 CTD 8.04 2.38 1.83 3.40 CTD 2.58 MCH P3 21.0 19.6 20.3 19.2 19.5 20.2 19.6 19.4 20.0 20.0 19.4 20.8 21.4 19.5 19.4 18.9 19.9 20.0 CTD 20.0 MCHC g/dL 35.3 35.3 35.1 34.6 34.1 35.7 34.9 35.3 35.7 35.3 34.7 36.0 35.1 35.2 34.8 35.2 36.4 35.3 CTD 35.7 MCV CL 59.3 55.5 57.9 55.6 57.0 56.5 56.3 54.8 56.1 56.7 55.9 S7.7 61.2 55.3 55.8 53.5 54.8 56.8 CTD 56.0 BBC Neutr ophil X10-9/L X10-9/L 21.75 16.51 11.16 15.68 19.11 1.62 1.08 1.49 2.13 2.72 16.21 14.04 11.45 13.77 18.88 1.67 2.14 0.97 1.63 4.26 12.01 19.98 16.54 13.28 12.59 1.73 1.61 1.35 0.91 1.00 16.93 14.59 18.58 CTD 14.44 2.12 2.92 2.24 CTO 1.59 MIN 252/004312 Group 1M 2M 3M 4M CTD Animal Lymph ocyte xlO-9/L 16 19 .41 17 14 .81 18 9,.26 19 13 .16 20 15 .67 6 13..99 7 11..25 8 10 .06 9 11 .71 10 13..99 11 10..04 12 17..80 13 14.,75 14 12..05 15 11..24 1 14..27 2 11..08 3 15,,57 4 CTD 5 12..13 Clotted sample APPENDIX 7 (Haematology - continued) Bob in ophil xlO-9/Ii Baao phil X10-9/L Mono cyte xlO-9/li LUC X10-9/L Pit X10-9/L PT BBC 0.26 0.15 0.17 0.14 0.10 0.13 0.06 0.03 0.05 0.08 0.21 0.20 0.12 0.10 0.28 0.13 0.21 0.10 0.08 0.26 654 1161 1091 1241 627 12.5 13.8 14.4 12.0 12.4 0.08 0.14 0.09 0.15 0.17 0.07 0.05 0.03 0.05 0.07 0.19 0.22 0.14 0.11 0.20 0.20 0.23 0.15 0.13 0.20 1026 1040 1221 1126 1167 12.9 12.6 13.0 13.3 12.8 0.08 0.09 0.08 0.08 0.11 0.04 0.10 0.0B 0.06 0.04 0.06 0.15 0.17 0.08 0.12 0.06 0.22 0.11 0.10 0.08 1028 1305 1013 1187 1085 13.4 13.5 14.5 12.8 13.5 0.07 0.11 0.08 CTD 0.13 0.06 0.06 0.09 CIO 0.05 0.22 0.21 0.31 CTD 0.29 0.20 0.21 0.29 CTD 0.26 954 773 1360 CTD 1004 13.7 13.5 13.9 11.9 13.5 APTT sac 15.7 20.5 19.8 18.2 19.2 13.3 18.4 19.1 20.9 18.9 18.8 18.2 19.3 20.6 20.5 19.6 19.7 16.1 13.3 15.1 MIN 252/004312 APPENDIX 7 (Haematology - continued) MEN 252/004312 164 : Group 1M 2M 3M 4M CTD Animal Aniso cyto Micro cyto Macro cyto Hypo chrom Hyper rfvrnm 16 17 18 19 20 6 7 8 9 10 11 12 13 14 15 1 2 3 4 CTD CTD CTD CTD CTD 5 Clotted sample Group IP 2F 3F 4F Animal 31 33 33 34 35 36 37 38 39 40 26 27 28 29 30 21 22 23 24 25 Bet I*/X* 0.455 0.417 0.432 0.441 0.418 0.396 0.414 0.415 0.408 0.412 0.387 0.408 0.386 0.412 0.412 0.390 0.392 0.417 0.441 0.414 Hb g/dL 15.6 14.8 15.2 IS.8 15.1 14.0 15.0 14.6 14.3 14.8 13.9 14.7 13.8 14.8 14.7 14.6 14.2 14.7 15.9 14.9 APPENDIX 7 (Haematology - continued) RBC Retie xlO-12/X. 8.00 7.53 7.77 7.96 7.29 2.51 2.80 2.87 2.14 2.26 7.29 7.25 7.66 7.35 7.38 1.95 1.94 2.08 2.33 1.87. 6.94 7.25 7.22 7.29 7.51 2.52 2.05 2.83 1.88 1.56 7.28 7.51 7.20 8.03 7.34 1.06 2.59 2.80 2.02 2.63 MCH P9 19.6 19.6 19.5 19.9 20.7 19.3 20.7 19.1 19.4 20.1 20.0 20.3 19.1 20.4 19.6 20.1 18.9 20.4 19.7 20.3 MCHC S/dL 34.4 35.5 35.1 35.8 36.1 35.4 36.3 35.2 35.0 36.0 35.8 36.0 35.8 36.0 35.7 37.5 36.1 35.2 36.0 36.0 MCV L 56.9 55.4 55.6 55.5 57.3 54.4 57.1 54.2 55.6 55.8 55.8 56.3 53.5 56.6 54.B 53.6 52.2 57.9 54.9 56.4 HBC Neutr ophil X10-9/L X10-9/L 9.91 9.90 13.13 7.17 10.51 1.13 1.47 0.88 0.56 0.72 6.12 13.33 6.66 6.47 6.18 0.75 1.22 0.90 0.66 0.66 6.21 4.21 4.79 7.31 5.99 1.39 0.70 0.76 0.89 0.67 6.48 7.03 8.73 9.24 7.34 1.18 0.81 2.18 1.05 1.04 MIN 252/004312 166 : Group 17 27 37 47 Animal 31 32 33 34 35 36 37 38 39 40 26 27 28 29 30 21 22 23 24 25 Lymph ocyte X10-9/L 8.20 7.94 11.72 6.35 9.21 5.08 11.63 5.49 5.63 5.26 4.66 3.35 3.79 6.14 5.14 4.96 6.02 6.31 7.98 6.08 APPENDIX 7 (Haematology - continued) Bosin ophil X10-9/L Baso phil xlO-9/Ii Mono cyte xlO-9/L LUC X10-9/1, Pit X10-9/L PT sec 0.27 0.21 0.12 0.08 0.21 0.03 0.02 0.05 0.02 0.03 0.09 0.15 0.17 0.08 0.20 0.19 0.11 0.18 0.07 0.13 1046 995 1262 838 1216 13.0 13.5 14.1 13.7 14.2 0.10 0.12 0.13 0.09 0.08 0.01 0.03 0.01 0.01 0.01 0.10 0.18 0.05 0.05 0.10 0.08 0.14 0.08 0.04 0.07 1095 1183 933 1008 1027 14.4 13.3 14.1 13.8 13.3 0.08 0.06 0.13 0.11 0.09 0.01 0.00 0.01 0.02 0.01 0.05 0.07 0.06 0.11 0.05 0.03 0.03 0.04 0.04 0.04 916 1162 886 1083 776 13.3 13.9 11.1 14.5 14.8 0.15 0.08 0.06 0.07 0.08 0.02 0.02 0.01 0.03 0.01 0.10 0.05 0.08 0.05 0.07 0.08 0.05 0.09 0.06 0.06 972 1059 1036 1194 1113 14.2 14.4 14.3 14.5 14.4 APTT sec 16.1 18.7 17.7 15.3 17.9 18.1 17.1 19.0 16.9 12.8 11.1 14.6 13.0 17.5 15.3 14.4 17.2 18.1 15.4 16.5 MIN 252/004312 Group IF 2F 3F 4F Animal 31 32 33 34 35 36 31 38 39 40 26 27 28 29 30 2221 23 24 25 APPENDIX 7 (Haematology - continued) Aniso cyto Micro cyto Macro cyto Hypo chrom Hyper chrom MIN 252/004312 APPENDIX 8 Blood chemistry - individual values Group 1M 2M 3M 4M Animal Aik. ALT AST Bill. Uraa Crt Glue Choi Trig Phos Total Total U/L U/L U/L umol/L nmol/L umol/L nmol/L nmol/L mmol/L 16 370 41 75 3 4.95 43 7.42 1.64 0.92 17 335 45 82 2 6.00 43 6.36 1.35 0.90 18 462 48 79 2 6.58 43 7.90 1.73 0.54 19 612 39 78 3 5.02 46 7.48 1.32 0.82 20 400 49 114i 2 6.19 41 7.86 1.44 0.37 6 498 41 79 2 6.05 42 7.88 2.01 0.88 7 406 47 79 2 4.60 38 7.28 1.86 0.92 8 396 43 70 2 5.87 41 6.94 1.67 0.50 9 518 37 77 1 5.44 40 6.31 1.38 0.77 10 416 45 68 1 5.90 44 7.67 1.54 0.96 11 534 41 71 1 5.39 39 6.85 1.52 0.46 12 466 36 71 1 4.91 43 5.83 1.36 0.59 13 457 46 81 1 6.13 40 6.12 1.49 0.70 14 386 32 66 2 4.91 42 7.99 1.49 0.42 15 385 36 73 2 4.78 42 7.28 1.06 0.35 1 389 39 88 2 6.69 45 6.86 0.99 0.26 2 379 42 87 2 5.39 42 6.70 1.36 0.46 3 337 52 100 2 5.06 42 5.84 1.31 0.84 4 430 39 95 3 5.89 51 8.18 1.08 0.66 5 372 39 99 1 5.84 40 7.77 1.05 0.81 MIN 252/004312 APPENDIX 8 (Blood chemistry - continued) MIN 252/004312 169 : Group 1M 2M 3H 4M Animal Na K Cl Ca Phos Total Alb al a2 Total Prot Glob Glob mmol/L mmol/L mol/L mmol/L mmol/L g/L g/L g/L g/L 16 142 3.4 101 2.78 2.43 63 32 13 4 17 139 4.2 99 2.75 2.45 64 32 13 4 18 140 4.2 100 2.81 2.36 65 30 15 4 19 139 4.0 99 2.79 2.34 65 33 14 4 20 138 5.2 101 2.59 2.25 64 30 15 4 6 139 3.9 100 2.56 2.23 60 32 11 3 7 139 4.0 101 2.63 2.31 63 31 11 4 8 141 3.7 102 2.52 2.02 59 32 11 3 9 141 3.9 101 2.70 2.15 62 31 13 4 10 140 3.5 99 2.72 . 2.09 64 32 11 5 11 139 4.1 100 2.66 2.48 62 32 13 4 12 140 3.9 100 2.68 2.45 60 30 12 4 13 140 4.1 100 2.69 2.94 61 32 11 4 14 138 3.9 98 2.56 2.11 59 29 13 4 15 139 3.7 101 2.54 2.17 61 29 12 4 1 141 3.7 102 2.45 2.20 60 32 11 3 2 140 3.6 101 2.61 2.18 63 34 11 3 3 139 3.7 101 2.63 2.35 60 30 12 3 4 139 4.2 101 2.56 2.26 59 32 13 3 5 139 3.5 102 2.64 2.20 64 34 12 3 APPENDIX 8 (Blood chemistry - continued) G roup 1M 2H 3M 4M Animal 16 17 18 IS 20 6 7 8 9 10 11 12 13 14 15 1 2 3 4 S Beta Slob g/L 12 12 13 12 12 11 14 11 12 13 1 12 11 11 13 11 11 12 10 12 Samoa Glob g/x. 2 3 3 2 3 2 4 2 2 3 2 3 3 2 3 3 3 3 2 3 A/G Ratio 1.03 1.00 0.86 1.03 0.88 1.14 0.97 1.19 1.00 1.00 1.07 1.00 1.10 0.97 0.91 1.14 1.17 1.00 1.19 1.13 MIN 252/004312 APPENDIX 8 (Blood chemistry - continued) Group 1M 2M 3H 4M Animal 16 17 IS 19 20 6 7 8 9 10 11 12 13 14 15 1 2 3 4 5 Alb % 51.5 50.1 46.1 51.1 46.8 53.5 48.8 53.9 49.4 50.0 51.8 49.8 52.3 49.5 47.1 53.6 53.4 50.1 53.8 53.7 al Glob 20.8 20.3 23.0 21.1 22.9 17.9 17.8 19.1 20.7 17.5 20.6 19.8 17.6 22.1 20.0 18.5 17.9 20.1 21.3 18.1 a2 Glob * 6.1 6.0 6.8 6.1 6.3 S.8 5.8 5.4 6.4 7.4 7.2 6.8 7.0 7.0 6.8 4.7 5.4 4.8 5.1 5.1 Beta Glob % 18.5 18.4 20.0 18.6 18.9 19.0 22.0 18.1 20.1 20.1 16.8 19.2 17.6 18.0 20.9 17.9 17.8 20.8 16.7 18.0 Gamma %Glob 3.0 5.1 4.1 3.1 5.1 3.7 5.6 3.6 3.4 5.0 3.7 4.4 5.6 3.3 5.1 5.2 5.5 4.2 3.1 5.0 MIN 252/004312 APPENDIX 8 (Blood chemistry - continued) Group IF 2F 3F 4F Animal Aik. ALT AST Bill. Urea Creat Glue Choi Trig Phos Total Total U/L U/L U/L umol/L tnnol/L umol/L nmol/L mmol/L naol/L 31 201 42 78 2 6.23 37 6.69 2.55 0.77 33 258 42 87 3 7.93 47 7.71 1.96 0.46 33 287 35 81 4 7.73 47 6.33 2.26 0.43 34 203 34 79 2 5.94 44 7.32 1.41 0.30 35 238 37 79 2 7.92 45 5.88 2.26 0.69 36 287 32 76 2 5.41 48 7.72 2.04 0.42 37 167 39 63 2 6.50 45 7.03 1.71 0.94 38 167 38 89 2 7.56 50 6.76 2.04 0.31 39 240 53 97 1 6.16 42 7.78 1.78 0.44 40 215 43 79 1. 6.96 48 7.95 1.97 0.37 26 246 31 83 2 5.74 43 6.76 1.61 0.44 27 230 27 71 2 5.13 43 6.03 1.39 0.45 28 320 36 100. 2 5.48 42 5.76 1.53 0.68 29 249 41 86 2 6.26 47 6.08 1.58 0.37 30 286 32 77 1 6.10 47 8.34 1.51 0.36 21 241 31 65 2 4.64 41 6.71 1.70 1.05 22 201 28 83 1 7.57 43 6.48 1.28 0.35 23 233 37 74 2 6.49 43 6.49 1.36 0.67 24 305 27 75 2 5.83 42 7.53 0.82 0.21 25 218 40 85 2 6.41 44 7.92 0.97 0.29 MIN 252/004312 APPENDIX 8 (Blood chemistry - continued) Group IF 2F 3F 4F Animal Na K Cl Ca Phos Total Alb al a2 Total Frot Glob Glob mmol/L maol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L 31 139 3.7 100 2.69 2.05 65 35 10 5 32 140 3.7 99 2.65 1.96 66 34 11 4 33 140 3.9 101 2.61 2.05 66 35 11 4 34 141 3.7 102 2.58 1.63 62 33 10 4 35 139 3.6 101 2.79 1.88 63 35 11 4 36 140 3.5 101 2.54' 1.51 64 32 11 5 37 140 3.7 99 2.79 1.95 68 39 12 4 38 140 3.8 102 2.58 1.92 61 32 11 4 39 140 3.3 102 2.42 1.47 63 35 11 4 40 138 3.4 99 2.62 1.81 65 36 11 4 26 140 3.5 103 2.45 1.49 55< 30 9 3 27 140 3.3 101 2.61 1.71 66 36 11 4 28 139 3.9 101 2.44 1.98 53 29 9 3 29 138 3.8 99 2.58 1.85 62 34 11 4 30 139 3.2 101 2.44 1.57 61 34 10 4 21 136 3.7 98 2.59 1.74 64 36 13 4 22 138 3.4 102 2.60 1.84 57 32 10 4 23 137 3.7 98 2.49 1.95 62 33 10 3 24 139 3.2 101 2.40 1.86 56 30 9 3 25 137 3.2 99 2.49 1.60 60 32 10 3 MIN 252/004312 Qroup IF -4*J. 2F 3F 4F APPENDIX 8 (Blood chemistry - continued) Animal 31 32 33 34 35 36 37 38 39 40 26 27 28 29 30 21 22 23 24 25 Beta Glob g/L 12 13 11 11 11 13 10 12 11 12 11 12 9 10 10 9 10 12 10 12 Ganna Glob g/L 3 4 4 3 3 3 2 3 3 3 2 4 2 3 3 3 2 4 3 4 A/G Ratio 1.17 1.06 1.13 1.14 1.25 1.00 1.34 1.10 1.25 1.24 1.20 1.20 1.21 1.21 1.26 1.29 1.28 1.14 1.15 1.14 MIN 252/004312 APPENDIX 8 (Blood chemistry - continued) Group IP 2F 3F 4F Animal 31 32 33 34 35 36 37 38 39 40 26 27 28 29 30 21 22 23 24 25 Alb \ 54.2 51.4 53.3 53.2 55.4 50.0 57.8 53.0 55.4 55.3 53.7 53.9 54.7 54.8 55.4 55.6 55.4 53.2 54.4 53.4 al Glob % 15.4 17.2 17.1 16.8 17.0 17.8 17.9 17.6 16.8 16.2 16.5 16.7 17.2 17.7 16.4 19.7 16.9 15.6 15.4 16.0 a2 Glob 6 7.0 6.S 6.2 6.1 5.8 7.1 5.9 6.1 6.0 6.1 6.1 6.2 6.4 6.1 6.1 5.9 6.2 5.6 5.7 5.1 Beta Glob % 19.2 19.2 16.7 18.3 17.2 19.9 15.1 19.1 17.7 17.8 19.4 17.8 17.5 16.9 16.7 14.7 17.9 18.8 18.5 19.3 Gannul Glob t 4.2 5.7 6.6 5.6 4.5 5.3 3.4 4.1 4.2 4.5 4.3 5.4 4.2 4.5 5.4 4.0 3.6 6.9 6.0 6.2 MIN 252/004312 APPENDIX 9 Absolute organ weights - individual values (g) Group ! Compound s Exposure level (ppm) t 1 Control 0 23 - T-7499 50 150 4 450 GROUP IN ANIMAL 16 17 18 19 20 TERMINAL BODY NT (9 ) 397.8 371.8 396.8 379.7 383.8 ADRENALS EFXDZDYMID 0.054 0.065 0.048 0.054 0.049 1.033 0.948 0.966 0.967 0.976 HEART 1.208 1.072 1.299 1.447 1.499 KIDNEYS 2.47 2.47 2.48 2.37 2.39 LIVER 14.00 14.36 14.62 14.20 16.69 LUNGS 1.254 1.217 1.167 1.310 1.347 Print No: 0024 Printed* 05-MAR-01 Xybion protocol number! MIN 252 TESTES 3.57 3.18 3.34 3.00 3.45 MIN 252/004312 176 2M 6 7 8 9 10 375.3 394.5 296.4 356.7 365.7 0.062 0.054 0.053 0.064 0.062 1.048 1.069 0.887 0.991 0.867 1.825 1.603 1.272 1.347 1.698 2.41 2.67 1.94 2.27 2.64 14.77 14.01 9.85 12.18 14.53 1.482 1.279 1.056 1.345 1.370 3.96 4.13 3.04 3.49 2.79 3M 11 12 13 14 15 341.2 360.0 395.4 357.8 336.6 0.063 0.060 0.060 0.055 0.054 0.813 0.920 0.977 0.868 1.020 1.088 1.403 1.574 1.170 1.148 2.24 2.21 2.64 2.22 2.45 12.15 12.00 13.55 11.39 13.07 1.115 1.188 1.334 1.333 1.271 3.19 3.42 3.36 3.25 3.24 4N 1 2 3 4 5 325.2 299.6 385.6 340.5 307.6 0.060 0.052 0.068 0.038 0.054 0.912 0.886 0.860 0.856 1.006 1.115 0.913 1.371 1.044 1.089 2.09 1.89 2.61 2.03 2.48 12.55 10.49 14.78 11.49 14.16 1.225 1.101 1.228 1.149 1.198 3.63 3.18 3.20 3.01 3.52 Epididyaid > Epididyaid APPENDIX 9 (Absolute organ weights - continued) Group Compound Exposure level(ppm): 1 Control 0 2 34 -- T-7499 - 50 150 450 GROUP IF ANIMAL 31 32 33 34 35 TERMINAL BODY WT (g) 211.1 234.3 198.8 212.0 217.7 ADRENALS 0.046 0.071 0.054 0.052 0.061 HEART 0.927 1.142 0.704 0.995 0.850 KIDNEYS 1.45 1.51 1.14 1.34 1.43 2F 36 37 38 39 40 188.0 224.4 211.0 211.7 219.1 0.063 0.071 0.082 0.067 0.062 0.825 0.868 0.966 1.036 0.797 1.53 1.61 1.37 1.40 1.50 3F 26 27 28 29 30 200.8 213.4 191.5 220.0 207.9 0.052 0.059 0.048 0.054 0.050 0.763 0.797 0.956 1.070 0.871 1.32 1.59 1.40 1.41 2.23 4F 21 22 23 24 25 194.2 177.8 222.1 220.2 210.5 0.075 0.059 0.064 0.084 0.064 0.904 0.722 0.721 1.003 0.849 1.52 1.45 1.49 1.54 2.31 LIVER 7.89 8.78 6.52 7.10 8.75 7.09 8.51 7.72 7.53 7.52 6.94 7.59 6.67 7.77 7.72 8.19 6.91 7.14 7.99 7.58 LUNGS 0.930 1.004 0.931 1.149 0.978 0.999 0.957 0.937 0.961 0.838 0.866 0.958 0.900 1.038 0.970 1.001 0.843 0.958 0.995 0.948 Print Ho: 0025 Printed: 05-MMI-01 Xybion protocol nuabar: MIN 252 MIN 252/004312 APPENDIX 10 Individual pathological findings MIN 252/004312 tToophieaniirononiutsitaionl feebxpoaetmehripnraaettvhioioenlwowgbiasystsu.ansdeecrotankdenpabthyotlhoegissttu.dyTphaethdoialoggniosst,etshreerpeosruteltds ohefrwehriecphrewseernet tthheencsounbsjeencsteuds Study pathologist: Peer review: DCShoeepniasluratlmEta.enBnttePgoagfth,PoMaltoh.Agoil.s,ot,gVye.t.M.B., M.R.C.V.S., F.R.C.Path., DDCoeapvnasidurtlJmt.aeLnntetPwoaitfsh,PoPalthoh.gDoils.ot,,gFy..R.C.Path., 178 APPENDIX 10 (Individual pathological findings - continued) Print No: 0026 Group : Compound Exposure level(ppm): i Control 0 2 34 -- T-7499 - 50 150 450 Printod: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0016 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 1 SACRIFICEI STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: F A TH O LOGY s0 B S E R V A T I O N 397.8 GRAMS NECROPSY HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group Compound Exposure level(ppa): 1 Control 0 23 4 -- T-7499 50 150 50 Print No: 002S Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0017 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 371.8 GRAMS NECROPSY PATHOLOGY OBSERVATIONS BISTOPATBOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LUNGS & BRONCHI : -ALVEOLITIS,-MINIMAL, FOCAL -ALVEOLAR OSSEOUS METAPLASIA,-MINIMAL, FOCAL NASAL TURBINATES : -TRANSITIONAL EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL >NOTE;>LESIQN IN LATERAL HALL TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL, FOCAL *** ANIMAL HAS N O GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group : 123 4 Compound Control Exposure level(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol nuabar MIN 252 ANIMAL NUMBER: 0018 DATE OF DEATH: U-DEC-00 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT : 396.8 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEY8 : -UNILATERAL PELVIC DILATATION, -SLIGHT LUNGS 6 BRONCHI ; -ALVEOLITIS,-MINIMAL, FOCAL NASAL TURBINATES : -TRANSITIONAL EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL >NOTE:>LESIGN IN LATERAL WALL AND VENTRAL PART OF NASOTURBINATE TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL, MULTI-FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group 12 3 4 Compound Control Exposure lavai(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol nuaber: MIN 252 ANIMAL NUMBER: 0019 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 379.7 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HI8TOPATHOLOGY KIDNEYS : -MEDULLARY TUBULAR BASOPHILIA,-MINIMAL, FOCAL -TUBULAR HYPERPLASIA, SIMPLE,-SLIGHT, FOCAL LARYNX : -EPITHELIAL HYPERPLASIA - ARYTENOIDS,-SLIGHT -EPITHELIAL HYPERPLASIA - VENTRAL POUCH,-MODERATE -INFLAMMATORY CELLS IN LAMINA PROPRIA (VENTRAL AND VENTROLATERAL),-SLIGHT LUNGS BRONCHI : -ALVEOLAR OSSEOUS METAPLASIA,-MINIMAL, FOCAL TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL, FOCAL *** ANIMAL HAS N O GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Print No: 0026 Group Compound Exposure level(ppm) : 1 Control 0 23 4 -- T-7499 - 50 150 450 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0020 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 383.8 GRAMS NECROPSY P A T H O LOGY O B S E R V A T I O N S HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LN MANDIBULAR : -CYSTIC ENLARGEMENT; RIGHT, ONE, 101*1. IK MANDIBULAR : -CYSTIC SINUSES,-MODERATE NASAL TURBINATES : -RESPIRATORY EPITHELIAL HYPERPLASIA,-SLIGHT, FOCAL >NOTE:>LESION IN DORSAL MEATUS MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group 12 3 4 Compound Control Exposure level(ppm) : 0 50 150 450 Print Mo: 0026 Printad: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0006 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY MEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 375.3 GRAMS * * * ANIMAL HAS N O GROSS OBSERVATIONS RECORDED *** Group Compound : Exposure level(ppm): ANIMAL NUMBER: 0007 DATE OF DEATH: ll-DEC-00 APPENDIX 10 (Individual pathological findings - continued) 123 4 0 SO 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 394.5 GRAMS *** ANIMAL BAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group Cpound Exposura lavel(ppm): 1 Control 0 23 4 -- T-7499 - 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0008 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY MEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 296.4 GRAMS *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group : 12 3 4 -J Compound 00 Exposure level(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0009 DATE OF DEATH: U-DEC-00 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 356.7 GRAMS *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Grotq> 12 3 4 0000 Compound Exposure level(ppe): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0010 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 365.7 GRAMS *** ANIMAL BAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group Compound Exposure level(ppm) : ANIMAL NUMBER: 0011 DATE OF DEATH: ll-DEC-00 123 4 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol nuabar: MIN 252 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 341.2 GRAMS *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 Group : Compound Exposure level(ppa) : ANIMAL NUMBER: 0012 DATE OF DEATH: ll-DEC-00 APPENDIX 10 (Individual pathological findings - continued) 123 4 0 SO ISO 450 Print Ho: 0026 Printed: 05-MAR-01 Xybion protocol nuaber: MIN 252 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 360.0 GRAMS MIN 252/004312 190 : +** a n u a l h a s n o g r o s s o b s e r v a t i o n s r e c o r d e d *** APPENDIX 10 (Individual pathological findings - continued) Group 123 4 Compound Control Exposure levai (ppm) : 0 50 150 450 Print Mo: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0013 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 395.4 GRAMS *** ANIMAL HAS M O GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) MIN 252/004312 192 : Group 123 4 Compound Exposure level (ppm): 0 50 150 450 Print Mo: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0014 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 357.8 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HI8TOPATBOLOGY KIDNEYS : -PELVIC DILATION, MINIMAL; LEFT. KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL -DILATED MEDULLARY TUBULES CONTAINING EOSINOPHILIC COLLOID,MINIMAL, FOCAL -UNILATERAL PELVIC DILATATION, "MINIMAL APPENDIX 10 (Individual pathological findings - continued) Print Ho: 0026 Group 12 3 Compound Cuiitrol *"* Exposure level(ppm): 0 50 150 4 450 Printed: 05-MAR-01 Xybion protocol numbar: MIN 252 ANIMAL HUMBER: 0015 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 336.6 GRAMS *** ANIMAL HAS H O GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group : 123 4 Compound Control Exposure level (ppa) : 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol n m b e r : MIN 252 ANIMAL NUMBER: 0001 DATE OF DEATH: 11-DEC-00 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 325.2 SRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LN MANDIBULAR -ENLARGED ; LN MANDIBULAR : -INCREASED CELLULARITY - GENERALISED,-SLIGHT LN PANCREATIC : -ENLARGED, MINIMAL; LN PANCREATIC : -SINUS ERYTHROCYTOSIS/ERYTHROPHAGOCYTOSIS,-MINIMAL MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group : 123 4 Compound Control Exposure level(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number MIN 252 ANIMAL NUMBER: 0002 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE V DATE OF DEATH: ll-DEC-00 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 299.6 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LN MANDIBULAR : -CONGESTED, MINIMAL; RIGHT, ONE LN MANDIBULAR : -SINUS EFYTHROCYTOSIS/ERYTHROPHAGOCYTOSXS,-SLIGHT LUNGS BRONCHI : -DARK AREA(S); RIGHT ANTERIOR LOBE, ONE, SUBPLEURAL, 2*91 LUNGS fi BRONCHI : -SUBPLEURAL INFLAMMATION,-MINIMAL, FOCAL MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group : Compound Exposure level(ppm): i Control 0 23 4 -- T-7499 - 50 150 450 Print No: 0026 Printad: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0003 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE OSVO DATE OF DEATH: ll-DEC-00 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 385.6 GRAMS PATHOLOGY OBSERVATIONS NECROPSY BISTOPATHOLOGY SKIN : -SCAB(S) ; TAIL, A FEW, 1M*. (ON TATTOO) SKIN : -SCAB,-PRESENT -EPIDERMAL HYPERPLASIA,-MODERATE, FOCAL -EPIDERMAL ULCERATION,-MINIMAL, FOCAL -DERMAL INFLAMMATION, -SLIGHT, FOCAL STOMACH : -ANTRUM WHITE NODULE (S); MUCOSA, ONE, NEAR TO LIMITING RIDGE, D M . STOMACH : -ECTOPIC N0NGLANDULAR EPITHELIUM IN GLANDULAR MUCOSA, FOCAL,PRESENT MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group Compound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 50 150 450 Print Ho: 0026 Printed: 05-MAR-01 Xybion protocol number: MIH 2S2 ANIMAL NUMBER: 0004 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 340.5 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LARYNX : -EPITHELIAL HYPERPLASIA - ARYTENOIDS,-SLIGHT TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL, FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group 123 4 Compound Control Exposure lavai(ppn): 0 50 150 450 Print Mo: 0026 Printed; 05-MAR-01 Xybion protocol nunber: MIN 252 ANIMAL NUfflER: 0005 DATE OF DEATH: ll-DEC-00 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 307.6 GRAMS ** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group 123 4 Compound Exposure level(ppm) : 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 VO ANIMAI NUMBER: 0031 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE VO DATE OF DEATH: ll-DEC-00 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 211.1 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LIVER : -MEDIAN CLEFT PALE AREA(S) ; ONE, SUBCAPSULAR, 2 m . -LOBE(S) NECROTIC; RIGHT ANTERIOR LOBE. LIVER : -SIDEROPHAGES, BILE DUCT HYPERPLASIA, FIBROSIS AND MINERALISATION - RIGHT ANTERIOR LOBE,-MODERATE LUNGS i BRONCHI : -ALVEOLITIS,-MINIMAL, FOCAL -EXTRAVASATION OF EOSINOPHILS,-MINIMAL, FOCAL MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group : 123 4 Compound Control Exposure level(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0032 DATE OF DEATH: U-DEC-00 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 234.3 GRAMS NECROPSY PATHOLOGY OBSERVATION HISTOPATHOLOGY TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL, FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group Compound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 - 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol nunber: MIN 252 ANIMAL NUMBER: 0033 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY KEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 198.B GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOFATHOLOGY LUNGS BRONCHI : -ALVEOLITIS,-MINIMAL, FOCAL TRACHEA : -SUBEPITHELIAL INFLAMMATION,-MINIMAL , FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) MIN 252/004312 : 202 Group 123 4 Compound Control Exposure lavai(ppm): 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0034 DATE OF DEATH: ll-DEC-00 SEX; FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 212.0 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -PELVIC DILATION, MINIMAL; RIGHT KIDNEYS : -UNILATERAL PELVIC DILATATION,-MINIMAL APPENDIX 10 (Individual pathological findings - continued) Group : Compound Exposure level(ppm): 1 Control 0 23 4 - T-7499 - 50 150 450 Print Ho: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0035 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 217.7 GRAMS NECROPSY PATHOLOGY OBSERVATIONS BISTOPATBOLOGY LARYNX : -EPITHELIAL HYPERPLASIA - ARYTENOIDS,-SLIGHT MIN 252/004312 : 203 *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group 123 4 Conpound Control Exposure level (ppsO : 0 50 150 450 Print No: 0026 Printad: 05-MAR-01 Xybion protocol numbar: MIN 252 ANIMAL NUMBER: 0036 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 188.0 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY STOMACH : -ANTRUM RIDGE, W2HmIT. E NOODLE (S) ; MUCOSA, ONE, NEAR TO LIMITING STOMACH : -ECTOPIC NONGLANDOLAR EPITHELIUM IN GLANDULAR MUCOSA, FOCAL, PRESENT UTERUS : -FLUID DISTENTION UTERUS : -LUMINAL DILATATION, -SLIGHT MIN 252/004312 Group Compound Exposure level(ppm): ANIMAL NUMBER: 0037 DATE OF DEATH: ll-DEC-00 APPENDIX 10 (Individual pathological findings - continued) 12 3 4 Print No: 0026 Printed: 05-MAR-01 0 50 150 450 Xybion protocol nvaaber: MIN 252 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 224.4 GRAMS MIN 252/004312 : 205 >* ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 10 (Individual pathological findings - continued) Group : 123 4 K> Compound O0\ Exposure level (ppm): 0 50 150 450 Print No: 0026 Printad: 05-MAR-01 Xybion protocol nuabar: MIN 252 ANIMAL NUMBER: 0038 DATE Or DEATH: 11-DEC-00 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE 8TUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 211.0 GRAMS * ANIMAL BAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group : Compound : Exposure level(ppm) : i Control 0 23 4 - T-7499 - 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol nuaber: MIN 252 ANIMAL NUMBER: 0039 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 211.7 GRAMS *** ANIMAL BAS NO GROSS OBSERVATIONS RECORDED **' MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group : Compound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 - 50 ISO 450 Print No: 0026 Printod: 05-MAR-01 Xybion protocol nunbar: MIN 252 ANIMAL NUMBER: 0040 DATE OF DEATH: 11-DEC-00 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 219.1 GRAMS ** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) MIN 252/004312 209 Group : Compound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 - 50 150 450 Print Mo: 0026 Printed: 05-MKR-01 Xybion protocol munber: MIN 2S2 ANIMAL NUMBER: 0026 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 200.8 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LN MANDIBULAR : -ENLARGED, MINIMAL LN MANDIBULAR : -INCREASED CELLULARITY - GENERALISED,-SLIGHT APPENDIX 10 (Individual pathological findings - continued) MIN 252/004312 : 210 Group 123 4 Compound Control Exposure level (ppm) : 0 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0027 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 213.4 GRAMS NECROPSY PATHOLOGY OBSERVATIONS BISTOPATHOLOGY UTERUS : -FLUID DISTENTION UTERUS : 'LUMINAL DILATATION,-SLIGHT APPENDIX 10 (Individual pathological findings - continued) Group Compound Exposure level(ppm): i Control 0 23 4 -- T-7499 - 50 150 450 Print No: 0026 Printed: 05-MAR-01 Xybion protocol number: HIM 252 ANIMAL NUMBER: 002B DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY MEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 191,5 GRAMS NECROPSY PATHOLOGY OBSERVATIONS BISTOPATHOLOGY UTERUS : -FLUID DISTENTION UTERUS : -LUMINAL DILATATION,-SLIGHT MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) 212 Gro\$> Coopound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 50 150 450 Print N o : 0026 Printed: 05-MAR-01 Xybion protocol nunber: MIN 252 ANIMAL NUMBER: 0029 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 220.0 GRAMS NECROPSY PATHOLOGY LUNGS C BRONCHI : -DARK AREA(S) ; LEFT, ONE, SUBPLEURAL, 2Mi OBSERVATIONS HISTOPATBOLOGY LUNGS t BRONCHI : -ALVEOLITIS,-MINIMAL, FOCAL -FOAMY ALVEOLAR MACROPHAGES,-MINIMAL, FOCAL VAGINA : -PALE AREA(S); SEROSAL ASPECT, (ME, RAISED, 2X2M4, VAGINA : -PROMINENT PROSTATIC TISSUE ON SEROSAL ASPECT,-PRESENT APPENDIX 10 (Individual pathological findings - continued) Print Mo: 0026 Group :1 23 4 tUo> Compound Exposure level (ppm) : Control 0 ---- - T-7499 --- -50 150 450 Printed: 05-MAR-01 Xybion protocol number: HIM 252 ANIMAL NUMBER: 0030 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: U-DEC-00 STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: S TERMINAL BODY WEIGHT: 207.9 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY STOMACH : -ANTRUM WHITE NODULE (S) ; MUCOSA, THREE, NEAR TO LIMITING RIDGE, DM. STOMACH : -ECTOPIC NONGLANDULAR EPITHELIUM IN GLANDULAR MUCOSA, FOCAL,PRESENT MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group 1 34 Print Mo: 0026 Printed: 05-MAR-01 Compound Control T-7499 ---- Exposure level (ppa): 0 50 150 450 Xybion protocol nuabar: MIN 252 K> ANIMAL NUMBER: 0021 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED/ TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 194.2 GRAMS NECROPSY PA THO tO GX OBSERVATION S HISTOPATHOLOGY NASAL TURBINATES : -OEDEMA IN STENO'S GLAND,-SLIGHT *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) : 215 Group Compound : Exposure level(ppm): 1 Control 0 23 4 -- T-7499 - 50 150 450 Print Ho: 0026 Printed: 05-MXR-01 Xybion protocol number: MIN 252 ANIMAL NUMBER: 0022 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY KEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 177.8 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY UTERUS : -FLUID DISTENTION UTERUS : -LUMINAL DILATATION,-SLIGHT APPENDIX 10 (Individual pathological findings - continued) Group : Coapound Exposure level(ppm): 1 Control 0 23 4 -- T-7499 - 50 150 450 Print Ho: 0026 Printed; 05-MAR-01 Xytoion protocol nu&bar: MIN 252 ANIMAL NUMBER: 0023 DATE OF DEATH: ll-DEC-00 SEX: jtkMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 222.1 GRAMS NECROPSY PA T H O LOGY OBSERVATIONS BISTOPATHOLOGY KIDNEYS : -INTERSTITIAL INFLAMMATION,-MINIMAL, FOCAL UTERUS : -FLUID DISTENTION, MINIMAL UTERUS : -LUMINAL DILATATION, -SLIGHT MIN 252/004312 APPENDIX 10 (Individual pathological findings - continued) Group Confound Exposure level(ppn): i Control 0 23 4 -- T-7499 - 50 150 450 Print Ho: 0026 Printad: 05-MAR-01 Xybion protocol nunbor: MIN 252 ANIMAL HUMBER: 0024 DATE OF DEATH: ll-DEC-00 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY HEIGHT: 220.2 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY HEART : -VALVULAR ENDOCARDITIS,-MINIMAL, FOCAL LARYNX : -INFLAMMATORY CELLS IN LAMINA PROPRIA (VENTRAL) ,-MINIMAL, FOCAL *' ANIMAL HAS NO GROSS OBSERVATIONS RECORDED Group Compound Exposure lave!(ppo): ANIMAL NUMBER: 0025 DATE OF DEATH: ll-DEC-00 APPENDIX 10 (Individual pathological findings - continued) Print Ho: 0026 23 4 0 SO ISO 450 Printad: 05-MAR-01 Xybion protocol number: MIN 252 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 29 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 210.5 GRAMS *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ** ANIMAL BAS NO MICROSCOPIC FINDINGS RECORDED ** MIN 252/004312 MIN 252/004312 ABDYMIINNHISATLRAATTIIOONNTOOFRTA-7T4S99 : 219 : Author Stuart Crackneil CONTENTS MIN 252/004312 Page TEST SUBSTANCE AND ADMINISTRATION..................................................................................221 EXPOSURE SYSTEM...............................................................................................................................222 PROCEDURE............................................................................................................................................. 224 TARGET CONCENTRATIONS..............................................................................................................225 EXPOSURE CHAMBER CONDITIONS...............................................................................................225 CALCULATIONS................................................................................ 226 RESULTS.....................................................................................................................................................227 VCHAAPOMUBRERCOTENMCEPENRTARTAUTRIOENA..N...D....R...E...L..A...T...I.V...E....H...U...M....I..D...I.T...Y................................................................................................. 222278 D ISC U SSIO N ............................................................................................................................................. 229 FIGURES ABC... SSSccchhheeemmmaaatttiiiccc oodiffattghhreeameaxuptooofmstuhareteevdsayspsaotmeumrplgiuensngeedsryatotsitoeenmxps..oy..ss.e.t.e.r.m.a..t...s...............................................................................................................................................................................................2.22333102 TABLES ABCD.... CNOChhopaamemmriabnbteiaenrlrgcctoeocmnnoccnpeedennirtttairroataunttirisooenonassfntoodhfferTTeel--xa77tp4i4ov99se9u9h-r.e.u.d.m.sa.y.ii.d.ls.yi.t.te.ym.m..-e...a..e..n..x....pv....o.a....sl...uu....er....es.......m..........e.....a......n.........v.......a......l....u......e......s...........................................................................................................................................................22..2233333456 APPENDICES AB.. MCheathmobdesrocfosnacmenptlreatcioonllsecotifoTn-7a4nd99an- ailnydsiivsifdouraTl -s7am49p9le...v..a..l.u...e..s..................................................................................................224367 : 220 : TEST SUBSTANCE AND ADMINISTRATION MIN 252/004312 TEST SUBSTANCE tTcsShuophelnoeftroneatnsnetosystsrtlaisfs3nlu5audboivkscratagiapdt/neocecdyue(lrtP,ihndBwaditlSehuTFrit)ce-)7hdaa4nnwi9dsd9itrw2hewaf-aaesirsr4.rr%seeudcfepftieiovcreifetldhunrotaolrtyuotgsshtuhealosbfeuoletllafatnohberios.urIasrtteeowprioniaerststhsaoiussnpspTt2lui-6ed7dy4A.9ap9sTr,iahliesl2iqT09u0-67i0d4.-9in99In8awf%osamrsmpaaealdlrtmficolyuinnloiirnfsordtoebemruret(datnnhteoeet ADMINISTRATION vTTgaehhpneeeorTcuahr-t7aigo4men9bn9seeyrrvsaatatepotmmorsuowrtshpawrhsoaefusrugerahstdhmwwereihnrdieicislhtupetrraeoedddsuwtrtcoeietahdtmhaebiyorrfatmotdsegrbitieevydreiwntahghierothlfweien-abaslloiqpdcuahyisadsemextdebp.soetrsuTscurohebnescitneaantncmtchroeaatsmiiponhbnteoesrresostfdapteirenosslcdterusaibsceeersoddtseobbelely.loctowhie.l tTgyehpneeeirgnaa-tilsoinmneesatyeirsrftledomuwriwntogastthhmeeovpnaripetlooiumreridngaethrnyreorpuahgtiahosoneuaotpfeptaahcrehatsuotusfdathyne.d eDthxueprodinsiugluretehsnetuasstiiurndsgyucptaphlleiybarwiartefelrdoewivnet-roliifntiheeedtaauptsmeirnoegsdpathudebrryee gas flowmeters. wTdchehelreriovemseeraydttteoirntagegtrresamspoihinnfiectodhr(edGedtCreu)strotiansmnguaabliysntsthtaiaesnincoepfcrhameatlmemimtoebrsieinpnrahgcreoyrsneycsgsetaenemntmreaprtlarieeotsiqno.usniMcreltoidrnsioaetolrtsoaodtbwajturaigstihtenmto.tuehntetstaawnriegmreeatlmsc,haadbmeabtsoeerdthceoontnecsetntmhtreaatteiogrinaassl Aasnuimseadlsfoarssthigengeednteorathtieoncoonfttrhoel gtersotuaptmreocsepivheedreas.n exposure to compressed air only, from the same source The duration weeks. of administration was a single 6-hour exposure on 5 consecutive days of each week for 4 The combined usage of T-7499 was determined, for each day of treatment, for the three test groups. : 221 : MIN 252/004312 EXPOSURE SYSTEM tvmEsdayaiapasslcptveteherleramesiydeaxlttfnophoseouareetspddaupweclrletheyeerfrpesglmuiyepicisndentoeweimnrfmaolgmotrokwtcohr.onecmaotntepondsrtitrasiolmnelld-alvgitnareaolervuiwaepal,hsiruodfwslilealoeu-gwrbeeeonm(dttwhyaoeeinriiginttcehhoostartnillnatmortgsoiasoltfneflvorraoiewalmxvlmpecsotehy,steleuairnrcsecd.yoelcmirnh,Cpdaaomremerlbs)opseaaeordnd,ndecaanteihtrlvsleasapuosntopfadupitnrlhyilteegsasegsnanedsesnstroeceaecortoalinatrttit,eroeosndalt FSicghuermesatAicadnidagBr.amTsheofcothmepvoanpeonutrpagretnseorafttihoenssyysstetemms aarneddaenscerixbpeodsuinrefucrhthamerbdeertasiylsbteemlowa:re presented in Vapour generation VtpTcoorhagemlegsvsmleveusaorvpins)aoelrutdveoresesvegreaetvsconosheeiarrlolalotfosifowuthnpliepqisslutyoieiesldsadttetTimcot-hhn7eaf4moo9trfe9bstG,ehtrwresos.htuueibpcTssshthtase2wnu,aclbi3sesqtumaatinondacdiensd4teeaspluii(anvLpreepaodrtlwyeya,tlamiInnanedetpteewtrrroemiasntsseghudaerilveasmtaoeolevfftaeie4ntsrt0dien(dpgNHswuiivgpiuathrhlnovddeaeosSrspweNaS)raitetscirrcoiofuemgilstaetpetnNred.isefweeiTlddithtelheear Ttttpc(eohshr-mteo7aasdm4ipeanu9ceiblcor9reeaensritvsdstnuha,alrspeeerttytoeheoeurtxdloofr,piualiou1nsgps5tthpuaiaoriprionPnermxlomeavilsmtpyeimsxsoTastorsteueeesttrleplrsesyeahilFzcef6erolrv0ouenamo)ss.Cp.rtoro.tFuhrEicleTsutthaehigdytdeielonpetoneneafsrseactgstoil(sonaPiurlgsTsbssstFwthathEanraano)dstcutefsgwuuthvbareiatirtnhnphelegoeeriusmmwdsrimiesaltutthneeetderreasildna,engidwaredvimxitjnahatilecxvaareteniunrsawrtleiwantodtwaetishtarahemsdebessealiuettinechvbhroesaemnroleqdaadfutaiiei6nodyntnimatrvlieeynmecxsetc.psldayeorlsarTsiuintehtrtodoeea rTehseiduaairl psuaprtpicliueldatetoanthdewvaaspdoruierdg(edneewraptoorisnta~n2dCse).condary dilution vessels was filtered to remove any Tlcoyhaledintdceeeslrtlbwmaeanrteedrmitahaleirnwetsaeeiinrgvehodtiraot(ftathhceeoonrresistgaeinnrvtaopl ircreysalsniundrdeecr(o4ns0tueppnpstils)iewudnebdreyerdtdhisrepielSdapynoeindtrsoocgor)ennwti.nausomusoluyn. tTedheocnoannteenltescotrfotnhiec Lmmastnieemqdetuelt,iirhnde7etdewvprbnaamyslavltpterdhosaiernateosmvpsapeiolztrvreeot)eret.sedcwwftreatohrsmeedmitenhlcfiervoobermpraeosdberaltootoecfdktthhaienegterovebaseypeaorcavrhniosyliairqtpituaoorindttihccdeuoelimlalistveeettehtrhyrrioanltuignmgevhiagbPlhevTttewFhs.eEaevPnteuatbbrhteiienecgnudlipoasrttfereisabfepiunlptttier.oornsxTi(mmhsetaaantlieiniqlflyouesildd2s A line drawing of the generation system is given in Figure A. NHMuuopnortrlose,iCgCoha,rIWdnidfiflu,losSturogiauhltbhyCG,oOlnathmriooolrs4g4aL0nt9d, 4,C,FLU2oS2aAd2HCBell Division, Portman Moor Industrial Estate, East : 222 : MIN 252/004312 Fpiscnuhaotfsaorfsmieactbhdlileeengtrh.btrroatousouepgesohnefsxufdlprieeeoxsiasebedltceoottoadnludTcac-trh7iyan4mg9d9ibtl,ouerttihaoaenitravfvnlaoegpwseosnueotlri/faaalainrpdinpmwlreioxatxtsimummroeauitxpenerltoydeddw1u5acit0tehdt1ha/iemnfuiatnhrp.teehxeTvrahopsefuotptuehprseltgyaaetponmpef rrocoasltpeporahrinsaetraweenawedsxadppsroaystshsuaeerinder The control group received clean air only at a rate of approximately 150 1/min. Inhalation chamber Tbcf0ih.eht7talee5omdwembxewt3phr.o.iitsshAuwrtaeathscpehayaarpmpaemebrxfeiodrorsafalwttehbedearsecueaponpfaiesnsrtdteapri,ytnorwlapemhs. siicdshaTtelehenfelisgauiunrnredtedergwnelaaqaslsusatvhlcoeodlnuitsasmtntrreigubecuonttitifoioannellaayocnhfmdthocceuhonantetmsesidsbtteeaardtimrwodofausspcath.caeuprIebpmorwomidxietaihmdl iinbaattoeetdhllyyye Astacicnelsessstostteheel cfrhaammeb. eTr hweasdothorrowugashstehaelefdrounstinogf tmheoublodxedseructbibonervsieaalainhginsgtreidp.door with a glass panel and pE4Ele.oaxvtcpeeholnst2liueaarvleneadlcnaiiasmgireaassblacleemoxntpposlotesrhsuuowcrletdeecdr4aepowcafacisgtithteaydsi,nroalwfews6inst4hsfroteaeratescal.hnmaIcnleyastgshhieiswscfeiranorpvemaebssutltheisgipsoaetflnieodhvneoe,dul.s1oNi0nnoaganc4ifamrgraaaemltssecwwoinmeodrrpeikvapiradtrrmeursaaeelnnlngyttse. odwnToehlnreeivs4uegsllseeavdv1eeolosnar. Athewgeltasasn-dpadnreyllaeldcodhooolr banudlbwthaesrumseodhytogrmomoneitteorrwchaasmsubsepretenmdepderiantuthree acnhdamreblaetri.vTehhiusmwiadsityv.isible through Tdrhaeinpaygreamsyisdtaelmbavsiae aofbeaallcvhaclvhea.mber was fitted with a 2-inch drain. The drain connected with a common Asitusaqtueadreinttuhbeuplayrraemxhidaaulsbt apslee.nuTmhi,s3coinnncehcetsedintodtihaemmetaeirnaenxdtrapcetrfsoyrsatetemd. along the ventral surface, was Tasstiihemreefillltoaotrrwtoaflllwlocewahysamommenbteeewarrshmuaiircorehfudlnotuthwesediwnsogaencsaot1hnt5ead0paverlaiyrpterodedusitl/ruumgbtiieeonnnufetlreova.wetisoAsmneilretretwoenlratlesesryiemt.udoatuthenedtecadht.atmhGebeefnrroethrnarttoiooufngahaptihruerwpianoslseemt-bdeuuaicsltut.rseDtdaiinloulneesnast TpArheMisssauwgrneaesihnmeslioidcuenptthreeedscsuhoranemtgbhaeeurg,sereec(lo0ant-i2dv5aemrytomdthiwleuatetiexorpnogvsauuersgesee)rlowotrmaos.llceoynnaencdtewdawsituhseedacthocmhaomnbiteorr btyhea antymloonspthuebree. Ebexuxtitrlradacicnttgiwonawsoitvhfetdnhrteaewdchitnaogmabtameirrostshwprhoaesurgaehcvciaoammapnalniesixhfhoealddubsttyosmtwaechkain.cshoaflla cshinagmlebefarsn mweoruentceodnonnecttheed.ouTthsiedechwaamllboerf tahier : 223 : MIN 252/004312 aEfimlxttebrriasec.nttTfplhoreewsisnwutreaersnwaalhdepjunresostespduerreuatswiioningtahlig.nateeacvhalcvheasmmboerunwtaesdminaitnhteaineexdtrainctthdeucratinnggeb-e2twtoee-n4 tmhemcwhaamtebrebrelaonwd Tonhley.control animals were exposed using a similar system to that used for the test groups, but received air PROCEDURE Aiadbeosnveteipc.aarlateexpeoxspuorseucrehacmhbaemrbtoerthwaatsusuesdedfofrothr eeatecsht ggrroouupps.. TThhee icnohnatlraotiloannsiymsatelsmwwearseseextpuopsaesd duessicnrgibeadn cpTcaohvrhemaoevmipdiroabauterastsrnm,lwyyteehanvresteassrpitigrooaantsfnieiotstdhifnoeessnrereqieoxnudpfetoftnhhrscoeeuemr.adenotchismaeegaherloses.lcdweiiTnivtghheedicnaadgnthueimees aactnolhsdatmwphleebarecesrepdlwaoticainaasltaecdahprarroenandgneegltdeeevmrametelin7n2t-esddoaofysftehqitnehuteeecnrhavcanaemilmsibneaatrclh.sceoIwnirndidtoihnirvigdniedtrouthtaoeal TTinhhteeerncdahillaupmernebtsesraunMrdeabggeenlneoehwrealttihocratgaoairuffgtlhoeewwrsoaowsmecr.heecskweidtchtoedenosnuraendthtahteocphearamtiboenr odfoothrse wchearembcleorsteodokanpdlasceecuwreitdh. aTnhde ttheestesxupbosstuanrecestsaurtptpimlyetodgogcluemveanlvteeds.between the pressure vessel and the metering valves were opened mThoevedmeleinvteroyf eonfttreasint emdavtearpioaul rtobutbhbelevsapinouthregPeTneFrEatodreslivceoruyldtubbeinmg.onitored during the exposures by the wwDcsesaehqaaumaauslrlmileiplnpdilabbgee.nerrdartffT.htooehferr,lTemufa5hsisenrheadamsimtlnsyblnagpsyomilinetsfihpse.ntelhwseeAeessxeryfprctwtireoheniesnarigrunmgearjteeehbcdcseewo,terellissldataeyahyicmrdritainoewprdgeflmaeecasisitnpnlnydwtpieomireiesond2pruxte0lemoeinmcmsstooaheallmdtrfl,eeotgplcbuhyaltagreesceh5dlketoiasagoivetnhposctateolo,opupnpfpmttduhotrsheomel,ysexetpisogoxmreaopfaaaspolcltcysleholuhlloaeywrrmcenoahembttcoheehsauedryatrolmrapiyignirnbrregiatesnbheprsteesheua.f.rronivenrdSaehilatnsahmdlfaertprhatoilsweoemynisrnciyeongcrahlgilcnuaeehgpmctethtbiteoehtesnonert ttIcshhnaoeemltlhe6psec-tlhaetpirosoetunfrtrriiooomedmfxepafretofmoaosmcurohrsteephcxhephpeeaoerrmiesxoupbdsroeae.rsmu1ipnr0elet,ausnardwnsta(ehGsqrousrowueungiptchcioh4nueg-tdtdGhoeenrvoriacuenempdaa1tin)hndetdoetsirtamhmoeefprGtlhriCneesgsestytpu.sudtmTyem,hpii,mstahdmtaeet1vd5icci-oamenteditnrleyoullitalveefetdiernerttdhedreoavctaamulusmotosdempunhrtaiientnriggce mThineuCtehainmtebrevraalsirtfhlorowu,gtheomuptetrhaetuerxepaonsdurreelpaetriivoedh. umidity were monitored and recorded at approximately 30- : 224 : MIN 252/004312 Atohftetteahsnetimmenaadltseorwifaetlrheperreeexmsspouovrseeudrv.eesthseeltwesatssruebcsotradnecde. sTuphpelvyatpoogugrleinvtahlevetseswt cehreamswbeitrcshwedasoaflfloanwdedthteo fcilneaalrwbeefigorhet Acatgtehse. Tenhde cohfatmhibsetrismwe,ertheewraasthsewdewreithunhlootawdeadtefrr.om the chambers and returned to their respective holding A summary of the operating conditions used is presented in Table A. TARGET CONCENTRATIONS The target concentrations of T-7499 were as follows: Group Designation 423 (((IHLnoitgewrh,dddooossseee))) Con(cpe415pn550mt00ra)tion aTvhaeilatabrlgeedtatcao.ncentrations were selected in consultation with the Sponsor, following the review of EXPOSURE CHAMBER CONDITIONS Analysis of chamber concentrations of T-7499 Fdacauoniortalreollecymtcthsltaieyistoefinadnirrtesmostydetnsehtthsieencomersdiabsemfexowdppr eloieonrsenulA-orlteopiasnppk,eeeosnnsafdamitimxhnpepAlselgie.snaqgsouefcannhccdrheoamimfnrajboetemocrgtiroaGaniprr.howT.uhepTreseh4aecirorteolasleaf2tcme.trepdlmMesionenctihogtoolalrsdeiscnttigegodhwf tbassasyymprbipeonlrtegfhoecrsomofalelntedhdcetuiisonsianjnemgcaptnaeldnde TIanrheheaglimavteieonthnionTdAoxopifpceoanlnodagilxyysADis.epwaarstmaednatpeteqduipfrmoemntaanmdepthroocdedsuurpeps.lieDd ebtayilsthoef tShpeoannsoalryttiocaalcpcroomcemdoudreasteusthede Airflow and pressure Tadenpihsspepurloraeaxyiitermhfdelaoctdweoilnsyptiilnna3tuy0ooe-mduesvialnyaculhuatenedcwihnaaatsemdriovbncaeutlrhsmewtethnaartrsoegudemgtchrohaonenuicgttkoer.eweadcahs upesexirnpfgoorsmutareepd.eraetdTahpetpurbcoehxaimmroabttaeemrlyent3ee0grsamtivianenudtperceihnsseteucrrkveeadlws ataost : 225 : MIN 252/004312 Temperature and relative humidity Taphperotexmimpaetrealtyur3e0o-mf tihneutgeeinneterarvtiaolns dwuartienrgbeaatchhaenxdpaolssuoret.he air in each exposure chamber was recorded at Taupphpetraobwxleeimtsauantpedplylide3rdy0-wmbiutihlnbuthtteeemiinnptseetrrrvuaatmulserentsht.roouf gahothuetremaochhyegxrpoomsuerteer. pRlaeclaetdiveinheuamcihdictyhawmabsefrouwnedreusriencgoradelodokat CALCULATIONS Ipdannraidtnoatrriednincegar.tlhtcoCuislomanrtieisnopeinoqmrumtiesanehytaltyslhe,eabtdheceeutnosmesucrmaloalauctilnuvldleaaeptdeeprdanrorucermnostnbtthdeiairnsstucowrreeuispslulalylnitncufcirelsousinmdwgeirtreuhopnuoernoathduteeinndrdgderaodetruarnnoiudnrmsinthibgneeortrshefepnaonluradmts.tbosenirglsyn, imfriocuuacnnhdteofdifgutfhoreer Chamber concentration Maavdeejuarasntgmechecnaomtnscbteeonrtthrcaoetnigocenennwetrraaasttiicooannlscsuwylasettereemd.cwalecruelanteedcewsshaeryreaneqdusaalmspamlepilnintegrvianltserwvaelrse wvaerrieede,maptliomyeedw. eiWghhteend : 226 : RESULTS MIN 252/004312 VAPOUR CONCENTRATION Analysed concentration of T-7499 Trehsepeecxtpivoesluyr.e mean and individual sample concentration values are presented in Table B and Appendix B, Tgohoedeaxgproeseumreenmt weainthctoanrgceetnctroanticoennstrfaotironeas.ch group exposed to T-7499 are presented below and were in Group 342(((IHLnoitegwrh, dddooossseee))) TargeCthamber 41555000 concMe41n4e56t7a92rantionA(pnap22(l9ms20y.d..0s1)6)ed (C114V93.5..%37 ) wh2Tcsbsyuoiuhagosbnbeshtscfbeetlemlhaeesennsixtsgt.scprwhewamoMetasfiirasutolihornrntksiweohsnwear.dtrnitoaethadcfelmethjeuxtetcihsepdntteseeimtmdcoHetnfivelin.sgudaethirsdiTcnaadothdunoieicedossfllnefirivspeaecechtxiiiroeneopynnnnottfeopsismuogiotpruffeeefrnlvsavucoataioaipndronniondacdwusteeirwanolwistbnavreusaefrbtorseeiyebuovmelnntienoddaswedtfhtneahooetnertdortvbhebeatraothpohsteaoushetouggtdrcrtheihiebongaelueuetievrnttLaadeetbtrohriyliaoweentnrotsaetrastotrsuyenrdusodtothypnefetImiftenohoafstneroecsdrrhatmmueltolsroeaitctntdceitsigohasuaentstbWiegsolotreinaeqofenxuutkvhcpipdaeao1sptswatobutenhauursdeesrtt Nominal concentration of T-7499 The data are presented in Table C and are summarised below: Group an2d,3Haignhd d4os(Leocwom, Ibnitneerd. ) Study mean no(mp2p1inm1a)l concentration A/N ratio 17006) A. //Nt= (V-AN--no--am-ly-i-ns-e-a-dl--cc-o-o-nn-c-c-ee-nn--tt-rr-aa-tt-ii-oo--nn^/ x 1, 0. 0. ITcainttohmcsmerhoeboasniupsnolehedmdedrbi,tinhecfaerlnpoaormnceteosastdlnhyuctesreheenamdottratfauos7tssine6ooo0nomfmfTfitnomh-ar7els4Heer9aga9vtciaahodnl.uedeleixusvpsemeordesaudytrhehienatmpvoeeetrahiaonerdtcigfhiewcanimaaeslrblayetcoradrtlesec.pmurlepaTsethesreedadt,uctarfheloecrfuonltraohtmteihoiennthtaharlserceseouenmttrcreeeedaanttteeardadctigoogrnnroosutauapnpnsds.t : 227 : MIN 252/004312 Tsyhsetecmalwcualsatpeedrfvoarlmueinogfatshedeasniganlyesde.d to nominal ratio was sufficiently close to 100% to confirm that the CHAMBER TEMPERATURE AND RELATIVE HUMIDITY The daily mean chamber temperatures and relative humidities are presented in Table D. Tm6fhsrt0ahuoed%edmaysncuR.aohrHeacddm)oismbrrceeeepfrllrraenettceisibvtmsiloeneprgehesrtufyahfmsteeutceirdgtmeiesutnipieaenosnrcandotrierdohpcinuoeomrraaadintnedidindimtgidedaiaslluusrrwtieianoefngrnrdiegtoehissfreiamtnnhsoitelttaudrcdcrhiyofeaonrmrw.sibdaeTelrelerrhegiadslrtoomdtwuoeopevssrhpiaahottvheinoearenneasaffctfwrhehoiecmttdhetadatayhriegtrhoetedfthetaorshtaiuegnwtncgsaoetscumod(seny4ud.0poi%ptfiTlotihhentdoees : 228 : DISCUSSION MIN 252/004312 mgCgrroeoenaautntpreoscrlotvhnoaacfrneitanhwtteiroaaTnsti-o7sbne4ees9tnw9(1eav9eta.n3tph%ooeuc, Hrc1adi3sge.i7holi%nvdseoaisoynedf.tog4Te.t5nhh%eeisraerwtxeisopapnosesrcwuetrifaevlseecclehytveafdimdoerbinneGtrtsrhfoeowurpcatosshe2gef,efLi3nceoiaerwnandltlsay4no)ds.fatIvinsatfreairacmttiooerdnyiafatoletrhetohxuepgohdsautihrlyee fTohrealsltugdroyumpse.an concentrations of T-7499 in each chamber were within 8% of die target concentrations Gccohonaomcdebneatrrgacrteoioennmcsee.nnttrTawthioaesndwceataelscrmualtiatnrtieabdtuiotbenedtowtofeaaencnootmhmebiinnaaavlteirocaongneocfeafnnatcaratloytirsosendastahfnoadtllowtwhaess: csolimghbtilnyedbenloowmitnhael acnhaalmysbeedr - the use of a single barometric pressure (760 mm Hg) in the calculations; - the calculation of an average nominal concentration for all groups; - use of the average chamber temperature for all three test groups in the calculations; - rtheseultteinndgeinnctyheodfetlhiveerteysot fmaamteirxitaulretoofvvaappooriusreainnd tlhiqeuidde.livery pipe-work to the vaporisers : 229 : FIGURE A Schematic diagram ofvapour generation system MIN 252/004312 V kV V V \/ b I y y 3 d n 4 e --f Tkl ? 4 Li _ 8 lc F ront view Side view Key: a Air supply 1 15 pm filter fbcde hgi j k NARPCTPTTSTrhoeeooiietrussgFntrsatthtogtEsmrmseogluoueaterfealebetfnvttisvseevnagtt(arrlaagpamilvalnavurvcenlceaegeodestteei(aslgcirun/nievorkanenapl)eptrdrreoioaensultlsliirevuovtgarneielrsvtnyereeodprliarlpeetoeysrearvirosiur pply) mno P qr tus V VMTTWTTLL7 heooeeapiamsseaapcmtttrddroepmomsruccefu-roibaeermbl/a-ltaltaslelestetiturthrdtraiierrniamergrlcieiinpeftrxagertfuoleurvnbdodraieemitilsnva(pttepnlooardeycasdhisriaugsmriutiabspleepddrl)yirselpislneaeryvoir : 230 : FIGURE B Schematic of the exposure system used to expose rats MIN 252/004312 Key: ABDFCE AEAGDSxanliiasrmpipsmfopselsoalreuislwnlriueegotcxnrcpopihadonoatetrsormtvuorirlbceeeacrnad(g0ce.7hs5ammb3e)r monitoring HGIJKL EGPPMDroxaearwhta-ieanfieinuvrleetsaxetdlrhvpeaelxeuntsrtuamct filter : 231 : FIGURE C Schematic of the automated sampling system MIN 252/004312 232 : MIN 252/004312 TABLEA Operating conditions of the exposure system ATatmrTgoeetsstpcmohneacrteeerngitarelantfeieorenadt(ipopnm) Test material flow control C h TGDamCCmehheielsnaaaubtmmeseemurnbbrartateeeaotmrreirrrpeefixrlaneotltrswauscusstar(egIe/(mmimn) water) (Con1trol) 0 N/A N/A N/A -2N11t55/oA00-4 (Low520dose)Grou(pInte1r,350dose) (Hig4h450dose) cWnoAenTeettaidr4gopl0heletrpvfellsaodoilswvdseifrreocmctgnofaernlTcseootyadrmaclopdliyenletlcrhidvfneeleeaodldrllwSvemrepeoansscuonorreneTestdadruoplupelesprvifleniladeogldwvae 50 -211t05o00-4 50 -211t05o00-4 50 -211t50o00-4 : 233 : MIN 252/004312 TABLEB Chamber concentrations of T-7499 - daily mean values sCdV ECxsCFMtVpNooa12111s111111o2435er6798n1e0od0135892467(asfdEa%.unfaxir)crepdioednsuetvroeifastVi1oa0nrti(oaLtG2ioo0r44444644444334w56353551no,13530555494267855499679uc=.d.30poo(ns2scede)ntrmaCteiaohnna)msxabme1r0p(0CIlenGo22st1111111111111111111ne2r0276755654647244644471worc32113613402849978898473,eue..d1n7rpeotr3saceto)ilolnec(tpepdmb)y (HG444444444444444444445ig575r5255575463395614490oh13883570049897906466742u..d56po4se) an automated sampling system : 234 : MIN 252/004312 TABLEC Combined nominal concentrations of T-7499 and analysed to nominal concentration ratio AsEd/xNMpN1l21111s1111o34256789e10lod023549786saddA.SuntrnaeanldyaTsre-dd7W/d40n00000000000000000000(0e.9ko.....................0v4444434444444444444449tgm2o11i1111111183858544444)au6iftnsioeadnl coMnTceeeamnn222222222222222222222t(pr1111111111122222122220caeCh.tr3ia)aomtnubrreaetrioTe-xV7p4or9el333333333333333333333(u9s2522065282422522522585lms)......................ve8294288828828802904824beda5poaosfuar Concentration N222222222222222222221o1000201001103010013281m1333722227727272277822i.n7alc percentage (ppm) A222222222222222222222n2111311250322142112211a1119333959060902447790ly.7sed Calculated from the following equation: A/N 111111111111111119999101100000012000011153572235722857446698.5 V = W xTxR Mw xlOOO Where WTRMw WAMGvaoeseilrgecachogutnelosactfrahtwneatsme(ti0gbm.he0art8ttoe2erf0miP5aplBLeuSrsAaFetdut(mr3ien0m2(tKh.o0e)l9'1(eg=Kx/pmm'o1)oeslua)nrete(kmgp)(C) + 273) c Calculated from the following equation: Nominal concentration ---V--1-0--^---(p--p-m---)-- va +v Where Va = Chamber airflow (1) for die exposure (162000 litres). d Nmoemaninoarlsctaonndcaerndtrdaetivoinatiionncluocdceudrreevde.n though a discrepancy in weight loss was noted. No change in the : 235 : MIN 252/004312 TABLED Chamber temperature and relative humidity - exposure mean values CsdVECxMVp2s11111111114oe23567981d01358267049(as%unCsr)teoanefdfaiTcr222222222222222222222die3222322222222222222022e1md.n.34(eGtCpvoroiofanutVtMirpoaoen222222222223332333333rl1Rai)19185709996789220600026an.H.t09icohna=m(bsT22222222e2222222222222d1re2222222222222222222021(m-t.*.L2e0Gpmmorwpeoaeudnrpao)t23233222333333332333s32uxRe11188920400099030000901r).H.e102s00(C()ITan222222222222222222222n111111t1111111111110e1201edm..r35mGrpereloadtuiiapvte222e2222222222222222233R6885557487667875767760h1d.H.uo93msei)dityT222222222222222222222(1111111111122221e202222%2(m..H54RGpiHgroh)udpo22222222222222222234111sR111120732688645224e46984).H.44 236 : APPENDIX A Methods of sample collection and analysis for T-7499 MIN 252/004312 SAMPLE COLLECTION Chamber concentration Aeececoxxxhllppprllooooesmcsssauuutmairrroteeeponsslg.c,erhwsaaapwenmhree.bareSeumrtatoamainmnkptueoalantienallfdgyrpolossimlnaaymempasprpowslliaepnemdyrglepeblpsnyiynuesgrwstgeyipemtrdohinrdwtgwraeliaotwhsfciiattnuthtgeeseddeccdwohhnaaittmomhlebbtavereearsrlneaa2sattflmmieonroofgsstsphpvahehameleveprrxeeele.pfsfoorFsrovuom1irra2ethacmPehTsiantaFemumEntbtephestlriua.npbngrTiidnohpgrseoutrfotbtoisrssoeataqnmnugtipehnanleeest A line drawing of the automated sampling system is given in Figure A. Tcsfablaoyhosmnpewutapprsaolpvsiulnlatefgclolmvohmlewaeosamnotmtwepbadereewyrtresseetarriomewmandpphejaluriuilrnses1gtgt5eoeaddnrsmtlyewysisdonti2etut0hthmtheemtahtcsGleyw/mmCccilthoiecnaas.hntmped(aDsb6sseu0ssorere0idaqinntumgtmjheelneroc/omatstieupcaidhghnllhe)ytcrhtyeoheb.cfeelcetAtGow,hnteCtethaeecepinnnhptsjasrteamohcmoxetbfiipomeHtlnrhianiegvatgehatsmll,alyvimonIe1nsep2(ptvle(hiVmrineamigri"ne1eluTd/o8ptieoe"aafpstslTe.soi,eenndfTt)LloohoatnheenwardtoscuaahubGmngicCndhpyglcA)gitl.nhaeigesr, wccWgohaanrhssocebmenoanpagttehtronaetogetiadrotauhnpttoe,ohmaaathplanlpetoderwdofpopsturrhamidmeaitpspetselispiwmntogairclstoas.myrTsscpwhtoeoeinmutccpnehwudnematdtrospatiobiwnonenauds,sfre6ota,h0rweinsnnaejecascolwptoneiinodrtcgnisohwtdoehfdaesgoosaufnassme3bad0pnalgdteossielpwmicnuaomersngse.decdsotiFhantoedberuelsyfcaiontmaerjeefdptcelbtirsiny,otgantarhslttieitnnaoecggfsha.tishtnbhegaegseaagtmacaphes METHOD OF ANALYSIS Cdenehdtaamiolefbdteh,ritsoatgampetpohesenprdhwiexri.ethsaamsupmlesmwareyreofanthaelymseedthboyd gvaaslidcahtrioomn,aitnogtrhaephInyh.aTlahteiomn eAthnoadlytoicfaslaPmrpolceedaunraelyastisthies : 237 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) CALCULATIONS GC analysis Tvmahapesossuaurmsseptdalnetosdaoprrfdescphaparrmeepebaaercrehdatvmianpogosapushr ebsrtaeagnwsd. eaTrredheiinsmjgeeicvttheeodndbinfetoloor wacagilnacsuelqcahutiranotgimotnahsteo1gcraoannpdche,2n.wtrhaitciohnwoafsTc-a7l4ib9r9atferdomusitnhge Concentration = V Va + V x 1,00 0 ,0 0 0 ppm (1) V = -W-----x--M-R----x---T- x -7-6--0-A-m--t-mm---H--g- (2) where VMARTWtm v. ======= tvamm0geto.amm0oasl8sulepoes2moesc0ouprue5asfhltoaTevmurfro-riwl7ealc.ua4ie(rpmt9Kim(rg9mee)h/s(motlsm)uofmgrTfeo)P-l7(B.mK4S9mF9H((3mg0)l2).09 g/mole) pIdannraidtnoatrriednincegar.ltchtoCuislomanrtieisnopeinqomurmtiesanheytaltyslhe,eabtdheceeutnosmesucrmaloalauctlinulvdlaeaeptdeeprdanroruecrmnostnbtdtheiiarnsstucworreeuipslsulalylnitncufcireslousimndwgeirtreuhopnuoernotahduteeinndrdgderaodetruarnnoiudnrmsinthibgneerotrshefepnaonluardmts.tbosenirglsyn, imfriocuuacnnhdteofdifgutfhoreer : 238 MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) COMPOUND SPECIFIC INHALATION ANALYTICAL PROCEDURE FOR T-7499 The analysis of T-7499 in air sample substrate Tmhoenitmoreitnhgodtesotuattlminoesdphinerethsisin daoncIunmhaelnattiohnasTobxeeicnolvoaglyidsattueddy.and is considered fit for the purpose of aTStthmuidsoysdpsohpceeurcemifsi.ecnaTtmhdeeentradeimslsueltnthitnsegabnsadasmiacdpdlpeirstio,oconefsdawupriepllsrobfxeoidrmetathateeilecadonnwacliyethsniitsnraaotifsounTp-p17l04e09m9teons5taa0mr0yppdlpeomdcu,bmayreenstya.nrianlgyesefdrobmy GteCs.t NprOesTenEt,TbhurtowuigllhobuetinthcilsuddeodcuinmaenStt,utdhyesspyemcibfoicl supinpdleicmateenst.that the relevant information is not available at EFFE CTIVE~DATE ; [ 2l~May lOQC Test substance T-7499 is mainly perfluorobutyl sulfonyl fluoride (PBSF), which has the formula C4F10O2S. ASupbpseaamrapnlceeStorage of Test Mixture Aliqnuaidpproximate temperature of +4UC. Protected from moisture. Equipment Balance Sartorius BP4100 Syringes HHaammiillttoonn 5100000sesreireisesgagsa-st-itgihgth(t5(01000maln)d 25 ml) Gas sample bags SKCINC Tediar 232-series (1 and 3 dm3 capacity) Syringe valve Mininert Push button valve Vacuum pump AEG ADEB 56 (or equivalent) Flow meter J & W Scientific ADM1000 (acoustic displacement) General laboratory glassware Consumables Syringes Sigma Aldrich 20 ml polypropylene : 239 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) Preparation of samples for analysis ssSyyarrmiinnpggleee.snToehefdethlveeaitlsevseitniasstecmrltooessdepdhineatrnoedtahtrheeecssaoymlrlienpcgltieendgreupmsoionrvtgeadandf2r0o1mm0 tlmhselysroianfmgteepsliftnitagttempdoowrstpifthoherraien"jiMescitwnioiinntheodrntr"atowvtanhlveienG.toCT.thhee Tpasacohmtreitvpgaoltaifenstdghs.eavmavlpavllevin.eg. TvThahelvevevaoalvflvetehiesoGfthtCehneisssywseriittnctgoheetdhisetoo"pltoehanede"d"ipnaonjesdcittti"hoenpocasonindtitoethnnetsasnoydrfitsnhigmeeusiylstrapinnleagoceeudasrlyeinpttohaestshreeudnininsjettaocrttitohines Preparation of calibration standards dSetatanidleadrdisnatrheepsrteupdayresdpeucsifinicgstuhpepfloemlloewnti.ng method. The actual standard concentration ranges used are as AoprfovTvoi-ld7ue4m9se9taon(fd4a0Tr0-d7p4Sl)9I9a. niTdshdeiinsTjpe-ec7nt4s9ien9dtoifsraovmagpaotshreibsaecgdy,lbinmydigexerntihtnloteorwoauagsrmchilniyntiglalnuadtsiionmngavakieahlo.utWpaiteroibglhsoewatpevpro.rlouxmimeawteiltyh 6a7i0rpt.og tcEohofvenaacciTreu.-na7ttU4rea9sta9iionsngvearrgipaeaonssugoterifgdighneattsoscsrystiahrbimeendgpgeliea(nssb)thsafageimtsstetpoudlfdewaypbisptaphrgeosacpivrsfiiieaacatelstiuhncpgeappvliaenacmjlievtecyent,iatoa.nncdcpuinoratrrttoetdloyucdpeirsobpdyeunscsyeerismntgaeneadsmuarredeadssuvcrooelvduemvrioenl(gusm)theoesf Storage of standards and samples The maximum storage periods for the various sample types are detailed below Sample type GSyarsinsgtaensdaamrdpsles Storage conditions RRoooomm tteemmpp..,, alimghbtient Storage period 515d0aymsinutes : 240 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) Calibration and quantification Cactaaltlihibberraabttieeognbinysntiainnnjgdeacortidfngseoadlcuuhtpiloainncasaltayenstidcoadfleersaiecvqheuctehanelcibeli.rnaetMioonefabssetuasrntedfiatthrudes, ipanesgadkaentaaurielneawdreeiinsgphthoteendssetlueidansytessapqceuhcairifneicsjemscuteipotphnloedom.fetnhte, Fsaomr pelaechusiinnjgectthieoneqoufatthioensabmelpolwe:measure the peak area response and determine the amount present in the Amount (ppm) = ^ ^ Where A IS = == IPcSnheltoraeopkrmeceaoaprtfoetcagoarflaricembasrlpaibtoirnoasnteiolinnoeflidnPeeerridvfeleurdiovrfeordobmufrtoycmlaliscbuarllafiobtinroayntliodfnaltuadoartiade (PBSF) in the sample Chromatographic conditions Analytical column C30PmSxIL0.535CmBm i(.d1.050%pm fdilimmethyl polysiloxane), Carrier gas Helium (4.25 ml/min, head pressure 18kPa, 3psig) Split vent Helium (20 ml/min) Septum purge Helium (1 ml/min) Split ratio 1:4.7 Make up Helium (31 ml/min) Oxidant Air (450 ml/min) Fuel Hydrogen (45 ml/min) Injection volume 250 pi via gas valve injection loop Gas valve temperature 60C Injector temperature 60C Detector temperature 100C Column temperature 35C Retention time PBSF approximately 2.5 minutes : 241 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) Quality assurance measures vWerhiefyn tpheerfmoremthaondceisoefsthtaeblsiysshteedmo. nTahechpraormamateotgerrsapahnidc ascycsetepmtanscixe cinrijteecrtiiaonasreosfeat osutatnbdealordww; ill be used to RQeTpPCaelialatQiottPnaeClgbaecrritfolaoaainutlmcyecntreoet(aCrt(naeU(VtcrUeSL,SPOnP)=Q)6) Typ<<<1i3c.106a.95224l5%4%%v8alue Accep>ta8b0l%e limits <<2550%%% < 10% iTnhdeepheingdheensttlyca. liTbhraetiroantiostoafnrdeasrpdonwsiellfabcetocrosmwpiallrebde aacgcaeinpstat balestiafnwdiatrhdinotfhesirmanilgaer 0c.o9n5cetont1ra.0ti5o.n prepared Avalqidu.aTlithyecrheseuclktssotafntdhaerqdumaluitsyt cfhoellcokwsteavnedrayrd6s cmounsctelnietrawtiiothninsathmepQleCs tfoolrerthanecaenlaimlyistiss. to be regarded as Arunquwailliltybechreegcakrdsteadndaasrtdheofcolonwcecnotrnacteionntraotfiothnewloilwl besetraucncetoptvaebrleifyquthaelitLyOchQecfokrstthaendraurnd..The LOQ for the Summary of method validation The raw data for the method validation is located in study MIN/244. pCootmenptaiarilsionnteorffetreisntgbplaenakk.s, standards and test samples showed that the analyte was well resolved from any P1r0e,0c0is0iotno d5a0t0apsphmowaenddcloeessfftihcaienn2ts.0o%f vtoarsiatatinodnarfdosr tPoBlSOFOoppfmle.ss than 1.4% with standards in the range of Utras5oaoc9nncl.uw41get05peei,oti0agan10bnh00oldt,ee0fpd0cc1p0oh7lmene.8tac)co3skeptnr1psstoqr0tpaad0umntuaidcoprareenpersdmds1rp,a.0ee0ghccTrootpehiwrpvsersmeeeivlLlo)yae.nitrmi(,oacinntathaleccolyuofLsleaiQOfstfeuiQocdafineapstnntieatfdaitokcisLaftatiii0rmcoe.a9nailt9lry9(eoL9sfu9pOsD1oiQnnea)sgtenefcdtoahtigreroeanTlisant-(tas7Litnv4Ocde9oa9Denrdr)cwreoadinrrlestlervalbipetaeiostoitsonesentnhottiafoabnsblylttya2ant.i1hand%eseadrldloionfw(wo1tre0has0east sSutabnsedqaurdesntloyfaTna-7ly4s9e9d aingatihnest frraensghest1a0n0datrods 1s0h,o0w00edpcpomncesntotrraetdionast wroiothmin t5e%mpoefrtahteuirrenofomrin5al.days and aaSgmaambinipeslntetsfrecoosfhnaldyistitinaonjnedscatre(drdo(soctmaand1tea,0mr0dp0sesprhapotmuwr)eedoufcnoTdn-e7cre4n9nt9orarsmtitooanrlesdlwigiinhthttiihnnge5i%cnojenocdftiittohioneinrssyn)roiamnngidenasfloucrbosn1ec5qe0unemtnratilntyiuotnaesns.aulynsdeedr : 242 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) GAS CHROMATOGRAPHS IN INHALATION TOXICOLOGY AT 25 SEPTEMBER 1997 ---S--yN-s1to-e.-m---- Hewlett Packard 2 ------5------ 4 5 6 1 8 HHHTTPPTTTPTPPSSSTPTSHSSSSSyyyyyhhhyhhhlhhhhhhhhhheeeeieeeeeiiieieiiieeeiieweeewwwmmmmmmmmmrrrrrrrrUUUUUUmlllmmmmmlmmaaaaeaaaeeaaennnnnndddddtddddttotoooooootttitiiiiizzzzzzzzzQQQQQQQQccccccPPPPuuuuuuuuuaaaaaauuuuuuuauaaammmmmmecceeeececeeskssskskssksttttttttaaaa*rrrrdddd ------5------ HHHTTPehheeereewwwkrrilllmmneeetttooEtttQQPPPlmuuaaaccceeekskksrttaaarrrddd Components of gas chromatography system 5890A AGPPPPGAPPPSS33GMCGMSSSC511E00CCUUU8PPCPPP8RROO1CCCGG44$55444445444411144---CC99SS5545411115777000--36--00000-AA-2444500000011B44C00000AA00AA0000044X0X0 GGPAXS1C8PuL1154t551o95110s60320yC0AA0sXtXem AhCACAAAIAAAIICACICIC}}ICAIgAIh}CC}}}nnnnnnnneaehhhhhhh/uu//hhuuuuu/76ttttttttasDDaDDeeeeeeeerrrrrrrttrrtttt6t8ttooooooooooooogoggoogggggse7e9ssmmimissimmmmammiirmmrrrrrrrdd3n0nnnnaaaaaaaaaaaamtaattajaaaataatmmtttaammtgtttaAeSeeeeiiioooiottptttttttttaooouorrroocreooooeoooepprrpprrulsffffnnntndtdggiggggrgggllallaaaotonieeeesrrorrrrrcrrcccsssesmggrgarraarraaaaaaaseoeeooeosaampppppppppaafvfffssAhhhhhhhmthhttttapwwwiwvvuclwwlwwwwwwwpvaaaiaaatnlgoliliriiieiriiirrevtvttttttttegaeeeshhhhhhrhhheaesavnmcFgPFFFcpsadaaaaIrIIIllpDDDDDpopsvmFligie....e.vIllpr,lDlraaalaEli.mrrvnyyCegmDiianvnanlalbaeedlnlttvessdFe,,cIFhaDaInep.Dadilt.leFadIrDya.uintolemt,atic * formerly Spectra-Physics : 243 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) PSBTUSFDY(T-S7P4E99C)IFIC SUPPLEMENT TO THE INHALATION ANALYTICAL PROCEDURE FOR Tobhtiasinsuedppflreommeanitrdseatmaiplsleasdcdoitliloenctsedanodnatmheenadbmoveentsstutdoyt.he procedure to be used for the GC assay of PBSF Tcohneceansstraayt,ioinnscowrpitohriantitnhge rtahnegaedodfit2io5ntso alOndOOamppemn.dments, is suitable for the analysis of PBSF, in air, at Details given in this supplement supersede those in the compound specific IAP. ["EFFECTIVE DATE f |~5 November 2000 Analytical standard NEBPSuuxaarpptmicpithreylyinedruamteber NT9SF6Cp-o7-oSt94n0S89st%9oa,rteP0((df1BPi8leSl2reF2fdluboyroMsualnfochlaensete2r-T4a%n)k) Chromatographs The analysis is performed using chromatograph 8. Carrier gas Helium (2.75 ml/min) Split vent Helium (24.5 ml/min) Split ratio 1:8.9 Detector temperature 150C Detector Range 0 Retention time PBSF approximately 2.7 minutes r 1" .*" i *a t l t 1 l Summary of method validation Manual injection Prarnegciesioofn1d0a0t0atsoh2o5wpedpmco. eTfhfiicsidenattsa oisflvoacraitaetdiownifthorthPeBMSFINo/f25le2sssttuhdayn d0a.t8a%. with standards in the : 244 : MIN 252/004312 APPENDIX A (Methods of sample collection and analysis for T-7499 - continued) Lah0Lrego.ei9smawap9isenie9ttcvs9tsteo8iqrvf4c,ueQoaatlyhnrnue.ecdasenLrnrteetOilrfgaaiQrctteiiavostaesinnoieodnronrLf(oLairmnssOtaailQltenys)odssiaffstroh,DdrawenPt(ie2tB1hc5.St3iaFo%tnonwiu(nni1Lltw0lOh0ebeD0iegr)ahpsnatepegrtmedeb)p1lyio0npt0teerh0aonerdttiourlaoecl5lgwey0rdeepassptsasimaolcocnoawc,nrerfpdaeotsla4rab.tp13ilo.ee%6an9ckahactaeno2rcdee5kfaf0pisr.pct5eaim1senpn.doptTanprhsomdee,f EFFECTIVE DATE : 10 November 2000 Calibration and Quantification Calibrate using standards with nominal concentrations 500 and 50 ppm. EFFECTIVE DATE : 16 November 2000 Preparation of standards Prepare standards in the nominal range 25 to 600 ppm. EFFECTIVE DATE : 17 November 2000 Calibration and Quantification Calibrate at 4 concentrations across the standard range. EFFECTIVE DATE : 2 December 2000 Summary of method validation Automated injections Prarnegciesioofn6d0a0tatos2h5owppedm.coTehfifsicdieantatsisolfovcaarteiadtiwonithfothr ePBMSIFN/o2f52lessstutdhyanda4t.a3.% with standards in the tLacroherofeeeseaQpafsefLuticracOtseniiqsevQtpuniefoatliyrcnaoe.ansfstedi0or.ean9Lgg9i(ram9Lei9snOis4tsiQ0toon)acfnfoaodnDnrcarePeeltyneBlastcSrtitaisiFv,otienowwneiir(ltlrLhoobOrfaesDssl1)ete/asctnsaobdrtnyeahcraetdhnpneotr1(tal2e.o3t5niw%otineatos2ilnltywat6hce0acei0sgerhpalttpnoeapgdwbmelel)i6anc0sehp0aeror1cto2dkr.ue22sgc85treaednpsadpsnamiadord.nc,,3Toh.hf6roore8ewrlLaeppitvempieoamriknt, : 245 : MIN 252/004312 APPENDIX B Chamber concentrations of T-7499 - individual sample values ExNpoos.ure 1 2 3 TWA tTimimee) i(SnThato2W04035me1u------r-113546pv2Arslae)l 1-2 T4W2305-----13546A 1-2 cwaleTciug24W35lh----a3546Atteedd faovreeraagceh:(LsGasoumrwmo4466665444486555554888611p---8u244443333389656424494776dlpioons2fge)o`wcceaigsCihohtneadmasbcfeoorn(lIclcnooGetwnner2222511111111111111111c111tors825r503896,6662688768687--68e8:7u5a31552804287796297457276n48dpttioor3sanetsi)o'n(s(apmpmpl)e (HGigr44444445oh66366675u38696d709po4se) - 4540- 55 444443655653- 562970 - 42- 8 conce44444n21111t- r19876ations weighted for Weighted concentration (ppm) = Concentration (ppm) Exposure x "Time weighting"(min) duration (min) tWhehemreidtphoein`ttismbeetwweeiegnhtcionngs'eicsutthieveressapmepctliivneg ionctcearvsiaolnbseitwfneoenadajdujsutsmtmenetnwtsatsomthaedeexposure system or : 246 : MIN 252/004312 APPENDIX B (Chamber concentrations of T-7499 - individual sample values - continued) Weighted concentration (ppm) Concentration (ppm) x "Time weighting"(min) Exposure duration (min) tWhehemreidtphoein`ttismbeetwweeiegnhtcionngs'eicsutthieveressapmecptliivneg ionctcearvsiaolnbseitwfneoenadajdujsutsmtmenetnwtsatsomthaedeexposure system or : 247 : MIN 252/004312 APPENDIX B (Chamber concentrations of T-7499 --individual sample values --continued) TW-----E---A---x----N-p----Too?89O.-s-tT*-S--u-i---m-ti-r--a-m-e---n-e----e-c)-- L---caw-ar-iS(dl-enhTMTTMc-aidtou22g04034523424W03535W5meW11s11seeeu^-l---------h------r-dd------vaaa13135535451p466434v62262r2AAItAtnnis-elaeae--)dl"dt-i-ofa'onvrereaagceh(:LsGasoumrw1om424244444446464554734323853563513p---u11135335168970657434847824730783d.l8pioons2fge,)o`wcceaigCsihhotanemdasbcefoor(nlIclcnooGetwnnerc1111111111111111111111111115ro11ts,r6e295522554222525454---25455355---4.:u.a671n867655869380897877d709206009pttoiro3saneti)so'n((spapmmp)le (HGigr444oh654u5d68po4se) 444423- 799 415240- .726 44444443127094- 2887204 444453- 304 444444444497375356788-.13889907240 concentrations weighted for Weighted concentration (ppm) = Concentration (ppm) Exposure x ''Time weighting" duration (min) (min) tWEhxehpemorseidutprheoesin`1tt0ismtboeet2ww0eeiwegnehrtceionmngs'oeincsiutdotiireveerdesbsapymeacputliitvnoegmioancttecedarvssiaaolmnbspelitiwfnngeoenadajdujsutsmtmenetnwtsatsomthaedeexposure system or : 248 : MIN 252/004312 APPENDIX B (Chamber concentrations of T-7499 - individual sample values - continued) ExNpoos."ure ll 12 13 '14 TsdWA tTimimee) iS(nMTMTTMhatI42o244WW24035203W350355me1s1s1s1Feee--u-------d---dd-------r----aaa15313133555644646246Fvp222AAArnnnslae)l cwaleciuglhatteedd (LGorwoudpos2e) 7444443544445435454333.3313455555540912607985 434334--588557 Chamber(IcnoGtner121111111R111111T1c111or40,554373344464e555455u.31550635097897n7609d779ptor3saet)ion (ppm) 1111111155544-54.91118209 64-.10 13-4.59 faovreeraagceh: sasummplionfg o`wcceaigsihotnedascfoonlcloewntsr:ations' (sample (HGigrmohudp-o-4s-e-)-------344443434434454443553458876309586943674566.113361850725592523606 34445449987479--3269692 -- concentrations weighted for Weighted concentration (ppm) Concentration (ppm) x "Time weighting"(min) Exposureduration (min) tFWEhixerhpsemtorsseiudatrpmheoespinl`1ett0ismotbfoeeet2waw0ceehwieggnehrrtcoeionumngps'oweincsaiutstothimrveeeadrnesbsuapyamelapcutliitvnoegmioancttecedarvssiaaolmnbspelitiwfnngeoenadajdujsutsmtmenetnwtsatsomthaedeexposure system or : 249 : MIN 252/004312 APPENDIX B (Chamber concentrations of T-7499 - individual sample values - continued) ExNpoo.s*ure 15 16 17 18 T1WA tTSimtiamneec) o1 acwraidS(lneThTTMMTcaitdou4420230234W55304WW52gWem351111sseeeu-----l------r-----d-dh------v11aa1a355334453vp66456224622rAAAAttnnisaleeae)lddtiofaonvreeraagceh(:LsGasoumrwo44m44444445465535324553453119p----u551331888052907746890252490dlpioons2fge)o`wcceaigCshihotaenmdasbceforo(nlIcclnGooetwnner221111111111c111111111211111o1rts900,578r7e455897559545555555-.555--u:63a111325n535335242253046045d760pttoiro3saneti)so'n((spapmmp)le (HGigr44464444444434444444445554441o5h26175555966585267578668408837--.u631313.868585d72760723903484866p2o4se) concentrations weighted for Weighted concentration (ppm) = Concentration (ppm) Exposure x "Time weighting"(min) duration (min) a WtEhxehpemorseiudtrpheoesin`1tt0ismtboeet2ww0eeiwegnehrtceionmngs'oeincsiuttothireveedresbsapym:eacp2utliit5vno0egmioan:cttecedarvssiaaolmnbspeltiiwfnngeoenadajdujsutsmtmenetnwtsatsomdiaedeexposure system or MIN 252/004312 APPENDIX B (Chamber concentrations ofT-7499 - individual sample values - continued) ExNpoos.1ure 19 20 TWA TtSitmiamnece) acwradiS(lenMMTThcaidtou24234505gWW03m1se1seeeul----d-----hd-r--vaaa13513564642vp2rAAttinnselaeae)dldtiofanovrereaagceh:(LsGasomurwmo24434233433335pu13218790090998dlpioon2sgfe)o`wcceaigsCihohtneadmasbcefoorn(lIcclnooGetwnner421c111111111t1ors2r904,6e66457535:5u6a13356n2849449d6pttioro3sanetsi)o'n(s(apmpmpl)e (HGigr444444oh254657u880d864po4se) 1444444474159435.11336643 2454.76 concentrations weighted for Weighted concentration (ppm) = Concentration (ppm) Exposure x "Time weighting"(min) duration (min) EWsyxshpteeormseurotehrseth1`e0timmtoied2p0woweiniegtrshetbimentgow'neiitesonrtehcdoenbrsyeescapuuetticovtmievasetaemidnptselarimnvgaplloibnccegatwsieoenns iafdnjuosatmdjeunsttsmetonttwheasemxpaodseure : 251 :