Document kDDDdMwM8qwGwED4BnX9RwGrV

Zoe nemN Sid eAA: ! - HAZLETONuasonsronies ue. ne SERIE IESPPR, cS RE 22S6-Reoe 01 RECENEDS DEC 27 1983 TO ey RAT TERATOLOGY STUDY T-3351 FINAL REPORT 0 Submitted to 3M Company $1. Paul. Minnesota Decenber 19, 1983 000532 ; _ @HAZLETON uaonronesavesioa ne se Sn OFFICE OF QUALITY ASSURANCE Project Title: Rat Teratology Study Project No.: 154-160 Quality Assurance inspections of the of the above referenced project were study and conducted review of according the final report to the standard operating procedures of the the general requirements of Office of Quality Assurance and according to the Good Laboratory Practice regulations that were issued on December compliance on and after 22, 1978, by the Food and Drug Administration June 20, 1979. Findings from the 1nspections for and final report review were on the following dates: reported to management and to the study director Inspections/Review Findings Reported Inspector/Reviewer Protocol 6/15/83 Study 7/6,8,11,12/83 Final report 12/6-9/83 6/15/83 15/83 12/16/83 E. Burroughs E. Burroughs P. Runge TL AO SO : Office of Quality Assurance 000533 EE Its EE @ HAZLETON Leonsrones auemon ne SUBJECT: Rat Teratology Study Project No. 154-160 under We, the undersigned, hereby declare that our supervision, according to the procedures the work was performed herein described. Study Director: Aus LAWRENCE T. WETZEL, Ph.D. Staff Scientist Department of Toxicology Laboratory Supervision: 4 Lads GEORGE A. BURDOCK, Ph.D. Rodent Toxicology I Department of Toxicology Teratology: Lod SD seo RUTH S. "DURLOO, B.S. Research Associate Department of Toxicology Report Preparation: BARBARR R- CSTPE, BAY, B.S. TseccihennitciaflicWrRietseourrces Department us . 000534 LT en HAZLETON wisonsrones sencanc ei NN RATR ThBLE oF conrenTs sumaRY ha\se wTRuCTION . CONTROL AND TEST MATERIALS 5 TEST antuaLs ` werians 5 Mating Period CGornopuapusndandPreDpoasargaetioLnevaenlds Adatnistration :5 5 VCMiaestsceaerrrneaaalln ESObxasacemrriivnfaaittciieoonnsofandFetRuesceosrds :;6 Skeletal Examination of Fetuses Tissue Preservation 8 Statistical Analyses Specimen, Raw Data, and Final Report Storage 8 9 RESHSaoUdrLytTaSlWie~tiygMhAtaTsnEdRNanACdLlinFOioAcoTadA) CoGnbssuemrpvtaitoinans Uterine Heights n ji32 i Gross Pathology RESULTS ~ CESAREAN OATA 213 FFPeertteaagllnanDVcieyzvbeilR1oaipttmeyse.,ntWCeotrapnotr,a aLnudteaS,exand Uterine Implantations i1hsd noes Table 1 - Summary of Maternal Clinical Observations Table 2 - Mean Maternal Body Weights and Body Weight Changes n 18 Table 3 - Mean Maternal Food Consumption Table 4 - Mean Terminal Body Weights, Gravid Uterine Weights, 19 Table 5 - and Terminal Body Weights Summary of Maternal Gross Minus Gravid Uterine Pathology Findings Weights. 20 21 Key to Tables 6 - 14 Table 6 - Summary of Mean Ovarian and Uterine Data 23 25 000535 @HAZLETON viomurones avec ne ee ada TABLE OF CONTENTS - coRTINED Table 7 Table 8 - Summary of Mean Litter Data - Summary of External Findings Table 3 - Summary of Visceral Findings Table 10 - Summary of Skeletal Findings Table Table 11 12 - Incidence Incidence of of Externa) Visceral Findings Findings Per Per Litter Litter Table Table 13 14 - Incidence of Skeletal Findings Per Litter Mean Incidence of External, Visceral, and skeletal Findings Per Litter ApvenDices Appendix 1 - Individual Clinical Signs Key to Appendices 2, 3, and 4 Appendix 2 - Individual and Mean Maternal Body Weights and Body Weight Changes Appendix 3 - Individual and Mean Food Consumption Appendix 4 - Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid uterine Weights Appendix Appendix 5 6 - Individual Individual and and Mean Mean Ovarian and Litter Data Uterine Data Appendix 7 - Individual Visceral Findings in Fetuses Appendix 9 - References Appendix 8 - Individual Skeletal Findings in Fetuses ne Page 26 27 28 29 30 3 32 33 534 39 43 47 51 755 59 67 000536 -- tae omital Sra HAZLETON Lsonsrones averios, ne, SPONSOR: 3K Company DATE: December 19, 1983 SUBJECT: FINAL REPORT . Rat Teratology Study Project No. 154-160 SUMMARY T-3351, suspended in corn ofl at concentrations intended to deliver 1, 5, or 10 administered by oral mg/kg/day (roups 2, 3, and 4, respectively), was intubation to mated female Sprague-Dawley CD rats on presumed gestation Days. 6-15. A fourth group (Group 1) of mated females served as the control group and received the vehicle only. Two Group 4 females died. Treatment-related clinical observations noted fn Groups 3 and 4 {included thinness, hunched appearances, rough haircoats, alopecia, and anorexia. Food consumed by Groups 3 and 4 animals was significantly less than control both during treatment and throughout gestation. Compared to control body weight gains, significantly less body weight was gained by Groups 3 and 4 females throughout gestation. Actual loss of body weight occurred in Groups weight 3 and minus 4 animals once treatment had been initiated. Terminal body gravid uterine weight was also significantly decreased in : 000537, Groups 3 and 4. Treatment-related gross pathology in Group 4 animals included incidences of gastrointestinal tract effects. Statistical analysis revealed a positive treatment-related trend in fetal loss, attributable primarily to an increased incidence of late resorbing fetuses and dead fetuses in litters from Group 4 dams. Mean male and female fetal body weights were significantly decreased in fetuses Groups 3 and 4 litters. Significant increases in external and visceral anomalies occurred in fetuses from Group 4 dams and included subcutaneous edema, cleft palate, and cryptorchism. Anomalies were observed primarily fn fetuses from Group 4 dams exhibiting signs of . severe maternal toxicity, i.e., decreased food consumption and weight loss. Fetal toxicity manifested as increased numbers of late resorbing fetuses and dead fetuses, decreased fetal body weights, and delays in skeletal system ossification occurred in litters derived from that group as well. The specificity and frequency of these results indicate that administration of T-3351 to pregnant rats during Days 6-15 of gestation at a dose of 10 mg/kg results in maternal toxicity, fetotoxicity and an increased incidence of external and visceral anomalies and skeletal T-3351 did not appear to be teratogenic in rats at dose levels Tess than or equal to mg/kg. 000538 + - cakes @HAZLETON sonurones menor ne. 15ers se INTRODUCTION This study was designed to evaluate the embryo/fetal toxicity and teratogenic potential of T-3351 when administered by oral gavage to pregnant rats during the period of fetal organogenesis. The rats were placed in breeding on June 27, 1983, and cesarean sections were completed on July 26, 1983. The study was conducted according to a protocol designed to satisfy existing guidelines of the Food and Drug Administration as well as Good Laboratory Practice Regulations. This report presents and discusses the methods and results from the study. CONTROL AND TEST MATERIALS The vehicle and control material, Duke's Corn 011 (Lot No. 80275), a slightly viscous, yellow liquid, was received from the C. F. Sauer Co., Richmond, Virginia, and was stored at room temperature. The test material, T-3351, a white powder, was received from the Sponsor on January 5, material was assumed methods of synthesis, sponsor. characteristics which 1983, and was stored at room temperature. The test to be 100% active compound. Information on the stability, as well as data on composition, or other define the test material, are on file with the : 000539 - SAG, ae. --a- TEST ANIMALS Tuo hundred sixty (one hundred thirty/sex) sexually mature, cesarean-derived Sprague-Dawley Cr1:CDe (S0)BR rats were received from Charles River Breeding Laboratortes, Inc., Kingston, New York, on June 8, 1983. The rat was selected for use fn this study because historically this species has been used in teratology studies and Is required by appropriate regulatory agencies. The animals were housed individually in elevated stainless-steel cages and acclimated to laboratory conditions for approximately three weeks prior to initiation of the study. During this tine, two male rats were euthanized due to an unthrifty appearance. Following an examination by a staff veterinarian for health status, one hundred twenty eight rats/sex were placed in breeding. The rats were housed one male and ane female por cage during breeding. Following confirmation of mating, the females were uniquely identified by ear tags and individually housed tn elevated stafnlesssteel cages. Food (Purina Rodent. Laboratory Chawe 5001) and tap water (via an automatic watering system) were available ad 1ibitus. The mean room temperature + the standard deviation was 72.9% + 1.36 and the mean relative humidity + the standard deviation vas 69.1% + 3.16 during the study. A tuelve-hour Mght/dark cycle was maintained. 000540 . I - Gn ha RRR + @HAZLETON somsrones mercrc -- -5- ETHODS Hating period During the mating period, one female was paired with one male until mating was confirmed or until two weeks had elapsed. A sufficient number of mated females, during the course of this study, were obtained after nine days of cohabitation and mating was discontinued. Daily vaginal examinations of each female were performed to detect the presence and viability of sperm or the presence of a copulatory plug. The day of observation of sperm or copulatory plug was designated as "Day 0% of gestation. Groupsand DosageLevels Upon confirmation of mating, each female was assigned to one of the following groups by use of a table of random numbers (Cochran and . Cox, 1987). Group. Number of Females 1 RT 2 2 3 2 4 2 #Bdausreidng otnhe idnodsiivnigduapleriaondi.malAnibmoadlys wweeirgehtdsoseadt presumed gestation. ma/Dkogs/ed?ay 0 1 5 10 oenacDhayswei6ghtihnrgougihnt1e5rvaolf All males and all remaining females were sacrificed via carbon dioxide asphyxiation and discarded without necropsy once a sufficient number of matings were confirmed. 000541 4 EEL EE SEEN --6- Compound Preparation and Administration Suspensions of the test materfal in the corn ofl vehicle were prepared on a weight per volune basis. The required amount of compound for each level was weighed on an electronic Arbor 126 balance and trans ferred to a 7 ml tissue grinder. Approximately 3 ml of corn ofl was added and the test material was ground into a fine suspension. This suspension was transferred to a precalibrated beaker and any remaining omouNd in the tissue grinder was rinsed into the beaker. Addition) corn ofl was added to the suspension to produce the desired concentration and the suspension was mixed ona magnetic stirrer. Prepared suspensions were mixed with a magnetic stirrer during dosing. Females were given the appropriate dosing suspension or vehicle by oral intubation on a daily basis beginning on Day 6 and continuing through Day 15 of presumed gestation. The dose administered to each female was based on the most recently recorded individual body weight. The test material was administered orally because of the relative ease and accuracy of dosing. Fresh dosing suspensions were prepared weekly and stored under refrigeration (approximately 40F). Samples of each test mixture as well as the vehicle were sent to the sponsor for analysis. Maternal Qbservations and Records All animals were examined twice datly for mortality and moribun- dity (from day of recetpt through animal sacrifice) and once daily for clinical signs of toxicity and pharmacologic effects (throughout gestatom. Tndividual body wetahts were recorded at- initiation and on Days 000542 sre 6:8. 12, 16, and 20 of presumed gestation. Food consumption during gestation was determined on Days 6, 8, 12, 16, and 20. Cesarean Sacrifice A gross necropsy was performed on all females found dead or terminally sacrificed. The uterus and ovaries from found dead females were removed and examined for the number of implantation sites and corpora lutea. Prior to terminal sacrifice and cesarean section on Day 20 of gestation, all surviving females were assigned random numbers so that all persomnel performing cesarean sections and/or subsequent external, visceral, or skeletal examinations of the fetuses were unaware Of the dose level from which the animals were derived. On Day 20 of gestation, all surviving females were weighed and sacrificed by carbon dioxide asphyxiation. The uterus from each female was excised, weighed, and examined for the number and placement of uterine implantation sites, number of live and dead fetuses, number of early and late resorbing fetuses, and any abnormalities. The uterus was not reueighed after the contents were removed. The ovaries were examined for the number of corpora lutea. Each female was examined grossly for any abnormalities. Each fetus was sexed, weighed, examined externally, and tagged for identification. Beginning at the ovarian end of the left uterine horn and continuing past the cervix into the right uterine horn, the' first available fetus was Fixed in Bouin's solution for visceral evaluathon. The next two fetuses were processed for skeletal evaluation. This 000543 process was repeated until all available fetuses were selected for evaluation. Dead fetuses were discarded without examination. Approximately one-third of all live fetuses from each litterwas fixed in Bouin's solution and sectioned by the Wilson Technique for assessing soft tissue development (Wilson and Warkany, 1965). During visceral examination of the fetuses, particular attention was paid to examination of the eyes, palate, and brain. Stainless-steel razor blades were used for sectioning procedures. Seton) cramnatton of resizes The remaining fetuses were eviscerated and placed in 95% ethyl alcohol. After proper fixation and dehydration, the skeletons were macerated and stained in 1% KOH with an alizarin red solution, and cleared in 30% glycerol (modified Staples and Schnell, 1964). This evaluation included examination of the skull, long bones, vertebral column, rib cage, extremities, and pectoral and pelvic girdles. .Bone alignment and degree of ossification were assessed. The following are the procedures for preservation of the various tissues: (1) The uterus and ovaries of each animal were preserved in 10% neutral buffered formalin; (2) The fetuses examined by Wilson's `technique were placed in tissue capsules, preserved in 95% ethyl alcohol and sealed in plastic bags after sectioning; (3) Fetuses stained for skeletal . 000544 nt @HAZLETONusourones mescanc Se- Se RRR 156-160 examination were preserved in alcohol (1:1) solution with bacterial and mold growth. plastic several bags containing a glycerol:ethyl crystals of thymol to retard Statistical Analyses } Maternal body weight changes (Days 6-8, 6-16, 16-20, and 0-20), total food consumption (Days 7-8, 7-16, 17-20, and 0-20), gravid uterine weight, and terminal body weight minus gravid uterine weight data of the ccoonmtprooulnd-tgrreoautpedwegrroeupscoomfpartehde sasmteatsiesxt.ically against the data of the Analyses of the above data were performed in the following order: Levene's test and Hunter, 1969) was for homogeneity of variance (Levene, 1960; Draper performed and if the variances proved to be homo- geneous, the data were analyzed by one-way classification analysis of variance (ANOVA) (Winer, 1971). If the variance proved to be hetero- geneous, a series of transformations was performed until variance homo- genefty was achieved. These transformations were logy, square (x), square root (x3), rectprocal (1/4), angular (arestne x72), and rank, in that order. If rank transformation was ineffective in removing variance heterogeneity, ANOVA of ranked data was completed. If ANOVA of untransformed or transformed data was significant, Dunnett's t-test (Dunnett, 1955 and 1964) was used for control vs. compound-treated group mean comparisons. If ANOVA was not significant, the analysis was complete. Data transformations are designated throughout this report as presented on the following page. 000545 -- RRR. "v0 - A = Data analyzed following logyg transformation. 8 = Data analyzed following square (X2) transformation. = Data analyzed following square root (x1/2) transformation. 0 = Data analyzed following reciprocal (1/X) transformation. = tDraatansfaonramlaytzieodn.following angular (arcsine x1/2) F = Data analyzed following rank transformation. Mean fetal body weights per litter were statistically analyzed as follows: Bartlett's test for homogeneity of varfances (Bartlett, 1931) was performed by one-way classification of covariance (ANCOVA) (Winer, 1971). If ANCOVA was significant, control vs. treatment group comparisons were analyzed using the Games and Howell modification of the Tukey-Kraner honestly significant difference test (cames and Howell, 1976). Tests for homogeneity of variances, ANOVA, and ANCOVA were evaluated at the 5.0% one-tailed probability level. Control vs. compound-treated group mean comparisons of the above data were evaluated at the 5.0% two-tailed probability level. In addition to the above data, percent fetal viability, percent fetal loss (dead and resorbing fetuses), percent early, late, and total. resorptions, and the number of dead fetuses were analyzed by nonparametric one-way ANOVA (Kruskal and Walls, 1952, 1953; Miller, 1966) and the Terpstra-lonckheere test for trend (Terpstra, 1952; Jonckheere, 1954). The 1itter was used as.the experimental unit. The teratology data listed on the next page were analyzed using the 000546 : RRR @Hf AZrLETONweomrones auenca ric 154-160 -n- Cochran-Armitage test for linear trend in proportions (Cochran, 1954; Armitage, 1955; Fleiss, 1981) and Fisher's "exact" test (Fisher, 1950). Visceral Anomalies Skeletal Vartants SCluebfctutapnaeloautse edema CrNyumpbteorrcohfislmitters affected Number of fetuses affected Pskeucltloral girdle VReirbtecbargael column LPiemlbvsic girdle SNounnebearliofgnn1einttters affected Nunber of fetuses affected If a significant trend was noted, the results of Fisher's "exact" test were evaluated at the one-tailed, 5% level. If 2 significant trend was not observed, or if there was a significant trend with severe departure from 1t, the results of Fisher's "exact test were evaluated at the two- tafled, 5% level. When using these statistical tests on indices, the actual proportions prior to conversion to percentages were the data that were analyzed. Statistical references are appended to this report, and statistically significant differences, as indicated by the aforementioned tests, are designated throughout this report by the term "significant and/or as follows: $+ = Significantly higher than the control value. : $= = Significantly lower than the control value. Specimen, Raw Data, and Final Report Storage Al specimens, raw data, and the final report.are stored in the archives of Hazleton Laboratorfes America, Inc. 000547 . na CRE en SERA @HTArZLESrTOeNnveeonssroneSsAsMeEnca1nic 154-160 Size RESULTS ~ MATERNAL ATA Mortality and Clinteal observations Individual clinical observations are presented in Appendix 1. A summary of clinical observations is presented in Table 1. Two Group 4 females were found dead section on Day 20 of gestation. Mo other sacrificed in extremis during the study. on Day animals 17 prior to were found cesarean dead or Treatment-related clinical observations were noted in several of the Groups 3 and 4 animals during the treatment and posttreatment phases. These observations included hunched and thin appearances, anorexia, bloody discharge bloody crust. from vagina, urine stains, alopecia, rough haircoat and B06y vetahts and Food Consumption Individual and mean body weights and body weight changes are presented in Appendix 2 and Table 2, respectively. Individual and mean food consumption values are presented in Appendix 3 and Table 3, respectively. Statistical analysis revealed a significantly lower than control mean body weight gain in the Groups 3 and 4 females for Days 6-8, 6-16, and 0-20 of presumed gestation. These findings were considered to be treatment related since, compared to the treatment phase, there was a tphreoptorretaitoendallfeymaglreesatdeurritnhganthceonptorsotltrienactrneeanstephianseme(aBnaybsod1y6-w2e0i)g.ht gain in Ot 000548 + a - Aa Fe SRE a @HAZLETONuisonsrones memo ne "a3 . 15610 Mean total food consumption was significantly Tower in the Group 4 females during Days 1720. Significant decreases fn mean food consumption were also noted in the Groups 3 and 4 females during Days 7-16 and 0-20 of presumed gestation when compared to control, concomitant with the decrease in body weight gain during the same intervals. Uterine veights Individual and mean terminal body weights, gravid uterine weights, and terminal body weights minus gravid uterine weights are presented fn Appendix 4 and Table 4, respectively. The mean gravid uterine weight of the Group 4 females was sig- nificantly Tower than control. The mean terminal body weights minus the gravid uterine weights were lower in the treated groups when compared to the control group, with significant decreases noted in the Groups 3 and 4 females. 3 GrossPathology A summary of maternal gross pathology findings is presented in Table 5. An increased incidence of stomach lesions was noted in the Group 4 animals. Remaining gross pathology findings were considered incidental. 000549 nt o ENR @HAZLETON eonrones semen nc. 154150 cue RESULTS ~ CESAREAN DATA Individual and mean ovarian and uterine data are presented in Appendix 5. Individual and mean litter data are presented in Appendix 6. Individual visceral and skeletal findings in fetuses are presented in Appendices 7 and 8, respectively. Summary ovarian and uterine data are presented in Table 6. Summary litter data are presented in Table 1. Summaries in Tales of the external, 8 through 16. visceral, and skeletal findings are presented rasnancy Rates. Corpora Lutes, and uterine Implantation Pregnancy rates, number of corpora lutea, and implantation efficiencies were comparable for all groups. Feta] Visht11ey, etahe, ang sex The mean incidence of early resorptions was comparable between the control and treated groups. Nonparametric analyses of the mean incidence of late resorptions, total resorptions, number of dead fetuses, and fetal loss did not Indicate statistical significance. However, there Was a significant linear trend towards an increased `incidence in these data with respect to control. The primary trend component was contributed by the high-dose group. The mean fetal weights of the Groups 3 and 4 males and females were significantly lower than theirrespective controls. The percent of male fetuses was 52.29%, 54.24%, and 60.38% for Groups 2, 3, and 4, respectively, and 44.01% for the controlgroup. RA 000550 - AAFe AN A HET Emr RR STNEA @ HAZLETONveorronessencane Ae as - Fetal Development Statistically significant findings in the teratology data are presented in the following chart: Statistically Significant Teratology Data Teratology Observation Group: VisScuebrcaultanaenooumsalieedsema CClreyfpttopraelnaitsem . Number of litters affected Number of fetuses affected skeletal variants Skull Pectoral girdle VRPUeiiernbttbvseicbcargaeglirdcloelumn Bone alignment NNuummbbeerr ooff fleittutesress aaffffeecctteedd 234 se sS+ S+ s+ S+ S+ S+ soS+ sSss+e s+ se Irend s $S $ S s S Sss s S s$s DfreopmarTturreend suSs NS s s S S3s S NS sNS S = Significant NS = Not significant 000551 HAZLETON somrones aera ne. 156-150 Sv- Significant treatment-related increases in the incidences of visceral anomalies and skeletal variants were noted in the Group 4 The number of Group 4 litters containing anomalous fetuses was also significantly increased compared to control. External and visceral anomalies observed included cleft palate, subcutaneous edema, and cryptorchism. Those anomalies occurred primarily in fetuses derived from litters whose mean male or female pup body weights were significantly below those of the corresponding control litters. Dams producing affected litters had significantly decreased food consumption throughout treatment and decreased body weight gain or actual body weight loss during gestation. In these animans. Numerous clinical signs of toxicity were also present 000552 [ umeLor mr sn es eto po i 3 1 cna [a Grow: 2 34 Jte eerr oofFETe tlSls Sl,sert spoungms soos ed e-- EE T3335 msmguuwnm soos N uut y r EEEds monaamnma. boo | tCH[BuaCnoechesed . ---- . hLBBEooergsSi tn tom vein a, rekrs, Rough haircoat 12!! \ 2 Ppr.eVHE lL ! voy,A I,pk35 , 6, 6 1Po2en13ldF arts fcte the mer of snes ith St prea Fito. i oggga : g ! Wea Haterana ToedrystWotTeoayeeStuo2dnyd oofdy5e5i8ght Changes 1ses0 : fod betant nen Meta (9) wuts GSoamii wToamh: sfmaem gWeens : o " ma mae w3ma w3w ; 6. 3298.47 3296.05 5202.80 H291.72 i* . " won m2e m5a m5e I{ 2 . niea w%en m3ee aies i" " " ms w2a w%w sin = 20 " se395.41 %389.69 5wr. %301.91 i Mean Change (g) Lo0o-6 2w8B.a3n8 2o5h.u0ml3 239S.h54s a2a8.ee43s :; 3sg wr 6-16F 6-20 0-20 Ra be om aw & 40.06 96.95 33.15 93.64 12.545 -32.605- 4.32 5.40 1 i Wa Nee 0S ass a0 aa fr et $e 2 te hos sr st, rn csr it 1 mn . Mean MaternalTaFboloed 3Consumption(a) Rat Teratology Study of 1-335) [CE i 5 IngBaersr) GTreoauprs1 GVroouipr2 rSooruts3 GTrooues 33 os w 15"2.6 1a.0 1a.92 1=9.33 i] u3.48 a35.09 za5 z6 :; 512 aws w5 n 255.50 iFr) . 136 w 52 0 5 x51.09 i25 i -20f " 91.25 a7 m 0i.87 3W9.335 g5 268 " 1H9F0.58 mnes i0h.05 h9i1.655 i ou? " a]a w5o sTenes aiz). it 3Sg $i && . 4: cIanlgciuctaatetsiontsh.e smamapllelsifzoeodocnonWshuimcphtitohnevamleuaenscwaelrceulnaottionisnclwuedreed bIansemde.an Ocnaltycutparteigonnasn.t feasles were used fn sean i Group and BoseLevel 0 mk) g 1 ak2g 3 5 mang 10 miog Mean Terminal Body Weights, Gravid TUtaereine4 Welahts, and Minus Gravia Uterine Weights Terminal Body Welghts Rat Teratology Study of 1-335) BoTdeyraMtentaalnt w 125.0 n 329.69 mn u 2 [3 02.9 uterGirnaeivgein u3s 125.50 7.46 2 02.515 vTeeirashitna)Minsuosdy Grautd Uterine vetans' E2)1.00 324.10 293.305 2 240.405 150-160 :: y :i 85 2N:1hIngdircaavtieds uttehreinSeamwpelleghtsizoef calculations. oonne WGhriocuhp t3hefemmeaalne wCaaslcuilnaatdivoenrstenwtelrye nboatserde.corOdneldy. pregnant females were used fn mean i 2g 3 4i aa ; Ld 3 Sumirycootf tTeerranvaatlaTaayGtreeSsts5syPaotfho1lo-g5y1Findings ops ONrmaesrnranoSdfhDeensrcierimoptticsoannpiantehsatony aarses we Focal areas - dark red Dose Level sw ow Group: _1 (mg/kg): 0 2 T BoE ): 3 o8w5 HEARRaTnt striem enarges snuEreeinairges -- hrraeagunarty shaped PnCeolenvsitsicainsadtsaacross midline RFaOriIiEed an sHnormetrned coranes &$S&rsoToivsesnue mss (3 = Finis tn antsts found esd during the stay ) '}} : )voi}} ' 1500 3 4i o0Rmga i Jb7 i 3 1 ) w. \ i. em fi:3 E&!B Orasn and Description omyiotf Tboersatta-- GeSetytofo1a35,riatoas [ . ops 1 2 3 a 4 Dose Level (ma/kg): 0 T 5 0 I STOeMAaCHo.ar mesa orator moss aESpipalthttieeenldeteasereeharse a-rraGh section tn ail) T - ooneee aprre e- tretas soStoeetnatnLeeddopwiotahtearcko7ind emat)eri alls ae , \ w8m 7 !Wmok oi ,1 w[UR PEYhEiR'eSpPrAToCmHEnSent [trind ---- . , \ 5B \! okie ALL LYNPH NODES \ * gS URmWINiAsRaYmiBiLAeDDeERe wie owwrass ute 3ag HE Buried ost (3 = Flies ta ates found ded during the sty, \: : , i:f i & SA RDA SAIS Ee... EP Key to Tables 6-14 154-160 Pregnancy rate (percent) - (number of pregnant rats/number of rats mated) x 100. Survival Rate (percent) - (number of rats surviving to Day 20/number of rats placed on study at initiation) x 100. + Mean Implantation Efficiency percent' ~- Group mean of ([implantations per Titter/corpora lutea per litter] x 100). Mean Incidence of Resorptions (percent) x 100). - Group mean of ([resorptions per 1itter/implantations per litter] Mean Incidence of Feta' Mortality (percent: Mean Incidence of Fetal Viability (percent) = Group mean of ([dead and resorbing fetuses per litter/ implantations per litter] x 100). ~ Group mean of ([1ive fetuses per litter/implantations per litter] x 100). Mean Incidence of External Variants Mean Incidence of External Anomalies Mean Incidence of Visceral Variants (percent) (percent) percent) - Group mean of ([number of fetuses per litter with external variants/number of fetuses per litter examined externally) x 100). - Group mean of ([number of fetuses per litter with external anomalies/number of fetuses per litter examined externally] x 100). - Group mean of ([number of fetuses per litter with visceral variants/number of fetuses per litter examined viscerally] x 100). values calculated separately for early, late, and total resorptions. 000559 TPT AEE EA EL ~2 - TE har 156-160 Key to Tables 6-14 - Continued Mean Incidence ofVisceralAnomalies(percent) Mean Incidence of Skeletal Variants (percent) Mean Incidence of Skeletal Anomalies (percent) - Gfvirestocuuespreaslmeaanpneormoaflile(istn/tnueumrmbbeerrwitoofhf fveitsucseersallyp]er.x 1l0i0t)t.er examined -Grfeotuupsesmeanperof l(it[tneurmberwitofh fsekteulesteasl pevrarilaintttse/rnumebxearmineodf skeletally] x 100). - Gfreotuuspes meanperof l(i[tntuemrberwitofh sfkeetluesteasl pearnomalliitetse/rnumebxearmineodf skeletally] x 100). 000560 SummaEroy ofTresatantoDjnayrioSatnyandofU1te4r8in1e uta 154100 : NPNraebrgenranocoffypaFratemgetnhst)wifehmrmeastingcon raed SKoesr'eaTteem0t1ssurviving to tay 20 WetTnoorntponmraestrtiuootfen:s ECTaiortelyrrHeesisoronprtpriiononnss TIRneeegiieinnenstiencaccointtdectonltaaettneoiodcffnosnwJaratraereypsoersesypMotreist5toe)irnosnbsus(t(5s5: Rein chieonf tothe soe tevaaGyro;w: _18 --2F _ 3 _4 J snwooon8m 8o5m @oOBa fp Wsoown oownoonBfb Wwehoamslease ogeNe | 1owso0 woos 0oia e0Hsokmno 0T09o%0 aainn wUoamh eNiEnk Pei A . F ne ate resorption was nated for femsle No. 21066, therefore, the actus] sen vas 0.04, g &g t i iL RastummTaerrystaoftoTgaMybelaenSt7Luidtyteofr aT3t3e5d1 ery . NeaFnetunsuessb.er oidf:eead IndeRieacinens tIncnaclidcdeueblnedcteedooffonffeettaaallpemvroiraJtbiaitlltiiettryybu((s31t)1s6: Reon percent mies (5) CL veHeaHnefso"bt!osdeysvetigRhatdsi(s9t1e0d mean Femsles ~~= ccuoomvndarjrtuiasantncecede amaeddajjnuusstteedd sean mean sos Level (mGaoawr:: --0 0 2 W00 Ss00 WSt0oe0 Rssems sssmmhoasdmm si she 3Ea]o wose a0es t: aasrmm w1sa aw am : . gaameoss s zfeeesf@ BScoPenexer",cdeenpatedrmcoefrnetttauslmiatlveyss,fnooactlneudddmesefaonrrebfsooerdmpyltewieolnhosg.htas27dv1ae0t1la,] Intchleurdeeforvea,luetshefaocrtuTailvesefaentusveos as dead fetuses. o0n.l0y. % SSEatistican analysts performed on covarlance adjusted means 5 '$ a28e i1 8 4% MNNuoimsnbbeehrr ooofff fffeeetttuuussseeesss wewxiianhmiinnveadrnioarmnat)s Mats Mmber of fetures wth anomie usSPhioenrleandeZGdtreyemprettoonf ancasites RSaummTaorrytootfoTnaEybxtteeSrtn0uaaly Fofind1i-n3g9s0 dose Level (mmGrrkoauyp:: 5a-t60 33 10 2 s is h ww0 moomomoowomos W wm b LEE ooo 5 o ws dFf 8g ; ag k | RaStummTaerryatoofloTgVayibslceSetr9uadny Fofind1i-n33g5s1 NMMoiumssbbbeeerrr aGofff Tffeeetttuuussseeesss wweiixttahhmiinnveadfroiramnats) atts Number of fetuses with snomaties MusBEbrEeari)na--ndccetoryneptbeariaonlfedhvaedraattrasknpthsmeraetecroinatained dark area KUriehdtenerery(ss))gi--rdpieaslrtveeianssdeddilated ~ nduiated MaPSseabilenarteande"dctelymapureteoonf anomslies Eryptorehisn tose Level (oGako)w: 10 2f mpmH ooowss 0v 1"we a `5i o 1) BeFolurefetfuesteussewsitwhithcleefdteaapatloautse.sksienenseexetnerenxatlelrynawlelryewecroenficromnefdirsaetdvaitscevrieslcerraxlamteaxsotmiionnation. o 232g2 . 154-160 ` b: 3 Ea a]? om n5s now1 .3 :a i 5) ') 5:i :2 =: s TowmES LE 2 s i> 8 : aomTaerrioaftweSyeeeStat1st0)s iofns1i-n3g9s1 serio 5 ose we mponaryos: 6 P + -- --4T -- RwRooemmeerrr o1off ffFeeEttuuEsseeSss WeSiatshEivnagrSioamnat)s tates nwwoooomwmoom nmow ww i Nerf (stus with nomics 5% nh WoTeSahpandiottnyitpeeisaotftaiatonertcosse car50 CIR HE IE scomtetemanstres wom ew 4csBHon ow ~CpEaiirpireratcaecslteaimasteriobetiiauionctndairornesniocroslee/noncsesd vara ssh acton esmtete nosed F)A E FE FE. Feiccatgoe 2SBrSthierra-bacraabppaaoraes3tsiusTetsion cows ) vow Poe = ernebras Sashes oa g3gg Z-E rTibs S 3 opse stir rfiesddmreear p---- ` 1 oe : verter cotaC ahrocrecslcarCcchaeennsttassmsmmeostniittcisisetsion ticomiete/morsites w0; ooRmoo1%Powon i; Pelvic ate - iebonnaussnto5contoimainirniecossleernoosesd B:: oh PHEE a ::: sonToeriaeo:evn-t <Saarsiiied - \Poa i Po a i HSeotsmaraameaoattehsckers Ve to Pessble preparation ds at tae of evisceration ; InctdenRcaet oTferaEtxotleorTgnayarleStFu1i1dnydinofgs1p-e3r35Litter 150160 : HlWauasmbbbeeerrr ooaffndt1pitetertecrressnteUoixftahmiiMnntetdnoorrsmaWitMhatveasriants Nasr 30 percent of Hters with anomties MaSsPhaieanrteaned aCplteerircnten:t of 11tters with snomstfes sos Level (egrrohauy:: 0 a1 aaow 0" woo0 a 00mm0n00oo0wmb woe 00% 00% Rm | smsa | & is g8a } . a nctdepRcate ToferVaitsocleoargvayleStFui1dn2ydoifngsT-e3r381Litter 154160 5i Bose Level (agG/rkoew):: 01 2T 33 Two s} NMMuoumssbbbeeerrr ooff and tiptteterecrressntweoixftahmii1nnietdtneorrsss)witMhafvtasriants Number and percent of Titers with anomalies 22 As 2ie 12s 13043 ANI 15 vss 0000 1438 13 ems NumEBbryaeeirsn)an-d-cecproeenrbtcraeainlntedwhiedtaahsrpkvhaamrraietaenrtcisoanltained dark area wan 5.56 KLlidvneery(-s)darpkelarveiass dilated Ureter(s) - distended mss 3280 N20 52 20 |5 ~ wdviated 14135 NuSskbienr andedepmeartcoeunst with anomalies Palate cleft was ass s73m.8s9 &V Cryptorchisa . was ama 2 8a J: H3 3 nceor oTrSekeeeentrsSsaeigoFs ur iter [Pe x\ ne tev rts ----3 --%; sfoe, ii REa A eyag p-- se gmN orw l Yle,, wemw1ha wTwee3 mowmmi7se s"eom3wawm shaeTaEndL oprearatcetpnitiL oswniencalorw suiraes(re m) s Weaoh awmma upmee syemse paren sieaion reir CP reaE mone ete octane Spal ors non esmene Wwe{ahsnist AnTs wma wCass wma wVaanan n.8 my Tn u= ha lemriaE eLreeen SIRE ven ngs moamm vVeisk ams mm Sa wee ass S VeCrcTtoTEnenBbsnacerratlaasraclthpiessaponre ified pVvaaln as VwAss ema 8g2 Pelvic strate- pcnorrastliSeeonaginesIntoamrleestemossiftes vvSviaeoemmm) wWeeSns naa meanmn mmummmneme PTOTotg es - 2 Bevin eres somite Ho Fated VveaEhnR ws wdWahEn oTowRmeSns | wen mn wie wma Sternsbrae -- checkered es 4 i Wea nctdence of ExtHeernaTle,atVeistTEoaenrtaeS.ha14ksyndofSk5el5e0ta1l Findtogs rec Litter 154160 i soe Lev (arGireosu:p: 1 2t $PEPwEf extermt HeNNniambbaeelrruewisthhcaivlneaacainttesesson a per Mtter busts: nctence of rian (1) 0o go0 opo0b0 8o0p0 waWn no Incidence of nami th oes oh awe viscera KeMNanaievrarluWweisRthcatlvcnaurclimaattnietcdsson a per Mtter hast: Tnctence of `variants (3) 0" a2 o sw ee3024 oa may g cience of nematic th os en fan Scere i LoMceMMaItncaihievraretlnucieweesitthohocffailvancvnruacaalriatnmtatteneesdssssocnn()a1per 1itter basis: 3o3s2 os aaowee Ne w8ays te 6a as oi g8 r a tn Low wes RRR MNautmebrenral Group "a Appendix 1 Individual Clinical Signsa Rat Teratology Study of T-33s51 Observation Body Area 158-160 Day(s: 217032 27034 Swollen Neck 3-20 27042 27046 27049 21087 221700529 22 AThlionpecia Hip-right 610-14 27063 27065 27066 27082 27083 1 Alopecia 3 Urine stains Bloody crust 1 Alopecia 1 Alopecia 1 Alopecia Alopecia 2 Alopecia Alopecia Alopecia Alopecia 2 Alopecia 2 Alopecia 2 Alopecia 3 Alopecia 3 Alopecia Paw-fore-right Nose Paws-fore-both Paws-fore-both Abdomen Paw-fore-right Paw-fore-left Paw-fore-both Leg-hind-right Side-right Paw-fore-right Paw-fore-right Abdomen Hips-both Hip-right 15-17 18-20 20 8-20 10-11 18-20 19-20 15 16-20 15-20 10-20 17-20 17-20 15-20 15-20 14-20 2088 3 HTuhnicnhed 1105,-115-19 Alopecia Alopecia Rough haircoat Leg-fore-right Leg-hind-right 10-14 10-12,14 10-12 27085 3 Rough haircoat 7-12 27090 27091 3 Rough haircoat 3 Alopecia Paws-fore-both 9-11 . 14-20 27095 Rough haircoat 3 Hunched. 8-11,13 27099 Rough hairceat 3 Alopecia Hips-both 8-9, 10-12,19-20 13-20 2700 Alopecia 3 Hunched Legs-hind-both 16-20 Thin 16-17,20 16,20 Anorexia Urine stains 16,20 18-20 Rough hafrcoat 16,20 Bloody crust Eyelids-both 20 Bloody crust Mouth 20 Bloody crust Nose 20 mo Bloody crust 3 Rough haircoat Paws-fore-both 20 8-9 . small Eye-left 15-20 Animals not included in this table appeared normal throughout the study. 000570  anki Sp Qn eben reREHd Za MNautmebrenral MOT 2109 mm ame m3 m4 2ms 21Ms6 am? . Appendix 1 - Continued Individual Clinical Signsa Rat Teratology Study of T-3351 Group Observation 4 Thhuinnched 4 THRhouiunngchhedhaircoat Anorexta Sensitive to touch Alopecia AlRooupgehcihaaircoat Bloody crust Bloody crust a Alopecia Alopecia 4 ThHuinnched Anorexta Urine stains Rough haircoat Bloody crust Bloody crust 4 Hunched Thin Anorexia Urine stains Alopecia Alopecia Alopecia Alopecia Rough haircoat Bloody crust BBllooooddyy ccrruusstt Bloody crust 4 Hunched ATnRhooiurngehxihaaircaat 4 Alopecia Alopecia. _ . 4 Alopecia = 4 Hunched Thin Anorexia Body Area Hips-both Hip-left Mouth Nose Abdomen Hips-both Mouth Nose Abdomen Hips-both Hip-left Neck Mouth PNaowsse-fore-both Vagina Sides-both Side-right Paw-fore-left 154-160 Day(s) 1183-270 311-031--2220,014-20 16,18-20 10-14,16-18,20 n-20 1103-20 20 20 10-20 10-20 111578---222000 19-20 16-20 20 20 20 12,20 12,17-18,20 14-18,20 n-19 9-18 8,19-20 8-18 15-20 16-20 1166--2200 18-19 13-20 111872-,122900 8-13 14-20 19-20 14-20 - 17,19-20 16-20 Animals not included 1n this table appeared normal throughout the study. . 000571 ci Ba Seine ARENA ~ 36 - InAdpivpiednudailx 1Cli-niCcoanltinSuiegdnsa Rat Teratology Study of T-3351 MNautmebrenral Group Observation Body Area m7 ame ams 2n20 ma M22 ma Ms 4 Urine stains Rough haircoat Bloody crust Bloody crust Bloody crust Bloody crust a Hunched Thin Anorexta Rough haircoat 4 Hunched Thin Anorexia Ataxia Soft feces 8loody discharge AlUoripneecisatains Alopecia AlRooopupegehcciihaaaircoat 4 THhunicnhed Anorexia Bloody discharge RFoouungdh dheaaidrcoat 4 THhunicnhed 4 hRuonucghhedhaircoat 4 AAllooppeecciiaa Alopecia huTnAhlcoihpneecdia Anorexia Alopecia Rough haircoat 8loody crust Genitals Mouth Nose Paws-fore-both Vagina Hip-left Hip-right sIindgeu-irniaglh-tright Vagina HAibpd-ormiegnnt Leg-hind-right Side-right Abdomen Paws-fore-both 2Animals not included in this table appeared normal throughout 154-160 Dav(s) 91-62-020 20 16-20 18-20 16-20 16-20 16-17,19-20 1316-18 12-20 16-20 16-18,20 18-20 8-10 19-20 n18-20 12-20 211053--2108 1166 16 116 16 77198,1117 1-6n-17 1120--2200 20 111866--,221008-20 16-17,20 15-20 8-3 19-20 the study. 000572 ition weal TARR RIA s(tTHR AGRE 3 RatInATdipevprieadntudoaillxogy1CliS-ntiuCcdoaynltoifnSuieg1d-n3s3a5) MNautmebrenral Group Observation Body Area anzs 4 THhuinnched AAlnoopreecxiiaa mar 4 Hunched Hip-left T8hlionody discharge vagina RFoouugnhd dheaiardcoat. ans 4 THhuinnched AAnloorpeexciiaa Paw-fore-right ang 4 HRuonucghhedhaircoat TAnhoirnexia \ URrouignhe hsatiaricnsoat Bloody crust, Paws-fore-both 154-160 oay(s) 1198,20 1176--2107,20 1166 .e16 1" 1144--2200 1151--1178,20 1148--2200 1166-20 1196-20 % PAnimals not included fn this table appeared normal throughout the study. 000573 . AR AR dD oArBHB ur S Siar sc&eWis - 38 - Key to Appendices 2, 3,'and 4 154-160 P= Pregnant NP = Not Pregnant C= 0= MS = NO = () = Cesarean Section Performed (preceded by day ofgestation) Death (preceded by day ofgestation) Moribund Sacrifice (preceded by day of gestation) Natural Delivery (preceded by day of gestation) Data excluded from meancalculation. Explanation of Footnotes . only data fron pregnant animals `surviving section were used in mean calculati to Day 20 cesarean ons. PInvalid food consumption value not fncluded fn meancalculations. "Gavid uterine weight inadvertently not recorded. 000574 ~ ten ct saEngTE ra i com S_e weeS eeeow S ----se e a paew wweaa e BORE MIB EIEmI ES Donon omomomom Boon Gadd d EooGEOBDE ENED ENED me nme mE Nomi om: ome daaad Bi Bom of 38 BW Boom BY oBdo@lom dmOoEoEs + BMLEEDEEL REDEEDIE REDD Hsae,BtenifnnidandBnn Bdnn BOBogOH OoBm OLEoRm OBRoMn Bfno thnebBeodBse d Bn dBe ddn ggg83g a me Hi pea vi tv JE Srl tn eo Cost maternal = "Sux siGnitatlen -- Mtr mln 0 6 8 uw aw_ et on -- oe ee en ew en wd = ont BE dae ns daa aang BOGE MBB EAR BU REEEE BONE BMEEIEE on WIC MS) RI) @B) (E85) (Bes) (0) BOER ELD @D en cen (In en Cle (en = ds Tole Tan am Cn Team Ban wie Ban wen noe Se wes Fy :g:g3s | i ten ctmgHELE Aopendt2x- Continued pa on ee ww" eo BheOrEtRhSei; 0 OWI % BIE8 IEw I w RIoEmD BOGOETE BOWaEDdEIEEIBEIWED ddoOR EE EODMEEN BEE RBEoEm per pie ome md dom OMG wd --------wemoema e OM OH aOw08 W e waon m wma ow &W0 WW0gEwEEar Eom oBp u ooSu BEOfE oBp oomw BDO3 Gd Boar owl amBfmOooGRPRECEEOmDYE omERIIEoBmID EBEIdI BEEOTIEWR BOpBooaworu BwoWEmOWEwoBmOBWE : mR aE dh awh dh EBS GRD @D db a Gh &0 dB lod TE US CREOWLORNOCUN BUS am ua um anon # 3gs f: & f is mgweSII ri chu _ Sol hr belie Sb aw Ww eprom eaea sm aw sw wi en doonRDOEDOED ED OBDOBDOED dOTH EPErerararE OED WOW oWomowmoW og ddbdrdp dp db doogE Eb ALE EDA OL of Bom om nH m= E} RG OR} A TR CUL AL a Sw dw Be de Bm Bw oo : i g a 3 i3 ii . oe ARE Maternal umber 2211003310 2277008332 2277003358 2211003376 2217003398 2277000810 2727008423 2727004454 2271008467 2127004889 2211005510 2127005523 21058 MeNan s.0. Sa3- IndivRiadtuaTleraatndolAopMgepyaenndSitFxuodoyd3 oCfonTs-u3m3p5t1iond Group 1 - 0 mg/kg InterFvooadl(C0oanvssumopftGioenst(aat)ion) 06 1:8 _992 13-16 17:20F 106 1M00.80 190.6 4B 2.8 aw S25 45.5 86. eN1E 19 017 2i22e.4 82.2 106.4 213.8 1820..76 166.0 1382..60 467 478588 6918 5810.69 691..58 1s1e9.e3 448 1091 15903 1133.9.82) 13.90 (360.)2 wa (5820.92) 95s (5856:.28) ls 1(0669..14) (123013..59) 107000 ier 112425..34 129.9 3413.38 3.2 679678 722 sNl1l0e 08748s 211712.016 7300 865 ise 1J6a..1 M2 442e.69 32 7819955 606 880.s1 787 9953.08 221050.055 990 1735 NMsE.S 123.4 33960 297 719518 418 678914 B26 s7s8l4 1e9l1es M8 1541 11434..35 N95 4S21.34 35 9901078 76 9973.57 111163.44 222379.86 T20 92.5 ise 112312..17 125.0 3396..86 3.2 871439 411 785a15 11026.24 128056..96 97.3 96.7 1a0.6 126.5300 784 752 98 183.7 122.67 15.924 32.48 6.812 724.58 13.416 728.52 12.317 927.25 1920.58 12633 25.916 156-160 _op0 daesasno d4so0g.s8 423647..47 (342419.s0) daessee 331%9..e4 443541..40 4411327 43409m.g3 54805..58 4a3s0..28 444002..13 406.3 4220.49 47.701 000579 Svat MaNtuemrbnearl 221100556 2127005587 21059 2127006610 2211006623 2121006685 2217006676 2211006698 2211007m0 2277007723 2120170574 2727001776 2211001798 HeNan 5.0. "a IndivRiadtuaATlpepreaantnddoilxoMgeya3n -StFouCododyntiCofnounesTd-u3m3p5t1iond Group 2 - 1 mg/kg 154-160 06 3-8 112326..00 3425..12 11306..85 3295..12 W732 (1108.52)31(306.2) 13$55..67 338..26 114.3.90 33970 19494..39 3335..62 N14L5.S8 240..74 11393..60 432.38 11085..46 4261.94 114271..30 4401..97 112.6.24 C228118 (115268..85) (2333..82) 1228.01 2335.03 20.444 5.735 TnterFvoaold 9-12 7634.07 6780.20 645 (6621..98) 8611.48 96315. 6603..36 67225.6 660164 6621.8 9923027 8632..28 (5653..14) 6238.38 8.893 C(DoanyssumpoftioGnesta(gtion) 1316 11-200 196 769.70 6499.60 M2 (6369..80) 76030s3 851533 7623..08 7579.20 4e1lss 10608.26 geazls 7509..00 65.8 (56.3) 10s821.95 8988..06 M2 (9117..39)b 97801.73 1969.094 8982.15 9974.07 10918.54s 110079.12 18143.63 9759.095 96.4 (64.9) 1i6s9s..9 116562..38 1a (116307..94) 116869..50 117697..19 115761..69 119406.10 11377..80 117872..82 21213..69 11671.43 164) (133.5) 2633.45 2937.17 12731.85 12.065 11.147 17.584 0-208 348340..48 430122..80 a2 (338236..20)b 430105..80 44a1l0i.g3 344085..30 432595..83 430s3a..30 440206..58 439s24..9s 338853..36 389.0 (355.2) 39237.03 35.568 000530 Maternal dumber 27080 22170088)2 27083 2277008845 217086 21087 21088 27089 221127000909120 27003 27094 217095 217096 27097 21098 21099 27100 2210 02 2703 2n04 N Mean $.0. 0-6 139.2 113398..95 1A 014211.57) M38 146.0 18.1 134.4 11349217...339 10.5 168.5 120.0 136.2 145.1 147.3 1m.4 141.8 1N1S8..49 130.8 (138.8) 23 134.92 17.356 Sas - Appendix 3 - Continued Individual and Mean Food Consumptiond Rat Teratology Study of T-3351 Group 3 - 5 mg/kg nferFvoaodl(CboanyssuompftGioensta(tq)ion 1-8 9-12 13-16 12-207 2-168 154-160 0-208 36.9 3246..53 42.7 (2213.34) 28.3 32.2 (156.2)0 35.0 322668...283 24.4 40.4 24.9 34.6 40.9 3209 26.6 28.9 3208..14 ana (24.5) 60.3 4486.73 646 (553.S2) 53.5 (N6.4)> (86.5)D 61.0 1454634...906 510 2.5 56.5 (119.9) 51.8 75.0 67.0 34.2 4456..05 42.4 (27.8) 70.0 6308.s9 395 (1e1e.l2r) 59.6 72.5 (88.1)b 61.3 665102...153 1908 69.0 55.2 (160.8)P 53.6 89.4 (110.3)P 1.5 6S5.119 43.4 (36.9) 104.7 8823.18 134 (s1o4.l2s) 88.0 123.9 (109.3) 871.8 798455...470 10009 96.1 N46 (100.6)b 100.9 (135.8) (108.9)b 0.0 9910.59 95.3 (64.3) 167.2 1143s.7 146s l(8e5.s8) 141.4 (221.1)b (330.8) 157.3 121253161...539 155.3 181.9 136.6 (315.3)b 152.3 197.3 (203.9)b 64.6 113396..28 116.9 (89.2) an 336363.04 356.3 (a30s1.a5) 343.2 (491.0) (558.2)b 3719.5 3a049098...16 386.7 446.5 mn.2 (552.1)b 398.3 (480.4)b (490.2) 206.4 335402..70 343.0 (292.3) 22 31.53 5.567 20 59.58 24.302 20 57.89 18.350 19 87.87 24.771 19 18 147.915" 363.145 38.131 59.846 000581 MatNuemrbnera.l 2211100s6 221n1008 2a1n0o9 a27mmz mma3 ams 2mm7e 221m1s9 2n20 na nz ana 221n1a2s4 2a1n2a6 aanneas N M5e.a0n. - a6 IndividuaAlppeannddixMea3n - FoCoodntiCnounesdumptiond Rat TerGarotuoplog4y-St1u0dyma/ofkgT-3351 154-160 06 3-8 M1286.75 43000.06 1108.0.3134e3s 11430.25 33.37 112.879 33.s7 215900 021802 128.7 29.7 1N6164 a29l.s0 (111250.68) (23821.950) 1507 42.3 98.5 24.3 120.6 33.5 158.2 4.4 n13s.o6 z34w.5 2W0I5 22056 (M132.2.01)) ((2371..95)) 22 13136..32309 2 3247..56741 TnterFvoaold _912 548165 243062 (sssd a408)3 2134 392 18060.65 (5175.22) 3s (48.00% 44.3 (66.2D (11290.3) s390a9 ((145..17)) 18 2306..18769 Co(Dnasyus mpoftioGnesta(gtfony 136 17-20 116 6177..49 258420 305.s0 I1561 0o.l0 26:3 S01.10 (0412.1) 0.0 (126.3) (164.9)D 545 (1906..030)P n1e5 ((200..04)% 9890..50 (29010..3200 825.03 10000.90 1000 905 (1100..00) (240.09) - (96.8)b (152.0)0 (119.8)b (11070.08)b 0- 0 (7(02.24)) 16960..00 212020.025 (117212.82)% 17850515 5222536 85.2 17525.15 (1"3a0.38) 73.8 (198.6) (242.7)b (162.1)b (3a6s0i.21) 8718.01 ((4333'.60)) 19 2226..73580 11 4369..839135 9547..362315" 000 421046..35 (533320.14)0 (33s200..80)b 329078..74 1197620 309.4 (2410901.20)b (2le7a1-l.s5)b (333.9)D (515.3)b (aa0.1)0 (151650.25)> 9-5 ((222186..81))0 wo 29602..486945- 000582 ditnins mmoi ~ar- WelghtIsn,diavnidduaTlermainndalMeaBnodyTeWremiiAngpahpltesndBioMxdinyu4sWeiGgrhatvsi,d UGrtaevriidne UWteeirgihntes 154-160 (g)a Rat TeGrraotuoplog1y-S0tundgy/kogf T-3351 MaNutmebrenra.l 2211003310 2127003323 221100335 2277003376 2127003398 2277000410 2127008423 2277004658 2217004876 2727004498 2120705510 2127005532 27054 MeNan 5.0. TBeordymiWneailght 38282..25 03647..38 4340610508 Not40P9r.e0gnant 23713.11 3374481.18 363881.029 333801.88 3a8n4a5 44378..08 a3s929.4s 349262..22 n26 3952.41 28.666 UteribnreavWieaight 3200.94 86781..28 6637..63 86.9 6796..22 8254..47 3080..81 8052..64 6142:53 8741.0 6074..38 8743016 82.1 742.35 14.704 WTeeirgmhintalMinBuosdy Gravid Uterine Weight! 230327..13 330106.00 328194183 3221 335003..99 229941.71 333071.84 3321900.42 330261.26 336624..98 3a3r9.y1 : 333291.016 330.5 32241.07 2s 000583 "as Individual and MeanAppTeernmdiinxal4 B-odCyonWteiinguhetds, Gravid Uterine 154-160 Welghts, and TerminaRlatBoTdeyratWoeliogghytsSMtiunduys ofGraTv-i3d351Uterine Weights (g)2 Group 2 - 1 mg/kg MaNtuemrbenra.l 2211005556 2217005578 2211005690 2211006612 2211006643 2211008656 2277006687 2127007609 2271001712 2271007783 2211001756 2211007787 21079 TBeordymiWneailght 338219.030 0az0iz Not38P8re.g4nant, 337596..48 383188..77 39376.28.4 3388560.58 3937076 0373173.8 443067..38 313102..83 338786..84 Not Pregnant UteriGnreaviWedight! 8782..55 8673..53 69.1 6649..02 74670.64 3810..90 8706..41 8330..70 8757..38 8864..87 607..73 8n.2s WTeeirngihntalMinBuosd)y i Sravid Uterine Weight 334008..58 23948.69 9.3 209.2.28 3a4s 312.4.85 330069..44 330370..60 332080..00 338213..15 22905.00 330055..22 HeNan 23839.69 752.59 5.0. 20.073 1.458 31234.10 14.093 000584 | - 49 - : WelghtIsn,diavnidduaTlermainndalMeaBnoAdpypTeeWnremdiiingxahlt4s -BMoidCnyounsWteiinGgurheatdvsi,d UGtrearviindeUWteeirgihntes 154-160 (g)2 Rat TeGrraotuoplog3y-St5umdgy/kogf T-3351 MatNuemrbenral 2277008810 2271008832 2211008845 2127008876 2727008889 2211009010 2121000932 2127009945 2277000976 2127009998 22110010 227110023 208 N M5e.a0.n TBeordymiWneallgnt 342555..30 335621..71 No3t8P2r.e6gnant 307520.71 a3s7280l79 336008..96 33913.23 341102..40 393s8a..76 36n6.s1 253547..10 337609..10 Not Pregnant 23 33567..01316 Uterirnaeviwdeight! 10132..92 6782..58 8799..48 7763.s6 6n1.z6 8785..03 n-.2 6587..41 685..02 - 681..26 7732.89 0.2 22 1734.8486 TWeerimgihntalMinBuasdy ; Gravid Uterine Weight! 238221..84 229820.36 229922..78 323699..14 230073..50 320835..36 300-.8 322933..32 330333.55 2300541.95 229663..11 299.9 22 22983..233855- 000585 S50 - WetahtIsn,divaindduaTlermainndalMeaBnoAdpypTeernWmdeiiinxgahlt4s B-MoidCnyounsWteiiGngruhaetvdsi,d GUrtaevriidneUWteeirgihntes 154-160 (g)a Rat TerGartouoplog4y-St10udmyg/okfg1-335) MatNuemrbnera.l 2211110065 221n1o078 2217110190 n2mm2 2211a3 a2m6s 2ammis 21a1290 annaze aamnaas 22n226s 2ann2a18 an2g N M5e.a0.n TBeodrymiWneaflaht 3376840.47 manna 3220590.93 382262.178 223450.53 334501..06 Not 21P.r2egnant, Foun1d80.D7ead 343391..63 3646.98 23a2n.s2 NFootundPreDegandant Not Pregnant 20 30713..92153 UteriGnreaviWedignt* 8528..39 5149..68 [172.]2 8460..58 4423.33 8525..44 38s 32.4 6n6.g0 7169..04 394.376 20 1670..753195 WTeeirgnhintalMinBuosdy A 6ravid Uterine Weight! 32692.54 229770.14 2163175 310802..20 1199821.20 226894.62 208.7 148.3 2261511.39 323950..58 1m88a.6 . 20 25491..541075 000586 HE il wife I Siet DE wie wien wie wien wie wie @ oF of oo oom rou @ op, Foam oH bE DW oF @ oF fF om FE ob omoroow room ro ow rH o dow FWn ouFw &us fio i $fom8 f i Hts : @o0 oF Er og fo 4H Cw EE ow h Hoge R.ToTw h; ome 2B hlws gggg< g mgs SGEELEIR ss he Sa RR we oI J ew Bw, um .: = oa HE E Bt i BEB i Igi oo8&J 31 &88 Ii 22 I5 }i: 2 I3 mooF on KH FE i 0s Fou 7 5 [I $ os iE | 3 8 ] 8 mBologn oF aE; o:H i I. 8 gg ga a FEEESe Boy mo ow 8 dios i opm + o# " ns H ws i we 8 a Ww kd ? 5 os : 5 ? 9 $ o8 1m : wi $ o $ i" Ei dm w, i 03 i +4 3 a 1 I me i oe " 20m W Ny 3 wa W na i ui : & 3 oe 3 wh i kb =o = 2 no oo oo oe ' iw k 1 a 3 a a n 3 a 8g:gg< ay EER, A wn#oeo 8i j%iFE8wl r i gii 8f doww6? "d 0d Hwi moo Hwdoo OeMdE hE row roar FEB wr wLNew e n w k 2 so afm MA etpregmat FE - rH Fm w 2%wwRw B3 oa B3 oa 18 om dn nt i si ggggs reBTot ue EEaterm) wie Tota) we Ga Total, ew atm NEpercent. dei ===2109 w .Pow Pas oF BE wonp wowRs BoE fO4.H d1LomoB "wmz HaoonmonoaoHn &B PPoopEo oH EsoB Oro F 8EHBH ou ` = boobs FE gg mBammemre 8 an BaGo odhteodowt,ae NHBoomme.e fHeeiEra,Rtt.RoS t iir n a stp 1 ss 0B whHooOmW foma, HoH _ town, ow, ona rE BE wie we Glee ou owem mmm HE en EB Pbpb om ow on Fo Poor oE og oor or om oan . eS$ M2 HeE w ooo EE PHOEBE Boe dom Boa, i 13 42 ue t HeeRd,t ttt is i SC a aacr onn o ch sn ete den SpE ow om onon EBB NM a a Hoot i a de wm aae rnWM eA =a wif es msa oGsioe wowe voemem mtmtm HmeE Sam RAa = he od EEE ED Eon OH iB fdooff oa E ow oE E FFrbmEnouo n Z Pood gos 1b o&m non ; o i&E gS iii fPo ood oEw onEEom fboFb iSdo. mobbooomw oEoE Fbbb E En mo on n oE@om H on Bon ' 3a spBwaumnee BBWyo04,GWe oomUm, m BHoeonwBm BhBs oMhhh BEaxn aGGee adZen SHEE ooa EET E tS eRlt yf tes. a SEro Teraielon ShueyotT31 BE ule oe Cha ee oa mmm GE EM % Po AEE Dr EHH Eb i bobo yo 41 PoE od bE ror muon Bou on mn WL me ew . % oi TPioPOHOoF EoBRDFbEoE B E on oH : 2sg is mmm H.8Goi04RBxs uBBolwa uHwWoRh1 oEu n wwoah a RE - ra HEErR E an aRsti no . J mE e PE EPEE PEEEE T EEPE EaEEaE EaE E EF FEE EEE T ET00 ffe u bbEe e dfEeuddeEe fbe o fEfe e ddEe e Efbe e dtEe e lbEe u bbeEe bte foRL an Hv Bo EeEfiRioe ee dsesnstmiafliicaetsio(n3)lettofefartus(esswi)th: * | ltt ree ee 8, 0.00 0.0 0.00 0.00 0.00 0.00 " 0.00 - om 00 00 - oe am ` en ro. gg2 73 ndgETE - mE e EERE EEEEEEE EEEE EE TefeGur sem s el HL 0 Sm Ne Hw Go ne om om 600 bw 6x bw ooo fh nr BIL an ares Hoe 3 ow , Bfcmmie i RI ALN Sar estnorsre censmi 0 0 8 eo 0 0 so 0 0 0 0 8 &522 8g oie TVEmEn te IS mmew p$3d P PEELE EE EP EE EEE E EE EEE EEE EAE TS Thae ter Si F ects s wine eT 0 fe fe fe fe de fe de de be be be te HH S E T g ) A . , pBBJnomohsiXnohmy 0or oe cast 0m om 00 0m om 00 om 00 ow ow ow om et < 8 88 wg T i E A - mLEeH RET 0 dentification letters) of fetuses with: hi from era fk ares $0NHPAEEE E 0 a fede du de be de so de de | --"- " , F ETEth at offetes win: Coptoraine ost g8ggg2 on oh on a " on ' eTES RRA } E[ E ---- mE ee E E oE ddEE iE ErEsE ErE Ed E E E d RE d E EST0 enti ication Tettar(s) of fetuses VHD: - dn fede fe be fe be te de te fe te S fl. atn pare cate re i ro, . mrts tn EEE aitEveE nte ttre of fours tn: fre oe oo 28 oh Si om 2 bm ony etrn. 3g g8 Eiri D iE dLbSa ; mmRe3 i0 ar3 rdi 0 p d3 Ld8 A HE et as Tr Tietonwthoa eae sen rth (5) i fm i Goo ` Se i Sw i Sw i bm 4 Soo : bm iol bee Seo i shen " fStnn m art m ts rcontd eo sm HE M F Jach a ies (3i)e hgormr sc, r aati 000 0000 00.00 00.00 00.00 00.00 00.00 M2 N 00.00 00.00 a0.00 2. red + etre. sg23S song UCTEASt MEd L3 eo 8 348 d ERTLIAEET I 0 STLon we Eid ee da fe fe fe fw be be te te te te te Teeatmiafitciantiogny+Netter(so) ofofetusrees vwhicthe:ral LTah le om om 0m oe - 0.00 0.00 15.00 10.00 10. " am s8mm EHiR a7o TE 5 L2F th 1 ot rH a crs 1 1 ttn, Te ot or 0 AASSAED SS A 2g8 . 22 TLLe tn, ces 13, veeE EEEES Fm Jopendta 1 Cantinand we F EE EET H n otmeycd mel lid didi Pmtfn fn fo fo ffu feee Eh oe FE Jor fetes thonormre visceral asmeles Ancmaties (3) mre what Looe 0 ss 8 Meathcation etter(so)ffotuseswith: bh fi ! ERi ursma i g.gg . 8g ASS a oe tn, rt. HT ERIS TI wd PEEP EEE 0 de de de de dude dn de fede de de ET sen Too. . rtTS eRsnEe pRlmi ecronntSwiend naw - moo " 3 an 2 lt oo nr TM oc mt tv om! E2gTdmEuam: FE 8 . . . og HERE c ---- mr iii PE EEE TRA ST or du da da de se da sn de he de Ho EE, oes : j a CR, Rn ; Be fm ' - wreCE1o8LhTEaItE iErrAegnEie a ovee mas Sg2 mic ave FREE, -paiSL thoracic contra noatused ., Brwaies: EcoEil wm. ` me :. ' fanem . " RSA A ne me ioii tpa bia Es E TE ETT 0 de de de dade he dee da de fe de f HT ET mpietmeastrie ' Sax fon x vr ERE Vr pe - or s fect) Shetle - scapals enification comple " " ` RE I Bnet rte he ARL mr Core ot ar SEEos e:t Td F SSo rapasom:.Fciii ee , aw Gg oto BSCE Semaration doses a ie of visciation eeldiacos a ERLE ITA mea pdid iie 0 de de de diode de dee de de i ptt vr Fn = : SA eri : Fuprascc pital sssification tncomplete/nefnoerds Nomex Wo wer won 0c ama 3 ! BHiE naeti ' .: 2S ic atte 22 tom -piJ ns sirc cS atinm-- onsite Ei 2 Ten or ae Th, wSET THE i melt id iii EEE HR A EE 0 de de de de dn de de ba dn be I EE EREE emiasi 4 FN x "eg. :g i E ETS E n Co toa wo gL wwc SEi ESs ' on ATEE EEe sfzt:- er em; : 0s - apeiates STes nas 1.1 Possible preparation dase at thee of evisceration. ' } :! wr i Fa mr iii iiiii yeg EERIE, Ident ication etter(s) of feteses w LL LLL LLE LL] Es T ten p hens sud x " a iRIVER SI foe 3H - . wed EREto mst . - "ow- . : Be oe - " Q wean mE 2 SE *5 - peric airate - pnts istication tcmpletes in BREET Bratton doe otteof esti BER "Hipare Sees 1 meld IEEE EEEEEE didi iid EES ME de fede fe du deeded de due SLcon Cpe mp nterparietalassfication Incoplate/mmassid . Gan te ay rTA rteenstris rr Bettys woo wm 0 3 ne Com $gn E EERE. : - Ser LEEEe we a 2S iia eietien teeters = Camus. @ wow "owen an PIC ated- pls enitiation ncompeter " * . leF la E it wee do 088 o ddf ERE, berof fetus coins wo mowdud 0bo odw ed1 adsem ow ow SLE E ecm e------ + ow EB Ii oSi F EER R Le "wr wmFou aw - am gus x:: g88vownn SA E EEiEE a EntCTsy wom Ca 25 < JC n 5 v meon is Pedic pedi - puts ne entiation tacmlete/ r= Pom : -5- RApepfeenrdeinxces Rat Teratology Study of T-3351 STATISTICAL METHODS/LABORATORY PROCEDURES 154-160 BArimoimteatgrei,cs,P.1,1:3"7T5e-s3t8s5, f1o9r55.Linear Trends in Proportions and Frequencies. ABgarritcleutltt,ureM.andS.A,ppl'iSeodneBioElxoagmyp.l"es 1937. J.of RoSytalatiSsttaitciaslt. MSeotch.odSsuppolf., ReIsVe:a1r3c7h-170i,n BCioocmhertarni,csW,. 1G0.:,417*-S4o5m1e, M1e9t5h4o.ds of Strengthening the Common Chi-Square Tests. SCooncsh,ranI,ncW.. NGe.w, Yaonrdk,Co1x9,57.G. N., Experimental Designs, 2nd Ed., John Wiley and RDervaipseir,ted.N". TeR.c.hnomaentdricsH,unt1e1r:,23-4W0., 1G9.8,9. "Transformations: Some Examples TDruenanettmte,ntsC.withW.,a Co"nAtroMlu.ltiplJ.e AmC.ompStaarti.sonAssoPcr.o,ced5u0r:e1086f-o1r121C,omp1a95r5i.ng Several R BDiuommmeetttr,ics,C. 20W:.4,62-4"9N1e,w 1T9a6b4l.es for Multiple Comparisons with a Control. EFidsihnebru,rgh,R. 19A5.0,. Statistical Methods for Research Workers, Oliver & Boyd, NFelweisYso,rk,J.`chL..,'9,St1a9t8i1s.tical Methods for Rates and Proportions, 2nd Ed., Wiley, AJlotnecrkaheteirvee,s." AS.iomRe.t,rikaA41:D1i3s3t-r4i5b,ut1i9o5n4-.free k-sample Test Against Ordered AKrnuaslkyasli,s.W. ).H.'Ama.ndStaWta.llfAss,socW.., A4.7,:58"3U-s6e21,of19R5a2n.ks in One-Criterfon Variance VKraursikaanlc,e AW.nalHy.siasn.dWalJl.isA,n. W.StaAt.!, A"sEsrorca.t,a t4o8:9U0s7e-9o1f1,Ran1k9s53.in One-Criterion CLoenvetnrei,butHi.o,ns Alto, 1960. `RtoobuPsrtobTaebsitlsityfoarnd EqSutaaltiitsyticosf, VaSrtfaannfcoersd,*UnifvnerIs.ityOlkpirnesse,ditpeadl,e aL W1i9l66l,er,Eh.R.6.G1.., Jr., Simultaneous Statistical Inference, McGraw-Hill, New York, 000611 v er "76 AppendiRxefe9 re-ncCeosntinued Rat Teratology Study of T-3351 STATISTICAL METHODS/LABORATORY PROCEDURES 154-160 Staples, R. E., "Detection Teratology, 9(3):A-37, 1974. of Visceral Alterations in Mammalian Fetuses.* 1564. Staples, R. E. and the KOH-Alizarin Red Schnell, V. S Method L., for "Refinements Fetal Bone." in Rapid Clearing Technic in Stain Technology, 39:61-63, 1AT4ge:ar3ip2ns7ts-rt3a,33T,rTe.n1d9J5,.2,. W"hTehne ATsiyemsptoatriec NPorremsaelnitty iannd OCnoensiRsatneknicnyg.*of KeInnddaalgl.'s MaTtehs.t Wilson, J. Techniques, G6. and The Univ. Warkany, of Chicago J., editors. 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