Document dDavJGvEwB766r7jd957yZVMQ

CORNINGHazleton Sponsor. 3M SL Paul,Minnesota 19@ 190 F9 5 FINAL REPORT Study Title: 5 DailyDose OralToxicitSytudywithT-6669inRats Author: SusanM. Henwood, MS, DABT Study CompleflonDate: April30, 1997 PerformingLaboratory: Coming HazletonInc. 3301 Kinsman Boulevard Madison,Wisconsin53704 LaboratoryProjectIdendrication: CHW 6329-197 Page I of90 CHW 6329-197 QUALrff ASSURANCE STATF2AENT Thisreporthas beenreviewedby theQualityAssuranceUnitof Coming HazletonInc.,in accordancewiththeFood and Drug Administratio(nFDA) Good LaboratoryPractice Regulations2,1 CFR 58 and theOrganisationforEconomic Cooperationand Development(OECD) Principleosf Good LaboratoryPracticeC,(81)30(Final)T.he fbllowinginspectionwsere conductedand findingsreportedtotheStudy Directorand management Writtenstatusreportsofinspectionasnd findingsareissuedtoComing Hazletonmanagement accordingtostandardoperatingprocedures. InspectionDates From TO 12/17/96 01/21/97 02/28/97 03/26/97 04/22/97 12/17/96 01/21/97 03/04/97 03/31/97 04/23/97 Phase ProtocolReview Ne=psy Data Review Report Review ReportRereview Date Reportedto Date to StudyDirector Management 12/17196 01/21/97 03/04/97 03/31/97 04/23/97 12/17/96 01/21/97 03/04/97 03/31/97 04/23/97 :sh4A"t@ Representative QualityAssuranceUnit Date 2 STUDY EDENTIFICATION 5 Daily Dose Oral Toxicity Study with T-6669 in Rats CHW 6329-197 Test Material Sponsor Study Monitor Study Director Study Location Study Timetable Study InitiatioDnate In-LifeStartDate In-LifeEnd Date Study Completion Date T-6669 3M Toxicology Services Building220-2E-02, 3M Center St Paul,Nfinnesota,55144-1000 Roger G. Perkins,PhD, DABT 3M ToxicoloU Services Building 220-2E-02, 3M Center St.Paul,Minnesota, 55144-1000 (612)733-3222 Susan M. Henwood, MS, DABT Coming Hazleton Inc. P.O. Box 7545 Madison, Wisconsin 53707-7545 (608)241-7221 Coming Hazleton Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 December 20,1996 January 2, 1997 January 21, 1997 April30, 1997 3 Toidcology SusanNL Henwood, MS Diplomate,ABT StudyDirector IloniasRyan StudyToxicologist SandraL Haley Study Coordinator To)dcologyOperations Pamela D. Gabris Supervisor Small Animal Toxicology Dose Formulation Dixie K Bushee Supervisor Quality Assurance SherryP, W. Petsel Manager CHW 6329-197 KEY PERSONNEL Laboratory Aninud Medicine Cindy J.Cary,DVM DiplomgLteA,CLAM Supervisor cuwcal Pathology RobertL Hall,DVK PhD Diplomate,ACVP ClinicaPlathologist Ronald Markevitch Supervisor Anatomical Pathology Thomas E. Palmer,PhD AnatomicalPathologist Jack Serfort Supervisor Necropsy 4 CHW 6329-197 CONTENTS Page QUALITY ASSURANCE STATEMENT ...........................2 ....... STUDY IDENTIFICATION ..................................3........ KEY PERSONNEL .......................................4........ ABSTRACT ............................................8....... OBJECTIVE ...........................................10....... REGULATORY COMPLLANCE ...............................1.0 ....... MAINTENANCE OF RAW DATA AND RECORDS ...................1.0...... TEST MATERIAL AND CARRIER ..............................10....... Identificatiaonnd Characterizatio.n............................10........ StorageConditions .....................................I.I....... Reserve Samples .......................................I.I....... Disposition..........................................I.I ....... SafetyPrecautions......................................I.I....... TESTSYSTEM .........................................I.I....... TestAnimal ..........................................II........ Justificati.on.........................................I.I ....... Identificati.on........................................1.2 ....... STUDY DESIGN ........................................1.2....... Dose LevelSelectioCnriteria................................12....... PROCEDURES .........................................1.3 ....... Acclimation..........................................13........ Housing and Maintenance ..................................13........ Placementof Animals intoGroups .............................14........ Dose Preparation.......................................1.4 ....... Dose Analyses ........................................1.4....... Dose Administration.....................................1.5....... AntemortOebmservati.o.n.s..............................1.5....... BodyWeight.s.......................................1.5....... 5 CHW 6329-197 CONTENTS (Continued) Page PROCEDURES (Continued) Blood Sample Collection.s .................................1.5..... PWmitoCyolAOxidasAectiviAtnyalys(iDsay6)....................1.6...... ScheduledSacrific(eDay 20) ................................16...... StatisticAanlalyses ......................................17...... RESULTS ............................................17...... Antemortem Observationsand Survival..........................1.7 ..... Body Weights and CumulativeBody Weight Gains ....................17...... Pal@toylCoA OxidaseActivityAnalysis.........................18...... AnatomicalPathology....................................1.8 ...... CONCLUSIONS ........................................1.8...... SIGNATURES ..........................................1.9 ..... REFERENCES .........................................2.0...... PATHOLOGY COMMENTS REPORT ....................................2.1 ...... ON THE DATA ................................2.3...... CODES, ABBREVIATIONS, AND UNITS .........................2.4...... GeneralCodes and Abbreviations.............................2.5...... Codes forClinicaPlathology ................................27...... Abbreviationasnd UnitsforClinicaClhemistry.....................2.8...... Codes forAnatomicalPathology .............................2.9...... TABLE I Summary ofAntemortem Observations........................30...... 2 Summary ofBody WeightData (g)..........................3.1...... 3 Summary ofClinicaClhemistryData .........................3.4 ...... APPENDIX A ..........................................35...... ProtocolDeviations.....................................3.6...... ProtocolTP6785 .......................................37....... MaterialSafetyData Sheet .................................5.0...... 6 CHW 6329-197 CONTENTS (Continued) page APPENDIX B ..............................................5.7........ IndividualAntemortem Observations ..............................5.8...... IndividualBody Weight Data (g) ................................5.9....... IndividualBody Weight Gain Data (g) .............................6.2...... APPENDIX C ..............................................6.6........ IndividualClinicalChemistry Data ................................67....... APPENDIX D ..............................................6.9........ IndividualAnimal liverWeight Values (g) ..........................7.0..... IndividualAnimal PathologyData ................................71....... 7 ABSTRACT CHW 6329-197 The purpose of thisstudy was to assessthe oml toxicityproduced when the testmaterial, T-6669, was administeredby the oralroute(gavage) to male ratsfor5 consecutivedays. Male Crl:CD*(SD)BR VAF/Plus'oratswere assignedto fourgroups (10/group).Each group receiveddose preparationscontainingthecarrier(reverseosmosis water) or 5.0, 14.0,or 42.0 mg of testmaterial/kgof body weighttday(mg/kg) for5 consecutivedays at a dose volume of 5 niLJkg. Five animalstgroupwere terminatedon Day 6 (theday followingthe finaldose)forhepaticpalmitoylCoA oxidase activitydeterminations; remaining animals were sacrificedand necropsiedon Day 20. Food and water were provided ad libitum.The animals were observed twice daily (a.m.and p.m.)formortalityand moribundity. Additionallye,ach animal was removed from itscage and observed forclinicalsignspredose and atapproximately 1,2.5,and 4 hours aftereach dose administrationand dailythereafter.Any abnormal or unusual findingswere recorded.Body weight data were collecteddailyon Days I through 20. Blood samples were collectedon Days -2 (pretest)6,,9, 15,and 20; serum and cellular fractionswere separatedand shipped to the Sponsor. On Day 6,fiveanimalstgroupwere sacrificeda,nd the liverwas removed from each animal and weighed. The rightlateral lobe of the liverwas collectedfrom each animal and weighed, then analyzedfor palmitoyl CoA oxidaseactivityt;heremaining livertissuewas alsocollectedand weighed. On Day 20, the remaining fiveanimals/group were sacrificedand subjectedto an abbreviated necropsy;the liverwas collectedfrom each animal and weighed. Remaining livertissues collectedatthe Day 6 sacrificaend the whole liverscollectedat the Day 20 necropsy were shipped to the Sponsor. All animals survivedto the respectivescheduled sacrificeT.here were no test material-relateadntemortem observations.Mean body weight gains forniid-and high-. dose animals (14.0and 42.0 mg/kg, respectivelyw)ere significantlryeduced duringthe dosing period;Day I to 6 body weight gainswere 23, 23, 0,and -4 g for the control,5.0-, 14.0-,and 42.0-mgtkg groups respectively.In general,rrlida-nd high-doseanimals showed a recovery in body weight gain over the firs3t days aftercompletion of the dosing period.The overallmean body weight gains(Day I to20) were freeof significant intergroupdifferencesand were similarfor allgroups. 8 CHW 6329-197 Adn3Wstudon ofthetestmaterw atalldoselevelswas associatewdithhigherhepatic palmitoylCoA oxidaseactivityM.ean palmitoylCoA oxidaseactivitioensDay 6 for controlanimalsand animalsgiven5.0,14.0,and 42.0mgtkg were 5,24,39,and 39 IRJ/G, respectively. There were no testmaterial-relatmeadcroscopicfindings. Based on significanitncreaseisnthelevelsof hepaticpalmitoylCoA oxidaseactivitya,n indicatioonf peroxisomeproliferatioant,alldoselevelstestedt,heno-effecltevelof T-6669 islessthan5.0mg/kg when administeretdomale Crl:CDO(SD)BR VAF/Pluse ratsfor5 consecutivedays. 9 OBJECTIVE CHW 6329-197 The purposeofthisstudywas toassesstheoraltoxicitpyroducedwhen thetestmaterial, T-6669,was administeredby theoralroute(gavage)tomale ratsfor5 consecutivedays. REGULATORY COMEPLL4NCE AU aspectsof thisstudywere inaccordancewiththeUnitedStatesFood and Drug AdministratioGnood LaboratoryPracticReegulationsforNonclinicaLlaboratory Studies,21 CFR 58,and theOrganisationforEconomic Co-operationand Development Principleosf Good LaboratoryPracticeC,(81)30,withtheexceptionthatanalysiosfthe testmaterialmixturesforconcentrations,olubilithyo,mogeneity,and stabiliwteyre not conducted. MAINTENANCE OF RAW DATA AND RECORDS Originalpaperdataand a copy of thefinalreportwillbe retainedinthearchivesof the WisconsinfacilitoyfComing HazletonInc.(CHW), forI yearfollowingthesigningof thefinalreporl One yearafterthesigningofthefinalreportt,heSponsor willdetermine thefinaldispositioonf thematerialsM.agneticallyencodeddatawillbe retainedat CHW. TEST MATERIAL AND CARRIER Identificatiaond Characterization Test Material.The testmaterialT,-6669 (FC-143),LotNo. 235,isa whitepowder and is93% to97% ammonium perfluorooctanoatIet.was receivedatCHW on September 25, 1996. Informationon synthesismethods,stabilitcyo,mposition,or othercharacteristitchsat definethetestmaterialison filewith theSponsor. 10 CHW 6329-197 Carrier.Ile carriewras reverseosmosiswaterand was providedby CHW. Informationon sourceand treaunentof thereverseosmosiswaterison filewith CHW. StorageConditions The testmaterialwas storedatroom temperature.The carriewras obtaineddirectlfyrom thein-housewatersystem underambientconditions. Reserve Samples Reserve sampleswere notrequiredtobe taken. Disposition Remaining testmaterialwas returnedtotheSponsoron March 10,1997. SafetyPrecautions Personnelinvolvedwith thestudywore theapparelrecommended by theCHW TestMaterialSafetyCommittee (TMSC) basedon theMaterialSafetyData Sheet (MSDS; Appendix A). CHW StandardOperatingProcedures(SOPS) appliedforany apparelnotcoveredby theTMSC recommendations. TEST SYSTEM TestAnimal Male Crl:CDO(SD)BR VAF/Plus* mts were obtainedfrom thePortage,Michigan,facility of CharlesRiverLaboratoriesI,nc.,on December 5,1996. The animalswere approximately8 weeks oldand weighed from 303 to395 g atinitiatiofntreatment (seeProtocolDeviationsA;ppendix A). Jusflrication The mt isfrequentluysed insafetyevaluatiosntudiesasa representativoef a rodent species. I CHW 6329-197 Identification Each animalwas assigneda temporarynumber upon arrivalB.eforeinitiatiofn treatmenta,n individuallcyoded passiveintegratetdransponderwas implantedintoeach animal.Afterrandomizationforplacementon teste,ach animalwas assigneda permanentnumber, and thetransponderwas coded withthepermanentnumber. Alldata foran animalarerecordedunderthesenumbers. STUDY DESIGN Animals were assignedto thestudyaccordingto thefollowingdesign: Group I (Control) 2 (Low) 3 (Mid) 4 (High) T-6669 Dose Level" (mg/kg) 0.0 5.0 14.0 42.0 Number ofAnimals Male' 10 10 10 10 a The controlgroup was giventhecarrie(rreverseosmosis water)only.Ile dosevolume was 5 ml]kg forallgroups. b Fiveanimalsineach group were sacrificefdorhepatic palniitoCyolA oxidaseanalysiosn Day 6. The remaining animalsineach group were observedforreversibility, persistenceo,r delayedoccurrenceof toxiceffectusntil sacrificoen Day 20. Dose Level SelectioCnriteria Doses were selectedbasedon theresultosfan acutetoxicitsytudy.Intheacutetoxicity study(CHW 61001760),animalswere dosed withT-6669 atconcentrationusp to 500 mg/kg. Mortalitywas notedforanimalsgiven500 mg/kg. At 250 mg/kg,clinical signswere limitedtored-stainefdaceand wet urogenitaalreafortwo offivefemales;all 12 CHW 6329-197 animalssurvivedtothescheduledsacrificeT.he highdose of 42.0mgtkg selectedforthis studyisone sixthoftheacutetoxicitnyo-adverse-effelcetvelof250 mglkg. ne low- and mid-doselevelswere selectedtoprovidean adequatedose response. PROCEDURES This studywas conductedinaccordancewith CHW ProtocolTP6785 dated December 20,1996. The protocoland protocoldeviationasreinAppendix A. Acclimation Fiftymales were receivedon December 5, 1996,and acclimatedinAnimal Room 512A for28 days beforeinitiatioofntreatment.In generala,nimalsinthisshipmentappeared healthy.During acclimationt,heanimalswere examinedforabnormalitieisndicativoef healthproblems,and body weightswere recordedforallanimalsatrandomization. Housing and Maintenance Animal Room 512A was used forthisstudy.Environmentalcontrolsfortheanimalroom were settomaintain19* to25*C (66a to77 OF),a relativheumidityof50% +')0%,and a 12-hourlight/2I-hourdarkcycle. The animalswere housedindividuall(yexceptforthefirsstixdaysfollowingarrival when theanimalswere group-housed)instainlesssteels,creen-bottomcages. CertifieRdodent Diet#5002 meal (PMI* Feeds,Inc.)was providedad libitum.The lot numbers arerecordedinthedata.The dietisroutinelaynalyzedby themanufacturerfor nutritionaclomponents and environmentalcontaminants. Water was providedad libitum.Samples of thewaterareanalyzedfortotaldissolved solidsand specifiemdicrobiologicaclontentand forselectedelements,heavy metals, organophosphates,and chlorinatehdydrocarbons.71beresultsareon filewith CHW. There were no known contaminantsinthefood orwaterthatwould have interferewdith thisstudy. 13 CHW 6329-197 Placement of Animals intoGroups ne animalswere examinedby a laboratoraynimalveterinariaonn December 31, 1996, and found tobe suitablfeorstudyconsiderationO.n Day -2,animalswere rankedby body weighl Animals withbody weightsexceeding:t2standarddeviationsof themean body weightand thosewithsignificapnhtysicalabnormalitiewsere excludedfrom selectiorTlhe number ofanimalsavailablfeorselectiownas reducedtotherequired number by excludinganimalsattheendsof thebody weightrange.The animalsselected forthestudy(40males)were assigneda computer-generaterdandom number and allocatetdogroups (I0/group)accordingtotherelativreankof therandom numbers. Group mean body weightswere analyzedusingBartletftsestforhomogeneity ofvariance atthe5.0% probabilitlyevel(Winer,1971)and found tobe homogeneous. Animals notused forthestudy(10males)were sacrificeadnd discarded. Dose Preparation Carrier.The carriewras reverseosmosiswater. TestMaterial.Dose concentrationwsere basedon thetestmaterialassupplied.Dose preparationwsere mixed dailyfordose administration. Each dose levelwas preparedindependently.The specifieadmount oftestmaterialwas weighed. A portionof thecarriewras placedintoa labeledcontainerand thetestmaterial was added and then mixed with a polytronhomogenizeruntilthetestmaterialwas suspended.The mixturewas thenplacedintoa calibratecdontainerand theappropriate volume ofcarriewras added toachievethedesiredconcentrationD.ose preparations were mixed foratleast10 minutesusinga magneticstiprlateand stirbar.Allcompleted dose preparationsappearedtobe solutions. The dose preparationwsere keptatroom temperatureuntildosing. Dose Analyses Dose analyseswere not done. 14 CHW 6329-197 Dose AAmini tration Gavage was usedbecausehistoricaltlhyeoralroutehasbeen therouteof choicefor administerinagknown amount oftestmaterial. The dose preparationwsere administereodnce dailyfor5 consecutivedays. The dose preparationwsere givenata dose volume of 5 mLJkg ofbody weight.Individuadloses were calculatebdased on themost recentlyrecordedbody weights.Animals were dosed atapproximatelythesame timeeach day. During dose administratiohno,mogeneous test materialsolutionswere maintainedusinga magneticstiprlateand stirbar. Antemortem Observations The animalswere observedatleasttwicedaily(am. and p.m.)formortalitaynd moribundity.Signsofpoor healthor abnormalbehaviorwere recordedas theywere observecl Predose and PostdoseObservations.Each animalwas removed from itscage and observedforclinicaslignspredoseand approximately1,2.5,and 4 hoursaftereach dose administratioannd dailythereafterA.ny abnormalorunusualfindingwsere recorded. Body Weights Individuablody weightdatawere recordedon thefirsdtay of treatmentand daily thereafter. Blood Sample Collections Blood samples were collectefdrom eachanimal on Days -2 (pretests;eeProtocol Deviations)9,,and 15 and atthescheduledsacrifice(sDays 6 and 20). Animals were not fastedbeforebloodsampling.Blood samples(approximatel1y.5mL) were collected from a jugularveinofallanimalspretesatnd on Days 9 and 15. At thescheduled sacrifice(sDays 6 and 20),as much bloodas possiblweas collectefdrom theposterior vena cava from each animal.At sacrificteh,eanimalswere anesthetizewdith sodium pentobarbitableforebloodsample collection. 15 CHW 6329-197 Followingcollectiont,hebloodsampleswere heldatroom temperatureuntilcentrifuged. The serum and cellulahrwdons were separatedand storedina freezersetto maintain -20*C:tlO* untilpacked on dryiceand shippedtotheSponsor on January2Z 1997. 71be Sponsorisresponsiblefortheretentioonr dispositioonfthesesamples. PalniitoyCloA OiddaseActivitAynalysis(Day 6) At theDay 6 scheduledsacrificfei,venonfastedaninmlstgroupwere anesthetizewdith sodium pentobarbitawle,ighed,and exsanguinatedinrandom orderfollowingtheblood sample collectionsT.he abdominalcavityof each animalwas opened,and theliverwas removed and weighed. The rightlaterallobeoftheliverwas collectefdrom each animal, weighed,and was flash-frozeinnliquidnitrogent,henstoredina freezersettomaintain -70*C 10* untilanalyzedby CHW forpalmitoylCoA oxidaseactivity. The remaininglivertissuewas collectedw,eighed,and storedina freezersettomaintain -20*C l0 * untilpacked on dryiceand shippedtotheSponsoron January13,1997. The Sponsor isresponsiblfeortheretentioonr dispositioonf thesesamples. Animal carcasseswere discardedwithoutfurtherexaminationafterlivertissuecollection. Scheduled Sacrifice(Day 20) On Day 20,theremainingfiveanimals/groupwere anesthetizewdithsodium pentobarbitalw,eighed,exsanguinateda,nd subjectedtoan abbreviatedgrossnecropsyin random orderfollowingthebloodsample collectionsT.he necropsyincludeda macroscopicexaminationoftheexternalsurfaceofthebody;allorificesa;nd the cervicalt,horacica,nd abdominalcavitieasnd viscera.Allabnormalitieosbservedwere recorded. 'Me whole liverwas collectedw,eighed,and storedina freezersetto maintain -20*C:tlO* untilpackedon dryiceand shippedtotheSponsoron January27,1997. ne Sponsorisresponsiblefortheretentioonr dispositioonf theselivers. No additionatlissueswere collecteda,nd theanimalcarcasseswere discardedafter necmpsy. 16 CHW 6329-197 Stadsfical Analyses Only data collected on or afterthe firstday of treatment were analyzed statistically. One-way analysisof variance[ANOVA (Winer, 197 1)]was used to analyzeDay I body weights;cumulativebody weight gains(Day I to 6 and Day I to 20 ),and pahnitoylCoA oxidase levels. Levene's test(Levene, 1960) was done to testforvariancehomogeneity. In thecase of heterogeneityof varianceatp :50.05,transformationswere used to stabilizethe variance. ANOVA was done on thehomogeneous or transformeddata. IftheANOVA was significantD,unnetes multiplecomparison t-tes(tDunnett,1964) was used forpairwise comparisons between treatedand controlgroups. Group comparisons were evaluatedatthe5.0% two-tailedprobabilitylevel.Groups 2 through4 were compared with Group I (control). RESULTS Antemortem Observations and Survival Antemortem observationsaresummarized inTable 1;individualdataarein Appendix B. All animals survivedto therespectivescheduled sacrifice.There were no test material-relateadntemortem observations. Body Weigbts and Cumulative Body Weight Gains Body weight data are summarized in Table 2;individualdata are in Appendix B. Cumulative body weight gainsover thedosing period(Days I to 6) and over the study duration(Days I to 20) areincludedinTable 2. Individualday-to-daybody weight gains and individualcumulative body weight gainsforDays I to 6 and Days I to 20 arealsoin Appendix B. Mean body weight gainsformid- and high-doseanimals(14.0 and 42.0 mg/kg, respectivelyw)ere significantlryeduced duringthe dosingperiod;Day I to 6 body weight gains were 23, 23, 0, and -4 g forthe control,5.0-,14.0-,and 42.0-mgtkg groups 17 CHW 6329-197 respectivelyI.ngeneral,themid- and high-doseanimalsshowed a recoveryinbody weightgainoverthefirs3t daysaftercompletionof thedosingperiod.The overalblody weightgains(Day I to20)were freeofsignificanitntergroudpifferenceasnd were similarforallgroups. PaimitoylCoA OiddaseActivityAnalysis Day 6 palmitoylCoA oxidaseactivitdyataaresummarized inTable3;individuadlataare inAppendix C. The PathologyReportcontainsa discussioonf thedata. On Day 6.palmitoylCoA oxidaseactivitwyas significantilnycreasedfortreatedanimals atalldoselevels.Mean hepaticpalmitoylCoA oxidaseactivitifeosrthegroupsgiven 0.0,5.0,14.0,and 42.0mg/kg were 5,24,39,and 39 IIJIG,respectively. AnatoniicalPathology Resultsof theDay 20 necropsiesand theDay 6 and Day 20 liverweightdataarein Appendix D. The PathologyReportcontainsa discussioonf thenecropsyfindings. At necropsyon Day 20,one animalfrom thegroup given5.0mgtkg had multipled,ark redfociof variablesizeintherightlobeof thethymus,and thepelvisof theleftkidneyin one animalfrom thegroupgiven42.0 mglkg was enlarged.These were considered incidentaflindingsand unrelatedtothetestmaterial.There were no macroscopic findingsin theremaininganimalsexamined. CONCLUSIONS Based on significanitncreaseisnthelevelsof hepaticpalniitoyCloA oxidaseactivitya,n indicatioonf peroxisomeproliferatiaotna,lldose levelstestedt,heno-effecltevelof T-6669 islessthan5.0mg/kg when administeretdomale Crl:CDO(SD)BR VAF/Plue mts for5 consecutivedays. 18 SIGNATURES CHW 6329-197 Sandm L Haley Study Coordinator Toxicology Coming HazletonInc. . A- Susan M. Henwood, MS Diplomate,ABT Study Director Toxicology Coming HazletonInc. f7 Date Date 19 RYYERENCES CHW 6329-197 Dunnett,C. W., "New TablesforMultipleComparisonswitha Control," Biometrics2,Q.482-491(1964). Levene,H.,"RobustTestsforEqualityof Variances,C"ontributiontsoProbabilila3n@d Statistic(se,ds.)L Olkinetal.C,h. 25,pp.278-292,StanfordUniversitPyress:Stanford, Californi(a1960). Winer, B. J.,"Designand Analysisof Single-FactoErxperiments,S"tatisticParlincillies inExpcrimentalDesigilSlecond Ed.,Ch. 3,pp. 149-260,McGraw-Hill: New York, New York (1971). 20 PATHOLOGY REPORT CHW 6329-197 SUNEMIARY The purposeofthisstudywas toassesstheoraltoxicitpyroducedwhen thetestmaterial, T-6669,was administerebdy theoralroute(gavage)toratsfor5 consecutivedays.The testmaterialwas administeredatdose levelsof0.0,5.0,14.0,and 42.0 mg/kg of body weight/day(mglkg). Administrationofthetestmaterialatalldose levelswas associatedwithhigherhepatic pahnitoylCoA oxidaseactivityM.ean palmitoylCoA oxidaseactivitioens Day 6 for controlanimalsand animalsgiven5.0,14.0,and 42.0mg/kg were 5,24,39,and 39 IU/G, respectively. There were no testmaterial-relatmeadcroscopicfindings. METHODS Four groupsof male Crl:CDO(SD)BR VAF/Pluse rats(10/group)were administeredthe testmaterialby oralpvage for5 consecutivdeaysata doselevelof 0.0(controglroup; receivedreverseosmosiswater),5.0,14.0,or42.0mg/kg. Fiveanimalsfrom each group were sacrificeodn Day 6 forhepaticpalmitoylCoA oxidaseanalyses.The whole liver was removed and weighed,and therightlaterallobeof theliverand theremainingliver were weighed. The analyseswere performedon therightlaterallobeof livera,nd the remainingliverwas frozenforshipmentto theSponsor.Allremaininganimalswere sacrificeadnd subjectedtoan abbreviatedgrossnecropsyexaminationon Day 20. At necropsy,macroscopicobservationwsere recorded,and theentireliverfrom each animal was weighed and frozenforshipmenttotheSponsor. StatisticalsliygnificandtifferencecsitedintheResultsand Discussionsectionarebased on comparisonsbetween thecontroland treatedgroups. 21 RESULTS AND DISCUSSION CiiW 6329-197 Mortality AD animalssurvivedtotherespectivsecheduledsacrifice. ClinicalPathology IndividuavlaluesforhepaticpalrrlitoCyolA oxidaseactivitayreinAppendix C. Mean valuesand theresultosf statisticcaolmparisonsareinTable3. On Day 6,palniitoyCloA oxidaseactivitwyas significandiyncrmed fortreatedanimals atalldose levels.Mean palmitoylCoA oxidaseactivitifeosrthegroupsgiven0.0,5.0, 14.0,and 42.0mgtkg were 5,24,39,and 39 IIJ/Gr,espectively. AnatomicalPathology Individualanatomicalpathologydatafortheanimalsnecropsiedon Day 20 arein Appendix D. Individualliverweightvaluesfortheanimalssacrificeodn Day 6 arealso inAppendix D. Day 20. At necropsy,one animalfrom thegroupgiven5.0mg/kg had multipled,arkred fociof variablesizeintherightlobeofthethymus,and thepelvisof theleftkidneyin one animalfrom thegroupgiven42.0 mglkg was enlarged.These were considered incidentaflindingsand unrelatetdothetestmaterial.There were no macroscopic findingsintheremaininganimalsexamined. Pathologists: RobertL Hall,DVM, PhD Diplomate,ACVP (ClinicaPlathology) @77 Date OR i-A 19@6mai-EP.almer,-P-Eg' Date Pathologist 22 CONUKENTS ON THE DATA CHW 6329-197 Variousmodels of calculatorcso,mputers,and computerprogramswere used toanalyze datainthisstudy.Becausedifferenmtodels roundoffortruncatenumbers differently, valuesinsome tables(e.g.m,eans, standarddeviationso,rindividuavlalues)may differ slightlfyrom thoseinothertablesf,rom individuallcyalculateddata,or from statistical analysisdata.Neithertheintegritnyor theinterpretatioofnthedatawas affectedby these differences. Tle unitsfordoselevelson theHazletonPath/ToxSystem (BPTS) summary tablesare otm&gt&y..v The number of animalslistedintheheadingof thesummary tablesforantemortem observationsreflecttshenumber of animalsassignedtoeach group atthestartof the study. The summary tablesforantemortemobservationisndicattehenumber of animalsfor which a conditionwas observedwithoutregardtothespecifincature,severity, reversibilintuym,ber ofincidencesperanimal,or thelengthof timethecondition persisted. Only observationfsoreach animalotherthannormalareindicatedon thesummary and individuaalntemortem observationtsables. HPTS considerstheday ofinitiatiofntreatmentas'Day 1,Week 1. Body weightdata areenteredatthestartofa studyweek (e.g.a,body weightrecordedon Day I is considereda Week I body weight,a body weightrecordedon Day 8 isconsidereda Week 2 body weight).Dailybody weightgaindataarecalculatefdrom day today and areindicateidntheappendixby theend pointday (e.g.b,ody weightgainfrom Day I to2 isshown asDay 2 gain,Day 2 to3 as Day 3 gain,etc.)C.umulativebody weightgain dataarecalculatefdrom thefirsdtay of thestudytotheappropriatdeay (e.g.D,ays I to6 and Days I to20)and areindicateidn thetablesas I - 6 CHNG and I - 20 CHNG. 23 - CODES, ABBREVIATIONS, AND UNITS GeneralCodes and Abbreviations Codes forClinicaPlathology Abbreviationasnd UnitsforClinicaClhemistry Codes forAnatomicalPathology CHW 6329-197 Note: 71befollowinglistosf codes,abbreviationsa,nd unitsareused by CHW. Some, butnotnecessarilayllo,f thisinformationmay be needed forthisreporl 24 CHW 6329-197 General Codes and Abbreviations wx weell N Number ofmeasurementisna group. Mean; MEAN Arithmeticmean. SD; S.D.;STAND DEV; STANDARD DEV; sd Standarddeviation. CHNG Change. PRE/DLY Predoseobservationd;ailyobservation once dosingiscompleted. Group mean issignificantdliyfferenftrom themean of thecontrolgroup(Group 1)at p:s 0.05. t Indicatebsody weightdatathatwas analyzedstatistically. NA No value;notapplicablen;otpresenl p PresenL 25 CHW 6329-197 General Codes and Abbreviations(Continued) DISPATCH Observationsdispatchedfrom thein-life module oftheHazletonPadi/ToxSystem (EM) tothenecropsymodule fordata collectiopnurposes.Observationsare duplicateosf thelastin-lifoebservations. T'BW Terminalbody weighl #,No. Number. Animal Death Codes: I TenninalsacrificaetDay 6. T TerminalsacrificaetDay 20. 26 NS QS/QNS NR Ti TE RE EE SE CEW 6329-197 Codes for CHWcal Pathology GENERAL CODES No sample Quantitynot sufficient No repeat(sample volume not sufficienftorrepeatanalysis) Technicianjudgment torepeattest Technicalerror(instrumentor technicianei,oxthatresultsin unacceptabledata,e.g.,unacceptableinstrument output,sample spillede,ntryof invaliddata) Recording error(recordedincorrectdata,e.g.,wrong number, speuing error,incorrectdate) Entry error(incorreckteyboard entry) Sampling error 27 CHW 6329-197 Abbreviations and Units for CHWeal Chemistry Test Abbreviation(Units) PalmitoylCoA oxidase PCOAO (IU/G) 28 - Codes for Anatomical Pathology CHW 6329-197 Code Ddinition ANDAAL DEATH CODES I Terminal sacrificaetDay 6 T Terminal sacrificaetDay 20 TISSUE ABBREVIANONS IN GL STOMACH, GL STOMACH, NONGL SALJV GI.,MANDEB LN, ANT NIIES/PANC AUDRMRY SEB GL LACRDAAL GLAND, EX HEMATO NEOPLASIA LACRDAAL GL INT CAV=,ABDOM SALIV GIPAROTID LN, TRACHEOBRON Lymph node Gland Glandular stomach Nonglandular Mandibular salivarygland Anteriormesenteric/pancreatilcymph Auditory sebaceous gland ExorbitEdlacrimalgland Hematopoietic neoplasia Internallacrimalgland Abdon-dnal cavity Parotidsalivarygland Tracheobronchiallymph node node 29 Corning Hazleton Inc. Madison, Wisconsin USA Table 1 Summary of Antemortem Observations ---------------------------------------------------------------------------------------------------------------- DAYS 1-20 NLMBER OF ANIMALS AFFECTED CATEGORY SEX: GROUP: DOSE; ------M-ALE ------- 1 2 3 4 0 5 14 42 KEYWORD QUALIFIER NUMBER. 10 10 10 10 ---------------------------------------------------------------------------------------------------------------- *** TOP OF LIST APPEARANCE SWOLLEN HOSE 0100 END OF LIST C> Corning Hazleton Inc. Madison, Wisconsin USA Table 2 Summary of Body Weight Data (g) --------------------------------------------------------------------------------------------------------------- SEX: --------------------M-ALE --------------------- GROUP: 1 2 3 4 DOSE: 0 5 14 42 DAY UNITS: MGIKG MG/KG MG/KG MG/KG --------------------------------------------------------------------------------------------------------------- it N MEAN S.D. 2 N MEAN S.D. 3 N MEAN S.D. 4 N MEAN S.D. 5 N MEAN S.D. 6 N MEAN S.D. 10 349 27.5 10 357 31.1 10 360 30.5 10 365 31.7 10 366 32.8 10 372 34.6 10 359 22.5 10 362 23.4 10 367 23.2 10 373 23.4 10 375 24.7 10 382 25.2 10 372 16.5 10 378 16.7 10 382 18.8 10 302 18.8 10 376 23.1 10 372 28.1 10 349 15.0 10 352 13.8 10 349 13.8 10 343 15.6 10 339 21.4 10 345 23.6 1-6t CHNG N MEAN S.D. CRNG AS t OF CONTROL 10 23 8.3 - 10 23 6.3 99 10 0* 22.1 0 10 -4* 20.6 -17 t Indicates data was analyzed statistically. Denotes statistical significance at ps 0.05. Corning Hazleton Inc. Madison, Wisconsin USA Sumary Table 2 of Body Weight Data (g) ---------------------------------------------------------------------------------------------------------------- SEX! --------------------HA-E.9 --------------------- GROUP: 1 2 3 4 DOSE: 0 5 14 42 DAY LINITSI 14G/KG MGIKG MG/KG MG/KG ---------------------------------------------------------------------------------------------------------------- 7 N 5 MEAN 375 S.D. 33.2 8 N 5 MEAN 374 S.D. 36.1 9 N 5 MEAN 389 S.D. 35.9 10 N MEAN S.D. 5 394 36.5 N MEAN S.D. 5 386 36.1 12 N MEAN S.D. 5 403 37.9 13 N 5 MEAN 407 S.D. 40.3 14 N s MEAN 413 S.D. 40.9 5 375 25.8 5 373 27.0 5 389 26.2 5 385 29.1 5 388 28.0 5 402 28.8 5 407 29.9 5 412 31.4 5 376 35.9 5 374 37.6 5 395 34.0 5 392 29.4 5 396 27.7 5 412 27.9 5 415 28.3 5 422 28.8 5 351 27.4 5 355 19.5 5 374 16.1 5 373 14.2 5 376 13.1 5 390 12.6 5 394 12.1 5 400 12.6 Corning Hazleton Inc. Madison, Wisconsin USA Table 2 Summary of Body Weight Data (g) --------------------------------------------------------------------------------------------------------------- SEX: --------------------MA-LE --------------------- GROUP: 1 2 3 4 DOSE-. 0 5 14 42 DAY UNITS: MG/KG MG/KG MG/KG MG/KG --------------------------------------------------------------------------------------------------------------- 15 N MEAN S.D. 16 N MEAN S.D. 17 N MEAN S.D. Is N MEAN S.D. 19 N MEAN S.D. 20 N MEAN S.D. 5 414 39.7 5 413 43.1 5 421 42.0 s 420 44.3 5 431 44.1 5 438 43.9 5 414 30.8 5 411 32.4 5 421 32.6 5 426 33.5 5 431 36.0 5 434 33.9 5 425 28.1 5 423 26.6 5 436 26.1 5 443 28.9 5 445 27.7 5 450 28.8 5 40S 13.2 s 404 11.3 5 414 10.8 5 419 9.6 5 424 11.6 5 431 14.3 1-20t CHNG N MEAN S.D. AS t OF CONTROL 5 ea 20.2 - 5 83 10.2 94 5 al 13.1 92 5 83 8.0 94 t indicates data was analyzed statistically. * Denotes statistical significance at p-g0.05. Table 3 Summary of Clinical Chemistry Data Males Day 6 DOSE mg/kg ------ PCOAO IU/G -------- 0.0 5.0 14.0 42.0 MEAN S.D. N MEAN S.D. N MEAN S.D. N MEAN S.D. N 5 1.4 5 24 1.8 s 39 3.3 5 35 5.4 5 CHW 6329-197 I APPENDEX A ProtocolDeviations Protocol1?6785 MaterialSafetyData Sheet 35 ProtocolDeviations Protocol.Animals. Weight at Initiationof TreatmenL "210 to250 g" CHW 6329-197 Actual Procedure. AD animals assigned to study exceeded 250 g at initiationb:ody weights at study initiatiornanged from 303 to 395 g. Protocol.Blood Sample CollectionsF.requency.'?redose(I day beforetheinitiadlose administratiotnoDay 1),on Days 9 and 15,atthescheduledsacrificienterval(sDays 6 and 20),and atunscheduled sacrificientervals." Actual Procedure. The predose (pretestb)lood samples were collectedon Day -2. These deviationsarenotexpectedtohave affectedtheresultosf thestudy. 36 CORNINCH22ICtOn Sponsor. 3M StPauLMmesotiL PROTOCOL TP6785 Study Title 5 DaUy Dose Ond To@iicitSytudy withT-6"9 inRaft Data Doo=ber 20,1996 Perforudng laboratory-. Coming HazletonLm 3301 Kinsman Boulevard Madison, Vuem2sin 53704 Laboratory ProjectI CHW 6329-197 37 STUDY I]DENTIFICANON 5 DailyDose OralToidcityStudy withT-6669 inRab CHW 6329-197 IP6785 Page 2 TestMaterial Sponwr Spomr Reprmntadve StudyDkwtor StudyLocation ProposedStudy rmwtable In-LifPeqmimental) StarDtate In-LifTeerminationDate TerminationDate T-w9 3M ToidcologySwAces Building220-2E-02,3M Center St.PauL NG=csou, 55144-1000 Roger0 Pakms, PhD, DABT 3M Tc"dcologyServices Building220-2E4Y2,3M Center St Pmd, Kmnesota, 55144-1000 (612)733-3222 FacimileNo. (612)733-1773 SusanK Bmwood, MS, DABT coming HazletonInc. P.O.Box 7S45 Ma&wn, VI 53707-7545 (608)241-7221 FaedmileNo. (609)242-2736 CorningHazletonInc. 3301 Kinsman Boulevard Mmfison,Vmonsin 53704 January 2@ 1997 January21, 1997 To be determined 38 L Study 5 DailyDose OralTox:tctyStudywithT-6669 m Rats CEN 6329-197 7?6795 Page 3 2. Purpose To a-t theord toideitpyroducedvhm thetestmateriailsadministerebdy the oralroute(gavaget)omts for5 consecufm days 3. Pegdatory Compliance ThisstudywillbeconductedinaccordancewiththeFood and Drug AAmWsbadon Cmod LaboratoryPracticReegulationfsorNonclh&d LaboratoryStacs, 21 ut@y,58 and theOrganisatiofnorEconomic Co-operatioannd Development PrincipleosfGood LaboratoryPtacticcC6(g1)30,wM theexceoon thatanalysiosf theftamataw mbchm fbrC 3obMW. bomogencity,and stability willnotbe conducte& 4. QuaUityAssurance The protocolaudy cc)nduc@m.d End reporwtillbe mdited by ffiQcuality AssuranceUnitinaccordancewiththeStandardOperatingProcedures(SOPS) ofthe Wmonsin &dUV ofComing HzdctonI= (CEW@ TestMaterld A. Identification T-6669 PhysicalDescription To be documentedintheraw data C. Parityand StabMW Respom-bilitoyf*c SponsorCmcludingundertestconditions)S.amplesof testmateriaalnd carriemrbftm forconcentudon.homogencit3rs,olubility. and SWEV analyseswillnotbe takenbeforesarninktration, D. Storap Conditions Room tempuattm IL ReserveSamples Ruerve samplc3oftestmillerisatrlenotrequiredtobe takem 39 CEN 6329-197 TP6785 Page 4 F. DbpositionofTestMaterW Any ummd testmateriawlM be mtzaed totheSpon= aftercompletionof ill-Utfsath* G. SafetiPrremations As req*ed by CHW SOPs and policies 6. Carrier A. Identification Revaw Osmosis water(RO wata) B. PbysicalDescription acar, colorlesHsqtnd C. StorageConditions Ambied 7. ExperiinentiDdesign A. Animals (1) Species Rat (2) Str3@ Cd:CDO(SD)BRVAFIPbiO (3) Source CharlesRiverLaboratorieIs=, (4) Age atInitiatioonfTreatment Young adult (5) Wei&t atInitiationfTreatment 210 to250 g (6) Number and Sex 40 males (7) Identirication MviduaUy codedpassivehdcgmted bmsponder implaub 40 (8) Husbandry CHW 6329-197 776785 Page 5 (a) Housing Mvidual (may be grow-houseddming acchmtion) (b) Food CardfiedRodentDiet#5002meW (PMID Fwds, Inc.)ad libkwn, unlesso%erwisespecified7U foodisroudmly emdyzed by &c rfkstnufiutmvr fornutritioncdomponentsand containinants. (c) water Ad libitumS.amplesofthewaterarenab-zedfortotadliaolved solidasndVmfied mcrobiologiccaolntenatndfor selected elements.hmvy mews, orgenophovhatcs.and cblodnated hydrocwbonl (d) Cont=Wants Ibm we no lmovvn rll*nsinntfsficfood orwaterthatwould bftdae withthisstudy. (e) Environment Eaviro=enW controlfsorthe-qnl*mraolom wM be setto rnmintai1n9'tD 25'C (66*to77*F),a rcmvc humift of 50% *20Y% anda 124xna HSIW12-bour dark cyclc M Acclimtion At leastI week (9) Randon3iization Via computcrlpnemted madom numbes forassigmmenttogmupl The linirmlwsillbe weighe& The wei&t vwiationofthemin,*rnal selectedforthestudywillnotexceed+-2standarddeviationosfthe mean weigbts. (10) Justification lle ratisfivqmtly usedinsaf@tyevaluatiosntudiesas a ofa rodentspeciel 41 B. Group DesigaiLdons CHW 6329-197 776785 Pap 6 CUOIUP Dose Levela (MWW Number of Malai@ I(Control) 0.0 10 2 (Low) 5.0 10 3 Ofid) 14.0 10 4 (Iggh) 42.0 10 a IU controglroupwillrcmve RO wateronly Ihe dose volumewiU be 5 mL/kg. b Fmc -nimmltineachgroupwM besacrificefdor pdmitoyi-CoAo)iidasaenAys6 on Day 6. rae an;mnlineachsroupwillbe observedforreverdbility. pmbtme, ordelayedoccmtenceoftomc effectusntil sacrificoend Day 20. C Dodng Protedwvs (1) DosingRoute Oralpvage (2) Remon forDosing Route Eastcxicfttheoralroutehasbeentherouteofchoicefor akwm mount oftestmaterial. (3) DosingDuration Fiveconsecutivdeaysofdow skdm;nisftdalTU firsdtayofUcatmat willbe designateadsDay 1.The do= wM be kArnl*nistaetred approximateltyhesame timeeachday. (4) Dose Preparation Mz contromlhnws willreceivReO wateronly.Testmktu= wM be preparedon eachdayof uilrntn6tratiMoniLtea materiawlillbe mixed inPo vata ata specificcm=tradon foreachdon lwel Mm dosefor eachanimskfloreachday ofdosingwillbe basedon themost recently recordedbody weighl The prepued testml-ctum wW be storedatroom temperaturuentilnrlm;nistmtioDnj-uingdose *Am;n;sbmfiak homogeneoustat materiamlktura willbe maintgned usinga magnedc stiprhde and stirbar. 42 CEW6329-197 TP6785 pap7 D. ObstrvatioonfAnhnah (1) ClWcal Observadons Twicedafly(&= andpzL)formmuhty thmugh Dq 20(a.=mortality onlyon Day 20. Animalswillbe observedpredow, atamoxu@y 2.5a,nd4 hoursafteerachdosea;nddailtyhereafftoerd&*A *gm Obswadonsmaybeextendewdhendirectbeydthedudy(firectm. (2) Body Weights Forravn% 1 9b4m initiatioofntmatinen(tDay 1),dany ffiereaftaetrs.cbeduledsacrific(eDsays6 and 20),and atdeath(when =vW exceedsIdaA (3) Blood Sample CoBeedons (a) Frequency Predose(Iday beforeffiWcU dose adrnini*-cbutfoiDoany l@ an Days 9 and 15,atthewbeduledsacrificienwvals (Days6 and 20), andatunsduxucd sacrifibcgecrval3 (b) MethoodfCallectionioNfuAmnbiemrah Animawlislnlobte&steodvcniifgobsrtcheducloeudecfion& Bloodsmnples(awoxbmately1-5mL) willbe collectefdiuma jugularveinofallanimalattheprodosefitwid andon Days 9 and 15. A bloodsample(asmuch aspossiblew)illalsobe collectefdrom &e posteriovrenacavaofeachnn;msislacrificeidna manund conditio(nifposm*blca)nd fim each stnl*rantislclheduled sacrific(eDsays6 mA 201 Tne bloodsampleswillbe storedatroom tempewm andthen centri&gedt;heseparatwe= and cellulafrmctionswillbe stored ina fmc= settomlemwn -20*C:ilO*.The 3e= and ccDWw fiwdons willbe pwked on dryiceand senttotheSponsorwithinI week afkeirn-Ufetemination.ne Sponsorisresponsiblfeorthe rctentioannd dispositionfthesamples. 43 7be sum CHW 6329-197 TP6795 Page a and cellulafrmcfionsampleswillbe shippedto-. James D. John= 3M F.T.& S Bldg.2-3E-O9 935 Bush Avenue SL Paul,Mbutsota 55106 James D. Johnsonorhisalternatweillbe notifierdegardintghe sbipmat ofthesamplel E. PaWtoyi-CoA 03ddaseAnalysh (1) Frequency and Number ofAnintab Fiveanimawgroup on Day 6 inrandom order (2) Method ofCollection On theday ofdw whodwed sacrificneo,nfiawtnirnal willbe anesthetizweidthsodiumpentobarbitwde,ighed,and or-mngll;nated TbL@abdomimw cavityofea& awmitlwillbe opened,andthelivaviU be removed andv&igbe& IU rightlataalobeoftheliverwillbe collected from eachmmitnelweighed,andflash-froziennliquidnitrogenI.he liver tsm willbe sun-edina ficem settomaintain-70OC:Lloeuntil analyzedby CHW forpdmitoyl-CoAoxidut acbvity. 7U rema*m*mgliverwillbe coYlect4weigbed,and placedintoa ficem settom-q;nta;-n20*C *100. The livenwiU be packed on dryiceand shippedtoJames D. JohnsonwithinI week afka in-hfcftminsfion.7be Sponsorn responsiblfeortheretentioannd dispositonofthelivers AniTn.glwsillbe dis@ afterscheduledliverfissm cou=don. Any,an;r"l selectefdorpalmitoyl-CoAoxidaseanalysitshatdiesor is sacrificeidna mon*bu@ conditiownillbe subjectedtoan abbreviaw grossn=opsy examinationa,nd allabnormalitiewsillbe recorded. Tissueswillbe discardeadftenr=opsy. 44 CHV6329-197 IP6795 Page 9 F. Termination(animahnotdesipatedhr pabWtoyi-CoAoxidosean*sis (L) Unnheduled SacriftaandDeaths Amy gn;malthatisfounddeadorb sacrificienda monlund condition willbe subjectetdoan abbreviategdma nemopsy examinationand an wM be r=orded.Anknob tobe ncri:BcedwM be anesthc6Z4cwdithsodium pentobwbita4weigb4 bledviatheposterior vena cava,and ;naW 7be wholelivewrillbecollect4 weighed,and placedintoa fiec= settoinik;nta-;2n0*c*ioe. 7be Hverswillbe packedon dryiceandshippedtoJames D. Johmm within I week aft in-liftemmaation.The Sponsorisresponsiblfeorthe retentioonrdispositionf&c livem The animalswillbe dscardedafter n=opsy- (2) ScheduledSacrifice On Day 20,iheanitnalwillbe amsde6zed wilhso&=.pemobwbitd, weigh4 bledviatheposterivomr cava, mmiinated,a,nds*ected toan abbreviategd= namom The vminialwillbe necropsiedinrandom order,and all wM be rewrded.71w wholelivewriU be coUect4 wtigb4 andplacedintoa fium setto rnolinta-i2n0'Ca:10*.The Hvem willbe packedon dryiceandshipped toJam= D. JohnsonwithinIwe& afteirn-M t=dnatioilThe Sponsorb responsiblfeortheretentioonrdispositionftheHvaL The animal WM be discardeadftenrecropy. G. StatistieAdnalyzes Statistiactawlym willbeperformedo@ Day I body weigbtkcumwmdve body wci& gains(Day2 throughtemiins6cm)a.ndpdmitoyl-CoAwddaselevel& A descriptionfstatisticaanlalyseissinAUachment I- Groups 2 *zougbl4 wM be comparedwithGroup I (controd. L Report A finarleporitncludintghoseitemsHsW belowwillbe submftte& Descriptioonfthecontroland testmatetials Descriptioonfthetestsystem Procedums Datesofcqmimental initiatiaonndummination T"ation ofmortalitdyataby doselevel Descripdonofany toidceffects Tabubfionofmean body weightsby doselevel Macroscopiocbr@ons 45 Pathologyreport PWmitoyl-CoA oxidaselevels Statisticfailndings CHW6329-197 7*P6795 Pap 10 9. LocationofRjtwData,Rewrds, and FinalReparl Originaldata,orcopiesthe=4 wfflbe avaimle atCHW tofar-Mtataeuditintghe su* duringitspwpm and beforewcapb= ofdw finarleporlVA= thefinal reportiscomplet4 alloriginaplaperdati6includintghoseitemsfistebdelow will be retaineidntheamhivesof CHW fora periodof I yearfollowingsigningof ft finalreporlOne yearaftersigningof&c finalreporta,llof theafoi .a-n!bd materials wM be sent to the Sion@, and a M= fee will be charge& The Spon= may electohne thematerialrsetaineidntheCHW uchives foran additionatlime and CHW willchargea storagefm Ifdw Sponsorchoosestohave CHW disposeof thematerial%a disposalfeewillbe chwged. Protocoland protocolamendments Dose prvarationrecords In-lifreecords Body weights I *$,-*@ data Dose adininwration Antemortem v Anatomicalpathologyrecords PWmitoyl-CoA oxidastanalysirsecords Statisticraelcords Sample collectiornecords Shippingrecords Studycomspondem FM repor(toriow signedcopy) Ile followinsgupportinrgecordswillbe rewned atCHW butwillnotbe sveved withthestudydall An;msilreceipatnd acclimatiornecords Water analysirsecords Anl*m-4r1oom temperatureand humidityrecords Refiignatoarnd fr"= temperaturmecords Instrwnentcalibratioannd maintenancerecords 46 PROTOCOL APPROVAL CHW6329-197 IP6785 Pap 11 Roger 0. Paidns,PhD Diplomatc6ABT SponsorRepmmutive 3M ST=VH= Henvmod, M Diplomatc6ABT SuidyDirac= TaxicokW CorningHazlewn I=. Rzpmmbtiw QualityAmurance Unit Coming HazletonI= t2, Date Date -TC Data 47 AITACHUIENT I StatisticMaeldiods CEW 6329-197 T?6785 Pap 12 Ile statisticmaelthods thatwillbe used me de="ood below. Lcvcne!stest (Lcvme, 1960)willbe dometotestforvariaumhomogeneity.Inthecm of heterogeneitoyfvariameatp:c0.05, - willbe used tostabiliztehe variance. AnalysisofvarianceJANOVA (Wumv, 1971a)]willbe done on dw homogeneous or transformedall IftheANOVA issipfficanDtu,nnetest-tes(tDumett, 1964)willbe wed forpdmw betweentreatedand controlgroupl One-way ANOVA willbe usedCifapplicablet)omdyze Day Ibody weigh% cumumve body weightgains(Days2 tmgh Urminafion)a,ndpdmitDyi-CoA oxidm levell IftheANOVA shows sipfficancfeorbodyweightsatinitiatioofnftatment(Day 1). one-way argdydsofcovaia= JANCOVA (W'ma. 1971b)]wiU be usedtoamalyzebody weigbt%withtheWU bodywtigbn a thecovariateA.ltboughLzvcnds testfor vuiar" bomoguicitywillbe done (sm abovelno h onSformafLowwillbe usedbecame cov&dancea&justmentremovesextrmcoushctmgcnck. N&e ANCOVA isS!Bwncukt, lengsquam means t-teg(SAS,1989)willbe usedforpdrwin compwiwtis bdvtm treatedand controlgroupl Group comparisonswillbe mhuw atthe5.00/Mox@.tOcd probabilitlyevel References Dunwd, C.W, "New TablesforMultipleCompaisons witha Control,B"iQmsWm 2T.482-491(1964). Levem R, -RobustTeft forEq=Hty ofVwiaw^w r-ontnbiftiotnosProbabiritLod sudgiGL (eds.1).Olidnetal.C,h.25,pp.279-292,StanfordUniversitPy=L- Stanford, Cal-forni(a1960). SAS InstitutIenc.,SAS/STAr Usees Guide,Version6,FourthEd.,Vol.2,p.909,SAS hmstiwt1c=: Cary,NorthCaolina (1989@ 48 AUacbment I (Confinued) CHW 63291-197 7P6785 Page 13 Vinm B. J,Memip andAnalysiosfSWgle-Factor StaflaticPa6lnciples inExpcdmogdj2ciLVL SecondEdL,Ch.3,pp.149..260M,oGmw-MU: New York. New York(19714 Vmcr, B.J.,"AnalysiosfCovadance,S"tatistiPcrailndW= inE=sdmoW nesign,' SecondEd.,CL 10,pp.752-812M,cGraw-MR: New York,New York (1971b). 49 MklLhi^L SAFETY DATA SHEET aM 3M Center St. psul, 14innesola $5144-1000 (612) 733-1110 CHW 6329-197 t3 1--6669 Copyrigbtl 1996, Minnesot2 Mining and Manufacturing Company. All rights r$36rved, Copying and/or downloading of this information for the purpose of properly utilizing 3M products i3 allowed provided that: 1) the information is copied in full with no changes unless prior agreement is obtained from 3M, and 2) neither the copy nor the,original 13 resold or otherwise di3tributed with the intention of earning a profit thereon. DIVISION,. SPECMTY CHEMICALS DIVISION TRADE NAME: FC-143 FLUO,M Brand fluorochemical Surfaciant ID NUMBER/U.PdC.: 98-0211-0006-0 00-51135-09138-8 98-0211-0891-9 98-0211-5489-7 00-51135-02941-1 98-0211-6581-0 98-0211-7394-7 00-51135-10762-1 ZF-0002-0378-4 ISSUED: August 23, 1996 SUPERSEDES: M4y 20; 1996 DOCUMENT: 10-3808-2 00-51135-09365-8 00-51135-10405-7 ...........4........................................................... 1. INGREDIE?IT C.A.S. NO. PERCENT .................................................................... AMMONIUM PERFLUOROOOTAROATE .............3825-26-1 93 AMMONrum PEAFLUOROliEPTANOATE ............6130-43-4 1 MMONIUM PERFLUOROPENTMOATE ............68259-11-0 1 AMDNIUM PERFLUOROHEXANGATE ..............216iS-47-4 0.1 - 97 .3 .3 .I .................................................................... 2. PHYSICAL DATA .................................................................... BOILING POINT: ...............N./A. YAPDP, PRESSURE: ....... R/A VAPOR DENSITY! ...............M.IA. EVAPORATION RATE: .............N/.A SOLUBILITY IN WATER: .....0.011.a0ppree, SPEOIFIC GRAVITY; .............0..4 - O.S Wateral (Bulk) PERCENT VOLKRILE: .............N/.A pH ........................c.a....s. (0.5% Aqueous) VISCOSITY: ..................N./.D MELTING POINT: .............:..N./A APPEARMCE AND OWR: Light colored powder; slight odor. ............% .......................................................... Abbreviations: R/O - Not Determined N/A Not Applicable 50 MWS% FC-143 FLUORAD brandFluorothesiesSlurfattant AugU3% 23, 1996 CHW 6329-197 PAGE 2 ................................................................... 3. FIREAND EXPLOSIOKNAZARDDKTA ............................................................ FT.ASH POINT: ............. t4on-flasmable FLANUBLE LIMITS - LEL: ........MIA FLNWBLE LIMITS - UEL:.......M.IA AUTOIGN1710H TEWEPATURE: ......N/A EXTINGUISHING MEDIA: Water, Carbon dioxide, Dry Chemical, Foam SPECIAL FIRE FIGHTING PROCEDUREST Wear full protectiveclothing,includinghelmet,sell-colitaiiied, positive pressure or pressure demand breathing appat,atualbunker coat and Pants, bands around arms, waist and legs, face mask, and protectivecoveringfor exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decompositionsection for products Of combustion. ......................a..............................a ............... 4. REACTIVITY DATA ........................................................... STABILITY: Stable INOWATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not Applicable KkZAFiDOUS POLYMERRZATION: Hazardous polymerization will not occur. FAZAADOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Nitrogen, Hydrogen Fluoride, Axmoni&. ............a................................................... S. ENVIRONMENTAL INFOWTION ..........a.................................................... $PILL RESPONSE: Observe precautionsfrom other sections. Collectspilled naterial. Use wet sweepingcompoundor water to avoid dusting. Clean up residue. PINCO in A C103ed container. REC"ENDED DISPOSAL: IncinerateIn an industrialor commercialfacilityin the presenceof a combustiblematerial. Combustionproductswill Include HF. Disposal alternativetDisposeof waste productin a facility permitted to accept chemicalwaste. ......................................................................... Abbreviations; N/D - Not Detei-minedN/A - Not Applicable 51 MSDS: FC-143 FLUORAD Brand FluoroChomical Surfactant August 23, 1996 CHW 6329-197 PAGE 3 ...................................................................... S. ENVIRONMENTAL INFORMATION (continued) ............................................................................. ENVIRONMENTAL DATA: Chemical Ox@gen Demand (C-00)- Nil(.00070 gfg); 20-Day Slochemicai Oxygen Demand (50020) o Nil; Theoretical Oxygen Demand (THOD) c 0.32 g/g; Fathead Minnow (Piosphales proselas) 96-hr LCSO u 740 mg/L; Water flea (Daphnia magna) 48-hr ECSO a 460 mg/L; Green Algae (Selenastruscapricornatut)14-day EC50 (call dry weight) z 73 og/L; Green Algae (Selenastrum capricornutum) 14-day ECSO (call count) = 43 mg/L bioconcentration factor (BCF) for ammonium perfluorooetanoate (PFO) = 1.8 REGULATORY INFORMATION: Volatile Organic Compounds: NIA. VOC Less H20 & Exempt Solvents: N/A. Since regulationsvary, consult applicableregulations or authorities before disposal. U.S. EPA Hazardous Waste Number = None (Not U.S. EPA Hazardous). The components of this product are in compliance with the chemical registration requirements of TSCAG EINECS, CDSLI AICS, MITI and KTCCL. EPCRA KAZARD CLASS: FIRE HAZARD: No PRESSURE*. No REACTIVITY: No ACUTE: Yes CHRONIC: Yes ............................................................. S. SUC-AESTED FIRST AID ............................................................. EYE CONTACT: Immediately flush eyes with large amount3 of water for at least 15 minutes. Bel immediate medical attention. SKIN CONTACT: Flush skin with large amounts of water. If irritationpersists,got medical attention. INHALATION: If 3ign3l$YgDtOMS occuro remove person to fresh air. 11 signs/SytptOM3 continue, call a physician. IF SWALLOWED: Do not induoe vomiting. Drink tWO glaSS$3 Of W8ter. Call a physician. ......................................a ....................... Abbreviations; N/D - Not Determined N/A - Not Applicable 52 msDS: FC-143 FLUORAD Brand Fluorochemical Surfactant August 23, 1996 CHW 6329-197 PAGE 4 ................................................................. 7. PilECAUTIONARY INFORMATION ................................................................. EYE PROTECTIOW: Avoid eye contact. Near vented goggles. SKIN PROTECTION: Avoid skin contact - Wear appropriate gloves when handling this material. A pair of gloves made from the following material(s) are recommended: butyl rubber. Use one or more of the following personal protection items &3 neceS3ary to prevent skin contact: head covering, coveralls. Protective garments (other than gloves) should be made of either of the following materials: polyethylenttpolyvinylidentchloride (Saranex). REC"ENDED VENTILATION: Use with appropriatelocal exhaust ventilation. Provide 3UIILCLGn't ventilation to maintain eM133ions below recommendedexposure limits. If exbaU3t ventilation 13 not adequate, use appropriater83piratory protection. RESPIRATORY PROTECTION: Avoid breathingof airbornematerial. Selectone of the following NI03H approved respiratorsbased on airborne concentrationof contaminantsand in accordancewith OSHA regulations: full-Tacohighefficiencyfilterrespirator,full-facesuppliedair respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not *at, drink or smoke when Using th13 product.Wash exposed areas thoro0ghlywith soap and watet. 14ashhands after handling and before eatinz g. RECOMMENDED STORAGS: Do not store containerson their 31de3. Store at room temperature. Keep container dry. Keep container closedwhen not in use. **PP,EVEN"rtiOISTURE CONTMINATION TO KEEP POWDER FREE FLOWING.** FIRE AND EXPLOSION "OIDANCE: Keep containertightly closed. NO smokingwhile handlingth13 material. tiMISHAZARD RATINGS: 14EALTli3: FLAJTWILITY: 0 REACTIVITY: 0 PERSORAL PROTECTION: X (See precautions, section T.) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN* -1........a.d ...............................v.........a ......... AYMNIU14 PERFLUOROOCTANOAIE .......... 0.01 MGtM3 TWA ACGIM Y AkimoNium PERFLUOROMEPTANOATE ......... 0.1 MGll13 TWA 3M y AMMONIUM PERFLUOROPENTANOATE ......... 0.1 MGtM3 TWA 3M y AMMONIUM PERFLUOROHEXANOATE .......... Oll MGIM3 TVIA 3M y _a ......v .............................a ..................... Abbroviationsi N/D - Not Determined N/A - Not Applicable 53 MsoS: FC-143 FLUOPAD brand FluoroChemicaSlurtactant AUgU3% 23, 19$6 CHW 6329-197 PAGE 3 EXposuRE LIMITS (contjnubd) INGREDIENT VALUE UNIT TYPE AUTH $KrN* ............0..............v ............................................... SKIN NOTATION,. L13ted substances Indicated with 'Y' under SKIN refer to the potential contribution to the overall exposure by tilecutaneous route including mucous membrane and eye, either by airborne or, more particularly, by direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: ACGIM: American Conference of Governmental Industrial Hygienists 3M: 3M Recommended Exposure Guidelines ..................................................................... 8. HEALTH HAZARD IDATA ............................................................... EYE CONTACT: Moderate Eye Irritation: sign3/3YMPtOM3 pain, tearing, andhazyvision. can lnclude redness, swelling, Airborbe product may cause eye injury consisting of corneal opacity. SKIN CONTACT: Product is not expectedto be irritatingto the skin. Mild Skin Irritation(after prolongedor repeated contact): signs/3ynptOMS can include redness,swelling, and itching. May be absorbed through the skin and'"Per3l:Sitn the body for an extended ties. INHALATION: Illness requiring medical attention may result from a single exposure by inhalation to moderatequantities of this material. May be absorbed by inhalation and persist In the body for an extended time. Single overtxposures above recommended guidelines, May CSU59; rrrita *ion (upper respiratory):3ign3/symptoms can include sorOne)3 of the nose and throat, coughing and sneezing. Prolonged oe repeated overexposures above recommended guidelines, may cause,. Liver Effects: signslsymptomscan include yellow 3kin(jaundice) and tonderne3s of upper abdomen. Repeated inhalation of airborne product above the exposure guideline can result In elevatedorganofluoridelevels in the blood. ............................................................. Abbreviations: NIO - Not Determined N/A - Not Applicable 54 MSDS; FC-143 FLUORAD Brand FluorochemicalSurfactant August 23, 1996 CHW 6329-197 PAGE 6 ............................................................... 8. HEALTH K&ZMD DATA (Continued) ----w ........................................................... IF SWALLOWED: lng&3ti0ft is not a likely route of exposure to this pi-oducl.. Illn9S3 may result from a single 3wallowing of a moderate quantity of this material. CANCER: A mixture ot ammonium perfluorooctanoate, ammonium perfluorohoptanoate, ammonium perfluoropentanoate and ammonium perfluorohexanoate, that was 93 to 97% AMMONIUM PERFLUOROOCTANOATE (382S-26-1) was fed to albino rats for 2 years, no compound induced carcinogenicitywas found in the study. There were statistically significant compound related benign Itsticular tumors. In a second two-year study there were statistically significant compound related benign tumot-ain the liver, pancreas, and testis when compared to ad libitum and pair-fed controls. Based on the current knowledge, these finding3 have no human health implications. (1983 and 1993 studies conducted j6intly by 3M and DuPont). MUTAGENICITY,., Not mutagenic in invitro mutagenicityassays. Did not cause cell transformation in a mammalian cell transformation &$say. REPRODUCTIVEIDEVELOPMENTAL TOXINS: Not teratogenic in rabbitsby oral 8dMln13lraiion.Not teratogenicto r3t3 by gavage or.inhalationexposures. OTHER HEALTH RAZARD INFORMATION'. A 3H Product Toxicity Sumeary.Sheet is available. ...........4.................................................. SECTION CHANGE DATES ......................v............v.........w........................... HEADING SECTION ChM ED SINCE May 20, 1996 issue ............................................................. Abbrovia*clon3: N/D - Not Determined N/A - Not Applicable 55 MSDS: FC-143 FLUORAD Sr&Ad Fluorochemical Surfaciant August 23, 1996 CHW 6329-197 PAGE .....................................a ............................ The informatiionthisMateriaSlafetyDataSheettliSD1S3)bellevetdo be correct as of 'thedate issued. 3M WE$ NO WARRANTIES, EXPRESSED OR IMPLIEDs INCLUDING$ BUT NOT LIMITED TOl ANY IMPLIED KARRANTY OF MEROMANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whetherthe 3M productis fit for a particularpurposeand suitablefor user's method of use or application.Given the varietyof factorsthat can affect the use and applicationof a 3M product,3020 of which are uniquelywithin the User'sknowledgeand control, It 13 essentialthat the user evaluatethe 3H productto determinewhetherit is fit for a particularpurpose and suitablefor user's method of use or application. 3M providesinformationin electronicform as a serviceTO its customers. Due to the remote P033ibilitythat electronictransfermay have resulted in errors, omis3ions or altsration3In th13 information,3H makes no representatioasas to its completenessor accuracy. In addition, information obtained from a databaseany not be &3 current as the informationin the MSDS availabledirectlyfrom 3m. 56 APPENDIX B IndividuaAlntemortem Observations IndividuaBlody WeightData(g) IndividuaBlody Weight Gain Data (g) CHW 6329-197 57 Corning Hazleton Inc. Madison, Wisconsin USA Appendix B Individual Antemortem Observations (only Findings Other Than Normal) ---------------------------------------------------------------------------------------------------------------- CATEGORY GROUP: M2 DOSE: 5 MG/KG ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH ()UALIFIER DAYS 1-20 DAY 'C' - COMMENTS LISTED AT END OF OBSERV@ AN CHECK PRE/D[,Y 2.5 HR PH CHECK 1.0 HR 4.0 C77265 T 3 APPEARANCE SWOLLEN NOSE 2 p p p p Lot 00 Corning H*Zleton Inc. Madison, Wisconsin USA Appendix B individual Body weight Data (g) ---------------------------------------------------------------------------------------------------------------- ANIMAL DAY DAY DAY DAY DAY DAY DAY DAY HUMBER 1 2 3 4 5 6 7 8 ---------------------------------------------------------------------------------------------------------------- GROUP: MALE I - 0 MG/KG C77251 300 392 392 400 399 408 C77252 376 304 389 396 397 404 C77253 359 377 379 381 390 390 391 397 40 C77254 344 350 355 361 361 367 371 367 38 C?725S 311 316 315 318 318 320 321 315 33 C77256 333 33S 34S 342 345 352 C772S7 342 352 3SS 364 365 374 C772S8 356 36S 366 374 371 301 383 386 40 C77259 303 304 309 314 311 312 C77260 382 392 395 399 401 408 409 406 42 GROUP: MALE 2 5 MG/KG C77261 390 390 397 405 405 410 C77262 384 389 393 397 399 410 411 410 42 C77263 34S 356 361 370 365 379 C77264 360 366 364 375 378 384 C7726S 341 342 350 3S7 361 363 369 365 30 C77266 351 352 357 362 361 368 375 367 39 C77267 386 392 397 404 411 417 C77268 320 319 325 331 330 33S 339 337 35 C77269 348 351 355 359 361 371 C77270 362 365 371 372 375 378 380 385 39 GROUP: MALE 3 14 MG/KG C77271 345 349 350 349 345 341 354 355 37 C77272 349 359 360 363 350 347 C77273 395 404 412 413 411 407 417 421 43 C77274 360 369 371 374 371 378 387 382 40 C77275 382 309 393 396 389 391 Corning Hazleton Inc. Madison, Wisconsin USA Appendix B Individual Body Weight Data (a) --------------------------------------------------------------------------------------------------------------- ANIMAE, DAY DAY DAY DAY DAY DAY DAY DAY NUMBER 1 2 3 4 5 6 7 8 ---------------------------------------------------------------------------------------------------------------- GROUPT 14ALE 3 - 14 MG/KG C77276 370 373 377 377 369 366 C77277 393 307 391 386 360 346 C77278 376 375 380 376 360 339 326 322 35 C77279 369 378 385 385 384 300 395 392 41 C77200 388 396 402 403 412 415 GROUP: HAIE 4 42 MG/KG C77201 359 366 361 359 362 367 371 369 38 C77282 324 326 322 312 295 316 C17283 345 352 354 331 308 294 303 321 34 C77284 353 350 351 352 347 356 366 367 38 C77285 332 335 331 328 332 337 C77286 352 354 341 340 343 352 357 359 38 C77207 331 342 345 347 349 3S6 358 357 37 C77288 362 367 357 358 3S2 371 C77289 367 365 364 359 356 347 C77290 361 360 360 347 342 351 Corning Hazleton Inc. Madison, Wisconsin USA Appendix B Individual Body Weight Data ig) --------------------------------------------------------------------------------------------------------------- ANIMAL DAY DAY DAY DAY DAY DAY DAY DAY NUMBER 11 12 13 14 is 16 17 is --------------------------------------------------------------------------------------------------------------- GROUP: MALE I - 0 NGIKG C77253 408 421 428 436 437 445 443 456 4 C77254 381 396 400 404 405 408 417 424 4 C77255 326 340 341 346 349 341 350 353 3 C77258 402 419 424 432 430 426 443 446 4 C77260 415 437 444 448 449 445 453 461 4 GROUP: MALE 2 - 5 MGIKG C77262 430 444 449 458 458 457 469 476 4 C77265 377 393 396 400 403 399 all 414 4 C77266 381 397 403 407 412 403 417 421 4 C77268 352 365 368 373 373 370 379 384 3 C77270 398 411 419 424 422 425 429 433 4 GROUP: MALE 3 14 MG/KG C77271 373 384 386 389 395 390 403 406 4 C77273 438 454 456 463 468 461 473 484 4 C77274 396 411 414 423 422 419 433 436 4 C77278 369 390 392 402 407 410 424 433 4 C77279 403 421 427 434 434 434 4411 455 4 GROUP: MALE 4 42 MG/KG C77281 383 399 404 410 417 410 424 432 4 C77283 360 375 382 386 393 391 402 409 4 C77284 394 40S 409 416 421 419 426 425 4 C77286 373 389 394 398 402 405 415 418 4 C77287 369 380 383 391 393 395 405 411 4 Corning Hazleton Inc. Madison, Wisconsin USA Appendix B Individual Body Weight Gain Data (g) Days 1 - 6 --------------------------------------------------------------------------------------------------------------- ANIMAL DAY DAY DAY DAY DAY TOTAL NUMBER 1-2 2-3 3-4 4-5 5-6 1-6 --------------------------------------------------------------------------------------------------------------- GROUPT MALE 1 0 MG/KG C77251 12 0 a -1 9 20 C77252 8 5 7 1 7 28 C77253 18 2 2 9 0 31 C772S4 6 5 6 0 6 23 C77255 5 -1 3 0 2 9 C77256 2 10 -3 3 7 19 C772S7 10 6 6 1 9 32 C77258 9 1 a -3 10 25 (3h C77259 1 5 5 -3 1 9 t%i C77260 10 3 4 2 7 26 GROUP: MALE 2 5 MG/KG C77261 0 7 8 0 5 20 C77262 5 4 4 2 11 26 C77263 11 5 9 -5 14 34 C77264 6 -2 11 3 6 24 C7726S 1 8 7 4 2 22 C77266 1 5 5 -1 7 17 C77267 6 5 7 7 6 31 C?7268 -1 6 6 -1 5 is C77269 3 4 4 2 10 23 C77270 3 6 1 3 3 16 GROUP: MALE 3 14 MG/KG C77271 4 C77272 10 C77273 9 C77274 9 C77275 7 1 -1 -4 -4 -4 1 3 -13 -3 -2 8 1 -2 -4 12 2 3 -3 7 18 4 3 -7 2 9 Corning Hazleton Inc. Madison, Wisconsin USA Appendix B individual Body Weight Gain Data (g) Days I - 6 --------------------------------------------------------------------------------------------------------------- ANIMAL. DAY DAY DAY DAY DAY TOTA]6 NUMBER 1-2 2-3 3-4 4-5 s-6 1-6 --------------------------------------------------------------------------------------------------------------- GROTJP: C77276 C77277 C77270 C77279 C?7280 MALE 3 3 4 -1 9 6 14 MG/KG 4 4 5 7 6 0 -8 -3 -4 -5 -18 -22 -37 -4 -16 -21 -37 0 -1 4 19 1 9 3 27 GROUPT MALE 4 42 MG/KG C77291 C77282 C77283 C77284 C77285 C77286 C77287 C77288 C77289 C77290 7 -5 -2 3 5 8 2 -4 -10 -17 21 -8 7 2 -23 -23 -14 -51 -3 1 1 -5 9 3 3 -4 -3 4 5 5 2 -13 -1 3 9 0 11 3 2 2 7 25 5 -10 1 -6 19 9 -2 -1 -5 -3 -9 -20 -1 0 -13 -5 9 -10 Corning Hazleton Inc. Madison. Wisconsin USA AppendiX B Individual Body Weight Gain Data (g) Days I - 20 ---------------------------------------------------------------------------------------------------------------- ANIMAL DAY DAY DAY DAY DAY DAY DAY DAY NUMBER 1-2 2-3 3-4 4-5 5-6 6-7 7-8 0-9 9 ---------------------------------------------------------------------------------------------------------------- GROUP: C772S3 C77254 C772SS C77250 C77260 GROUP: MALE I - 0 MG/KG is 2 6 5 5 -1 9 1 10 3 MALE 2 - 5 MG/KG 2 9 0 6 0 6 3 0 2 8 -3 20 4 2 7 1 6 10 4 -4 17 1 -6 is 2 3 18 1 -3 16 C77262 C77265 C77266 C77268 C77270 GROUP: 5 1 1 -1 3 MALE 3 4 8 s 6 6 14 MG/KG 4 2 11 7 4 2 5 -1 7 6 -1 5 1 3 3 1 -1 16 5 -3 16 7 -8 is 4 -2 17 2 5 13 C77271 4 C77273 9 C77274 9 C77278 -1 C17279 9 1 -1 -4 -4 13 8 1 -2 -4 10 2 3 -3 7 9 5 -4 -16 -21 -13 7 0 -1 4 7 GROUP: MALE 4 42 MG/KG C77281 7 -S -2 3 5 4 C77283 7 2 -23 -23 -14 9 C77284 -3 1 1 -5 9 10 C77286 2 -13 -1 3 9 5 C77207 11 3 2 2 7 2 1 16 4 18 -5 19 -4 30 -3 18 -2 18 is 27 1 19 2 21 -1 14 Corning Hazleton Inc. Madison, Wisconsin USA Appendix 8 Individual Body Weight Gain Data (g) Days I - 20 ---------------------------------------------------------------------------------------------------------------- ANIMAL DAY DAY DAY DAY DAY DAY DAY DAY NUMBER 11-12 12-13 13-14 it-is 15-16 16-17 17-18 18-19 19 ---------------------------------------------------------------------------------------------------------------- GROUP: KALE 1 0 MG/KG C77253 C77254 C77255 C?7258 C77260 GROUP: 13 is 14 17 22 KALE 2 7 4 1 5 7 5 MG/KG 1 8 -2 13 5 4 1 3 9 7 0 5 3 -8 9 3 s a -2 -4 17 3 -1 4 1 -4 8 8 6 c77262 C7?265 C77266 C77260 C77270 GROUP: 14 16 16 13 13 KALE 3 5 3 6 3 8 14 MG/KG 9 0 -1 12 4 3 -4 12 4 5 -9 14 5 0 -3 9 5 -2 3 4 7 6 3 5 4 3 5 0 4 11 C7?271 11 C77273 16 C77274 15 C77278 21 C77279 le 2 3 6 -5 13 3 2 2 7 5 -7 12 11 -1 3 9 -1 -3 14 5 6 2 10 5 3 14 9 0 1 6 7 0 0 13 8 1 GROUP: KALE 4 - 42 MG/KG C77281 16 5 6 7 -7 14 0 7 C77283 15 7 4 7 -2 11 7 2 C77284 11 4 7 5 -2 7 -1 a 1 C7?286 16 5 4 4 3 10 3 2 C77287 11 3 8 2 2 10 6 6 !.' CHW 6329-197 APPENDEK C IndividuaCllinicalChemistryData (Day 6 PalmitoylCoA oxidaseactivity) 66 Appendix C Individual Clinical chemistry Data males Day 6 ANIMAL NUMBER ------ PCOAO IU/G -------- Group: I C77251 C77252 C77256 C77257 C77259 Dose Level: 0.0 4 7 6 4 4 Dosage Unit: mg/kg MEAN S.D. N Group: 2 C17261 C77263 C77264 C77267 C77269 MEAN S.D. N Group: 3 C77272 C77275 C77276 C77277 C77280 5 1.4 5 Done Level: 5.0 24 24 27 24 22 24 1.8 5 Dose Level: 14.0 41 30 42 34 41 Dosage Uniti mg/kg Dosage Unit: mg/kg MEAN S.D. N 39 3.3 5 Appendix C individual Clinical Chemistry Data Males Day 6 ANIMAL NUMBER ------ PCOAO IU/G -------- Group: 4 C77282 C77285 C77288 C77209 C77290 MEAN S.D. Dose Level: 42.0 40 42 45 33 33 39 5.4 Dosage Unit; mg/kg 00 CHW 6329-197 i APPENDEK D IndividualAnixnalLiverWeight Values (g)-Day 6 Sacrifice IndividualAnimal PathologyData -Day 20 Sacrifice 69 Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Liver Weight Values (9) Day 6 Sacrifice ---------------------------------------------------------------------------------------------------------------- TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH-I;SUBSET=ALL ORGAN ABBREVIATION: LIO - WHOLE LIVER, Lll - LIVER, RIGHT LATERAL LOBE, L12 SEX DOSE AN114AL TERMINAL - ----- ----- ------ ------- ----- ----- ------ GROUP NUMBER BODY WT (g) LIO LII L12 ---------------------------------------------------------------------------------------------------------------- m I C77251 m I C77252 m I C77256 m I C77257 m 1 C77259 400.4 394.5 341.5 368.5 306.8 0.0 ma/ka 17.7972 16.4199 16.7503 17.8283 11.9291 2.0690 2.5533 2.4016 2.5079 1.7786 14.8583 13.8876 14.2603 15.1900 10.1344 m 2 C77261 m 2 C77263 m 2 C77264 m 2 C77267 m 2 C77269 403.4 370.1 378.9 408.6 363.6 S@O malka 24.1163 20.6942 21.0651 24.2954 18.5405 3.4006 2.8659 2.7795 3.7472 2.3964 20.5902 17.6999 18.2230 20.4106 16.0782 m 3 C77272 m 3 C77275 m 3 C77276 3 C77277 Nm 3 C77280 14.0 mg/ka 343.2 380.2 360.7 346.9 411.8 23.2873 24.3657 20.S304 16.1233 28.2782 3.1381 3.0794 2.7287 2.444S 4.67S2 20.0752 20.4197 17.7335 13.6463 23.4964 N 4 C77292 m 4 C77285 m 4 C77288 m 4 C77289 m 4 C77290 42.0 malka 306.8 328.8 363.5 344.7 345.7 17.2123 22.4106 23.7435 21.1160 24.1904 2.3432 3.6637 3.3208 3.2493 3.5439 14.8110 18.6428 20.3694 17.7983 20.5800 Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D rndividual Animal Pathology Data Day 20 Sacrifice --------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77253 SEX: 14ALE DOSE GROUP: I SACRIFICE STATUS: SCHEDULED, TERMINAL SAC DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:48 PROSECTOR: DEBBIE PIRKEL RECORDER: DEBBIE PIR -PO-S-T---FI-X---W-EI-G-H-ER-:---N-O-T--A-VA-I-L-A-BL-E------------------P-A-TH-O-L-OG-I-S-T:----D-R-. --T-OM---PA-L-M-ER-----------------W-EI-G-H-ER-:---T-R-OY---E-LL-E-NB---------- ORGAN NAME ---------------------- WHOLE LIVER (LIO) RT LAT LOBE LIV (LII) REMAINING LIV (L12) ABSOLUTE ORGAN WEIGHT (GRAMS) -------- 16.6610 -------- -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- 4.0453 t --------------- ORGAN TO BRAIN WEIGHT RATIO --------------- --------------- 0 s --- WEI NOI NOI G R 0 S S P A T H 0 L 0 G Y 0 B S E R V A T I-0-N-S --- -----------O-R-GA-N---N-AM-E-------------- S-E-V-ER-I-T-Y,----KE-Y-W-OR-D-(-S-) ---OR---PH-R-A-S-E -----------F-REE-TEXT COMMENTS AND NOTES -------------------------------------------GENERAL COMMENT (OC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77254 DATE OF DEATH: 01/21/97 SEX; MALE DOSE GROUP: 1 SACRIFICE STATUS; SCHEDULED, TERMINAL SACR STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:40 PROSECTOR: DEBBIE PIRKEL RECORDERI DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE4 ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- ST --- WHOLE LIVER (LIO) 15.3258 3.6171 t -------- WEI RT LAT LOBE LIV (Lll) -------- -------- -------- NOT RE14AINING LIV (L12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- . . * G R 0 S 9 P A T H 0 L0 0 Y 0 B S E R V A T 1 0 N 9 ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ---------------------------------------------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice --------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77255 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SAC DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TINE OF NECROPSY: 01/21/97 10:53 PROSECTOR: DEBBIE PIRKEL RECORDER: DEBBIE PIR POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TON PALMER WEIGHER: TROY ELLENB ---------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- WHOLE LIVER (LIO) RT LAT LOBE LIV (LIl) REMAINING LIV (L12) ABSOLUTE ORGAN WEIGHT (GRAMS) -------- 13.7530 --------------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT -------- 3.8708 --------------- ORGAN TO BRAIN WEIGHT RATIO --------------- --------------- 0 sI --- WEX NOT NOT G R 0 S S P A T H 0 L 0 G Y 0 B S E R V A T 1-0-N-S --- ---------- ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- -4 GENERAL COMMENT (OC) . . . . . . . . . w -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77258 SEX: MALE DOSE GROUP: I DATE OF DEATH: 01/21/97 STUDY DAY OF DEATHI 20 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:13 PROSECTOR.- NANCY DIEDRICH POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER RECORDER: NANCY DIEDR -------------------------------------------------------------------------------------WE-I-G-HE-R-:---T-RO-Y---E-LL-E-NB-E-I--------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO 0 ST -------- WHOLE LIVER (LIO) 17.8225 4.0060 1 -------- RT LAT LOBE LIV (LIl) -------- -------- -------- WEI NOT REMAINING LIV (L12) -------- -------- -------------------------------------------------------------------------------------------------------------N-O-T--------- G R0 S S PA T H 0 L 0 G Y 0 B S E R V A T 1 0 N S O-R-G-A-N--N-A-M-E------------- S-E-V-E-R*I-T-Y*-,-*--K-EY-W-O-RD-(-S-)---O-R--P-HR-A-S-E-----------F-REE-TEXT COMMENTS AND NOTES GENERAL COMMENT (GC) -------------------------------------------- .... ...... ...... .... ....... ....... ..... .... -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ------------------------------------------------------------------------------------------------------------------ Corning Hazleton Inc. Madison. Wisconsin USA APPENDIX D individual Animal Pathology Data Day 20 Sacrifice ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77260 SEX: MAIE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDUBED, TERMINAL SACRI DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: 4 DATE AND TIME OF NECROPSY: 01/21/97 10:20 PROSECTOR: DEBBIE PIRKEB RECORDER: DEBBIE PIRKE POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBEC ----------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- ABSOLUTE ORGAN WEIGHT IGRANS) -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- ORGAN TO BRAIN WEIGHT RATIO -------- 0R ST ---- WHOLE LIVER 4LIO) 17.6747 3.8191 t -------- WEIG RT LAT LOBE LIV (Lll) -------- -------- -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ----------------------------------------------------------------------------------------------------------------- * G R0 S S P A T H 0 L0 G Y 0 B S E R V A T 10 N S ORGAN NAME SEVER*ITY*. KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- LA GENERAL COMMENT (GC) . -NO 'tL@CiOiC6PIC'Lisiois' -ABBR NEC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Nadison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice --------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77262 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SAC DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHTT DATE AND TIME OF NECROPSYT 01/21/97 10:34 PROSECTOR: DEBBIE PIRKEL RECORDER: NANCY DIED POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER., TROY ELLENB --------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- WHOLE LIVER (LIO) RT LAT LOBE LIV (Lll) REMAINING LIV (L12) ABSOLUTE ORGAN WEIGHT (GRAMS) -------- 21.8525 --------------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- 4.5880 1 --------------- ORGAN TO BRAIN WEIGHT RATIO --------------- -------- 0 s WE w NOI G R 0 S S P A T H0 L 0 G Y 0 B S E R V A T 10 N s --------- ORGAN NAME SEVERITY. KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Day 20 Sacrifice Data ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C?7265 SEX: HALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10;34 PROSECTOR: NANCY DIEDRICH RECORDER: NANCY DZEDR POST-FIX WEIGHERs NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBO ---------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- ABSOLUTE ORGAN WEIGHT (GRAMS) -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- ORGAN TO BRAIN WEIGHT RATIO -------- 0 ST WHOLE LIVER (LIO) RT LAT LOBE LIV (LIl) 18.0251 -------- 4.5329 1 -------- -------- WEI -------- NOT REMAINING LIV (L12) -------- -------- ---------------------------------------------------------------------------------------------------------------NO-T ORGAN NAME ---------------------- GENERAL COMMENT (GC) G R0SS PATH0L0G Y 0B SERVAT 10Ns SEVERI*TY,* * KEYWORD(S) OR PHRASE --------------------------------------- FREE-TEXT COMMENTS AND NOTES --------------------------------------------- . ..... ...... ...... ...... ..... ....... ..... .. -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ---------------------------------------------------------- L------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77266 SEX: KALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 11:07 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE( ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 01 ORGAN NAME ---------------------- (GRAMS) -------- TO 13ODY WEIGHT (t) -------- WEIGHT RATIO -------- ST ---- WHOLE LIVER (LIO) RT LAT LOBE LIV (Lll) 20.5301 -------- 4.83S2 t -------- -------- WEIG -------- REMAINING LIV (L12) -------- -------- NOT -------- NOT ----------------------------------------------------------------------------------------------------------------- SL * * 'I G R 0 S S P A T H 0 L 0 G Y 0 8 S 9 R V A T 1 0 N S O-R-GA-N--N-AM-E----------------;-.-V-ER-I-T-Y.--KE-Y-WO-R-D-(S-)--O-R--PH-R-A-SE----------F--R--E-E----T--EX--T--C-O-MM-E-N-TS---AN-D--N-OT-E-S ----------------------- -4 GENERAL COMMENT (GCJ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00 -NO MACROSCOPIC LESIois -ABBR NEC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Nadison, Wisconsin USA APPENDIX D individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77268 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:20 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIRK POST-FIX WEIGHERs NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENB ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- ST WHOLE LIVER (LrO) 16.0923 4.2471 t -------- WEI RT LAT LOBE LIV (LII) -------- -------- -------- NOT RE14AINING LIV (LZ2) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- * * * G R 0 S S P A T H 0 L.0 G Y 0 B S E R V A T 1 0 N S ORGAN NAME SEVERITY, KEYWORD(S)OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- GENERAL. COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . -ABBR NEC PERFORM THYMUS (TH) . . . . .. RE@ FOCUS(I)/AREAIS)* . . ..... . . .RIGHT-LOBE:.MULTIPLE DARK-RED-FOCI,.UP TO-1. ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison. Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- AN114AL NUMBER: C77270 SEX.- MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:08 PROSECTOR: NANCY DIEDRICH RECORDER: NANCY DIEDR POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WRIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- ST --- WHOLE LIVER (LIO) 20.4477 4.6272 t -------- WEI RT LAT LOBE LIV (Lll) -------- -------- -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- .*. G R0S S PATH0 L00Y 0 BSERVAT 10NS ORGAN NAME SEVERITY, KEYWORD(S)OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- 00 GENERAL COMMENT (GC) . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NBC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D individual Animal Pathology Data Day 20 sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77271 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:26 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (1) -------- WEIGHT RATIO -------- ST --- WHOLE LIVER (LIO) 20.9805 5.1868 t -------- WEI RT LAT LOBE LIV (LIl) -------- -------- -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- * . * G R 0 S S PA T H 0 L 0 G Y 0 B S E R V A T 10 N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ---------------------------------------------------------------------------------- 00 GENERAL COMMENT (GC) . . . . . . . . . -NO MACROSCOPIC LESIO@S' -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBERT C77273 DATE OF DEATH: 01/21/97 SEX: MALE STUDY DAY OF DEATH: DOSE 20 GROUP: 3 SACRIFICE STATUS: STUDY WEEK OF DEATH: 3 SCHEDULED. TERMINAL SACRI TERMINAL BODY WEIGHT: 4 DATE AND TIME OF NECROPSY: 01/21/97 10:40 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIRKE POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBEC ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO ST -------- ---- WHOLE LIVER (LIO) RT LAT LOBE LIV (LIl) 24.5697 -------- 5.1091 1 -------- -------- WEIG -------- REMAINING LIV (L12) -------- -------- -------- NOT NOT ----------------------------------------------------------------------------------------------------------------- * * * G R 0 S S PA T H 0 L 0 G Y 0 B S B R V A T 1 0 N S ORGAN NAME SEVERITY. KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ------------------------------------------------------------------------------------ 00 GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ti -NO MACROSCOPIC LESIONS -ABOR NBC PERFORM -------------------------------------------------------L-:-------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice --------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77274 DATE OF DEATH: 01/21/97 SEX: MALE STUDY DAY OF DEATH: DOSE 20 GROUP: 3 STUDY WEEK SACRIFICE STATUS: OF DEATH: 3 SCHEDUE.ED, TERNINAI, SAC DATE AND TIME OF NECROPSY: 01/21/97 10:07 POST-FIX WEIGHER: NOT AVAILABLE PROSECTOR: DEBBIE PIRKEL TERMINAL BODY WEIGHT: RECORDER: NANCY DIED PATHOE.OGIST: DR. TOM PALMER -------------------------------------------------------------------------------------WE-I-G-"E-R-s---T-R-OY---E-LL-E-N-R --------- ORGAN NAME ---------------------WHOLE LIVER (LIO) RT LAT LOBE LIV (LII) REMAINING LIV (L12) ABSOLUTE ORGAN WEIGHT IGRANS) -------- 23.1173 -------- -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- 5.2912 t --------------- ORGAN TO BRAIN WEIGHT RATIO ---------------------- -------- 0 s WE NO N(y SL ORGAN NAME 'VERITY. KEYWORD(S) OR PHRASE S E R V A T r0 N S ---------------------- --------------------------------------P-REE-TEXT COMMENTS AND NOTES 00 GENERAL COMMENT (GC) . . . . . . . . ------------------------------------------- -NO MACROSCOPIC LiSiO@S' -ABBR NEC PERFORM ..... ...... ...... ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Nadison. Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77278 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:13 PROSECTOR: DEBBIE PIRKEL RECORDER: DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE ---------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- ABSOLUTE ORGAN WEIGHT IGRANS) -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT 1?4) -------- ORGAN TO BRAIN WEIGHT RATIO -------- 0 ST --- WHOLE LIVER (LIO) RT LAT LOBE LIV (LIl) 22.4307 -------- 5.0886 t -------- -------- WEI -------- NOT REMAINING LIV IL12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- * * * G R 0 S S P A T H 0 L 0 G Y 0 B S E R V A T 10 N S ORGAN NAME ---------------------- SEVERITY, KEYWORDIS) OR PHRASE FREE-TEXT --------------------------------------- COMMENTS AND NOTES -------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . .o@s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO 14ACROSCOPIC LESI -ABBR NEC PERFORM ----------------------------------------------------------- :1 ---------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77279 SEX: KALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRI DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: 4 DATE AND TIME OF NECROPSY: 01/21/97 11:07 PROSECTOR: DEBBIE PIRKEL RECORDER: DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBE( ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WRIGHT RELATIVE ORGAN TO BRAIN 01 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT It) -------- WEIGHT RATIO -------- ST ---- WHOLB LIVER (LIO) RT LAT LOBE LIV (Lll) 20.6890 -------- 4.5904 1 -------- -------- WEIG -------- REMAINING LIV IL12) -------- -------- NOT -------- -----------------------------------------------------------------------------------------------------N-O-T ---------- SEVE; * * G R 0 S S P A T H 0 L 0 G Y 0 B S E R V A T 1 0 N S ORGAN NAME ---------------------- -----I-T-Y-, --K-EY-W-O-R-D(-S-)---O-R--PH-R-A-S-E-----------F-REE-TEXT COMMENTS AND NOTES --------------------------------------------- 00 GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIONS -ABBR NEC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice --------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77281 SEX.- MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SAC DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 11:00 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIR POST-FIX WEIGHERI NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENB --------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- s WHOLE LIVER (LIO) 21.3918 4.8464 t -------- WE RT LAT LOBE LIV (Lll) REMINING LIV (L12) --------------- --------------- --------------- Nol -------------------------------------------------------------------------------------------------------N-O-I------ * * * G R 0 S S P A T H0 L 0 G Y 0 B S E R V A T 10 N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESI(iis -ABBR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Nadison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77283 SEX.- MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY: 01/21/97 10:53 PROSECTOR: NANCY DIEDRICH RECORDER: DEBBIE PIRK POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENB ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- 5T WHOLE LIVER (LIO) RT LAT LOBE LIV (Lll) 20.9047 -------- S.0568 t -------- -------- WEI -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- & 6 * G R 0 S S P A T H0 L 0 G Y 0 B S 9 R V A T 10 N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ----------------------------------------------------------------------------------- GENERAL COMMENT (GC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -NO MACROSCOPIC LESIO@s -ABOR NEC PERFORM ---------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ----------------------------------------------------------------------------------------------------------------- ANIMAL NUMBER: C77204 SEX. MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRI DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERNIHAI, BODY WEIGHT: 4 DATE AND TIME OF HECROPSY: 01/21/97 10:26 PROSECTOR: DEBBIE PIRKEB RECORDER: DEBBIE PIRKE POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLENBEC ----------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0R ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WEIGHT (t) -------- WEIGHT RATIO -------- ST ---- WHOLE LIVER (LIO) RT LAT LOBE LIV (Lll) 21.2416 -------- 4.8663 1 -------- -------- WEIG -------- REMAINING LIV (L12) -------- -------- NOT -------- NOT ----------------------------------------------------------------------------------------------------------------- * * * G R 0 S S PA T H0 L 0 G Y 0 B S E R V A T 10 N s O-R-GA-N---N-AM-E-------------- S-E-V-E-RI-T-Y-,---K-EY-W-O-RD-(-S-)---O-R--P-HR-A-S-E-----------F-REE-TEXT COMMENTS AND NOTES --------------------------------------------- 00 GENERAL COMMENT (GC) . . . . . . . . . . 00 -NO MACROSCOPIC LESIONS* -ABBR NEC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D Individual Animal Pathology Data Day 20 Sacrifice ----------------------------------------------------------------------------------------------------------------- ANIMAE, NUMBERT C77206 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRI DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH.- 3 TERMINAL BODY WEIGHT: 4 DATE AND TIME OF NECROPSY: 01/21/97 11:00 PROSECTOR: DEBBIE PIRKEL RECORDER: DEBBIE PIRKE POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY ELLEMBEC ----------------------------------------------------------------------------------------------------------------- ORGAN NAME ---------------------- ABSOLUTE ORGAN WEIGHT (GRAMS) -------- ORGAN WEIGHT RELATIVE TO BODY WEIGHT (t) -------- ORGAN TO BRAIN WEIGHT RATIO -------- 0R ST ---- WHOLE LIVER (LIO) 24.6307 5.8714 t -------- WEIG RT LAT LOBE LIV (LII) -------- -------- -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ----------------------------------------------------------------------------------------------------------------- * * * G R 0 S S P A T H 0 L0 0 Y 0 8 S E R V A T 10 N S ORGAN NAME SEVERITY. KEYWORDIS) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ------------------------------------------------------------------------------------ 00 GENERAL COMMENT (GC) . . . . . . . . -NO MACROSCOPIC LE"SIONS' ' ' * * ' * * * * * * ' * ' ' * * * " ' * * ' * ' * * * * ' ' -ABBR NBC PERFORM ----------------------------------------------------------------------------------------------------------------- Corning Hazleton Inc. Madison, Wisconsin USA APPENDIX D individual Animal Pathology Data Day 20 Sacrifice ---------------------------------------------------------------------------------------------------------------- ANIMAI,NUMBERs C77207 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED. TERMINAL SACR DATE OF DEATH: 01/21/97 STUDY DAY OF DEATH: 20 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: DATE AND TIME OF NECROPSY. 01/21/97 10.47 PROSECTOR: NANCY DIEDRICH RECORDERZ NANCY DIEDR POST-FIX WEIGNERs NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: TROY EBLENOW ---------------------------------------------------------------------------------------------------------------- ABSOLUTE ORGAN WEIGHT ORGAN WEIGHT RELATIVE ORGAN TO BRAIN 0 ORGAN NAME ---------------------- (GRAMS) -------- TO BODY WRIGHT -------- WEIGHT RATIO -------- ST --- WHOLE BIVER (LIO) 18.9698 4.6133 -------- Wol RT L.AT LOBE bIV (Lll) -------- -------- -------- NOT REMAINING LIV (L12) -------- -------- -------- NOT ---------------------------------------------------------------------------------------------------------------- * * * G R0 S S PA T H 0 L0 G Y 0 B S B R V A T 10 N S ORGAN NAME SEVERITY, KEYWORD(S) OR PHRASE FREE-TEXT COMMENTS AND NOTES ---------------------- ---------------------------------------------------------------------------------- GENERAL KIDNEY COMMENT (GCI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -ABBR NEC PERFORM (KD) . . . . .- . . . . . . . . . . . . . . . ..... . . . LARGE PELVIS(ES) -Liri ----------------------------------------------------------------------------------------------------------------