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ro Olay 202 3%i2iem From-Charles River Labs Arkus Div. i momar ARLE -- HYorosshaimn,By 108 10 Teter 215) 443.0587 asusnser oem pa 1096 ARGU;I S RESR EARC!H Charles River Laboratories Discovery and Development Services From: Raymond G. York, Ph.D., DABT=e -- ! To: John Butenhoff, Ph.D., DABT ComPphaonnye:: 3(6M51C)o7r3po7r-a1t9e62Toxicology Fax: (651)733-1773 Date: 01May 2002 Total Pages: 22 i RE: Protocol 418-020 iAttached is the information on covariant analysis for study 418-020. analyses were done for females and two for males. So =i 3S3R zpg2:8 = = Four different Females: 1. Sexual maturation w/ gestational age at sexual maturation as covariate 5 2. Sexual maturation w/ body weight at maturation as covariate : 23.. EEssttrroouuss ssttaaggeess ww// dgeasyta6t4iopnoasltawgeaenaitnsgtabrotdoyf wesetirgohutsatsesctoivnagriaastecovariate I 3 5&2 Males: . F285 21.." SSeexxuuaall mmaattuurraattiioonn ww// bgeosdtyatwieoingalhtaagtemaattusreaxtuiaolnmaastucroavtairoinataes covariate 2=S We are mating defining in the P "gestational age" in this context as generation until evidence of sexual the time in maturation days from evidence of in the F1 generation. Tdehleivreeraesdoanfitnegr 2is3rdatasysnaotfurgaelsltyadteiloinvetreyndafttoerw2e1igtho m2o3rdeayasndofagpepsetaartitoon.dePvuelposptfhaasttaerre tthheanfiprsutppsuopfwtohuelsdabmee2s3egxeasfttaetrio2n1aldadyasysofolgdesatnatdiotnh.e Both pups would be 1 day old second pup only 21 gestation but dmaoyrse oplud.psIfwaerteestdesluibvserteadnceearslhyo(rrteednuecdedprpeagrtnuarnitciyon(rleendgutche)dinleonngethgorfougpe,sttahteiopnuoprs ifmay reach sexual maturation a day gestation or parturition length. oItrwtowuoldbeafporpeeaprupthsatinthaegtreosutpsuwbhsitcahnhcaedwaaslocnaguesring a delay in the time sexual maturation, spent in utero. By when using it actually affected the matemal animal by each pup's age from time of conception to changing sexual maturation as the covariant, the length of the stay in utero Is factored out. M59 263 4 DieMar-200121102A:2G7oIm, SISFVr1omA-CAharias RivAaer LaVbEsDArUguIs EDivSu TENE bar Ssups SernOsTtr +po Feat patency of F1 generation male and female rats, respectively, using gestational age at sraetxsu)aolrmdaatuyroiftyvaagnidnatlhepabtoednycywe(ifgehmtalreecroatrsd)eadsotnhethceovdaaryiaotfesp.reputial separation (male `The Analysis of Covariance was also used to evaluate estrous cyciing of F1 generation bfeomdaylweeriagths,t uassinthgegecsotvaatriioantaesl.age at the start of estrous testing and day 64 postweaning hPelseiatsaeterteoviceownttahcetamtet.ached information and ifyou have any questions, please do not ALCO oa orwena (NET'S TEST) WATH 800VELGH AS 10126vont,sarc4, COURRIATE 702 1 ; { SEX GRP MW ean so 2 " IF1TPI oRHn60 uEd34.a8en8n33 2baR.0eR7e5e6.2 Fg : woTPowBuod sdsoaaiEmmmEs nsiiaummmmse Roane mab. :ii f Stansticntly ifferen fran Control sro 1 (950.05 ACOA (UNEIT"S TEST) WLTH GESTATION AGE AS COUNRATE C9 i seFx aGoIPveodga udhnmaemann jmosmoems 2 ER oon 5 fan b1 og2 sie 48.4615 mm 2.13708 3oDoHod57 4da90a3R3n33 3Ed14em29s5 E :g 0) Wo scaiseicaTly significans differences hen compared with Concro ra 1 50.05) 2 3 ACOA CHRDATES T73[WrE--ny 08 SY WGN 41502C6AoTnyE, arch 4, 200 2 ii wCo 1aoPronomon ww soeaun osumee i oPIRRonmoieam gE teesmo .i con OwoRMgEowTESTS Ton H ESwTRUSwAGEwAS COMATE : F HT1Poopno3209 mm 45..6063646478 sgeuem 00..3808546058 .Ei 5 8 539286 0.68526 *(1) 3 *G ( signS ifich antIly dEE ifSfEpeereennt, ffSroonmCConCTtOrSoAl008grouOp 01 0(p<0.05) atropa erso tshe esignmificaancegolf eshe gglouved, z AEECR ER SE SIRENSSA Sl lu Tt oly Hee i 5 I ANCOVA WITHDAYBOODFY WMAETIUGRHITTYAS COVARIATE 10:26 Monday, March 4, 2002 5 2 Sdehnsorrt:er : SW EShhrn inwe Fa iOTn8 7i t BBRYRR Inmy, Ta, :: de pteClass Level Information wr Tn ares :i SwEHrtprev 3}} bS F servatE iom -- ns vs. be. 28--28 Eii 2 < I ANCOVA WITHDABYOODFY WMEAITGUHRTITAYS COVARIATE 10:26 monday, March 4, 2002 6 < Th mixed procedure = Covariance Parameter Estimates. i tna i cov parm Estimate i Fit statistics i SM2 soees nLog maLitkeLlieheoo,d, 1B20i6.e4 i:i BIC T(semall3erTesitssboeftteFri)xed effect2s52.0 i& o-yEffect oFLPoommDF oEuvoalRwuee coPrm>mF : [PA ere ar eee UH `standard or cue ees agaomrr 311 mpEmHRssE osoaemmm Mowmmo moommeag ccouommn F g g 5 z ANCOVA WITHDAYBOODFY MWAETIUGRHITTYAS COVARIATE 10:26 monday, March 4, 2002 7 EH The wed procedure Lest savas ans 2 fre cow mcs scanUdaHed or cv negou = Per GROUP 5 wae guy om we com oi3ff6e2r4e1n0c3es of L0e.a2s8t71spares#2means 12680 <.0001 i :i= fect Gow ow escute stanCdaCrFd or tule reo fo Abeer Age ii - aaGxosew b32 o1l1 uBiomEsmEn oOEosRRReEMfmatom n0L23wHB0 feeUuomNmnsEes ahaommmmemewewmss 0eodonommem iiz: 2 : 3 8 E ACOA NIToarB0OoYf wWEmXuGHmTTyAS COWRIATE 10:26 onday, varch 4, 2002 8 i The Wied procedure Model tnforeation i= SCoSoupvtaaEirisnaestnecevwSaerraaicteure vruSoeauaaian Residual Variance vethad brofile 37 FDiergerdeesErroefctFsroSsCdonvovtehtohdod hRoeosti)duBaalsed 2:: Class Level. Information Cass Levels values : on 5 1234s i oimensions : cSCSoouvljaaoermtmi:sanscIen paraeters % :1i STRGshoietEerariOvaGatcuitspooeennrr:svastbvuioabtojlneicsutsed Bs2i2t0lo0 i= E q I acon wroussoobf wVuEnRrTryXs counzaTe 10:26 vontey, arch 4, 202 9 i: he uixed procedure : Covariapncie pearameter i vem etme 7 resin) 70m i Fit statistics 228s Lan Likelhood as iz eAScamGmaietorr 1siebebnteere) 13i i4e60d4 ii Tope 3 Tests of Fixed Effects i crrect WO eve war arour PTOoEm BpEa ocoom ! Least squares wears ef cow ode seaCndUarRd or tvle. ees i ii aadoawnr 33 wwsaaoesa oSeEmnEs mBoomOWo mEoDnhco ooRm lo EE Sp F aloe o r s :5:: ANCOVA WITH BODY WEIGHT AS COVARIATE. wee won or come BE Differences of `standard Least ce oes Squares Means 0 a te :- i 11 MEOH OB Boe Em : i 2 n - acon ver oSaERoErIsOaLry A5 count20:26 ondsy, ware 4, 2002 11 i= The Wises procedure : ode) nforsation Lf oeupsene dseence a variat ble wSiorik.adaran = SDpFeeErResEiEoIafEFItreIIesionwebtabtohdod ThRSeorsaailfdiubnaealses :: cisss Level formation i Aus reels ates cour s 1234s i cS GSioinvfamerce ietasnce parameters 3 83;: BBELer subtece 2n0 OTobtsaelrvaSthisoenrsuatHiootnsUsed 108 & 3 12 ancoun wer SoxEyEoRrTIwaSrLurHaErEy As comma10:26 onday, varch 4, 2002 32 io he wed procedure covariance armecer i= vemEstinatess me 4= Resta 0.2779 3 Fit statistics i MHA-L2ECReGesaaLotg mlerLi1k3eRlbiehEtotoed,r) ida45a7d3.]3 :iEI Type 3 Tests of Fixed effects i EsEfefeactc oF1POmBOeF OeFIvwalEeue PcarommF Lens wares means Effect Gow estimate StanEdramredr oF tvalue pro lc E3 a saaex}1 Rspauutegt mBdmMmsE m mps oW mwam ocomo 3 5 3 sco win aCayEoEr OwaLrnAGsE As comnaatt10:26 nonday,waren 4, 2002 13 i* he mined procedure Lesst squares means o fre wow ecimate staCndUaErSd 0 tule pro ltl = ara--4 mBaRmS Lsomme mme o@soRn ocoNm 3 oFfferences of Leas squares weans fret cor cow estimate StanCdUaRrRd or cule beslt austen Ade i:: wa Gaoawwn 231sd1d1 oS0C.mA1h6eE6e3 So0C.0RR9m7ME32 2BH B7EE6 H L1N7Gh1 SDG0.iRR08eR8N7 oMnMumnmmmercmemwss 0GES.2O6RA3R9 3z!x z i 1% I forme J LR-- yt ony, wre 4, 20 14 =i osoioetrreeerse :i i E Ba EedReL ri,n oirn i. EEE. EE i!: Class Level Information 2 ws Class Levels vs1ae2lu3ies4nss ii BE3Nr5kwaEdspe agi} . S Enrt. %2 zi 2 5 5 z coun 1TH GoxEySToArTIwOuNrAuLnArGrEy AS COURIATE20:26 vonday, varch 4, 2002 15 E2 The mixed procedure =o covariaincae ptaraneter ova estimate =: residual 0.3806 3 Fit statistics HceGsmiolgerik1e5rbtoeoesr) a 204 : ii ME ec Cae nter Tsebteettresr) 1Bi4e E Type 3 Tests of Fixed effects i cetece TOR rune esr F sa5ae 1POaWm TsEmeRw ocuosm Least squares means eofecc Gow eviete stanUdEamredr or tvae ers lel i a Gaoown i1 pamionames ccoaEmbRms gIBm owHsanEe ciom s i 5 1 I ou or mua 10:26 bonday, arch 4, 2002 16 = ANCOVA WITH GESTATIONAL AGE AS COVARIATE : he ised procedure 5 ] Lesst squares wens effect ow ee stanCdeadredr or twale pes lel - agonw 41 B mamEe EoRm B am osmme ocoNm i oifferences of Lease squares Hens I Elect Gow ow cima stanCdHarRd UE vila pro tl Austen AP :i Gaaarowaxve 21 3 o1 b i1 uB0C .e1m87a2 R 0Su.1H1e8RE 3 2IM6RE8I 1Sh.J5RI8 0MDi .MA14EE7J mnouammetmt-inesu 0S2.3A188R3 3:z 5 7 I | monoSmcEEcRn Bsoewe st et, rh 4,00 3 riy he as more : Model Information |] BBE RE BISSAEwE EoE AeeeR., HEEoih. iii:: ] ss eet rion : EwEn 3 nes :: pgSaiointsssptrmaosses ;}: BR we ul .3 E E EEm RyI. E 8i iii : z ig 7 wnTBNT ETITTBLoars 1 rs, Wh 3 38 I ens rate Covariance Parameter ctoiv otine i- Estimates E Fit Statistics 3 R S 2RensLooigC Lli ikeleihn oN o,d ] 3oW81Ol.0 iii Type 3 Tests of Fixed Effects i J owse "PBBB cuomm owurem : ase Spares sr coe op wie PIE Standard 0 cue mem i w = i Group 1 4Hf.74Zm08 0m o.16t45 3B3E9 2O8o.E8E3 o<Om.00m01 z 5 19 z confSmE-cosEus spiresae 10:26 end, rch 4, 2002 19 i. heeasmixaerd eprsoceadnurse i wre oge ne soommee soaT nUiHarRd eow e cue eo af Igoe mong om i7! | offFereSnacpeisarodf Least Squares Heans Ii Ecggafoafaaewrct G3t1ofROUoP11}G1ROUP Esosstsimmamaagamtne EeGesrarmdhomr aoooOummmF tvaotbnlummee aProo>onhuhlntl Aamaadjeummstwemeoenst AoooudnujlimnP ii? 2 z wo I ANCOVA WITHFoDAeYs64-30e0sYtiWoEuIsGHsTmASsCOVARTAT1E0:26 onda,"arch 4, 2002 20 = | i " | savin reste SEXF =sensemmeeasmsemeeessnamnssesneesseme anne &: ode) Tforaation SEoatpieiaettnonvahaercrhioee SEER Tartine ethos fgBaHarrgon ae Hata = :3z Siri oF Fron vathod esa : Class Level snformation ams lets values :: aw 3 trees i oisensions CSSSioohvujrraeicoeatnschelparaneters 1$; . IheSarRvaTSatcdietoptoneernsnstWWuibsjoeenct, b0i3] ii: i 2! = AnconaroOmAes64 scostyioWusERsTa5cescomarat1y0:26 wordy,sarch 4, 2002 21 i I ------ & The Mixed procedure 2 Gouramis CovarEisantciemaPtaersameter i = esi 01am Fit statistics 3 i MHH2ceeEGsatogTteLIie 1k5eTLihtee aeo,dr) hR230'ees6 :2: Tope 3 Tests of Fined effects i | oeeEfrofesscot oFPpoommOF oFsvaelRuwe ooprossmF ease stiares wens fre Gow emt StanCdCarHd or twee res saoanr}1 s coy f om Mom RmEa cooom g a3 mE te MER oS 22 2 i rues- coms smicts 10:26 vouday, arch 4, 2002 22 ANCOVA WITH DAY 64 BODY WEIGHT AS COVARIATE y i | sm : rec con Least eee SSqcuaCanrOaeHrsNaueans oe cvie seo : aaosr 3s sos OoMwEn aWM wBnE coamm :i || dre oor con somDeiffersetncaenUsdTaroEdf Least Squares Means 0 cue roi asm age :i= fgF worAn E 13 13 SSGOiMaEtE GGa2MmamEtmM da1RR pIg1ata osgsainenne nomsmmemeemen gogoupuwnemn i:i i 23