Document wqq59BpKaNRL3XkqxwRgb37j4

"'@HAZLETONLAWRATORAIMEERISCA,INC. DEEC:C211311199y8833] 7[ y R-ATTERATOLOSGTYUDY T-3351 FINARLEPORT Submittteod 3M Company St.Paul.-Minnesota December19, 19d3 ""@HAZLZTCNLABORATOARMEIIQEICSA.INC. OFFICE OF QUALITY ASSURANCE Project Title: Rat Teratology Study Project No.: 154-160 Quality Assurance insoections of the study and review of the final report of the above referenced project were conducted according to the standard operating procedures of the Office of Quality Assurance and according to the general requirements of the Good Laboratory Practice regulations that were issued on December 22, 1978, by the Food a'ndDrug Administration for compliance on and after June 20, 1979. Findings from the inspections and final report review were'reported to management and to the study director on the following dates: Inspections/Review Findings Reported Inspector/Revi ewer Protocol 6/15/83 Study 7/6,8,11,12/83 Final report 12/6-9/83 6/15/83 7/15/83 12/16/83 E. Burroughs E. Burroughs P. Runge derick G. dt* Ofrfeic OFficcee ooff QQuuaallii Assurance H A-ZLET (DNLABORATOFIIES AMERICA. INC. ......... '0 74.ANN@$CLvsgt4P4AVLIRGINIA3algCXU.SA. SUBJECTR:atTeratolSotguydy Project No. 154-160 We, the undersigned, hereby declare that the work was performed under our supervision, according to the procedures herein described. Study Director: LAWRENCE T.'WETZEL, Ph.-Y.Staff Scientist Department of Toxicology Laboratory Supervision: Teratology: GEOROE A. BURDOCK, Ph.7 Rodent Toxicology I Department of Toxicology RUTH S. OURLOO, B.S. Research Associate Department of Toxicology. Report Preparation: BARBARK R. COLPEAN, 8--.A."B,.S. Technical Writer Scientific Resources Department tls "'@HAZL=CNLABORATORAIMEERISCA.INC. TABLE OF CONTENTS Page SUMMARY 1 INTRODUCTION 3 CONTROL AND TEST MATERIALS 3 TEST ANIMALS 4 METHODS 5 Mating Period 5 Groups and Dosage Levels 5 Compound Preparation and Administration 6 Maternal Observations and Records 6 Cesarean Sacrifice 7 Visceral Examination of Fetuses 8 Skeletal Examination of Fetuses 8 Tissue Preservation a Statistical Analyses 9 Specimen, Raw Data, and Final Report Storage 11 RESULTS - MATERNAL DATA 12 Mortality and Clinical Observations 12 Body Weights and Food Consumption 12 Uterine Weights 13 Gross Pathology 13 RESULTS - CESAREAN DATA 14 Pregnancy Rates, Corpora Lutea, and Uterine Implantations 14 Fetal Viability, Weight. and Sex 14. Fetal Development 15 TABLES Table 1 - Summary of Maternal Clinical Observations 17 Table 2 - Mean Maternal Body.Weights and Body Weight Changes 18 Table 3 - Mean Maternal Food,Consumption 19 Table 4 - Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights 20 Table 5 - Summary of Maternal Gross Pathology Findings 21 Key to Tables 6 - 14 23 Table 6 Summary of Mean Ovarian and Uterine Data 25' 6"-."HAZtw=CNLABORATORIES AMERICA, INC. v.pa.f..^ 28,60. u.s..A. TABLE OF CONTENTS CONTINUED Table 7 - Summary of Mean Litter Data Table 8 - Summary of External Findings 26 Table 9 - Summary of Visceral Findings 27 Table 10 - Summary of Skeletal Findings 28 Table 11 - Incidence of External Findings Per Litter 29 Table 12 - Incidence of Visceral Findings Per Litter 30 Table 13 - Incidence of Skeletal Findings Per Litter 31 Table 14 - Mean Incidence of External, Visceral, and Skeletal 32 Findings Per Litter 33 APPENDICES AKpepyentdoixAppIen-diIcnedsiv2i,du3a,lCanldin4ical Signs' 34 Appendix 2 - Individual and Mean Maternal Body Weights and 38 Body Weight Changes Appendix 3 - Individual and Mean Food Consumption 39 43 Appendix 4 - Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid uterine Weights 47 Appendix 5 - Individual and Mean Ovarian and Uterine Data 51 Appendix 6 - Individual and Mean Litter Data Appendix 7 - IndividualVisceral Findings in Fetuses 55 59 Appendix 8 - Individual Skeletal Findings in Fetuses 67 Appendix 9 - References 75 VHAZL=CNLABORATORIES AMERICA,INC. 9200 L.EESSURG TURNPIKE, VIENNA, VIRGINIA 22'180. U.S.A. SPONSOR: 3M Company MATERIAL: T-3351 OATE: December 19, 1983 SUEBJECT: FINAL REPORT Rat Teratology Study Project No. 154-160 SUMMARY T-3351, suspended in corn oil at concentrations intended to deliver 1, 5, or 10 mg/kg/day (Grou.Ps2, 3, and 4, resp.qctively),was administered by oral intubation to mated female Sprague-Dawley COID 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 in 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 3 and 4 animals once treatment had been initiated. Terminal body weight minus gravid uterine weight was also significantly decreased in H.).L.*.a.L=.C.N.L.A.BORATORIES AMERICA. INC. 154-160 2- Groups 3 and 4. Treatment-related gross'pathology in Group 4 animals included incidencesof gastrointestinaltract effects. Statistical analysis revealed a positive treatment-relatedtrend in fetal loss, attributableprimarilyto an increased incidenceof 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 in 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 ossificationoccurred 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, fetotoxicityand an increased incidence of external and visceral anomalies and skeletal variants. T-3351 did not appear to be terato4enic in rats at dose levels less than or equal to 5 mg/kg. HAZLE-CONLASOF;ATOF;IES AMEFIICA,INC. ,a itse@oa rw. @,e N^. @,mooft,&aw... w.&^, -3 154-160 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, Dukels* Corn Oil (Lot No. 80275). a slightly viscous, yellow liquid, was received from the C. F. Sauer Co., Richmondi Virginia, and was stored at room temperature. The test material, T-3351, a white powder, was received from the sponsor on January 5, 1983, and was stored at room temperature. The test material was assumed to be 100% active compound. Information on the methods of synthesis, stability, as'well as data on composition, or other characteristicswhich define the test material, are on file with the sponsor. "'@HAZt.=CNLABORATORAIMEERISCA. ......... 4 154-160 TEST ANIMALS Two tyundredsixty (one hundred thirty/sex) sexually mature, cesarean-derived Sprague-Dawley Crl:CDo (SO)BR rats were received from Charles River Breeding Laboratories, Inc., Kingston. New York, on June 8, 1983. The rat was selected for use in this study because historic-ally 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 'approximatelythree weeks prior to initiationof the study. During this time, 'two male rats were euthanized due to an unthr*iftyappearance. Following an -examinationby a staff veterinarian for health status, one hundred twenty eight rats/sex were placed in breeding. The rats were housed one male and one female per cage during breeding. Following confirmation of mating, the females were uniquely identified by ear tags and individuallyhoused in elevated stainlesssteel cages. Food (Purina Rodent Laboratory Chow* 5001) and tap water (via an automatic watering system) were available ad libitum. The mean room temperature + the standard deviation was 72.90 F + 1.36 and the mean relative humidity the standard deviation was 69.1% + 3.16 during the study. A twelve-hour light/dark cycle was maintained. '"-')FAAZL=CNLABORATAOMERRIICAE. SINC. 154-160 METHODS Mating 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 04 of gestation. Groups and Dosage Levels 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, 1957). Group Number of Females Bose a mg/kg/day 1 25 0 2 25 1 3 25 5 4 25 10 a Based on individual animal body weights at each weighing interval during the dosing period. Animals were dosed on Days 6 through 15 of presumed gestation. All males and all remaining females were sacrificed via carbon dioxide asphyxiation and discarded without necropsy once a sufficient number of matings were confirmed. (IVDHAZL=CNL.ABOFIATORIES AMEFIICA.INC. 6 154-160 Compound Preparation and Administration Suspensions of the test material in the corn oil vehicle were prepared on a weight per volume basis. The required amount of compound for each level was weighed on an electronic Arbor 126 balance and transferred to a 7 ml tissue grinder. Approximately 3 ml of corn oil was added and the test material was ground into a fine suspension. ' This suspension was transferred to a precalibrated beaker and any remaining compound in the tissue grinder was rinsed into the beaker. Additional corn oil was added to the suspension to produce the desired concentration and the suspension was mixed on.a 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 400 F). Samples of each test mixture as well as the vehicle were sent to the sponsor for analysis. Maternal Observations and Records All animals were examined twice daily for mortality and moribun- dity (from day of receipt through animal sacrifice) and once daily for clinical signs of toxicity and pharmacologic effects (throughout gestation). Individual body weights were recorded -at,-initia:tionand on Days 6".."'HAZtn==NLABORAAMETRIOCAR,IINCE.S 7 154-160 6. B. 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 n.umbers so that all personnel 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 d@ad fetuses, number of early and late resorbing fetuses, and any abnormalities. The uterus was not reweighed 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 evaluation. The next two fetuses were processed for skeletal evaluation. This "'-"*'HAZL=CNLABORATAOMERRIICAE.SINi,@.gogeou-0 V.60V..0040A.a0m4'.sk&e0jA.. 8 154-160 process was repeated until all available.fetuses were selected for evaluation. Dead fetuses were discarded without examination. Visceral Examination of Fetuses Approximately one-third of all live fetuses from each litter was 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. Skeletal Examination of Fetuses 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. Tissue Preservation 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 H A Z LLt 0 ......... NLASOF;ATOF;IES AMERICA,INC. 9 154-160 examination were preserved in plastic -bags containing a glycerol:ethyl alcohol (1:1) solution with several crystals of bacterial and mold growth. 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 control group were compared statisticallyagainst the data of the compound-treated groups of the same sex. order: Analyses of the above data were performed in the following Levene's test for homogeneity of variance (Levene, 1960; Draper and Hunter, 1969) was 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 heterogeneous, a series of transformations was performed until variance homogeneity was achieved. These transformations were loglo, square (X2), square root (X1/2), reciprocal (I/X),angular (arcsine X1/2), and rank, in that order. If rank transformationwas ineffective in removing variance heterogeneity, ANOVA of ranked data was completed. If ANOVA of untransformed or transformed data was significant, Dunnett's t-test (Ounnett, 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. R"HAZL=CNLABORATORIES AMERICA. lt4C. 10 154-160 A = Data analyzed following loglo transformation. B = Data analyzed following square (X2) transformation. C = Data analyzed following square root (XI/2) transformation. D = Data analyzed following reciprocal (I/X) transformation. E = Data analyzed following angular (arcsine XI/2) transformation. F = Data analyzed following rank transformation. Mean fetal body weights per litter were statisticallyanalyzed as follows: Bartlett's test 'forhomogeneity of variances (Bartlett, 1937) was performed by one-way class'ificationof covariahce (ANCOVA) (Winer, 1971). I-f ANCOVA was significant, control vs. treatment group comparisons were analyzed using the Games and Howell modification of the Tukey-Kramer honestly significant difference test (Games 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 probabilitylevel. In addition to the above data, percent fetal viability,percent fetal loss (dead and resorbing fetuses),percent early, late, and totalresorptions, and the number of dead fetuses were analyzed by nonparametric one-way ANOVA (Kruskal and Wallis, 1952, 1953; Miller, 1966) and the Terpstra-Jonckheeretest for trend (Terpstra, 1952; Jonckheere, 1954). The 11tter was used as the experimental unit. The teratology data listed on the next page were analyzed using the AnhHAAZLLmtONLABORATORIEASMERI'%-,IAN.%@'. 154-160 Cochran-Armitage test for linear trend in proportions (Cochran, 1954; Armitage, 1955; Fleiss, 1981) and Fisher's mexact" test (Fisher,1950). Visceral Anomalies Skeletal Variants Subcutaneous edema Cleft palate Cryptorchism Number of litters affected Number of fetuses affected Skull Pectoral girdle Rib cage Vertebral column Pelvic girdle Limbs Bone alignment Number of litters affected Number of fetuses affected If a significanttrend was noted, the results of Fisher's Nexact" test were evaluated at the one-tailed, 5% level. If a significanttrend was not observed, or if there was a significant trend with severe departure from it, the results of Fisher's lexactltest were evaluatedat the two- tai'led.5% level. When using these statisticaltests on indices, the actual proportions prior to conversion to percentageswere 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 OsignificantN and/or as follows: S+ = Significantlyhigher than the control value. S- = Significantly lower than the control value. Specimen, Raw Data, and Final Report Storage All specimens, raw data, and the final report.are stored in the archives of Hazleton Laboratories America, Inc. -**)HAZL=CN.R"ATORIAEMESRICA. lt4C. 12 - 154-160 RESULTS MATERNAL-DATA Mortality and Clinical Observations Individual clinical observationsare presented in Appendix 1. A summary of clinical observations is presented in Table 1. Two Group 4 females were found dead on Day 17 prior to cesarean section on Day 20 of gestation. No other animals were found dead or sacrificed in extremis during the study. 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 from vagina, urine stains, alopecia. rough haircoat and bloody crust. Body Weights 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 significantlylower 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 proportionally greater than control increase in mean body weight gain in the treated females during the posttreatment phase (Days 16-20). ("@HAZLZM(=NLABORATOARMEIFIEICSA,INC. 13 - 154-160 Mean total food consumption was significantly lower in the Group 4 f emales during Days 17-20. Significant decreases in 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 Weights Individual and mean terminal body weights, gravid uterine weights, and terminal body weights minus gravid uterine weights are presented in Appendix 4 and Table 4. respectively. The mean gravid uterine weight of the Group 4 females wa-s significantly lower 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. Gross Pathology 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. ""@HAZLETCNLAi3OF;ATORIES ......... AMERICA, INC. 14 - 154-160 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 7. Summaries of the external, visceral, and skeletal findings are presented in Tables 8 through 14. Pregnancy Rates, Corpora Lutea, and Uterine Implantations Pregnancy rates, number of corpora lutea, and implantation efficiencieswere comparable for all groups. Fetal Viability, Weight, and 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 their respective controls. The percent of male fetuses was 5@.29%,.54.24%, and 60.38% for Groups 2, 3. and 4, respectively,and 44.01% for the control group. ""-@HAZLZMCNLA8OF4ATOF;IES Iw ......... AMERICA, INC. - 15 154-160 Fetal Development Statisticallysignificantfindingsin the teratologydata are presentedin the followingchart: StatisticallySignificantTeratologyData Teratology Observation Group: Visceral anomalies 'Subcutaneous edema Cleft palate Cryptorchi sm Number of litters affected Number of fetuses affected Skeletal variants Skull Pectoral girdle Rib cage Vertebral column Pelvic girdle Limbs Bone alignment Number of litters Number of fetuses affected affected -2 3 4 S+ S+ S+ S+ S+ S+ S+ S+ S+ S+. S+ s- S+ S+ S+ S+ Trend s t s s s s s s s s s s s s Departure from Trend s s NS NS s s s s s s s NS NS s S = Significant NS - Not significant r"')HAZLM70NLABOF;ATAMOEFF;IICIA,IENCS. a In* sauna tu. N..w 4. @'a @@a. @'no.ft'A ma'ac@ U.S.^. 16 154-160 Significant treatment-related increases in the incidences of visceral*anomalies and skeletal variants were noted in the Group 4 fetuses. 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. Numerous clinical signs of toxicity were also present in these animals. Table I Summary of Maternal Clinical Observationsa Rat Teratology Study of T-3351 Observation Group: Dose Level (mg/kg): Pretreatment Days 0-5 1 23 4 6- I- 5- 10 Number of females examined Number of females appearing normal Number of femd4es found dead or sacrificed prior to Day 20 25 25 25 25 25 25 25 25 0 000 Hunched Thin Anorexia Ataxia Sensitive to touch Soft feces Bloody discharge from vagina Urine stains Alopecia Rough haircoat Bloody crust Small eye Swollen neck Treatment Days 6-15 1 234 o- - -5 10 25 25 25 25 21 21 16 11 0 00 0 27 11 7 I I I I 33 59 66 anumerals indicate the number of animals with that particularfinding. Table 2 Mean Maternal Body Weights and Body Weight Changes Rat Teratology Study of T-3351 Body Weig t Mean Weight (g) Mean Change (9) Day(s) 0 N: 6 N: 8 N: 12 N: 16 N: 20 N: 0-6 6-8. 6-12 6-16F 6-20 16-20F 0-20F Group 1 0 mg/kg 270.08 24 298.47 24 301.39 24 318.31 24 338.53 24 395.41 24 28.38 2.93 19.84 40.06 96.95 56.88 125.33 Group 2 I mg/kg 271.03 23 296.05 23 298.04 23 308.18 23 329.20 23 389.69 23 25.03 1.99 12.13 33.15 93.64 60.45 118.67 Group 3 5 mg/kg- 263.25 23 292.60 23 288.22 23 * 290.09 23 305.34 23 367.11 23 29.54 -4.58S-2.70 12.54S74.32 61.77 103.86S- N: Indicates the sample size on which the mean calculationswere based. Only pregnant female calculations. Table 3 Mean Maternal Food Consumption(-g) Rat Teratology Study of T-3351 Interval Days 0-6 N: 7-8 N: 9-12 N: 13-16 N: 17-20F N: 7-16B N: 0-20B N: Group I 0 mg/kg 132.67 24 37.48 24 74.58 24 78.52 24 97'.25 24 190.58 24 420.49 24 Group 2 1 mg/kg 128.01 23 35.03 23 68.38 23 68.45 23 97.17 23 171.85 23 397.03 23 Group 3 G 5 mg/kg 1 134.92 1 23 31.53 22 59.58 20 57.89 20 87.87 19 147.9]S19 363.14S- 2 18 N:'Indicates the sample size on which the mean calculationswere based. Only pregnant females calculations. Invalid food consumption values were not included in mean calculations. Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg Table 4 Mean Terminal Body Weights, Gravid Uterine Weights. and Terminal Body Weight Minus Gravid Uterine Weights Rat Teratology Study of T-3351 Terminal Body Weight 395.41 M: 24 389.69 M: 23 367.11 M: 23 301.91 M: 20 Gravid Uterine WeightB 74.35 24 75.59 23 73.46 22a 60.51S20 Termina Weight Gravid Uteri 321.( 24 314.1 23 293. 22a 241.0 20 athe gravid uterine weight of one Group 3 female was inadvertentlynot recorded. N: Indicates the sample size on which the mean calculationswere based. Only pregnant femal calculations. Table 5 Summary of Maternal Gross Pathology Findings Rat Teratology Study of T-3351 Organ and Description Group: 1 2 Dose Level (mg/kg): 0 1 Number of animals examined Number with no gross pathology 25 25 22 24 THYMUS Firm I Enlarged 1 LUNG Focal areas - dark red HEART Right atrium-enlarged SPLEEN Enlarged Cystic KIDNEY(S) Enlarged I-rregularlythaped Pale Pelvis dilatpd Constricted across midline .Mottled tan and dark red Pelvis had a firm, white calculus ABDOMEN Tissue mass Findings in animals found dead during the study. Table 5 - Continued Summary of Maternal Gross Pathology Findings Rat Teratology Study of T-3351 Organ and Desgriplion Group: 2 Dose Level (mg/kg): 0 1 STOMACH Glandular mucosa - pitted areas - reddened - depressed area - red Nonglandular mucosa - raised area - dark (thin section in wall) - pinpoint, depressed areas - walls thin - thickened area red - dark foci Distended with dark red material Contained yellow mucous Walls thickened Walls thin PEYERIS PATCHES Enlarged/prominent I MANDIBULAR LYMPH NODES Reddened ALL LYMPH NODES Enlarged URINARY BLADDER Walls thickened Contained thick white fluid OVARY(IES) Clear fluid-filledcyst Findings in animals found dead during the study. 23 Key to Tables 6-14 154-160 Pregnancy-rate (percent) - (number of pregnant rats/number of rats mated) x 100. Surviva-IRate (i)ercent)- (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 litter/corporalutea per litter] x 100). Mean Incidence of Resorptions (percent)a - Group mean of (Cresorptions per litter/implantationpser litter] x 100). Mean Incidence of Fetal Mortality (percent) Mea n Incidenceof Fetal Viability(percent) Mean Incidence of External Variants (percent) Mean Incidence of External Anomalies (percent) Group mean o; ([dead and resorbing fetuses per litter/ implantationsper litter] x 100). G roup mean of ([live fetuses per litter/implantationsper litter]x 100). 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). Mean Incidence of Visceral Variants (percent) Group mean of ([number of fetuses per litter with visceral variants/number of fetuses per litter examined viscerally]x 100). avalues calculated separately for early, late, and total resorptions. 24 - 154-160 Key to Tables 6-14 Continued Mean Incidenceof Visceral Anomalies (percent) Mean Incidence of SkeletalVariants (percent) Mean Incidence of Skeletal Anomalies (percent) Group mean of ([number of fetuses per litter with visceral anomalies/number of fetuses per. litter examined viscerally]x 100). Group mean 'of ([number of fetuses per litter with skeletal vdriants/number of fetuses per litter examined skeletally]x 100). Group mean of ([number of' fetuses per litter with skeletal anomalies/number of fetuses per litter examined skeletally]x 100). Table 6 Summary of Mean Ovarian and Uterine Data Rat Teratology Study of T-3351 Number of females with mating confirmed Number of pregnant females Pregnancy rate (%) Number of females surviving to Day 20 Survivalrate (%) Mean number of: Corpora lutea Implantations Early resorptions Late resorptions Total resorptions Indices calculated Qn a per litterbasis: Mean implantationefficiency (%) Mean incidence of early resorptions Mean incidence of late resorptions Kean incidence of total resorptions Group: I Dose Level (mg/kg): 0 25 24 96 25 100 16.6 14.5 1.0 0.0 1.0 87.90 7.00 0.00 7.00 aone late resorption was noted for female No. 27086, therefore, the actual mean was 0.04. Table 7 Summary of Mean Litter Dataa Rat Teratology Study of T-3351 Mean number of: Fetuses - aead - live Indices calculdted on a per litterbasis: Mean incidedte.offetal mortality (%)c Mean incidenceof fetal viability(%) Mean percent males Live fetuses Mean body weight.(g)d Males - unadjusted mean - covariance adjusted mean Females - unadjusted mean - covariance adjusted mean Group: 1 Dose Level (mg/kg): 0 0.0 13.5 7.00 93.00 44.01 3.28 3.28 3.17 3.16 2 1 0.0 13.6 5.53 94.47 52.29 3.23 3.23 3.10 3.10 asex. percent males, and mean body weight data include values for live fetuses only. bone dead fetus was noted for female No. 27101, therefore. the actual mean was 0.04. cpercent mortality includes resorptionsas well as dead fetuses. dstatistical analysis performed on covariance adjusted means. Table 8 ,Summary of External Findings Rat Teratology Study of T-3351 Number Number Number Number of fetuses examined of fetuses within normal limits of fetuses with variants of fetuses with anomalies Number and type of anomalies Skin - edematous Palate cleft Group: 1 2 Dose Level (mg/kg): 0 1 325 313 325 313 0 0 0 0 0 0 Table 9 Summary of Visceral Findings Rat Teratology Study of T-3351 Group: 1 2 Bose Level (mg/kg): 0 1 Number of fetuses examined Number of fetuses within normal limits Number of fetuses with variants Number of fetuses with anomalies 115 113 111 105 4 a 0 1 Number and type of variants Brain - cerebral hemisphere contained dark area Eye(s) - containeddark-material Liver - dark areas Kidney(s)- pelvisdilated Ureter(s)- distended - undulated 4 10 I 4 3 5 I Number and type of anomalies Skin 7 edematous Palate- cleft Cryptorchism 0 1 I atwelve fetuses with edematous skin seen externally were confirmed at visceral examination. bfour fetuses with cleft palate seen externallywere confirmedat visceral examination. Table 10 Summary of Skeletal Findings Rat Teratology Study of T-3351 Group: 1 2 Dose'Level (mg/kg): 0 1 Number of fetuses examined Number of fetuses within normal limits Number of fetuses with variants Number of fetuses with anomalies 210 200 134 113 76 87 0 0 Number and type of variants 150 193 Lagging ossification Skull - incomplete closure (<75%) 6 15 - interparietalossifjcationincomplete/nonossified 30 40 - parietalsossificationirregular 1 14 - supraoccipital ossificationincomplete/nonossified 23 31 - supraoccipital nonfused I - hyoid ossification incomplete/nonossified 22 46 Pectoral girdle - scapula ossificationincomplete Rib cage - sternebrae bipartite I - sternebrae I and 2 fused I - sternebrae <3 ossified 5 1 - ribs <13 ossifieda I - ribs >13 ossified (cervicalregion) - ribs 13th pair smalia 4 - ribs ossificationirregular 1 2 Vertebral column - sacral arches nonossified 2 - thoracic centra ossificationincomplete/nonossified le 20 - thoracic centra nonfused 10 12 - caudals <3 ossified 13 5 Pelvic girdle - pubis ossificationincomplete/nonossified 8 --ischium nonossified I Forelimbs - <3 ossified I Hindlimbs - <4 ossified 2 1 Bone Alignment Ribs - angulated 1 4 Sternebrae - checkered apossible preparation damage at time of evisceration. Table 11 Incidence of External Findings Per Litter Rat Teratology Study of T-3351 Number Number Number Number of litters examined of litterswithin normal limits and percentof litterswith variants and percent of litters with anomalies Numher and percent of litters with anomalies Skin - edematous Palate cleft Group: Dose Level (mg/kg): 1 0 24 24 0/0.00 0/0.00 2 1 23 23 0/0.00 0/0.00 Table 12 Incidence of Visceral Findings Per Litter Rat Teratology Study of T-3351 Number of litters examined Number of litters within normal limits Number and percent of litters with variants Number and percent of litterswith anomalies Number and percent with variants Brain - cerebral hemisphere contained dark area Eye(s) - containeddark material Liver - dark areas Kidney(s) - pelvisdilated Uret- er(s)- distended - undulated Number and percent with anomalies Skin - edematous Palate- cleft Cryptorchism Group: Dose Leve.1(mg/kg): 1 0 24 2D 4/16.67 0/0.00 2 1 23 16 7/30.43 1/4.35 1/4.17 3/12.50 4/17.39 5/21.74 1/4.35 1/4.35 Table 13 Incidenceof Skeletal Findings Per Litter Rat Teratology Study of T-3351 Group: 1 Dose Level (mg/kg): 0 Number of litters examined Number of litters within normal limits Number and percent of litters with variants Number and percent of litters with anomalies 24 1 23/95.83 0/0.00 Number and percent with variants Lagging ossification Skull - incomplete closure (<75%) - interparietalossificationincomplete - parietalsossificationirregular - supraoccipitalossificationincomplete - supraoccipital nonfused - hyoid ossification incomplete/nonossified Pectoral girdle - scapula ossificationincomplete Rib cage - sternebrae bipartite - sternebrae I and 2 fused - sternebrae <3 ossified ribs <13 ossifieda ribs >13 ossified (cervicalregion) ribs 13th pair smalla ribs 'ossificatioinrregular Vertebral column - sacral arches nonossified - thoratic centra ossificationincomplete/ nonossified - thoracic centra nonfused - caudals <3 ossified Pelvic girdle - pubis ossificationincomplete/nonossified - ischium nonossified Forelimbs - <3 ossified Hindlimbs - <4 ossified Bone alignment Ribs - angulated Sternebrae - checkered 4/16.67 15/62.50 1/4.17 12/50.00 1/4.17 10/41.67 1/4.17 3/12.50 1/4.17 1/4.17 2/8.33 9/37.50 6/25.00 6/25.00 4/16.67 1/4.17 1/4.17 2/8.33 1/4.17 2 1 23 0 23/100.00 0/0.00 5/21.74 11/47.83 4/17.39 12/52.17 16/69.57 1/4.35 1/4.35 1/4.35 1/4.35 16/69.57 7/30.43 4/17.39 1/4.35 2/8.70 2 19/8 0/ 6/2 8/3 1/ 10/4 9/3 4/1 2/ I/ 1/ 1/ 11/4 2/ 5/21 1/4 114 1/4 2/8 2/8 .,@,-.i-Atindnamave at tirneof pitirrara+ip- Table 14 Mean Incidence of External, Visceral, and Skeletal Findings Per Litter Rat Teratology study of T-3351 Mean values calculated on a per litter basis: Number with variants Number with anomalies Incidence of variants Incidence of anomalies Mean values calculated on a per litter basis: Number with variants Number with anomalies Incidence of variants Incidence of anomalies Mean values"calculatedon a per litter basis: Number with variants .Number with anomalies @Incidence of variants Incidence of anomalies Group: Dose Level (mg/kg): External Visceral Skeletal 1 0 0.0 0.0 0.0 0.0 0.2 0.0 4,0 0.0 3.2 0.0 36.3 0.0 2 1 0.0 0.0 0.0 0.0 0.3 0.0. 7.0 0.7 3.8 0.0 44.6 .0.0 34 Appendix 1 IndividualClinical Signsa Rat Teratology-Study of T-3351 154-160 Maternal Number Group 27032 1 27034 1 27042 1 27046 1 27049 1 27057 2 27059 2 -27062 2 27063 2 27065 2 27066 2 27082 3 27083 3 27084 3 27085 3 27090 3 27091 3 27095 3 27099 3 27100 3 27101 3 Observation Alopecia Urine stains Bloody crust Swollen Alopecia Alopecia Alopecia Alopecia Alopecia Alopecia Alopecia Alopecia Alopecia Thin Alopecia Alopecia Alopecia Alopecia Alopecia Hunched Thin Alopecia Alopecia Rough haircoat Rough haircoat Rough haircoat Alopecia Rough haircoat Hunched. Rough haircoat Alopecia Alopecia Hunched Thin Anore'xia Urine stains Rough haircoat Bloody crust Bloody crust Bloody crust Bloody crust Rough haircoat Small Body Area Paw-fore-right Nose Neck Paws-fore-both Paws-fore-both Abdomen Paw-fore-right Paw-fore-left Paw-fore-both Hip-right Leg-hind-right Side-right Paw-fore-right Paw-fore-right Abdomen Hips-both Hip-right Leg-fore-right Leg-hind-right Paws-fore-both Hips-both Legs-hind-both Eyelids-both Mouth Nose Paws-fore-both Eye-left Day(s) 15-17 18-20 20 9-20 8-20 10-11 18-20 19-20 15 16-20 10-14 15-20 .10-20 6 17-20 17-20 15-20 15-20 14-20 15-19 10, 15-19 10-14 10-12,14 10-12 7-12 9-11 14-20 B-11,13 8-9,11 10-12,19-20 13-20 16-20 16-17,20 16,20 16,20 18-20 16,20 '20 20 20 20 8-9 15-20 aanimals not included in this table appeared normal throughout the study. 35 Appendix 1 - Continued IndividualClinical Signsa Rat Teratology Study of T-3351 154-160 Maternal Number Group Observation Body Area Day(s) 27107 27109 27111 27112 27113 27114 27115 27116 27117 4 Hunched Thin Rough haircoat 4 Hunched Thin Anorexia Sensitive to touch Alopecia Alopecia Rough haircoat Hips-both Hip-left Bloody crust Bloody crust 4 Alopecia Alopecia 4 Hunched Mouth Nose Abdomen Hips-both Thin Anorexia Urine stains Rough haircoat Bloody crust Mouth Bloody crust Nose 4 Hunched Thin Anorexia Urine stains Alopecia Abdomen Alopecia Alopecia Alopecia Hips-both Hip-left Neck Rough haircoat Bloody crust Bloody crust Bloody crust Bloody crust 4 Hunched Mouth Nose Paws-fore-both vagina Thin Anorexia Rough haircoat 4 Alopecia Alopecia. 4 Alopecia 4 Hunched Sides-both Side-right Paw-fore-left Thin Anorexia 18-20 13-17 13-20 9-12,14-20 10-20 16,18-20 10-14,16-18,20 11-20 10 13-20 20 20 10-20 10-20 18-20 15-20 17-20 19-20 16-20 20 20 20 12,20 12,17-18,20 14-18,20 11-19 9-18 8,19-20 8-18 15-20 16-2'0 16-20 16-20 18-19 13-20 12,20 17-20 18-19 8-13 14-20 19-20 14-20 17,19-20 16-20* a Animals not included in this table appeared normal throughout the study. 36 Appendix I - Continued Individual Clinical Signsa Rat Teratology Study of T-3351 154-160 Maternal Number Group 27117 4 27118 4 27119 4 27120 4 27121 4 27122 4 27123 4 27125 a Observation Urine stains Rough haircoat Bloody crust Bloody crust Bloody crust Bloody crust Hunched Thin Anorexia Rough haircoat Hunched Thin Anorexia Ataxia Soft feces Bloody discharge Urine stains Alopecia Alopecia Alopecia Alopecia Rough haircoat Hunched Thin Anorexia Bloody discharge Rough haircoat Found dead Hunched Thin Rough haircoat Hunched Alopecia Alopecia Alopecia Alopecia Hunched Thin Anorexia Alopecia Rough haircoat Bloody crust Body Area. Genitals Mouth Nose Paws-fore-both Vagina Hip-left Hip-right Inguinal-right Side-right Vagina Abdomen Hip-right Leg-hind-right Side-right Abdomen Paws-fore-both Da-v(s) 16-20 9-20 20 16-20 18-20 16-20 16-20 16-17,19-20 19 16-18 12-20 16-20 16-18,20 18-20 8-10 19-20 18-20 11 12-20 13-18 15-20 20 16 16 16 16 16 17 7-9,11-17 7-16 7-11 16-17 10-20 12-20 20 18-20 16-20 16,18-20 16-17.20 15-20 8-9 19-20 aanimals not included in this table appeared normal throughout the study. Maternal Number Group 27126 4 27127 4 27128 4 27129 4 37 Appendix 1 - Continued Individual Clinical Signsa Rat Teratology Study of T-3351 Observation Hunched Thin Anorexia Alopecia Hunched Thin Bloody discharge Rough haircoat Found dead Hunched Thin Anorexia Alopecia Rough haircoat Hunched Thin AnorexiaUrine stains Rough haircoat Bloody crust Body Area Hip-left Vagina Oaw-fore-right Paws-fore-both 154-160 Day(s) 18,20 19 16-17,20 17-20 16 16 16 16 17 14-20 14-20 17-18,20 15-17 18-20 14-20 16-20 16 19 16-20 16 aanimals not included in this table appeared normal throughout the study. 38 Key to Appendices 2, 3,'and 4 154-160 P - Pregnant NP - Not Pregnant C - Cesarean Section Performed (preceded by day of gestation) 0 - Death (preceded by day of gestation) MS - Moribund Sacrifice (preceded by day of gestation) ND - Natural Delivery (preceded by day of gestation) - Data excluded from mean calculation. Explanation of Footnotes a Only data from pregnant animals 'surviving to Day 20 cesarean section were used in mean calculations. b Invalid food consumption value not included in mean calculations. c Gavid uterine weight inadvertently not recorded. Appendix2 Individualand Mean MaternalBody Weightsand Body Weight Changesa Rat TeratologyStudy of T-3351 Group 1 0 mg/kg Maternal NumbeL_ 21030 27031 27032 21033 27034 21035 27036 21037 21038 27039 27040 27041 27042 27043 27044 27045 27046 21041 21048 27049 21050 21051 27052 21053 21OS4 k Kean S.O. Disposition 0 0 v eight g Dav of Gestation' 6 8 12 16 20 P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C NP. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C 217.3 261.9 24S.7 2S6.S 2SO.] 312.7 248.7 (266.4) 2S3.8 291.2 241.6 241.1 259.8 281.1 218.7 218.0 269.8 294.0 298.5 270.1 260.5 216.0 2?S.2 280.8 272.9 303.8 288.B 202.0 274.3 263.4 3S].S 279.5 (260.6) 283.4 321.4 273.0 277.3 286.6 310.9 297.4 316.0 293.6 323.6 327.7 310.1 28S.3 306.6 299.0 306.0 301.2 306.B 284.4 292.1 21S.S 2S9.0 351.0 283.3 (278.7) 284.7 330.6 212.0 281.2 .294.7 318.1 297.9 312.9 224.1 320.1 330.S 327.1 293.6 301.2 306.6 303.6 306.4 340.3 314.1 311.S 299.9 281.1 362.4 312.2 (212.8) 296.4 343.5 289.2 293.S 307.2 334.8 302.5 328.4 313.2 314.S 3S2.0 343.1 316.8' 321.4 327.1 294.2 333.4 3SO.6 329.2 338.6 314.4 300.9 353.6 340.2 (279.2) 309.1 36S.I. 310.9 319.9 333.2 34S.6 336.S 342.8 336.0 311.3 370.8 376.1 334.3 34S.4 3S6.8 329.6 351.8 422.5 386.2 404.3 367.8 341.4 446.6 409.0 (287.1) 373.1 427.1 340.8 374.1 361.9 366.2 390.8 381.8 384.S 371.1 430.8 441.0 392.S 399.4 422.2 396.2 412.6 24 270.8 18-790 24 298.47 20.S80 24 301.39 21.463 24 316.31 21.071 24 336.53 20.479 24 39S.41 28.666 0-6 26.5 26.9 36.3 17.0 13.3 38.8 30.8 (-5.6) 29.6 30.2 32.2 36.2 26.8 23.8 18.7 38.0 23.8 29.6 29.2 40.0 24.8 30.6 23.8 2S.2 28.3 24 28.38 6.658 Body Weiaht Chanae Days 6-8 -6-12 6-16F -6-2 3.0 -4.4 10.1 1.2 -4.4 -O.S 3.8 (17.9) 1.3 9.2 -1.8 3.9 8.1 7.2 O.S -3.1 0.5 -3.S 2.8 17.0 6.3 0.6 7.6 -2.4 5.2 36.5 25.3 3S.S 2S.6 17.7 10.9 33.4 (12.0) 13.0 22.1 IS.4 16.2 20.6 23.9 S.1 12.4 19.6 -2.1 24.3 33.0 31.S 14.8 28.1 -11.8 32.2 54.B 40.4 S6.6 40.1 37.5 2.1 60.7 (10.4) 2S.7 43.7 37.1 42.6 46.6 34.7 39.1 26.8 42.4 -6.3 Si.1 66.0 49.0 36.6 51.8 23.6 SO.6 lie 99 122 93 ?B gs 129 (26. es 105. 75. 96. 95. 51. 93. 65. 90. 47. Ill. 136. 107. 92. 123. 90. Ill. 24 24 24 24 2.93 19.64 40.06 96. S.364 12.SBS 16.908 21. Maternal Number-- 210$S 21OS6 21051 27068 21OS9 21060 21061 21062 21063 21064 2106S 21066 21061 21068 21069 21010 21011 21012 21073 21014 21015 21076 .21011 21078 21019 N mean S.C. LIU-OIULON P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C Np. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P, 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C UP. 20-C Appendix 2 - Continued Individual and Mean Maternal Body Weights and Body Weight Changesa Rat Teratology Study of 1-3351 Group 2 1 mg/kg Body Weight (g) Day of Gestation - 0 6 - -8 12 16 20 268.7 294.4 262.2 282.2 261.8 (2S3.4) 210.1 261.0 267.9 261.8 259.8 219.1 275.5 271.8 283.0 263.6 26S.6 286.2 275.6 278.4 265.1 259.5 262.8 264.9 (2SB.S) 290.8 318.8 286.5 3)2.0 304.9 (272.7) 301.0 228.1 298.0 296.0 299.4 296.2 307.7 202.0 3)4.4 209.4 291.9 310.4 301.6 324.3 280.3 290.7 293.6 283.2 (281.7) 294.2 323.2 282.8 300.S 302.4 (212.1) 296.4 271.8 294.7 301.8 294.0 304.7 304.S 219.8 317.6 299.7 293.7 316.6 290.1 324.8 292.0 285.9 292.S 290.6 (21S.7) 310.5 343.3 295.7 327.1 312.6 (289.S) 303.2 290.7 225.1 314.7 302.8 304.7 305.8 298.2 329.3 292.4 295.5 311.1 306.7 350.4 302.4 294.6 304.S 296.8 (215.5) 329.2 360.2 301.7 344.7 324.8 (269.3) 319.8 307.7 317.3 325.3 329.0 323.8 336.6 320.1 355.7 329.6 300.2 338.9 334.3 380.5 324.4 316.1 328.6 323.2 (214.S) 381.3 429.0 372.2 401.1 388.4 (272.1) 379.4 356.8 378.1 388.7 397.8 362.4 385.5 386.8 413.7 397.6 311.6 413.3 407.8 436.3 372.3 370.6 388.4 276.8 (273.1) 23 271.03 9.26S 23 23 296.05 298.04 18.650 13.221 23 308.18 15.771 23 329.20 18.312 23 389.69 20.073 0-6 -6-8 Body Weight Change (g) Days -6-12 b-16@- 6-20 22.1 24.4 24.3 37.1 (19.3) 30.9 -38.9 30.1 34.2 39.6 17.1 32.2 10.2 31:4 2@.8 26.3 24.2 26.0 4S.9 22.6 31.2 30.8 18.3 (23.2) 3.4 4.4 -3.1 -11.6 -2.5 (0.0) -4.6 43.7 -3.3 S.8 -5.4 8.5 -3.2 -2.2 3.2 10.3 1.8 6.2 -10.9 0.5 3.7 -4.8 -1.1 1.4 (-6.0) 19.1 24.5 9.2 15.1 1.7 (16.6) 2.2 62.6 -2.9 18.7 3.4 6.5 -1.9 16.2 14.9 3.0 3.6 0.7 5.1 '26.1 14.1 3.9 '10.9 13.6 (-6.2) 38.4 41.4 15.2 32.1 19.9 (-3.4) 18.8 19.6 19.3 29.3 29.6 27.6 20.9 38.1 41.3 40.2 8.3 28.5 32.7 S6.2 36.1 25.4 35.0 40.0 (-7.2) 90.5 110.2 85.7 09.1 83.5 (-0.6) 70.4 128.7 80.7 92.7 98.4 66.2 11.0 104.8 99.3 108.2 85.9 )02.9 106.2 112.0 84.0 80.1 94.8 93.6 (-8.6) 23 25.03 ls.egs 23 1.99 10.774 23 12.13 )3.617 23 33.15 14.547 23 93.64 14.i64 Maternal -Number 27080 210BI 21082 27063 27094 27085 21086 27061 27088 27089 27090 21091 27092 27093 27094 2709S 27096 21097 21098 27099 21100 27101 27102 27103 27104 N hean.. S.O. Appendix 2 - Continued Individual and Mean Maternal Body Weights and Body Weight Changesa Rat Teratology Study of T-3351 Group 3 5 mg/kg Body Weiaht (a) Day of gestation Disposition 0- 6 a 12 16 20 - P. 20-C P. 20-C P. 20-C P. 20-C NP. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P, 20-C P. 20-C P. 20-C P. 20-C NP. 20-C 211.9 267.1 256.1 309.2 291.4 285.0 305.7 266.9 273.2 310.1 289.2 279.5 343.9 299.9 28S.2 425.3 355.0 3S2.7 272.9 301.1 300.1 306.6 306.0 361.1 (258.7) (300.9) (276.1) (2B6.3) (246.5) (279.4) 271.1 295.2 284.7 307.1 329.1 382.6 261.3 286.3 282.7 296.2 312.2 372.1 214.0 229.6 305.9 251.5 294.1 2S].l 304.9 263.7 316.0 269.9 415.7 342.9 261.6 224.7 293.4 243.2 285.4 24S.8 298.2 242.7 317.1 255.1 314.7 308.6 2S2.6 287.0 260.5 275.3 302.8 360.9 276.0 309.1 302.2 290.6 316.9 391.3 265.5 297.3 206.9 291.6 312.3 373.2 251.7 281.2 289.4 299.6 312.2 372.0 276.7 210.6 305.1 301.2 297.7 298.7 300.1 216.9 336.2 278.3 410.4 358.6 290.0 319.7 320.5 314.7 335.6 398.7 213.0 299.8 292.0 293.6 313.4 311.5 2S9.1 290.0 273.1 287.4 302.7 366.1 282.8 320.2 310.5 225.4 269.9 254.1 216.5 260.4 294.0 280.0 293.3 281.7 280.3 289.9 305.1 308.9 357.0 369.0 216.8 287.6 292.8 216.5 292.0 310.1 (212.7)(202.9) (214.9)(251.1) (253.7) (275.4) 23 263.25 18.525 23 292.80 18.070 23 288.22 16.876 23 290.09 16.252 23 305.34 22.346 23 367.11 3S.036 -0-6 Body W*ight Chanue (a) Davs 6-8 6-12 6-16F 6-20 31.3 24.3 28.9 28.2 (42.2) 10.1 25.0 31.8 21.9 31.8 18.5 34.4 33.1 31.0 29.5 28.4 30.6 2B.9 26.6 30.9 37.4 17.S 19.6 70.8 (10.2) -3.5 -4.5 -11.8 -1.0 (-24.6) -10.5 -3.6 -11.1 -0.4 -8.0 2.6 -6.5 -6.9 -10.4 6.2 -7.4 -2.5 0.0 -7.0 -16.9 -9.7 -0.7 1.7 5.2 (-B.0) 0.9 -2.2 -5.5 5.5 (-14.6) 11.9 9.9 -0.9 12.2 4.B -0.5 -11.7 -18.S -5.7 18.4 -s.0 -22.3 -5.0 -6.2 -2.6 -24.8 -13.7 9.9 -11.1 (-31.8) 34.7 8.5 0.2 4.9 (-54.4) 33.9 25.2 12.2 10.4 23.1 11.9 IS.B 7.8 15.0 31.0 31.1 -22.9 16.1 13.6 12.7 -50.4 11.1 28.9 4.4 (-29.2) 116.1 63.6 67.7 60.0 (-21.5) 87.4 85.8 109.9 91.4 Bi.3 65.4 73.9 B2.2 75.9 90.8 105.3 57.4 79.0 71.7 76.1 -66.1 63.0 89.0 82.5 (-7.5) 23 29.54 10.523 23 -4.SBS6.103 23 -2.70 11.235 23 12.54S16.834 23 74.32 34.272 Maternal Number 2710S 21106 21107 21108 27109 21110 21111 21112 21113 .21114 21115 21116 21117 21118 21119 21120 21121 21122 21123 21124 21125 21126 27121 21128 21129 u Mean Appendix 2 Continued Individual and Mean Maternal Body Weights.and Body Weight Changesa Rat Teratology Study of 1-3351 Group 4 - 10 mg/kg Disposition 0 godv Welaht (a Day of Gestation 6 8 -'12 16 20 P. 20-C P. 70-C P. 20-C po 20-C P. 20-C P. 20-C P. 20:-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C P. 20-C UP. 20-C P. 20-C P. 11-0 P. 20-C P. 20-C P. 20-C P. 20-C P. 20-@c P. 20-C. P. 17-0 UP. 20-C UP. 20-C. 219.5 303.4 293.2 286.8 281.4 364.1 274.2 310.1 302.2 301.S 322.1 318.4 239.1 25S.2 255.3 226.0 212.1 271.7 28S.4 315.6 312.0 299.1 291.7 371.2 273.2 301.8 267.7 250.3 230.3 209.3 2S4.7 269.9 293.1 300.4 219.3 301.1 213.3 305.9 261.4 302.8 325.9 386.7 251.7 281.3 281.0 214.1 238.2 ?22.6 294.6 267.0 334.6 318.2 318.1 313.0 274.5 282.1 254.1 254.4 235.3 240.5 242.9 274.3 211.1 263.1 26S.6 351.6 241.8 215.6 278.7 214.7 210.3 340.0 303.0 339.4 339.1 289.8 211.4 247.2 (272.6) (273.3) (27S.3) (256.5) (224.9) (206.6) 255.6 29S.4 286.4 326.3 279.3 329.7 244.3 292.4 221.8 271.0 180.7 - 264.4 281.4 269.3 268.4 26S.2 349.6 253.4 310.8 264.3 275.6 332.5 283.2 277.7 338.2 292.5 270.4 319.2 218.9 26S.S 335.4 296.7 331.3 414.9 366.8 251.8 211.2 277.0 290.5 275.9 286.2 248.9 270.0 223.2 243.9 211.4 232.2 259.9 293.4 280.2 274.5 245.9 - (263.6) (286.2) (283.9) (270.1)(225.1)(189.1) (297.0) (299.5) (293.5) (259.4)(234.6)(256.8) 22 269.30 20.329 22 297.72 22.698 22 293.63 22.916 22 276.25 21.762 22 265.12 32.153 20 301.91 73.253 0-b 23.9 35.8 16.1 30.2 28.6 38.4 30.S 29.6 40.0 31.2 31.4 33.6 36.4' (0.7) 30.8 30.9 17.0 22.2 21.7 16.9 25.2 19.3 33.5 (22.6) (2.5) Body Weight Change (g) n.ys 6-8 6-12 6-16F 6-20 -10.2 -16.6 -8.S -3.2 0.1 -21.2 -3.6 -15.9 -14.1 -si.s -13.8 -19.8 0.7 5.5 -0.3 -6.6 -16.S -60.1 -S.2 -36.1 -3.2 -11.2 3.1 -0.9 -0.3 -49.6 (2.0) (-16.8) -1.1 -42.1 3.4 -33.9 -12.1 -13.0 2*.l -5.2 5.1 -13.3 9.3 - -4.3 -1.1 -28.1 -4.3 -20.S -13.2 -18.9 (-2.3) (-16.1) (-6.0) (-40.1) -22.0 12.0 -42.5 -23.9 -71.5 -31.7 9.4 -43.1 -80.5 -63.8 -8.1 -5.3 -68.0 (-48.4) -64.6 -55.3 -16.2 -10.1 2.9 13.5 -53.8 -46.6 -41.5 (-61.1) (-64.9) 61.3 61.7 16.5 61.6 -92.5 32.8 86.3 -58.5 -99.3 -77.7 77.3 64.4 -92.2 (-@6.1) -IOS.? - 68.2 55.7 82.4 83.6 -65.6 -58.3 - (-96.5) (-42.1) 22 28.43 6.951. 22 -4.OSS7.IIS 22 -21.48 11.769 22 -32.60S29.333 20 5.40 15.133 Maternal Number- 27030 27031 27032 27033 27034 27035 27036 27037 27038 27039 27040 27041 27042 27043 27044 27045 27046 27047 27048 27049 27050 27051 27052 27053 27054 N Mean S.D. 43 Appendix 3 Individual and Mean Food Consumptiona Rat Teratology Study of T-3351 Group I - 0 mg/kg 154-160 0-6 140.9 130.0 140.6 123.7 80.6 166.0 139.2 (113.8) 134.9 145.4 122.3 129.9 136.1 142.4 141.2 146.9 115.9 123.4 144.5 149.3 119.5 132.7 127.1 125.0 126.5 24 132.67 15.924 Food-Consumption (a) Interval (Davs of Gestation) - 7-8 9-12 -13-16 17-20 F 7-168 42.8 33.1 45.5 32.0 18.6 44.7 36.2 (30.1) 33.4 43.3 31.8 37.2 42.9 44.6 34.2 39.1 37.6 29.7 42.4 51.3 36.5 36.6 39.8 36.2 30.1 92.5 73.2 86.1 75.8 48.8 69.8 80.9 (52.2) 75.9 76.8 69.7 72.2 79.5 89.5 60.6 79.1 75.8 41.8 91.7 90.8 77.6 74.9 81.3 47.1 78.4 87.1 71.3 82.2 80.6 51.9 44.8 86.8 (55.2) 77.9 91.6 71.1 73.0 80.1 81.5 78.7 69.4 78.1 82.6 93.5 97.7 72.1 74.4 85.5 97.3 75.2 101.7 91.1 106.4 96.6 67.8 109.1 106.4 (69.7) 107.0 107.8 84.5 86.5 95.8 93.0 99.0 78.4 95.4 71.8 113.4 116.4 92.5 112.2 106.4 96.7 98.1 222.4 177.6 213.8 188.4 119.3 159.3 203.9 (137.5) 187.2 211.7 172.6 182.4 202.5 215.6 173.5 187.6 191.5 154.1 227.6 239.8 186.2 185.9 206.6 180..6 183.7 465.0 398.7 460.8 408.7 267.7 434.4 449.5 (321.0) 429.1 464.9 379.4 398.8 434.4 451.0 413.7 412.9 402.8 349.3 485.5 505.5 398.2 430.8 440.1 402.3 408.3 24 37.48 6.812 24 74.58 13.416 24 78.52 12.317 24 97.25 12.693 24 190.58 25.916 24 420.49 47.701 Maternal -Number 27055 27056 27057 27058 27059 27060 27061 27062 27063 27064 27065 27066 27067 27068 27069 27070 27071 27072 27073 27074 27075 27076 27077 27078 27079 N Mean S.D. 44 Append ix 3 - Continued Individual and Mean Food Consumptiona Rat Teratology Study of T-3351 Group 2 - I mg/kg 154-160 Food Consumption (g) Interval Mays of Gestation) - 0-6 7-8 9-12 13-16 122.0 136.0 130.8 136.5 141.7 (108.2) 131.5 55.7 135.6 143.0 145.9 99.3 144.9 111.5 145.8 133.0 119.6 145.4 105.6 147.3 127.0 131.2 126.4 128.5 (156.8) 35.2 42.1 35.2 29.1 38.2 (30.2) 31.6 38.6 34.2 39.1 34.7 33.6 35.2 22.7 40.4 42.8 34.9 41.9 24.4 41.9 40.7 27.8 28.1 33.2 (23.8) 63.7 74.1 68.0 70.2 64.5 (67.9) 62.8 81.8 61.4 73.5. 63.1 60.3 63.6 64.2 72.6 61.4 60.6 62.1 62.8 93.2 72.7 87.8 63.2 65.1 (53.4) 71.0 69.7 49.6 69.0 71.2 (39.8) 66.0 68.6 70.9 55.3 81.3 63.0 72.8 59.2 77.0 67.6 41.5 68.2 100.6 88.8 68.2 59.0 70.0 65.8 (56.3) 92.5 169.9 108.9 185.9 88.6 152.8 98.0 112.1 168.3 173.9 (77.9)b (137.9) 91.3 160.4 70.3 189.0 98.7 166.5 99.4 167.9 116.9 179.1 92.1 156.9 88.5 171.6 97.7 146.1 94.0 98.5 190.0 171.8 101.4 109.2 137.0 172.2 107.1 187.8 113.6 223.9 84.3 181.6 79.5 174.6 95.9 161.3 96.4 164.1 (64.9) (133.5) 384.4 430.8 372.2 402.8 427.7 (324.0)b 383.2 315.0 400.8 410.3 441.9 348.3 405.0 355.3 429.8 403.3 358.0 426.8 400.5 484.8 392.9 385.3 383.6 389.0 (355.2) 23 128.01 20.444 23 35.03 5.735 23 68.38 8.893 23 68.45 12.065 23 97.17 11.147 23 171.85 17.544 23 391.03 35.568 Maternal Number- 27080 27081 27082 27083 27084 27085 27086 27087 27088 27089 27090 27091 27092 27093 27094 27095 27096 27097 27098 27099 27100 27101 27102 27103 27104 N Mean S.D. 45 Appendix 3 - Continued Individual and Mean Food Consumptiona Rat Teratology Study of T-3351 Group 3.- 5 mg/kg 154-160 ---Food Consumption (g) Interval CDays of Gestation) 0-6 7-8_ 9-12 -13-16 17-20 F 7-16 B B -0-20 139.2 139.9 138.5 136.1 (141.5) 127.7 113.8 146.0 118.1 134.4 97.9 132.3 147.3 130.5 168.5 120.0 136.2 145.1 147.3 177.4 141.8 118.9 115.4 130.8 (138.8) 36.9 34.5 26.3 42.7 (21.4) 23.3 28.3 60.3 48.3 46.7 64.6 (53.2) 55.7 53.5 70.0 60.9 38.5 39.5 (11.2) 66.7 59.6 104.7 82.8 83.1 73.4 (74.2) 80.5 88.0 167.2 143.7 111.5 146.8 (85.8) 145.7 141.4 411.1 366.4 333.1 356.3 (301.5) 353.9 343.2 32.2 (116.4)b 72.5 123.9 (221.1)b (491.0)b (156.2)b (86.5)b (88.1)b(109.3)b(330.8)b (558.2)b 35.0 28.3 26.8 36.2 24.4 40.4 24.9 61.0 46.9 44.6 153.0 51.1 72.5 56.5 61.3 52.3 60.5 67.1 79.8 69.0 55.2 87.8 74.7 85.4 95.0 100.9 96.1 114.6 157.3 127.5 131.9 256.3 155.3 181.9 136.6 379.5 300.1 349.6 498.6 386.7 446.5 371.2 34.6 (1,19.9)(b160.8)b(100.6)b(315.3)b(552.1)b 40.9 57.8 53.6 100.9 152.3 398.3 32.9 75.0 89.4 (135.8)b 197.3 (480.4)b 26.6 28.9 30.1 67.0 34.2 44.0 (110.3)b(108.9)b(203.9)b(490.2)b 1.5 0.0 64.6 206.4 65.1 91.9 139.2 350.0 28.4 31.1 (24.5) 56.5 42.4 (27.8) 51.9 43.4 (36.9) 90.5 95.3 (64.3) 136.8 342.7 116.9 343.0 (89.2) (292.3) 23 134.92 17.356 22 31.53 5.567 20 59.58 24.302 20 57.89 18.350 19 87.87 24.771 19 18 147.91S- 363.14S38.131 59.846 Maternal Number 27105 27106 27107 27108 27109 27110 27111 27112 27113 27114 27115 27116 27117 27118 27119 27120 27121 27122 27123 27124 27125 27126 27127 27128 27129 N Mean S.D. 46 . Ap pendix 3 - Continued Individual and Mean Food Consumptiona Rat Teratology Study of T-3351 Group 4 - 10 mg/kg 154-160 Food Consumption (g) Interval (Days of Gestation) 0-6 7-8_ 9-12 -13-16 17-20 F B - 7-16 126.5 148.7 110.0 138.3 140.5 143.2 136.7 128.9 150.0 142.9 123.7 117.4 164.1 (115.8) 124.6 150.7 98.5 120.6 158.2 137.6 115.0 123.5 133.7 (122.7) (113.0) 30.6 40.0 144.4 37.3 31.7 31.3 36.7 33.8 21.2 30.8 29.7 29.0 44.5 . (32.5) 28.9 42.3 24.3 33.5 41.4 34.5 28.1 26.6 27.5 (31.5) (27.9) 41.5 58.6 17.9 67.4 80.0 99.5 90.0 166.0 296.5 414.2 23.6 40.2 54.2 23.0 (200.2)b 222.2 91.3 100.5 (532.4)b 330.1 (145.5)b 55.7 48.3 40.1 1.4 21.3 39.2 86.5 10.6 (57.2) 15.2 31.5 0.0 2.3 * (177.2)b (320.0)b 35.6 85.0 122.6 350.8 75.1 100.9 160.1 397.7 1.6 0.0 75.5 204.4 0.0 0.0 22.6 172.6 b.o 1.1 52.3 196.1 26.3 90.5 95.2 309.4 57.1 (1'10.0)b 172.6 (400.0*)b 0.0 0.0 55.1 219.2 (41.1)b (24.9) (130.8)b (271.5)b 0.2 0.0 44.3 168.9 0.0 - 73.8 - (48,O)b(126.3)b (96.8)b(198.6)b(393.9)b 44.3 (164.9)b(152.0)b'(242.7)(b515.3)b (66.2)b 54.5 (119.8)b(162.1)b(440.1)b (129.3)b(196.3)b(117.8)b(360.1)b(615.5)b 17.1 39.9 48.4 0.0 7.5 11.8 0.0 45.2 160.2 0.0 74.0 197.5 - 87.7 - (41.1) (20.4)b (0.4) (93.0)b(216.1)b (15.7) (0.0). (72.2) (43.6) (228.8) 22 133.33 16.209 22 37.64 24.571 le 36.86 20.179 19 22.75 26.380 14 17 13 39.33S- *97.63S- 262.89S46.891 54.321 90.464 47 Appendix 4 Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Rat Teratology Study of T-3351. Group I - 0 mg/kg 154-160 (g)a Maternal Number- 27030 27031 27032 27033 27034 27035 27036 27037 2703B 27039 27040 27041 27042 27043 27044 27045 27046 27047 27048 27049 27050 27051 27052 27053 27054 N Mean S.O. Terminal Body Weight 422.5 38B.2 404.3 367.8 341.4 446.6 409.0 Not Pregnant 373.1 427.1 348.8 374.1 381.9 368.2 390.8 381.8 384.5 371.1 438.B 447.0 392.5 399.4 422.2 396.2 412.6 24 395.41 28.666 Gravid Uterine WeightB 90.4 90.9 8B.2 67.8 66.6 97.3 86.9 69.2 76.2 54.7 B2.4 80.1 30.8 80.6 52.4 72.3 .64 .5 74.' 0 B4.1 74.8 60.3 B3.1 74.6 B2.1 24 74.35 14.704 Terminal Body Weight Minus Gravid Uterine WeiahtF 332.1 297.3 316.1 300.0 274.B 349.3 322.1 303.9 350.9 294.1 291.7 301.8 337.4 310.2 329.4 312.2 306.6 364.B 362.9 317.7 339.1 339.1 321.6 330.5 24 321.07 23.215 48 Appendix 4 - Continued Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Rat Teratology Study of T-3351 Group 2 - I mg/kg 154-160 (g)a Maternal Number. 27055 27056 27057 27058 27059 27060 27061 27062 27063 27064 27065 27066 27067 27068 27069 27070 27071 27072 27073 27074 27075 27076 27077 27078 27079 N Mean S.D. Terminal Body Weight 381.3 429.0 372.2 401.1 388.4 Not Pregnant 379.4 356.8 378.7 388.7 397.8 362.4 385.5 386.8 413.7 397.6 377.8 413.3 407.8 436.3 372.3 370.8 388.4 376.8 Not Pregnant 23 389.69 20.073 Gravid Uterine WeightB 72.5 88.5 73.3 86.5 69.1 69.2 64.0 67.4 74.6 80.0 37.9 76.1 80.4 83.7 90.0 77.8 85.3 84.7 84.8 77.3 60.7 83.2 71.6 23 75.59 11.458 Terminal Body Weight Minus Gravid Uterine WeiqhtF 308.8 340.5 298.9 314.6 319.3 310.2 292.8 311.3 314.1 317.8 324.5 309.4 306.4 330.0 307.6 300.0 328.0 323.1 351.5 295.0 310.1 305.2 305.2 23 314.10 14.093 49 Appendix 4 - Continued Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Rat Teratology Study of T-3351 Group 3 - 5 mg/kg 154-160 (g)a Maternal Number Terminal Body Weight Gravid B Uterine Weight Terminal Body Weight Minus Gravid Uterine Weight 27080 27081 27082 27083 27084 27085 27086 27087 27088 27089 27090 27091 27092 27093c2.7094 27095 27096 27097 27098 27099 27100 27101 27102 27103 27104 425.3 355.0 352.7 361.1 Not Pregnant 382.6 372.1 415.7 342.9 374.7 308.6 360.9 391.3 373.2 372.0 410.4 358.6 398.7 371.5 366.1 254.1 357.0 369.0 370.1 Not Pregnant 103.9 72.2 72.4 68.5 89.8 79.4 76.6 73.5 71.2 61.6 75.3 88.0 - 71.2 87.1 65.4 65.2 68.0 61.6 48.2 73.9 72.9 70.2 321.4 282.8 280.3 292.6 292.8 292.7 339.1 269.4 303.5 247.0 285.6 303.3 - 300.B 323.3 293.2 333.5 303.5 304.5 205.9 283.1 296.1 299.9 N Mean S.D. 23 367.11 35.036 22 73.46 11.486 22 293.38S- 28.235 50 Appendix 4 - Continued Individual and Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Rat Teratology Study of T-3351 Group 4 - 10 mg/kg 154-160 (g)a Maternal Number Terminal Body Weight Gravid B Uterine Weight Terminal Body Weight Minus F Gravid Uterine Weight 21105 27106 27107 27108 27109 27110 27111 27112 27113 27114 27115 27116 27117 27118 27119 27120 27121 27122 27123 27124 27125 27126 27127 27128 27129 364.7 378.4 271.7 377.2 209.3 325.9 386.7 222.8 235.3 240.5 351.6 340.0 247.2 Not Pregnant 180.7 Found Dead 349.6 331.3 414.9 366.8 211.4 232.2 Found Dead Not Pregnant Not Pregnant 82.3 58.9 54.6 79.8 72.2 64.4 86.5 40.8 43.3 42.3 82.4 55.4 38.5 32.4 71.7 66.0 79.4 76.0 @39.7 43.6 2B2.4 319.5 217.1 297.4 137.1 261.5 300.2 182.0 192.0 198.2 269.2 284.6 20B.7 148.3 277.9 265.3 335.5 290.8 171.7 188.6 N Mean S.D. 20 301.91 73.253 20 60.51S17.739 20 241.40S- 59.517 Maternal Number 27030 27031 27032 27033 27034 27035 27036 27031 27038 27039 21040 27041 21042 21043 27044 27045 27046 27047 27048 27049 270SO 27051 27052 27OS3 27054 Total u Kea0n S. Ovarian Corporl Lutea is 11 19 19 16 20 14 Not Pregnant 16 19 16 is 17 14 is 19 13 12 is 19 .17 16 16 is le 399 24 16.6 2.12 Appendix 5 Individual and Mean Ovarian and Uterine Data Rat leratology Study of T-3351 Group I - 0 mg/kg Uterlne Implantation loplaftt&tions LttltltKM_M Early Lesorptions I Early Resorptions Late Resorptions is 100.0 0 0.0 0 11 100.0 1 5.9 0 Is 78.9 1 6.7 0 is 70.9 3 20.0 0 Is 93.8 1 6.7 0 le 90.0 1 5.6 0 14 100.0 0 0.0 0 14 81.5 2 14.3 0 11 89.S 4 23.5 0 13 81.3 1 7.7' 0 Is 100.0 0 0.0 0 Is 88.2 1 6.7 0 7 50.0 2 28.6 0 16 se.9 1 6.3 0 10 52.6 0 0.0 0 13 100.0 0 0.0 0 12 100.0 1 8.3 0 is 100.0 1 6.7 0 le 94.7 1 5.6 0 14 82.4 0 0.0 0 13 81.3 2 15.4 0- is 93.9 0 0.0 0 is 83.3 0 0.0 0 17 94.4 0 0.0 0 348 24 14.S 2.45 24 07.90 13.472 23 24 1.0 1.04 24 7.00 8.024 0 24 0.0 - Late Resorptions 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 .0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 24 0.00 - Tot Posorp 0 1 1 3 1 1 0 2 4 1 0 1 2 1 0 0 1 1 1 0 2 0 0 0 23 24 1.0 1.0 ts's EZ L9*0 slo el st 00*0 010 ca cz 0 toe*t ts*s cz L9*0 9*0 cz t69*6 tilig cz 19*1 t,tt ca tte L*9 1 0*0 0 L"9 t 9*EG SL 0*0 0 L'L L 0*0 0 010 0 010 0 oloot et L'L I L*99 el L*g L 0*0 0 L*9 t C*tg SL E*9 1 0*0 0 E*9 t 6*00 9t cli 010 0 9'S t o*o 0 0'0 0 0'0 0 t*t6 9t 9*S 1 0"0 0 010 0 O*OOL OL 0*0 0 C*96 tlt 0*0 0 010 0 0*0 0 tlt6 9t O*tt 1 0*0 0 O*tt 0*" It E*9 010 0 C*9 zlte 9L slzt C*OL t 010 0 010 0 S*Zt O*OOL 9IL E'tt t 9*99 0'0 0 L*9 1 L'L t 010 0 0*0 0 010 0 0*0 0 t*t6 9L L*g t o*oot st L*L t 6*Z6 Et 010 0 0*0 0 010 0 L*99 EL 010 0 010 0 010 0 L*t6 tL E*ot L*9 0*0 0 E*9 t L*L 1 010 0 010 0 0*0 0 0*0 0 010 0 E'tt z O*OOL tt L'9 L o*oot st 0*0 0 O*OOL EL E*9 t L't6 9t L*L I L*90 Et suolidi litiol suolzaioso ltiol (9) SU0140josTo -su-0--t-Ilio-sed SUO zaiosav elyl siel AtJ13 Ajiv] A3uat3l@IT3 suotioluelm-1 uoll#4uwidwi oulialn 64/bw t - a dnoig iscc-ljo Apn4SA50toleiolits 2420 SUIJOIA PUW UIIJVAO UVOW PUe t2nptAIPUl PsnuilUO3 - 5 xtpuaddv ools cz 9910 L'O cz LL L69*t 61*0 cz 11*0 0'0 ca t 6tg*S OL't 91 00*0 L*O cz 9t LIO*L OL'16 cz Lt'i 9,tt cz SEC 0*0 0 010 0 0*0 0 C*E$ tt 0*0 0 L*9 L 0*0 0 0*0 0 0*0 0 0*0 0 0*0 0 S*Lg tt L*g L 9*c6 SL 0*0 0 alto st 1 0*0 0 c,tt z 0*0 0 0*0 0 0'0 0 O*SL ZL CV) tn C'WL E*cl a 0'0 0 O*LL tt e*t6 SL 9 6*S 1 0*0 0 0*0 0 010 0 0*0 0 O*BL t*t6 9t 6,S t o*oot Lt 0*0 0 6*26 EL L*L 1 6'9 L 0*0 0 010 0 010 0 010 0 L*L L t*96 tL 6*5 L S*68 Lt 0*0 0 o*oot SL 0*0 0 L*L 0*0 0 0*0 0 0*0 0 9*16 zt L*L I 0,00t EL t'L 0*0 0 010 0 0*0 0 t*L t t*C6 wt 0*0 0 0,00t st L*9 t L'9 L 0*0 0 O*OOL SL 010 0 010 0 0*0 0 O*So Lt 9*1,t L*L 0'0 010 (%) suot3dio LIPIOI z 1 0 0 S-U-0-ri-Tios-al tolot 0*0 0*0 010 0*0 suotigiosid0491 0 0 0 0 -su-oj-lwjo-sa-H flel E'tt I'L 010 0'0 Suoilid.A3usas 2 I 0 0 SU0140joses tJR3 o*oot @t E*cs WL 6*z6 et oloot 9t tx)-A 3::-u-t3kjj3 lifluttowl uotl ILdol au;ioln 09L-bSt 61/5ws - C dnoig tsEE-1io ApnisAbotolliaIlts 0190*UtJalfPlURUgtJOA0UVOWPUR tenplAIPUl penuiluo-3S xlpueddv Maternal Number 21105 21106 27101 27108 21109 27110 27111 27112 27113 27114 27])S 27116 27111 27118 27119 271204 21121 27122 27123 21124 21125 27126 271214 27128 27129 Total N mean S.D. Ovarian 17 11 14 17 is 21 11 14 is Is 16 14 22 Not Pregnant is 11 is Is Is 16 16 is is Not Pregnant Not Pregnant 365 22 16.6 2.09 Appendix 5 - Continued Individual and Mean Ovarian and Uterine Data Rat Teratology Study of T-3351 Group-4 - 10 mg/kg Uterine Implantation I!WJARLSA 1-O1-ORRS, Efficiency 1%) 15 86.2 11 64.7 13 92.2 16 94.1 11 94.4 14 66.7 16 94.1 13 92.9 is 100.0 15 83.3 15 93.8 12 85.7 14 63.6 13 86.7 16 94.1 is 100.0 is 100.0 is 83.3 16 100.0 13 $1.3 is 93.3 13 86.7 Early Early ttiqrgti-ons Resorptions 0 0.0 1 9.1 3 23.1 1 6.3 1 5.9 0 0.0 1 6.3 1 7.7 2 13.3 2 13.3 0 0.0 1 8.3 0 0.0 13 100.0 0 0.0 3 20.0 0 0.0 1 6.3 0 0.0 1 6.7 Late Late tnailli-oni ResorDtions 0 0.0 0 0.0 0 0.0 0 0.0 is 88.2 0 0.0 0 0.0 0 0.0 1 6.7 0 0.0 0 0.0 0 0.0 1 7.1 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 317 22 14.4 1.50 22 07.72 10.255 31 20 1.6 2.86 20 11.32 21.929 17. 20 0.2 3.34 20 S.10 19.674 4FOwW dead on ky 17. therefore. fetal data not available. Tot amom 0 1 3 1 16 0 1 1 3 2 0 1 1 13 0 3 0 1 0 1 40 20 2.4 4.2 Appendix 6 Individualand Mean LitterDatea gat TeratologyStudy of T-3351 Group I - 0 mg/kg Maternal Number 27030 21031 27032 27033 27034 27035 27036 27031 27038 27039 27040 27041 27042 27043 21044 2704S 27046 27047 27048 27049 27050 270S] 27052 27053 270S4 Total N Unadjusted Mean Unadjusted S.D. Covartance Adjusted Mean Covariance Adjusted S.D. Total Resorptions Dead 0 0 1 0 1 0 3 0 1 0 1 0 0 0 Not Pregnant 2 0 4 0 1 0 0 0 1 0 2 0 1 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0. 0 0 23 24 1.0 1.04 0 24 0.0 - Number of Fetuses Fetal LqtL-Mb Live Live 0.0 5.9 6.7 20.0 6.7 5.6 0.0 is 100.0 16 94.1 14 93.3 12 80.0 14 93.3 17 94.4 14 100.0 14.3 23.6 7.7 0.0 6.7 28.6 6.3 0.0 0.0 8.3 6.7 5.6 0.0 15.4 0.0 0.0 0.0 12 OS.7 13 76.5 12 92.3 Is 100.0 14 23.3 5 71.4 is 23.8 10 100.0 13 100.0 11 91.7 14 23.3 17 24.4 14 100.0 11 $4.6 is 100.0 is 100.0 17 100.0 24 7.00 8.024 325 24 13.5 2.64 24 93.00 8.024 Sax M_alts Fe 7 a 6 7 7 7 5 5 9 12 5 7 7 5 7 6 7 7 5 4 11 4 10 1 4 12 3 3 7 5 8 5 6 6 8 9 a 6 a s 6 7 a 2 13 6 11 146 24 6.1 2.59 179 24 ?.S 2.30 Sex. percentules. and mean weight data Includevalues for live fetusesonly. 6 Percent fetal loss includesresorptionsas well as dead fetuses. C Statisticalanalysisperformedan covarianceadjustedmeans. Percent -KgLIes 46.7 50.0 50.0 50.3 35.7 70.6 50.0 41.7 46.2 58.3 26.7 28.6 20.0 00.0 30.0 38.5 45.5 42.9 S2.9 42.2 45.5 46.7 13.3 35.3 24 44.01 14.828 Appendix 6 - Continued Individualand Mean LitterDataa Rat TeratologyStudyof T-3351 Group 2 - I mg/kg Maternal -Cumber 27055 27056 27051 27058 .21059 21060 27061 27062 27063 27064 27065 27066 21067 21068 21069 27070 27071 27012 27073 27074 27015 27076 27071 21018 27079 Total R-elq[RUO-ns 9!@a-d 1 0 1 0 0 0 1 0 2 0 Not Pregfiant 0 0 0 0 1 0 1 0 0 0 1 0 2 0 1 '0 2 0 0 0 0. 0 1 0 0 0 1 0 0 1 0 0 0 I 0 Not Pregnant Number of fetuses Fetal Loss (%)b Live Live 7.1 92.3 6.3 93.8 0.0 13 100.0 6.1 14 93.3 14.3 12 85.1 0.0 11 100.0 0.0 13 100.0 1.1 12 92.3 6.1 14 93.3 O.o 16 100.0 14.3 6 OS.7 12.5 14 87.5 6.3 is 93.8 11.8 is 88.2 0.0 16 100.0 0.0 14 100.0 S.6 17 .94.4 0.0 16 100.0 6.3 is 93.8 6.7 14 93.3 7.7 12 0.0 13 100.0 6.1 14 93.3 Sex I-Lfae-s---F _e_-m_-e&lT- Percent -Hales -M Ma 4 1 9 4 a 6 5 1 66 7 3 53'3 3 69'2 3 57*1 3 41*1 3 7 4 a 5 3 9 6 a a 8 2 4 s 9 7 8 10 5 )5 1 7 7 9 a 6 10 s 10 e 6 10 2 5 6 63.6 3 61.S 2 25.0 3 42.9 3 $0.0 2. 33.3 3. 3S.7 2. 46.7 3. 66.7 3. 93.8 3. SO.0 3. S2.9 2. 37.S 2. 33.3 3. 57.1 3. 83.3 3. 38.S 3. 42.9 3. Total N Unadjusted Mean Unadjusted s.o. Covarionce Adjusted Haan Covariance AdjustedS.D. Is 23 0.8 0.61 0 313 23 23 0.0 S.S3 - 4.SN 23 13.6 2.27 23 94.48 4.802 165 23 7.2 2.66 a Sex. percentmales.and mean weightdata Includevaluesfor live fetusesonly. b Percentfetal lossincludesresorptionsas wellas dead fetuses. c Statisticalanalysisperformedon covarianceadjustedmeans. 148 23 6.4 2.4S 23 23 52.29 3. 16.688 0. 3. 0.0 Appendix 6 - Continued Individualand Mean LitterData& Rat Teratology Study of T-3351 Group 3 - 5 mg/kg Maternal Number. 27000 21081 21062 27083 21084 27085 21086 21081 21086 27089 21090 27091 2.7092 21093 27094 27095 27096 27097 27098 27099 27100 27101 21102 27103 27104 Total N Unadjusted Mean Unadjusted S.D. Covarionce Adjusted Mean Covariance Adjusted S.D. Number of fetuses Total Fetal Resorptions Dead Loss Live Live 0 0 0 0 1 0 2 0 Not Pregnant 0 0 1 0 0 0 1 0 1 0 0 0 0 0 1 0 1 0 0 0 1 0 3 0 2 0 2 0 0 0 0 0 1 1 0 0 0 0 Not Pregnant 0.0 0.0 7.1 14.3 0.0 6.7 0.0 1.1 7.7 0.0 0.0 5.9 7.1 o.6 5.9 18.8 13.3 14.3 0.0 0.0 13.3 0.0 0.0 16 100.0 13 100.0 13 92.9 12 85.7 17 100.0 14 93.3 Is 100.0 13 92.9 12 92.3 12 100.0 is 100.0 1& 94.1 13 92.9 13 100.0 16 94.1 13 81.3 13 86.7 12 85.7 12 100.0 16 100.0 13 86.7 14 100.0 14 100.0 17 23 0.7 0.86 1 23 0.0 0.21 23 5.28 6.028 317 23 13.8 1.57 23 24.72 6.018 Sex Eg]-et Females a a 0 5 a 5 3 9 5 12 7 1 11 4 6 7 9 3 0 4 9 6 12 4 2 4 1 6 8 8 6 7 7 6 6 6 6 6 10 6 5 a 6 a a 6 172 23 7.5 '2.04 145 23 6.3 2.01 Sex. perciat males. and mean weight data Includevalues for livefetusesonly. Percent fetal loss 1-icludersesorptionsas well as dead fetuses. c Stattsticalanalysis performe.odn covarianceadjusted mans. Percent Males 50.0 61.5 61.5 25.0 29.4 50.0 73.3 46.2 75.0 66.7 60.0 75.0 69.2 53.6 50.0 46.2 63.6 50.0 50.0 62.5 38.5 42.9 57.1 23 64.24 13.293 Appendix 6 - Continued individualand Kean LitterData& Rat Teratology Study of T-3351 Group 4 - 10 mg/kg Maternal Number Number of Fetuses Total Fetal Resorptions Dead Loss Live Live Sex M4]es Female 27105 27106 27101 27106 27109 27110 27111 27112 21113 27114 27115 21116 27117 27118 27119 27120 27121 27122 27123 27124 27126 27126 27127 27126 27129 lot&) N Unadjusted Kean 'Unadjusted S.O. Covariance Adjusted Kean Covariance Adjusted S.D. 0 1 3 1 16 0 1 1 3 2 0 1 1 Not Pregnant 13 Found Dead 0 3 0 1 0 1 found Dead Not Pregnant Not Pregnant 48 20 2.4 4.28 0 0 0 0 1 0 0 0 6 0 0 0 2 0 0 0 0 0 0 0 9 20 0.5 1.39 o.0 9.1 23.1 6.3 100.0 0.0 6.3 7.7 60.0 13.3 0.0 8.3 21.4 100.0 0.0 20.0 0.0 b.3 0.0 6.7 20 12.43 30.771 is 100.0 10 90.9 10 16.9 15 93.0 0 0.0 14 100.0 Is 93.8 12 92.3 6 40.0 13 86.7 is 100.0 11 21.7 ]I 78.6 0 0.0 is 100.0 12 80.0 is )00.0 is 93.9 13 100.0 14 93.3 231 20 11.6 4.61 120 80.52 30.601 10 5 6 4 4 6 9 6 a 6 7 8 10 2 4 2 6 7 9 6 a 3 8 3 9 6 6 6 10 5 9 6 7 6 9 5 139 Is 7.7 1.90 92 le 5.1 1.68 Sex. percent ules. and man weightdata Includevaluesfor live fetusesonly. ba Percentfetal lossincludesresorptionsas well as dead fetuses. c Statisticalanalysisperforwd on covarianceadjustedmans. Percent Males 66.7 60.0 40.0 60.0 S7.1 46.7 03.3 66.7 46.2 60.0 72.7 72.7 60.0 50.0 66.7 60.0 53.8 64.3 18 60.38. 10.655 Appendix I IndividualVi:clralFindingsin Fetuses Rat Tora o ogy Study of T-3351 Group I - 0 MoAg Haternal Number: Number of fetuses examined Number of fetuseswithin normal limits Number of fetuseswith one or more visceralvariants Percentof f*tuseswith on* or more skeletalvariants Identificatiolnetter(s)of fetuseswith: Brain - cerebral hemispherecontaineddark area Eye(s)- containeddark mterial Liver - dark areas Kidney(s)- pelvisdilated Ureter(&)- distended - undulated Number of fetuses with-one or more visceralanomalies Percentof fetuses with one or more visceral anowlies (%) Identificatiolnetter(s)of fetuseswith:. Skin - edematous Palate- cleft Cryptorchism 2 2 2 2 2 2 2 7 7 7 7 7 7 7 7 0 0 0 0 0 0 0 0 3 3 3 3 3 3 3 3 0 1 2 3 4 5 6 7 5 6 5 4- 5 6 5 NP 5 6 5 3 5 6 5 0 0 0 1 0 0 0 0.00 0.00 0.00 25.00 0.00 0.00 0.00 I 0 0 0 0 0 0 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 2 7 0 3 8 4 4 0 0.00 0 0 0.00 0 up Not Pregnant.. Appendix 7 - Continued IndividualViscera]Findings in Fetuses Rat Teratology Study of T-3351 Group I - 0 mg/kg Maternal Number: Number of fetuses examined Number of fetuseswithin normal limits Number of fetuseswith one or more visceral variants Percentof fetuseswith one or more skeletalvariants Identificatiolnetter(s)of fetuseswith: Brain cerebral hemispherecontained dark area Eye(s) containeddark material Liver - dark areas Kidney(s)- pelvisdilated Ur*ter(s)- distended - undulated Number of fetuses with one or more visceralanomalies Percentof fetuseswith one or more visceral anomalies (%) Identi'ficatiloentter(s)of fetuseswith: Skin - edematous Palate- cleft Cryptorchiss 2 2 2 2 2 2 2 2 2 7 7 7 7 7 7 7 7 7 0 0 0 0 0 0 0 0 0 4 4 4 4* 4 4 4 5 5 3 5 6 7 a 9 o I 2 5 4 5 4 5 6 S 4 2 4 3 5 3 6 6 6 4 0 1 1 0 1 0 0 0 0 0.00 20.00 25.00 0.00 25.00 0.00 0.00 0.00 0.00 A i K 0 0 0 0 0 0 0 0 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Appendix 7 - Continued IndividualVisceralFindingsIn fetuses Rat Toratology Study of T-3351 Group 2 - I mg/kg Haternal Number: Number of fetuses examined Numberof fetuseswithinnorml limits Numberof fetuseswith on* or more visceralvariants Percentof fetuseswith one or wre skeletalvariants(%) Identificatiolnetter(s)of fetuseswith: Brain - cerebralhmisphere containeddark area (ye(s)- containeddarkmterial Liver - dark areas Kidney(s)- pelvisdilated Ureter(s)- distended - undulated Nwjber of fetuseswith one or more visceral anomalies Percentof fetuseswith one or more visceral anomalies (%) Identificatiolnetter(s)of fetuseswith: Skin - edmtaus Palate- cleft Cryptombism 2 2 2 2 2 2 7 7 7 7 7 7 0 0 0 0 0 0 5 5 5 5 5 6 5 7 9 0 4 5 4 5 4 NP 4 5 4 5 4 0 0 0 0 0 0.00 0.00 0.00 0.00 0.00 0 0 0 0 0 0.06 o.oo o.oo o.oo o.oo 2 7 0 6 1 4 4 0 0.00 2 2 7 7 0 0 6 6 2 3 8 4 a 3 0 1 0.00 25.00 A 0 0 0 0.00 0.00 0.00 NP Not Pregnant. Appendix 7 - Continued IndividualVisceralFindingsin Fetuses Rat TerstologyStudy of T-3351 Group 2 - I mg/kg Maternal Number: Number of fetuses examined Number of fetuseswithin normal limits Number of fetuseswith one or more visceral variants Percentof fetuseswith one or more skeletalvariants Identificatiolnetter(s)of fetuseswith: Brain - cerebral he@ispherecontained dark area Eye(s) - containeddark material Liver - dark areas Kidney(s)- pelvisdilated Urater(s)- distendlp - undulatit Number of fetuseswith one or mrs visceral anomalies Percentof fetvs*swith one or more visceral anomalies(S) identificatiolnette- r(s)Of fetusesWith: Skin - edematous Palate- cleft Cryptorchism 2 7 0 68 5 5 0 0.00 2 2 7 7 0 0 6 7 9 0 5 6 5 5 0 1 0.00 16.67 2 2 7 7 0 0 7 7 1 2 5 6 5 4 0 2 0.00 33.33 2 2 2 2 1 7 7 7 0 0 0 0 7 7 7 7 3 4 6 6 s 5 4 6 4 4 3 0 1 1 1 0.00 20.00 20.00 25.00 2 Q A A G A N A 6 A 0 0 0 0 1 0 0 0 0 0.00 0.00 0.00 0.00 16.67 0.00 .0.00 0.00 0.00 N NP Not Pregnant. Appendix 7 - Continued 'IndividuaVlisceralFindingsin fetuses Rat Teratology Study of T-3351 Group 3 - 5 mg/kg Maternal Number: Number of fetuses examined Number of fetuseswithin nonul limits Number of fetuseswith one or more visceral variants Percentof fetuseswith one or more skeletalvariants Identificatiolnetter(s)of fetuseswith: 2 7 0 6 0 6 4 2 33.33 2 2 2 2 7 7 7 7 0 0 0 0 a 8 a a 1 2 3 4 5 5 4 NP s 4 4 0 1 0 0.00 20.00 0.00 Brain - cerebral beimispherecontaineddark area Eye(s)- containeddark mterial Liver - dark areas Kidn*y(s)- pelvisdilated Ureter(s)- distended - undulated 3 A N Number of fetuseswith one or more visceralanomiles Percentof fetuseswith one or more visceral anomalies (%) Identificatiolnatter(s)of fetuseswith: Skin - edematous palate- cleft Cryptarchism 0 0 0 0 0.00 0.00 0.00 0.00 2 2 2 2 7 7 7 7 0 0 0 0 a a a 8 5 6 7 9 6 5 5 5 6 5 5 6 0 0 0 0 0.00 0.00 0.00 0.00 2 A 0 0 0 0 0.00 0.00 0.00 0.00 MP Not Pregnant. Appendix 7 - Continued IndividualVisceral Findings In Fetuses Rat Teratology Study of T-3351 Group 3 - 5 mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more visceral variants Percent of fetuses with one or more skeletal variants (9) Identificationletter(%)of fetuseswith: Brain - cerebral hemisphere contained Eye(s) - contained dark material Liver - dark areas Kidney(s)- pelvisdilated Ureter(s) - distended - undulated dark area Number of fetuses with one or more visceral anomalies Percent of fetuses with one or more.visceral Id:nnotimflicia*sti(o%n)loefttfeert(uss)es with: Skin - edematous Palate - cleft Cryptorchism 2 2 2 2 2 2 2 7 7 7 7 7 7 7 7 7 7 0 0 0 0 0 0 0 1 1 1 9 9 9 9 9' 9 9 0 0 0 3 4 5 6 8 9 0 1 2 5 5 6 5 5 4 4 6 5 5 5 5 6 5 5 4 4 4 5 5 0 0 0 0 0 0 0 0 0 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.0 0 0 0 0 0 0 0 2 0 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 33.33 0.00 0.0 A AG NP Not Pregnant. Appendix 7 - Continued Individual Visceral Findings in Fetuses Rat Teratology Study of T-3351 Group 4 10 mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fet-useswith one or more visceral variants Percent of fetuses with one or more skeletal variants (%) Identificationletter(s)of fetuseswith: 2 2 2 2 2 7 7 7 7 7 I I I I I 0 0 0 0 0 5 6 7 a 9 a 5 4 4 5 0 4 4 4 5 1 0 0 0 20.00 0.00 0.00 0.00 Brain - cerebral hemisphere contained dark area Eye(s) - containeddark material m Liver - dark areas Kidney(s)- pelvisdilated Ureter(s) - distended - undulated Number of fetuses with one or more visceral anomalies Percent of fetuses with one or more visceral- anomalies (%) Identificationletter(s)of fetuseswith: 0 0 0.00 - 0.00 0 0.00 0 0.00 Skin - edematous Palate - cleft Cryptorchism 2 2 2 2 2 7 7 7 7 7 I I I I I 1 1 1 1 1 0 1 2 4 b c d 5 5 4 2 5 5 5 3 0 0 0 0 0 0 0 0.00 0.00 0.00 0.00 0.0 0 0.00 0 1 2 5 0.00 25.00 100.00 loo.ot BE ADGJN A BE JN a All fetuses resorbed and/or dead. b fetus E was noted to have cleft palate during external examination. c All fetuses were noted to have edematous skin during external examination. Fetuses 6 and E were confirmed at visceral examination. noted to have cleft palate during external examination. fetus E was confirmed at visceral examination. d fetuses H and N were noted to have edematous skin during external examination. FettfsN was confirmed at visceral examination. Fetu to have cleft palate during external examination. e fetus E was noted to have cleft palate during external examination. f All fetuses were noted to have edematous skin duri external examination. Fetuses A. E, 3, and H were confirmed at visceral exami and L were noted to have cleft palate during exterInnal examination. Fetuses A and J were confirmed at visceral examination.. Appendix 7 - Continued IndividualVisceral Findings in Fetuses Rat Teratology Study of T-3351 Group 4 10 mg/kg 2 Maternal 1 Number: 8 Number of fetuses examined NP Number of fetuses within normal limits Number of fetuses with one or more visceral variants Percent of fetuses with one or more skeletal variants Identificationlatter(s)of fetuseswith: Brain - cerebral hemisphere contained Eye(s) - contained dark material Liver - dark areas Kidney(s)- pelvis dilated Ureter(s)--distended - undulated dark area Number of fetuses with one or more visceral anomalies Percent of fetuses with one or more visceral Anomalies (%) Identificationletter(s)of fetuseswith: Skin - edematous Palate - cleft Cryptorchiss 2 2 7 I 1 2 9 0 0a FO 2 7 I 2 1 54 0 0.00 2 2 7 7 I I 2 2 2 3 4 3 0 0.00 5 s 0 0.00 2 7 I 2 4 5 5 0 0.00 2 7 I 2 s b s 0 0 0.00 2 2 7 7 I I 2 2 6 7 5 FD 0 0 0.00 I 1 0 20.00 2S.00 0.00 0 5 5 0.00 100.00 100.00 AD@JM BENKU E Dim ENN G BHK NP - Not Pregnant. FO - Found Dead. a All fetusts resorbed and/or dead. b All fetuses were noted to have edematous skin during external examination. Fetuses A. D. G, J. and N were confirmed at F. J. and L were noted to have cleft palate during external examination. Fetus 3 was confirmed at visceral examination. visceral i Appendix a Individual Skeletal Findings in Fetusesa Rat Teratology Study of T-3351 6roup I - 0 mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more skeletal variants Percent of fetuses with one or more skeletal variants (%) Identificationletter(s)of fetuseswith: 2 2 2 2 2 2 2 2 7 7 7 7 7 7 7 7 0 0 0 0 0 0 0 0 3 3 3 3 3 3 3 3 0 1 2 3 4 6 7 10 5 5 50.00 10 6 4 40.00 9 6 3 33.33 a 7 1 12.50 9 4 5 55.56 11 a 3 27.27 9 'NP 7 2 22.22 2 2 7 7 0 0 3 3 a 8 5 8 3 0 37.50 0. Skull - incomplete closure (<75%) - tnterparietalossification incomplete/nonossifted Di i FH L JN - parietalsossificationIrregular - supraoccipital ossification incomplete/nonossifted - supraoccipital nonfused i N FK - hyoid ossification incomplete/nonossified Pectoral girdle - scapula ossification Incomplete .Rib cage - sternebrao bipartite FN 61 Di FIK H B i - vternebrac I and 2 fused -@sternebrae <3 osstflid EF - ribs <13 ossiftedb - ribs >13 ossified(cervicalregion) -- ribs 13th pair smallb CFHI ribs ossificationIrregular Vertebral column - socral arches nonosstfied - thoracic centra ossificationincomplete/ nonossified - thoracic centra nonrused - caudals <3 ossified E K B OF E Pelvic girdle - pubis ossificationincomplete/ nonossified FIK - ischium nonossified F Foreliabs - <3 ossified Rtndlimbs - <4 ossified E lotALigment Ribs - angulated Sternebrae - checkered NP - Not Pregnant. a There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. Appendix 8 - Continued IndividualSkeletal Findings in fetusesa Rat Teratology Study of T-3351 Group I - 0-mg/kg Maternal Number: Number of fetuses examined Number of fetuseswithin normal limits Number of fetuseswith one or more skeletal variants Percent of fetuses with one or more skeletal variants Identificationletter(s)of fetuseswith: Lagaina Ossification Skull - incomplete closure (<75%) - lnterparietalossificationincomplete/nonossified - parietalsossificationirregular - supraoccipitalossificationincomplete/nonossified - supraoccipital nonfused - hyoid ossification lncoinplete/nonossified Pectoral girdle - scapula ossificationincomplete Rib cage - sternebrae bipartite - sternebrae I and 2 fused sternebrae <3 ossified ribs <13 ossifiedb ribs >13 ossified (cervicalregion) ribs 13th pair somilb ribs ossificationirregular Vertebral column - sacral arches nonossified - thoracic centra ossification incomplete/ nonossified -.thoracic centra nonfused caudals <3 ossified Pelvic girdle - pubis ossificationIncomplete/ nonossifted * - 4schium nonossified Foreliabs - <3 ossified Hindliabs - <4 ossified Done Alignment Ribs - angulated Sternebrae - checkered There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. 2 7 0 4 3 3 I 2 66.67 2 7 0 4 4 10 a 2 20.00 2 2 2 2 2 7 I. 7 7 7 0 0 0 0 0 4 4 4 4 4 5 6 1 a 9 6 5 1 16.67 a 7 1 12.SO 7 3 4 57.14 9 s 4 44.44 11 7 4 36.36 2 7 0 5 0 9 a 1 11.11 2 7 0 5 1 7 6 1 14.29 2' 7. 0 5 2 10 9 1 10.0 D R DE K c L F IL 8 D i F I ILR R BCFL rii u F 0 R 3 R R Appendix 8 - Continued IndividualSkeletal findings in Fetusesa Rat Teratology Study of T-3351 Group 2 - I mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more skeletal variants Percent of fetuses with one or more skeletal variants (%) Identificationletter(s)of fetuseswith: taggina Ossification Skull incomplete closure (<75%) interparietalossification incomplete/nonossified parietals ossificationirregular supraoccipital ossification Incomplete/nonossified supraoccipital nonfused hyold ossification incompl*te/nonossified Pectoral girdle - scapula ossification incomplete Rib cage - sternebrae bipartite - sternebrae I and 2 fused - sternebrae <3 ossified - ribs <13 ossifiedb ribs >13 ossified (cervical region) ribs 13th pair smilb ribs ossificationirregular Vertebral column - sa.cralarches nonossified - thoracic centra ossificat4onincomplete/ nonossifW - thoracic centra nonfused ciudals <3 ossified Pelvic girdle - pu-bis ossificationIncomplete/ nonossified - lschim nonossified Foreliabs - <3 ossified Hindliabs - <4 ossified Done Alignment Ribs - angulated Sternebrae - checkered 2 7 0 5 5 8 6 2 25.00 2 2 7 7 0 0 5 5 6 7 10 9 1 10.00 9 7 2 22.22 2 2 2 7 1 7 0 0 0 5 -5 6 8 9 0 9 a NP 5 1 4 7 44.44 87.50 p p m p I 8 BEGHJK BEGJK BGHJ BEGHJK I F BFRO K i 2 2 2 2 7 7 7 7 0 0 0 0 6 6 6 6 1 3 4 7 2 5 71.43 5 a 4 0 1 8 20.00 100.00 9 6 3 33.33 E CEHLM EFK CE BCEFH SCEFHI m CE L BCEHK c 0 Cc" NP Not Pregnant. a There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. Appendix 8 - Continued Individual Skeletal Findings in Fetusesa Rat Teratology Study of T-3351 Group 2 - I nkg/kg Maternal Number: Number of fetuses examined Number of fetuseswithin norml limits' Number of fetuses with one or more skeletal variants Percent of fetuses with one or more skeletalvariants (%) Identificatiolnetter(s)of fetuseswith: Lagging Ossification Skull.- incomplete closure (<75%) - lnterparietal ossification incomplete/nonossilfed - parietalsossificationirregular - supraoccipitalossification Incomplete/nonossified - supraoccipital nonfused - hyoid ossification Incomplete/nonossified Pectoral girdle - scapula ossification incomplete Rib cage sternebrae bipartite sternebrae I and 2 fused sternebrae <3 ossified ribs <13 ossifiedb ribs >13 ossified (cervicalregion) ribs 13th pair umilb ribs ossificationIrregular Vertebral column - sacral arches nonossified - thoracic centra ossificationincomplete/ nonossified - thoracic centra nonfused - caudals <3 ossified Pelvicgirdle - pubis ossificationincomplete/ nonossified - 1schium nonossifted forellobs - <3 ossified Hinditabs - <4 ossified Done Alionmat Ribs - angulated Sternebrae - checkered 2 7 0 fi 8 10 8 2 20.00 2 7 0 6 9 10 9 1 10.00 2 2 2 7 7 7 0 0 0 7 7 7 0 1 2 10 7 3 30.00 9 11 7 0 2 11 22.22 100.00 2 7 0 7 3 10 8 2 20.00 2 2 2 2 7 7 7 7 0 0 0 0 7 7 7 1 4 5 6 7 10 9 a 0 2 2 20.00 100.00 a 3 5 62-50 a 6 2 25.0 CEFHIK L BECFHX SCEFHJ IL L LNR LMO CEFIKL H BCEFKL N F FJMO IL L MR I SHI H ECEFLO p CFHJLN EIL EL R R F 0 c I L f H 8 E3 HR F 0 R NP - Not Pregnant. a There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. Appendix 8 - Continued IndividualSkeletal Findings in Fetusesa Rat Teratology Study of T-3351 Group 3 5 mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more skeletal variants Percent of fetuses with one or more skeletal variants Identificationletter(s)of fetuseswith: 2 2 2 2 2 7 7 7 7 7 0 0 0 0 0 a a a 8 a 0 2 3 4 10 a 8 NP a 5 7 7 2 3 1 1 20.00 37.50 12.50 12.50 LLUIM-OlLif-ic_Ltjon Skull - incomplete closure (<75%) - lnterparletalossificationincomplete/nonossified - parietalsossificationirregular a H - supraoccipital ossif.1cationlncoinplete/nonossified - supraoccipital nonfused - hyold ossifit tion Incomplete/nonossitie4 8 Pectoral girdle - scaaptilaossification incomplete Rib cage - sternebrae bipartite - sternebras I and 2 fused - sternebrae <3 ossified - ribs <13 ossifiedb ribs >13 ossified(cervicalregion) i ribs 13th pair *aallb ribs ossificationirregular Vertebral column - sacral arches nonossified - thoraciccentra ossificationincomplete/ nonossified - thoracic centra nonfused I CH 0 caudals <3 ossified Pelvic girdle - pu-bis ossificationincomplete/ nonossified - ischium nonossified forelimbs - <3 ossified Hindlimbs - <4 ossified Done Alignment Ribs - ingulated Sternebrae -.checkered 2 2 2 7 7 7 0 0 0 8 a a 5 6 7 11 a 3 27.27 9 I 2 22.22 10 I 9 90.00 BE N 0 B. H F I E BEFHLO B HK INO BCF B 2 2 7 7 0 0 a 8 a 7 0 1 0.00 12.5 c RP - Not Pregnant. a There were no skeletal anomalies noted In any fetuses. b Possible preparation damage at time of evisceration. Appendix 8 - Continued IndividualSkeletal Findings in Fetusesa Rat Teratology Study of T-3351 Group 3 - 5 mg/kg Maternal Number: Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more skeletal variants Percent of fetuseswith one or more skeletal variants Identificationletter(s)of fetuseswith: Lag-gingOssification Skull - incomplete closure (<75%) - interparietalossification incomplete/nonossified - parietalsossificationirregular - supraoccipitalossification incomplete/nonossified - tupraocci,pita-Inonfused - hyoid oss fication iscomplete/nonossified Pectoral girdle - scapula osstfication incomplete Rib cage - sternebrae bipartite - sternebrae I and 2 fused -.sternebrae<3 ossified - ribs <13 ossifiedb - ribs >13 ossified(cervicalregion) - ribs 13th pair smallb - ribs ossificationIrregular Vertebral column - sacral arches nonossified - thoracic centra ossification Incomplete/ nonossified - thoracic centra nonrused - caudals <3 ossified Pelvic girdle - pubis ossificationincomplete/ nonossified - ischium nonostified Foreliabs - <3 ossified Hindlimbs -.<4 ossified Done Aiiqmwnt Ribs - angulated Sternebrae - checkered I NP - Not Pregnant. a There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. 2 7 0 9 3 8 3 5 62.50 NK CEI K IK 2 7 0 9 4 8 8 0 0.00 2 2 2 7 7 7 0 0 0 9 9 9 5 6 7 10 7 3 30.00 8 3 5 62.50 8 5 3 31.50 2 7 0 9 a 0 0.00 2 2 7 7 0 1 9 0 0 a 10 4 0 4 10 50.00 100.00 Fl I 0 JN OG N El BCEFHI KLNO 0 L BCEFHI KLNO HL LN 8EFIK BLN LM EK ILN 8CEFHI KLNO BILN LN LN BEIKL 2 2 7 7 1 1 0 0 1 2 8 9 8 7 0 2 0.00 22. N N L N Appendix 8 Continued IndividualSkeletalfindings In Fetuiesa Rat Teratology Study of T-3351 Group 4 - 10 mg/kg 2 7 1 Maternal 0 Number: 5 Number of fetuses examined Number of fetuses within normal limits Number of fetuses with one or more skeletal variants Percent of fetuses with one or more skeletal variants Identificationletter(s)of fetuseswith: taggina Ossification Skull - incomplete closure (<7S%) - interparietalossificationincomplete/nonossified - parietals ossificationirregular - supraoccipitalossificationincomplete/nonossified - supraoccipital nonfused - hyoid ossification Incomplete/nonossified Pectoralgirdle - scapula ossification incomplete Rib cage - sternebrae bipartite - sternebrac I and 2 fused - sternebrae <3 ossified - ribs <13 assifiedb - ribs >13 ossified(cervicalregion) - ribs 13th pair swilb - ribs ossificationIrregular Vertebral c6lumn - sacral arches nonossified - thoracic tentra ossificationincomplete/ nonossified - thoracic centre nonfused - caudals <3 ossified Pelvic girdle - pubis ossificationincomplete/ nonossified - ischium nonassified Forelimbs - <3 ossified Hindlimbs - <4 ossified 10 3 7 70.00 CFHILN 0 Igne Alionwnt Ribs 7 angulated HN Sternebrae - checkered WP - Wat Pregnant. a There were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. C All fetuses resorbed and/or OeAd 2 2 2 7 7 7 1 1 1 0 0 0 6 7 8 6 6 0 0.00 .6 2 4 66.67 10 10 0 0.00 CFI K 2 2 2 2 2 2 7 7 7 7 7 7 1 1 1 1 1 1 0 1 1 1 1 1 9 0 1 2 3 4 c 0 9 10 0 4 a 3 7 0 0 0 6 3 a 4 a 66.67 30.00 100.00 100.00 100.0 8 CH BEFHX BCEFHI CDFO KL CE COFO BCEFH Lm EFHI BFHK COO CEIL F H E BCFHIK CDO CH BCEFH LM BCEFH L CH FO I K CH FO el BH K BCEHIK COFO CEFI CH c BCEFHK CDFO CEHI L Ef COFO 0 Appendix 8 - Continued Individual Skeletal Findings in Fetusesa Rat Teratology Study of T-3351 Group 4 10 mg/kg 2 7 1 Maternal 1 Number: a Number of fetuses examined NP Number of fetuses within normal limits Number of fetuses with on* or more skeletal variants Percent of fetuses with one or more skeletal variants Identification)otter(s)of fetuseswith: Lagging ossification Skull - Incomplete closure (<7S%) - interparietalossification incomplete/nonossified - parietals ossificationirregular - supraoccipitalossification incomplete/nonossified - supraoccipital nonfused - hyoid ossification incomplete/nonossified Pectoralgird)* - scapula ossification incomplete Rib cage - sternebrae bipartite - sternebrae I and 2 fused - sternebrae <3 ossified ribs <13 ossifiedb ribs >13 ossified (cervicalregion) ribs 13th pair sg&llb ribs ossificationirregular Vertebral column - sacral arches nonossified - thoraciccentra ossificationincomplete/ nonossified - thoracic contra nonfused - caudals <3 ossified Pelvic girdle - pubis ossificationincomplete/ nonossified - ischium nonossified forelimbs - <3 ossified Hindlimbs - <4 ossified Bone Alignment Ribs - angulated Sternebrae - checkered NP - Not Pregnant. FO - Found Dead. a Yhere were no skeletal anomalies noted in any fetuses. b Possible preparation damage at time of evisceration. C All fetuses resorbed and/or dead. 2 2 7 7 1 1 1 2 9 0 c 0 FO 2 2 2 2 2 2 2 7 7 7 7 7 7 7 1 1 1 1 1 1 1 2 2 2 2 2 2 2 1 2 3 4 5 6 7 10 8 10 10 8 9 Fo 4 5 10 3 0 0 6 3 0 7 a 9 60.00 37.50 0.00 70.00 100.00 100.00 CIKO N CF FIKO N 8 BCEFHI CDFIJL KL No CJHP BFIKL I BCMP NK CIJLNO OCEFIL K BCEFHI L CKL CDFGIJ LNO . a CKL c BNP L IL el OCEFHI CJL KL BCHIKL KL KL 8 BCEFHI CIJLO KL 75 Appendix 9 References Rat Teratology Study of T-3351 STATISTICAL METHOOS/LABORATORY PROCEDURES 154-160 Armitage, P., "Tests for Linear Trends in Proportions and Frequencies.0 Biometrics, 11:375-385, 1955. Bartlett, M. S., *Some Examples of Statistical Methods of Research in Agriculture and Applied Biology." J. Royal Statist. Soc. Suppl., IV:137-170, 1937. Cochran, W. G., "Some Methods of Strengthening the Common Chi-Square Tests.U Biometrics, 10:417-451, 1954. Cochran, W. G., and Cox. G. M., Experimental Designs, 2nd Ed., John Wiley and Sons, Inc., New York, 1957. Draper, N. R., and Hunter, W. G., utransformations: Some Examples Revisited." Technometrics, 11:23-40, 1969. Ounnett, C . W., "A Multiple "Comparison Procedure for Comparing Several Treatments with a Control.0 J. Am. Stat. Assoc., 50:1096-1121, 1955. Ounnett, C. W., "New Tables for Multiple Comparisons with a Control.$ Biometrics, 20:482-491, 1964. Fisher, R. A., Statistical Methods for Research Workers, Oliver & Boyd, Edinburgh, 1950. Fleiss, J. L., Statistical Methods for Rates and-Proportions, 2nd Ed., Wiley, New York, Ch. 9, 1981. Jonckheere, A. R., 'A Distribution-free k-sample Alteratives." Biometrika 41:133-45, 1954. Test Against Ordered Kruskal, W. H. and Wallis, W. A., "Use of Ranks in One-Criterion Variance Analysis." J. Am. Stat. Assoc., 47:583-621, 1952. Kruskal, W. H. and Wallis, W. A., "Errata to Use of Ranks in One-Criterion Variance Analysis." J. Am. Stat. Assoc., 48:907-911, 1953. Levene, H., "Robust Tests for Equality of Variances," in I. Olkin edited, Contributions to Probability and Statistics, Stanford University Press, Palo Alto, 1960. Miller, R. G., Jr., Simultaneous Statistical Inference, McGraw-Hill, New York, 1966, Ch. 6.1. 76 Appendix 9 - Continued References Rat Teratology Study of T-3351 STATISTICAL METHODS/LABORATORY PROCEDURES 154-160 Staples, R. E.. 00etection of Visceral Alterations in Mammalian Fetuses." Teratology, 9(3):A-37, 1974. Staples, R. E. and Schnell, V. L..,"Refinements in Rapid Clearing Technic in the KOH-Alizarin Red S Method for Fetal Bone." Stain Technology, 39:61-63, 1964. Terpstra, T. J., mthe Asymptotic Normality and Consistency of Kendall's Test Against Trend, When Ties are Present in One Ranking.' Indag. Math. 14:327-333, 1952. Wilson, J. G. and Warkany, J., editors. Teratology: Principles and Techniques, The Univ. of Chicago Press, Chicago, 1965, pp. 251-277. Winer, B. J., Statistical Principles in Experimental Design, 2nd Ed., McGraw-Hill, N.Y., Chs. 3-9, 1971. Winer, B. J., Statistical Principles in Experimental Design. 2nd Ed., McGraw-Hill, N.Y., Ch. 10, 1971.