Document 7ORRNqeJ22z0j4MmeRLkD03jg

(OHAZLEOCC3NLABORATAOMRERIICEA,SINC.. 9200 LEESBURG TURNF31KE, VIENNA. VIRGINIA --2180, U.S.A. May 13, 1983 William C. McCormick, III Toxicologist Toxicology Services Medical Department/3M 220-2E 3M Center St. Paul, Minnesota 55101 CEI "IA-Y 6 83 Dear Bill: Enclosed are four copies of the final report for Project No. 154-158, Pilot Rat Teratology Study with T-3351. With copy of this letter, I am forwarding one copy of the report to Marshall Johnson. If you have any comments or questions, please contact me. Best regards. Sincerely, LTW:sm Enclosure cc: E. Marshall Johnson LAWWRREENNCCEE T. ETEKTZZEELL, Ph.D. Staff Scientist Department of Toxicology PHONE (703) 8S3-5400. TELEX 899436 CHAZLABS VINA), CABLE HAZLABS VVASH OC "a'HAZLL'tCNLABOPAAMTEFO41RC&IINEC.S fooo L .... I.. TLI..11... PILOTTERATOLOSGTYUDIYNRATS T-35514@000 FINARLEPORT Submittteod 3MCompany StP.aulM,innesota May 13, 1983 '@ahHAZLEMONLA.BORATORIES AMERICA, INC. SUBJECT: Pilot Teratology Study in RatS Project No. 154-158 We, the undersigned, hereby declare that the work was performed under our supervision, according to the procedures herein described. Study Director: W*R E NCE*. W4L-,-07. D-. Staff Scientist Department of Toxicology Laboratory Supervision: Teratology: AB.@KBU, K, *PP DD.. Rodent Toxicology I Department of Toxicology RAJTHS. [MtM,-T@T Research Associate Department of Toxicology Report Preparation- L4A Group Leader Scientific Resources Department tlh ""@HAZL=CNLABC)RATOiqlES AMERICA,INC. vioulowfa *also. Lu@@ TABLE OF CONTENTS Page SUMMARY 1 INTRODUCTION 3 CONTROL AND TEST MATERIALS 3 TEST ANIMALS 4 METHODS 4 Mating Period 4 Groups and Dosage Levels 5 Compound Preparation and Administration 5 Maternal Observations and Records 6 Cesarean Sacrifice and Tissue Preservation 7 StatisticalAnalyses 8 Specimen, Raw Data, and Final Report Storage 9 RESULTS - MATERNAL DATA 10 Clinical Signs 10 Mortality 10 Body Weights 10 Food Consumption 11 Gross Pathology 11 Uterine Weights 11 RESULTS - CESAREAN DATA 12 Pregnancy Rates, Corpora Lutea, Uterine Implantations, and ImplantationEfficiency 12 Fetal Viability,Weight, and Sex 12 Fetal Development 12 TABLES Table 1 - Clinical Observations During Gestation 14 Table 2 - Mean Maternal Body Weights and Body Weight Change 15 Table 3 - Mean Food Consumption Values 16 Table 4 - Summary of Gross Pathology Findings 17 Table 5 - Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights 18 Key to Table 6 19 Table 6 - Sumary of Ovarian, Uterine, and Litter Data 20 Table 7 - Mean Incidence Values for Visceral and Skeletal Findings 21 Key to Table 8 23 Table 8 Mean Incidence of Visceral and Skeletal Findings per Litter 24 "'-'@'HAZLETONLASOFAIMEARITCAO,IRNCI. ES TABLE OF CONTENTS CONTINUED Page APPENDICES Key to Appendix 1 25 Appendix 1 - Individual and Mean Maternal Body Weights 26 Appendix 2 - Individual and Mean Food Consumption Values 28 Appendix 3 - Individual Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights 31 Appendix 4 - Individual and Mean Ovarian, Uterine, and Litter Data 34 Appendix 5 - Individual and Mean Live Fetal Data 36 Appendix 6 - IndividualVisceral Findingsfor Each Litter 38 Appendix 7 - IndividualSkeletalFindingsfor Each Litter 39 Appendix 8 - References 42 (OHAZL=CNLABORATORIES 9200 LEESBURG TURNPIKE. VIENNA, VIRGINIA AMERICA,INC. 22180. U.S.A. SPONSOR: MATERIAL: SUBJECT: 3M Company T-3551 FINAL REPORT Pilot Teratology Study in Rats Project No. 154-158 DATE: May 13, 1983 SUMMARY T-3351, suspended in corn oil at concentrationsintended to deliver 1, 5, 10, or 20 mg/kg/day (Groups 2, 3, 4, and 5, respectively), was administeredby oral intubation to four groups of mated female Sprague-Dawley SOO rats on gestation Days 6 through 15. A fifth group (Group 1) of mated females served as the control group and received the vehicle only. No Group 5 females survived to cesarean section on gestation Day 20. Treatment-relatedclinical observationsin Groups 4 and 5 included hunched and thin appearance of the animals. In addition, tremors and convulsions as well as prostration and anorexia were noted in Group 5 animals. Body weight gain of the dams during gestation was less in Groups 3, 4, and 5 compared to control, with a significantreductionin weight gain occurring in Groups 4 and 5. A significantdecrease in food consumption occurred in Groups 4 and 5. Apparent treatment-related ulcerated areas on the glandular and/or nonglandular stomach mucosa were found during necropsy of Group 5 females. ""@HAZLEMONLABORATOAMREIRIECAS.INC. 2 154-158 Analysis of individual and mean uterine data demonstrated no effect of T-3351 on fetal mortalityor fetal viability. Mean fetal weight was comparablefor either sex in all groups. External, visceral, and skeletal abnormalitieswere generally similarfor all groups. One Group 3 fetus was found to have a small eye upon external examination. Another Group 3 fetus had fused hind limbs and anal atresia. Skeletal examination confirmed the external limb anomaly. "'-'HAZLEMONLAWRATORIES AMERICA,INC. 3 154-158 INTRODUCTION This study was designed to determine the maternal and embryo/ fetal toxicity of T-3551 when administered by gavage to pregnant rats during the period of fetal organogenesis for the purpose of setting dosage levels for an expanded teratology study. The rats were placed in breeding on January 11, 1983, and cesarean sections were completed on February 3, 1983. This report presents the methods and results from this study. CONTROL AND TEST MATERIALS The vehicle and control material, Dukels'M Corn Oil (C. F. Sauer Co., Richmond, Virginia), Lot 80235, was received on May 28, 1962 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 methods of synthesis and stability, as well as data on composition or other Characteristics which define the test material, are on file with the sponsor. r"-."HAZL=CNLASOFtATORIES VIEPWTMA. VMQ#f4M AMERICA, INC. 2*16,M UJLA. 4 154-158 TEST ANIMALS Sexually mature, cesarean-derived Sprague-Dawley Crl:CDO(SD)BR rats were chosen for this study because they are sensitive to a number of agents which are known to be embryotoxic and/or teratogenic. Rats have historicallybeen used in safety evaluationstudies of this type and are required by the regulatory agencies. Thirty-sixmale and thirty-six female thirteen-week old rats were received from Charles River Breeding Laboratories,.Kingston, New York, on December 22, 1982. The rats were held in quarantinefor two weeks, during Which time a health status examination was performed by a staff veterinarian. The rats were housed one male and one female per cage during breeding. Following confirmation of mating, the females were housed individuallyin elevated wire-mesh cages with food (Purina Rodent Laboratory ChowO 5001) and tap water available ad libitum. The females were uniquely identifiedby ear tag after mat.ingwas confirmed. Temperatures in the study room ranged from 71 to 760F with a relative humidity between 37 and 65%. METHODS Mating Period During the mating period, one female was paired with one male until mating was confirmed or until two weeks had elapsed. Daily vaginal examinations of each female were performed to detect the presence and '0'-'HAZL=CNLABORATORIES VIIEIVIVA@ VfftGW40A 20"M AMERICA. INC. LTMA. 5 154-158 viabilityof sperm or the presenceof a copulatoryplug. The day of observationof sperm or copulatoryplug was designatedas mday 00 of gestation. Groups and Dosage Levels Upon confirmationof mating, each female was assignedto one of the followinggroups. Dosea mg/kg/da-y Number*of Females 0 6 2 1 6 3 5 7 4 10 7 5 20 7 a Based on individualanimal body weights at each weighing interval during the dosing period. The females were placed in the dose groups one at a time beginningwith the high-dosegroup and continuingsequentiallythrough the control group until all mated females were assignedto a group. All males were sacrificedvia carbon dioxide asphyxiationand discardedwithout necropsy once a sufficientnumber of matings were confirmed. Compound Preparation and Administration Suspensionsof the test material in the corn oil vehiclewere prepared on a weight per volume basis. The required amount of compound "'@HAZLL'ZCNLAWFIATAMOERRIICAE.ISNC. 6 154-158 for each level was weighed on an electronic Arbor 126 balance and transferred to a 7 ml homogenizer. Approximately5 ml of corn oil was added and the test material was,ground into a fine suspensionwith a teflon pestle. This suspension was transferred to a beaker and any remaining compound in the homogenizer 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 stirrer for approximatelyten minutes. Prepared suspensions were mixed with a magnetic stirrer during dosing. Females were given the appropriate dosing suspension or vehicle by oral intubationon a daily basis beginning on Day 6 and continuing through Day 15 of gestation. The test material was administeredorally because of the relative ease and accuracy of dosing. Fresh dosing suspensions were prepared weekly and stored under refrigeration (approximately410F). 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 mori- bundity (from day of receipt through study termination)and once daily for clinical signs of toxicity and pharmacologiceffects (throughout gestation). Body weights and food consumption were recorded on Days 0, 6, 8, 12, 16, and 20 of gestation. "'-'HAZLZMCNLABOF;ATORIES AMEFIICA,114C. 7 154-158 Cesarean Sacrifice and Tissue Preservation A gross necropsy was performed on all animals (found deads, moribund sacrifices,and terminal sacrifices). The uterus and ovaries from found dead animals and animals sacrificed because of moribundity were removed and examined for the number of implantation sites and corpora lutea, respectively. Prior to gestation Day 20, survivingfemales were assigned random numbers and all personnel performing cesarean sections and/or external, visceral,or skeletal examinationsof the fetuses were unaware of the dose level from which the animals were derived. On Day 20 of gestation,all survivingfemales were weighed and sacrificedby carbon dioxide asphyxiation. The uterus from each female was weighed and examined for the number and placement of uterine implantationsites, number of live and dead fetuses, early and late resorbing fetuses, and any abnormalities. The uterus of each animal was reweighed after the contents were removed. The ovaries were examined for the number of corpora lutea. Each live fetus was sexed, weighed, and examined for external abnormalities. Findings were recorded. Beginning at the ovarian end of the right uterine horn, the first six fetuses, regardless of sex, were selected for further evaluation. After the external examinationwas completed, the first fetus was identifiedwith a tag and fixed in Bouin's solutionfor soft tissue evaluation(Wilson,et al., 1965). The next two fetuses were eviscerated, tagged, and processed for skeletal examination using a "-'HAZLL'CCNLAsoRAToRiEs AmERicA,iNc. 8- 154-158 technique modified from that reported by Staples (Staples and Schnell, 1964). This sequence was repeated until a maximum of six fetuses had been selected. StatisticalAnalyses Mean maternal body weight gains (Days 6-16 and 0-20), total food consumption, gravid uterine weight, terminal body weight minus gravid uterine weight, resorption incidence,percent males, and fetal viability were analyzed in the following order. Levene's test for homogeneityof variances (Levene, 1960; Draper and Hunter, 1969) was performed and if the variances proved to be homogeneous, the data were analyzed by one-way classificationanalysis of variance (ANOVA) (Winer, 1971). If the variances proved to be heterogeneous,a series of transformationswas performed until variance homogeneity was achieved. These transformations were: loglo, square 2 (X ), square root (Xh), reciprocal (I/X), angular (arcsineXii),and rank, in that order. If rank transformation was ineffective in removing variance heterogeneity,ANOVA of ranked data was completed. If ANOVA of untransformed or transformed data was signifi- cant, Dunnett's t-test (Dunnett, 1955 and 1964) was used for control vs. compound-treated group mean comparisons. If ANOVA was not significant, the analysis was complete. Mean number of fetuses with visceral variants per litter and skeletal variantsper litter were analyzedby Fisher'sexact test for proportions(Fisher,1950). "-"'HAZLL"tCNLABORAAMTEORIRCAI.IENCS. 9 154-158 In addition to the above data, analysis of covariance (ANCOVA) (Winer, 1971) was used to analyze mean fetal weights. The litter was used as the experimental unit. Levene's tests and ANOVA were evaluated at the 5.0% one-tailed probabilitylevel. Control vs. compound-treated group mean comparisons were evaluated at the 5.0% two-tailedprobabilitylevel. Statisticalreferencesare appendedto this report, and statistically significantdifferences,as indicatedby the aforementionedtests, are designatedthroughoutthis report by the term "significant"and/or as follows: S+ = Significantlyhigher than the control value. S- = Significantlylower than the control value. SRecimen, Raw Data, and Final Report Storage All specimens,raw data, and the final report are stored in the archives of Hazleton Laboratories America, Inc. '-@'HAZLETONLASOFIATORIES T@1.1l..It. VigtMNA. VIONOMIA AMERICA. INC ffl"X& UAA@ 10 154-158 RESULTS MATERNAL DATA Clinical Signs Summary clinical signs are presented in Table 1. Treatrnent-relatecdlinicalobservations were noted in the Groups 4 and 5 animals during the treatment and posttreatment phases. These included hunched and thin appearance and rough haircoat in both groups, and tremors, convulsions, prostration,and anorexia in the Group 5 females. Mortality All Group 5 females were found dead or sacrificed in a moribund conditionprior to cesarean section on Day 20 of gestation. Al1 other animals survived to Day 20 of gestation. Body Weights Individual and mean body weights and body weight changes are presented in Appendix 1. Mean weight values and body weight changes are presented in Table 2. Lower mean body weight values were noted in the treated groups. Decreased body weight gain during gestation was noted in Groups 3, 4, and 5, with significant decreases notedinGroups4 and5. (Dueto thenumber of deaths in Group 5, body weight change values could not be determined for Days 0-20.) '"""**'HAZLZMCNLABORATORIES 9=0 Laussupto TUI%NMKIL AMEFIICAI,NC. VIUMNA. V.046*.m0o4o.& 154-158 Food Consumption Individual and mean food consumption values are presented in Appendix 2. Mean food consumption values are presented in Table 3. Decreased food consumption was noted in all treated groups in a dose-related fashion. Significantly lower total food consumption values (Days 0-20) were noted in Groups 3 and 4. (Due to the number of deaths in Group 5, total food consumption values could not be determined.) Gross PathologX Summary gross pathology findings are presented in Table 4. Treatment-relatedfindings were noted in the Group 5 rats and included ulcerated-like areas on the glandular or nonglandular stomach mucosa. Uterine Weights Individual and mean uterine weights are presented in Appendix 3. Summary uterine weights are presented in Table 5. The terminal body weights minus the gravid uterine weights were lower in the treated groups when compared to the control group; however, no statisticaldifferenceswere noted. The gravid uterine weights were comparable between the control and treated groups. -@'HAZLL't=NUWRATOAIME;RIICAE,ISNC. 4ansouple TUOON"IL vispdn^. VORGIP4&& *also, AA- - 12 - 154-158 RESULTS - CESAREAN DATA Individualand mean ovarian, uterine, and litter data are presented in Appendix 4. Individualand mean live fetal data are presented in Appendix 5. Summaryovarian,uterine,and litterdata are presentedin Table 6. Pregnancy Rates, Corpora Lutea, Uterine Implantations,and Implantation 'Efficiency Pregnancyrates were 100 percent for Groups 2, 3, and 5; 83 percent for Group 1, and 86 percent for Group 4. The mean number of corpora lutea and uterine implantationswere comparablefor all groups. The implantationefficiencyfor the controlgroup was low; however,this may be attributedto two animalswhich had implantationefficienciesof 38.4 and 35.7 percent. Fetal Viability,Weight,and Sex The mean incidenceof resorptionswas similar for Groups 1-4. The mean incidenceof fetal viabilitywas comparablebetweenthe control group and Groups 2, 3, and 4, as were mean fetal weights and the percent males per litter. Fetal Development Individualvisceralfindingsare presentedin Appendix6; indivi- dual skeletalfindings are presentedin Appendix 7. Mean incidencevalues ""'@HAZLZMCNLM30MTOAMREIRIECAS,lt4C. 13 154-158 for visceral and skeletal findings are presented in Table 7 (based on number of fetuses) and Table 8 (basedon number of litters). External anomalies were noted in two Group 3 fetuses at cesarean section. These included a small right eye in one fetus, and short and thickened tail, no external genital/analopening and hind legs which were joined from the pelvis to the feet in the other fetus. All remaining fetuses appeared normal. There was no apparent treatment-related increase in the incidence of visceral or skeletal variants or anomalies. Incompletely descended testes were noted in two Group 4 fetuses. Skeletal anomalies noted in one Group 3 fetus included a small pubis, fused ischia, and cojoined hind limbs. This fetus was the one for which the tail and hind limb external abnormalities were noted. Observation Nmber which appeared nomal Alopecia Hunched Rough haircoat Bloody crust Thin Urine stains Tr s Convulsions Prostrate Anorexia Soft feces Bloodydischarge frm vagina Table I ClinicalObservationsDuring Gestation Pilot TeratologyStudy of T-3SSI In Rats Group 1 uazs 6-1!P I 65 6 I I 2 vayU2s D--T 6-15 lb-zu 65 6 Gro 3 ua@UsP U-5 6-15 77 7 1 4 vayU2s ri-15 16-2 7z 3 I I 2 1 I I 2 1 I I Table 2 Mean Maternal Body Weights and Body Weight Change Pilot Teratology Study of T-3551 in Rats Group and Dose Level 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. 0 231.67 13.831 216.53 8.640 231.16 10.085 228.75 19.401 228.29 14.039 Body Weight (g) @o Gestation b a 12 16 256.38 9.187 260.96 12.574 280.92 13.102 303.22 20.915 243.78 12.094 246.60 11.064 266.08 18.994 289.38 16.705 253.44 10.692 253-79 8.578 262.24 9.340 291.21 9.111 254.33 18.145 254.85 20-020 262.15 21.046 266.08 24.184 245.06 13.923 238.70 9.242 202.73 6.388 175.05 5.720 20 355.36 33.795 348.43 23.213 350.90 16.306 333.75 26.277 - Body Wei Days 6-16 46.84 16.308 45.60 9.297 37.77 9.800 4.37s22.593 -64.SOS17.593 Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. Table 3 Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 23.73 2.501 20.88 4.330 21.47 1.923 21.61 2.006 19.31 2.741 Food Consumption igm/day) ffa-oYf Gestation iz 16 20.13 4.132 21.55 3.597 21.10 4.145 18.50 4.159 19.85 2.874 19.27 2.430 16.47 2.735 16.60 1.880 17.66 1.269 16.90 1.615 17.46 5.336 10.91 3.879 14.13 2.743 4.74 4.028 0.32 0.240 20 26.05 4.284 27.00 2.211 24.81 1.585 22.11 3.021 - Total F ---Yays Table 4 Summary Gross Pathology Findings Pilot TeratologyStudy of T-3551 in Rats ORGAN AND DESCRIPTION Group: 1 2 3 4 Dose Level (mg/kg):--g -T -s T( Number of animals examined Number of animals with no gross lesions STOMDJ I Ulcerated-likeareas/darkfoci on glandularmucosa Smooth glandular mucosa Ulcerated-likeareas on nonglandularmucosa 6 6 7 7 6 5 7 7 VAGINA Red discharge HEART Enlarged atria LIVER Mass on median lobe adhered to diaphragm DIAPHRAGM Small hole Group and Dose Level 1 0 mg/kg 2 1 mg/kg 3 5 mg/kg 4 10 mg/kg 5 20 mg/kg Table 5 Mean Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats Mean S.D. Mean S.D. Mean S.D. Mean S.D. Mean S.D. Terminal Body Weight 355.36 33.795 348.43 ?3.213 350-90 16.306 333.75 26.277 Gravid Uteriat_@i@ht 62 82 28:982 72.45 8.097 74.71 12.846 74.78 12.312 - Terminal Body Wei Minus Gravid Uterine 292.54 9.634 281.93 .30.878 276.19 10.211 258.97 19.333 - 19 Key to Table 6 154-158 Pregnancy Rate (percenit)- (number of pregnant feinales/numberof females which showed evidence of-copulation)x 100. Survival Rate (percent)ai (number of females survivingto Day 20/number of females which showed evidence of copulation)x 100. mean ImplantationEfficiencX (Rercent) Group mean of ([resorptions per 7!tter/corporalutea per litterjx LOU). Mean Incidenceof Resorptions (percent) Group mean of ([resorptionsper litter/implantatio5nesr iitterix iuu). Mean Incidence of Fetal Viability (Rercenia - Group mean of fetusesper litter/implantations-pjeirtterix luu). C live Table 6 summaryof ovarian,Uterine, and LitterData Pilot TeratologyStudy of T-3551 in Rats Group: 1 2 3 Dose Level (mg/kg 0 -- r- 5 Number of females with mating confirmed Number of females pregnant Pregnancyrate (%) Number of females survivingto Day 20 Survivalrate 6 6 7 5 6 7 83 100 100 8 6 6 7 100 100 100 10 Mean number of: Corpora lutea Implantations Resorptions Fetuses - dead - live Indicescalculatedon a per litterbasis: Mean implantationefficiency(%)b Mean resorptionincidence(%) Mean incidenceof fetalmortality %) Mean incidenceof fetal viability1%) Live fetuses Mean body weight (g) Males - unadjusted - covariance adjusted Femal es - unadjusted - covariance adjusted Percent males 15.2 15.5 15.7 1 11.0 13.7 13.7 1 0.6 0.5 1.0 0.0 0.0 0.0 10.4 13.2 13.0 1 70.0 88.2 89.1 1 4.1 3.6 7.0 0.0 0.0 0.0 95.9 96.4 93.0 3.70 3.65 3.48 3.48 42.3 3.35 3.36 3.25 3.26 50.7 3.51 3.53 .3.34 3.34 54.6 a The ovaries of two animalswere too autolyzedto count corporalutea. b Data analyzedfollowingrank tr'ansformation. Table 7 Mean Incidence Values for Visceral and Skeletal Findings Pilot Teratology Study of T-3551 in Rats Group: 1 2 Bose Level (mg/kg): 0 Visceral Examination Number of fetuses examined Number and percent of fetuses having one or more visceral variants 10 12 14 1/10.0 2/16.7 2/ Number and percent of fetuses with: Dilatedureter(s) Undulated ureter Incompletely descended testis 1/10.0 2/16.7 1/ 0/ 0.0 0/ 0.0 1/ 0/ 0.0 0/ 0.0 0/ Number and percent of fetuses having one or more visceral anomalies 0/ 0.0 0/ 0.0 0) Skeletal Examination Number of fetuses examined Number and percent of fetuses having one or more skeletal variants 18 24 28 13/72.2 15/62.5 22 Number and percent of fetuses with: Skull incomplete closure (less than 75%) interparietalincompleteossification supraoccipital incompleteossification nonfused nonossified - hyoid - incomplete - nonossified - parietalsirregularlyossified - zygoma incomplete 0/ 0.0 3/12.5 0 6/33.3 13/54.2 13 7/38.9 13/54.2 14 0/ 0.0 2/ 8.3 2 1/ 5.6 0/ 0.0 0 0/ 0.0 0/ 0.0 4 0/ 0.0 8/33.3 0 3/16.7 9/37.5 0 0/ 0.0 2/ 8.3 0 Table 7 - continued Mean IncidenceValues for Visceral and Skeletal Findings Pilot Teratology Study of T-3551 in Rats Group: 1 2 Dose Level (mg/kg): @1 - Rib cage - sternebrae bipartite - ribs Irregularlyossified Vertebral column thoraciccentra - incomplete - nonfused lumbararch irregularlyossified sacral arch - incomplete - nonossified caudals 3 Pelvic girdle - ischitunincomplete - ischil#nand pubis irregularlyossified - pubis nonossified 1/ 5.6 0/ 0.0 0/ 0/ 0.0 0/ 0.0 0/ 1/ 5.6 0/ 0.0 0/ 4/22.2 0/.0.0 0/ -0/ 0.0 0/ 0.0 2/ 0/ 0.0 1/ 4.2 0/ 1/ 5.6 0/ 0.0 0/ 1/ 5.6 0/ 0.0 0/ 1/ 5.6 0/ 0.0 0/ 0/ 0.0 0/ 0.0 2/ 1/ 5.6 0/ 0.0 0/ Bone alISment Sternebrae cneCKerea Fused ribs, malaligned thoracic centra and arches Atigulatedrubs Extra rib(s) Femur short Number and percent of fetuses having one or more skeletal anomalies 1/ 5.6 0/ 0.0 1/ 5.6 2/11.1 0/ 0.0 0/ 0.0 0/ 0.0 0/ 0/ 0.0 1/ 3/12.5 17/ 1/ 4.2 1/ 0/ 0.0 0/ 0.0 1 Number and percent of fetuseswith: Bone aliqnment Fused iscnia and cojoined hind limbs; pubis small 0/ 0.0 0/ 0.0 1 23 Key to Table 8 154-158 Mean Incidenceof VisceralAnomalies (Rercentl Group mean of ([number of fetuseswith ancxnaliisper litter/numberot fetusesexamined viscerally per 11tterI x 100). Mean Incidenceof Visceral Variants (percent'= Group mean of ([number of fetuseswith variantsper litter/niber of Tle-@tuseexsamined viscerallyper litter]x 100). Mean Incidenceof SkeletalAnomalies (Rercerit-i Group mean of ([numberof fetuses with anamali per litter/numberot-fetusesexamined skeletally per litter]x 100). Mean Incidenceof Skeletal Variants (percent@= Group mean of ([numberof fetuses with variantsper litter/numberof ro,@usesexamined skeletallyper litter]x 100). Table 8 Mean incidenceof Visceral and SkeletalFindings per Litter Pilot Teratology Study of T-3551 in Rats Group: 1 Dose Level (mg/kg): 0 Visceral Number of litters examined Mean values calculatedon a per litter basis Number of fetuses with variants Incidence of variants Number of fetuses with anomalies incidence of anomalies Skeletal 5 0.2 10.0 0.0 0.0 Number of litters examined Mean values calculatedon a per litter basis Number of fetuses with variants incidenceof variants Number of fetuses with anomalies Incidence of anomalies 5 2.6 71.6 0.0 0.0 2 3 6 03 17:5. 0.0 0.0 7 0.3 14.6 0.0 0.0 6 2.5 62.5 0.0 0.0 7 3.1 78.6 o.1 3.6 25 Key to Appendix 1 154-158 p = Pregnant NP = Not Pregnant C - Cesarean Delivery, Preceded by Day of Gestation FD n Found Dead, Preceded by Day of Gestation MS = Moribund Sacrifice,Preceded by Day of Gestation Maternal Number 20736 20737 20138 20739 20740 20741 Mean S.O. 20743 20744 20745 20746 20747 20748 Mean S.D 20730 20751 20752 20753 20754 20755 20756 Mean S.O. Date of mating 1/11/83 1111/83 1/11@/83 1/12/83 1/12/83 1/13/83 1/11/83 1/11/83 1/11/83 1/12/83 1/12/83 1/13/83 1/11/83 1/11/83 1/11/83 1/12/83 1/12/83 1/13/83 1/14/83 Disposition 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 Appendix I individualand Mean Maternal Body Weightsa Pilot TerstologyStudyof T-3551 in IR&ts . Body Weig4t (grga-ms-)--Day of12Gestation 15 (201.3) 217.3 224.4 238.9 226.0 252.2 231.76 13.831 212.5 222.9 216.5 210.4 230.1 206.8 216.53 8.640 223.3 216.8 245.4 238.6 228.8 238.7 226.5 231.16 10.085 Group I - 0 mg/kg (230..1) 243.0 256.5 259.3 254.6 268.5 256 36 9:187 (230.0) 246.3 250.9 265.8 264.0 277.8 260.96 12.574 (234.5) 269.5 267.4 280.8 288.0 298.9 280.92 13.102 Group 2 - I mg/kg 232.4 255.4 245.0 240.4 260.7 230.8 240.7 260.9 239.3 240.6 260.7 237.4 243.78 12.094 246.60 11.064 262.1 281.1 259.1 252.4 296.2 245.6 256.08 18.9" Group 3 - 5 mgfkg 248.3 240.1 270.6 258.7 254.2 259.7 242.5 253.44 10.692 253.3 240.3 262.0 2S?.3 246.7 26S.0 251.9 253.79 8.578 267.1 26D.8 249.4 255.6 262.4 279.2 261.2 262.24 9.340 (239.9) 286.1 200.8 300.6 318.9 329.7 303.22 20.915 290.9 301.3 277.6 281.1 315.3 270.1 289.38 16.705 291.7 286.9 295.6 284.6 288.1 309.3 282.3 291.21 9.111 20 (2483) 322:4 318.0 361.8 385.8 388.8 355.36 33.795 339.0 364.8 336.6 329.0 388.2 333.0 348.43 23.213 345.4 342.5 344.5 349.3 353.3 385.7 335.6 350.90 16.306 Only pregnant animals were Includedin group mean. Numbers within parentheseswere excludedfrom man calculations. Data analyzedfolicuing ranktransformation. Maternal Number 20757 20758 20759 20760 20761 20762 20763 Mean S.O. 20764 2076S 20766 20767 20768 20769 20770 Mean S.D. Date of Noting 1/11/83 1/11/83 1111/83 1/12/83 1112/83 1/13/83 1/13/83 1/11/83 1/11/83 1/11/83 1/11/83 1/12/83 1/12/83 1/13/83 Disposition P, 20-C P, 20-C P. 20-C p 20-C Np: 20-C P. 20-C P, 20-C P. 14-FO P, 17-FO P, 17-MS P. 20-FD P, 19-FO P, 18-FO P, 18-FO Appendix I - Continued individual and Mean Maternal BodalWeights& Pilot TeratologyStudyof T-355 in Rats 0 214.6 206.4 237.9 246.2 (229-1) 253.0 214.4 228.76 19.401 233.7 226.5 224.2 211.1 224.9 221.4 256.2 228.29 14.039 Boly Weight torams)- Day of Gestation 6 a iz 15 Group 4 - 10 mg/kg 248.2 226.9 268.9 266.9 (239.S) 272.B 242.3 237.6 223.5 269.3 269.4 (2389) 272:1 257.2 260.3 225.7 283.5 274.1 (225.2) 276.3 253.0 254.33 18.145 254.85 20.020 262.15 21.046 Group 5 - 20 mg/kg 266.9 225.6 299.3 267.1 (199.6) 277.9 259.7 266.08 24.184 250.4 246.1 255.0 219.2 245.0 237.2 262.5 245.06 13.923 243.3 230.9 250.2 233.2 232.3 230.0 251.0 238.70 9.242 199.2 191.7 209.4 1".7 203.4 209.3 206.4 202.73 6.388 165.6 160.6 171.0 175.7 178.6 178.8 175.05 5.720 20 335.8 290.5 349.8 342.3 (229.1) 365.6 318.5 333.75 26.277 a Only pregnant animals were included In group mean. Numbers within parentheses were excluded from mean calculations. b Data analyzedfollowingrank transformation. Maternal Nwber 20736 20737 20738 20739 20740 20741 Mean S.D. 20743 20744 20745 20746 20747 20748 Mean S.D. Appendix 2 Individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 22.6 25.4 21.5 21.2 27.7 24.0 23.73 2.501 19.3 21.9 19.3 24.3 21.5 19.0 20.88 4.330 Food Consumetion (gm/day) ITay-of Gestation 8 12 -16 Group I - 0 mg/kg 18.8 22-6 15:1 19.2 27.0 18.1 16.9 22.2 19.5 20.2 27.4 23.1 14.2 24.6 19.8 19.4 23.6 25.0 20.13 4.132 21.55 3.597 21.10 4.145 Group 2 - I mg/kg 18.1 20.2 15.4 25.9 14.4 17.0 19.4 20.4 17.9 22.5 23.3 15.6 - 20.1 19.3 16;5 23.2 19.6 16.9 18.50 4.159 19.85 2.874 19.27 2.430 20 17.4 27.6 27.8 28.5 26..7 28 3 26.05 4.284 27.3 28.5 24.7 29.4 28.2 23.9 27.00 2.211 Total Constanpi ua 0 366 494 427 437 531 485 457 58 418 444 382 404 441 37. 41( 2@ Maternal Ntsnber 20750 20751 20752 20753 20754 20755 20756 Mean S.D. 20757 20758 20759 20760 20761 20762 20763 Mean S.D. Appendix 2 - Continued individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats 6 21.5 19.4 22.2 22.6 25.6 22.7 18.3 21.76 2.385 18.8 19.4 21.7 24.1 21.0 23.3 23.0 21.61 2.006 Food ConsumEtion (gm/day) Day of Gestation 8 12 16 Group 3 - 5 mg/kg 21.3 15.4 15.4 14.2 13.7 19.0 16.3 19.2 17.5 14.7 14.5 17.0 18.3 15.0 19.0 18.2 17.0 15.8 17.0 19.4 17.2 16.47 2.735 16.60 1.880 17.66 1.269 Group 4 10 mg/kg 17.0 15.4 19.9 17.9 15.3 16.8 16.0 16.8 28.2 18.0 17.6 10.6 15.6 15.4 12.6 10.9 15.9 8.9 3.6 11.4 13.1 16.90 1.615 17.46 5.336 10.91 3.879 20 24.8 23.7 26.5 26.2 23.8 26.3 22.4 24.81 1.585 22.8 23.4 23.3 20.7 15.9 23.8 24.9 22.11 3.021 Total Consumpt Day u- 423. 384. 396. 389. 400. 429. 360. 397. 23. 355. 396. 398. 368. 276. 376. 383. 365. 41. Maternal NLxnber 20764 20765 20766 20767 20768 20769 20770 Mean S.D. Appendix 2 - Continued individual and Mean Food Consumption Values Pilot Teratology Study of T-3551 in Rats Food Consumption tgm/day) Day Of Gestation 8 12 16 20 Group 5 - 20 mg/kg 15.3 18.7 1.3 - 20.5 11.1 1.2 0.2 21.2 14.5 3.8 0.6 15.8 16.8 12.4 02 22.0 12.4 5.8 0.3 19.0 12.1 6.8 0.6 21.4 13.3 1.9 0.0 19.31 2.741 14.13 2.743 4.74 4.028 0.32 0.240 Total Consumpt Da) 0- Maternal Number 20736 20737 20738 20739 20740 20741 Mean S.D. 20743 20744 20745 20746 20747 20748 Mean S.D. Appendix 3 individualTerminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats Terminal Body Weight (_qy Not Pregnant 322.4 318.0 361.8 385.8 388.8 355.36 33.795 Gravid Uterine Weight (g) Group I - 0 mg/kg 32.9 32.8 71.3 97.7 79.4 62.82 28.982 Terminal Minus Gravi 2 2 2 2 3 2 Group 2 1 mg/kg 339.0 62.7 3 364.8 80.9 2 336.6 72.0 2 329.0 80.9 2 388.2 63.3 3 333.0 74.9 2 348.43 72.45 2 23.213 8.097 Maternal Number 20750 20751 20752 20753 20754 20755 20756 Mean S.D. 20757 20758 20759 20760 20761 20762 20763 Mean S.D. Appendix 3 - Continued individual Terminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats Terminal Body Weight (IL 345.4 342.5 344.5 349.3 353.3 385.7 335.6 350.90 16.306 335.8 290.5 349.8 342.3 Not Pregnant 365.6 318.5 333.75 26.277 Gravid Uterine Weight (g) Group 3 5 mg/kg 73.0 77.6 56.1 68.8 85.0 95.5 67.0 74.71 12.846 Group 4 10 m4/kg 72.6 61.2 65.3 86.4 92.8 70.4 74.78 12.312 Terminal Minus Gravi 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Maternal Number 20764 20765 20766 20767 20768 20769 20770 Appendix 3 - Continued IndividualTerminal Body Weights, Gravid Uterine Weights, and Terminal Body Weights Minus Gravid Uterine Weights Pilot Teratology Study of T-3551 in Rats Terminal Body Weight (91 Gravid Uterine Weight (g) Group 5 20 mg/kg Terminal Minus Gravi - - - - - - - - Maternal Ntaber 20736 20737 20738 20739 20740 20741 Mean S.D. 20743 20744 20745 20746 20747 20748 Mean S.D. 20750 20751 20752 20753 20754 20755 20756 Mean S.D. Ovarian Corpora Lutea Uterine Impl antatIons Not Pregnant 13 14 16 16 17 15.2 1.64 5 5 14 16 is 11.0 5.52 14 15 is to 15 15 15.5 1.38 13 14 14 16 11 14 13.7 1.63 15 15 is 17 16 19 13 15.7 1.89 13 14 12 15 is 17 12 13.7 3.35 Appendix 4 Individualand Mean Ovarian.Uterine.and LitterData Pilot TeratologyStudy of T-3551 in Rats implantationa Efficiency(%) Early Resorptions Resorption incidences(%I Group 1 0 mg/kg Ntnber of Fetuses Dead Alive 38.4 35.7 87.5 100.0 88.2 70.0 30.47 92.9 87.5 93.3 88.9 73.3 93.3 88.2 7.71 86.7 93.3 80.0 88.2 93.8 09.5 92.3 89.1 4.82 0 0.0 0 0.0 1 7.1 0 0.0 2 13.3 0.6 4.1 0.89 6.00 Group 2 1 mg/kg 2 15.4 0 0.0 0 0.0 1 6.3 0 0.0 0 0.0 0.5 3.6 0.84 6.30 Group 3 - 5 mg/kg 0 0.0 0 o.0 2 16.7 3 20.0 1 6.7 1 5.9 0 0.0 1.0 7.0 1.15 8.28 0 0 0 0 0 0.0 0.00 0 0 0 0 0 0 0.0 0.00 0 0 0 0 0 0 0 0.0 0.00 5 5 13 16 13 10.4 6.08 11 14 14 is it 14 13.2 1.72 13 14 10 12 14 16 12 13.0 1.91 a Oat& analyzedfollaoingrank transfonnation. Maternal Nmber- 20757 20758 20759 20760 20761 20762 20763 Mean S.D. 20764 20765 20766 20767 20768 20769 20770 Mean S.D. Appendix4 - Continued Individualand Mean Ovarian.Uterine,and LitterData Pilot TeratologyStudy of T-3551 In Rats Ovarian Corpora Lute& Uterine Implantations Implantat Iona Efficiency 15 14 16 16 Not Pregnant 18 19 16.3 1.86 13 13 10 is 17 13 13.5 2.35 86.7 92.9 62.5 93.8 94.4 68.4 83.1 14.08 14 12 16 b b is 13 14.0 1.58 13 it 13 3 10 14 11 10.7 3.68 Early Resorptions Resorption Incidences(%I Group 4 - 10 mg/kg 0 0.0 1 7.7 0 o.0 0 0.0 0 0.0 1 7.7 0.3 2.6 0.52 3.98 Group 5 - 20 mg/kg Nmber of Fetuses Be-ad A] 0 0 0 0 0 0 0.0 0.00 13 12 10 Is 17 12 13.2 2.48 Data analyzedfollowingrank transformation. b Ovarieswere too autolyzedto count corporalutea. Maternal Ntaber 20736 20737 20738 20739 20740 20741 Mean S.D. 20743 20744 20745 20746 20747 20748 Mean S.D. 20750 20751 20752 20753 20754& 20755 20756 Mean S.D. Nmber of Males Sex Number of Females Not Pregnant 2 2 8 5 5 4.4 2.51 3 3 5 11 8 6.0 3.47 4 7 9 5 7 7 9 6 4 7 8 6 6.8 6.3 2.32 0.82 7 6 8 4 5 12 7 7.0 2.58 6 a 2 a 8 4 5 5.9 2.34 Appendix 5 Individualand Mean Live Fetal Data Pilot TeratologyStudy of T-3551 in Rats Percent Males Group 1 Males Unadjusted 0 mg/kg Mean Weight (grams) Adjusted %-a-d3u 40.0 40.0 61.6 31.3 38.5 42.3 11.39 4.1 3.3 3.3 3.8 4.0 3.70 0.381 3.7 3.4 3.1 3.5 3.7 3.65 3.4 0.127 0.24 Group 2 - I mg/kg 36.4 3.4 3.3 64.3 3.5 3.4 50.0 3.2 3.0 60.0 3.1 3.0 36.4 3.4 3.3 57.2 3.5 3.5 50.7 3.35 3.36 3.2 12.03 0.164 0.106 0.21 Group 3 5 mg/kg 53.9 3.3 3.3 42.9 3.4 3.3 80.0 3.4 3.4 33.4 3.6 3.1 38.6 3.8 3.5 75.0 3.8 3.6 58.4 3.3 3.2 54.6 3.51 3.53 3.3 17.90 0.219 0.099 0.1 a Sex of one pup could not be determined. Maternal Number 20757 20758 20759 20760 20761 20762 20763 Mean S.O. 20764 20765 20766 20767 20768 20769 20770 -N-mWreof Males Sex Number of Females 6 7 7 5 6 4 8 7 Not Pregnant 12 5 8 4 7.8 5.3 2.23 1.34 Found Dead - Day 14 Found Dead '.Day 17 Moribund Sacrifice - Day 17 Found Dead - Day 20 Found Dead - Day 19 Found Dead - Day 18 Found Dead - Day 18 Appendix 5 - Continued individualand Mean Live Fetal Data Pilot TeratologyStudy of T-3551In Rats Percent Males ga-les Un-adJusted Group 4 - 10 mg/kg 46.2 3.7 58.4 3.2 60.0 3.8 53.4 3.4 70.6 3.2 66.7 3.3 59.2 8.83 3.43 0.258 Group 5 - 20 mg/kg Mean Weight (grns) AOjust ui-a-d 3.3 3.2 3.6 3.2 3.3 3.2 3.47 3.31 0.111 0.1 Appendix 6 IndividualVisceralFindingsfor Each Litter Pilot TeratologyStudy of T-3551 In Rats G2roup 1 0 mg/k z i z 0 0 0 0 0 7 7 7 7 1 Animal 3 3 3 4 4 Ntaber: 7 8 9 0 1 Number of fetuses examined by Wilson'stechnique Number of fetuses within normal limits Number of fetuseshaving one or more visceralvariants Percent of fetuses having one ormore visceral variants(%) Number of fetuseswith: Dilatedureter(s) 2 2 2 2 0 0 0.0 0.0 2 2 2 2 1 2 0 1 0 0.0 50.0 0.0 I Number of fetuses having one or more visceral anomalies Percent of fetuses having one ormore visceral anomalies 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 Gro z z 0 0 0 7 7 7 4 4 4 3 4 s 2 2 2 2 2 1 0 0 1 0.0 0.0 50. I 0 0 0 0.0 0.0 0. OUP 3 5 mg/k -7 z GZE i i z z 0 0 0 0 0 0 0 7 7 7 7 7 7 7 Animal 5 5 5 6 5 5 5 Ntaber: 0 1 2 3 4 5 6 Number of fetuses examined by Wilson's technIque 2 2 2 2 2 2 2 Numberof fetuseswithin normal limits 2 2 2 2 2 2 2 Number of fetuses having one or more visceralvariants 0 2 0 0 0 0 0 Percent of fetuseshaving one ormore visceralvariants(%) 0.0 100.0 0.0 0.0 0.0 0.0 0.0 Number of fetuseswith: Dilated ureters tfndualted ureter I IfK@mpletelydescended testis Number of fetuses having one or more visceral anomalies Percent of fetuseshaving one ormore visceralon &lies(%) 0 0 0 0 0 0 0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Group 4 z ic 4e 0 0 0 7 7 7 5 s s 1 8 9 2 2 2 2 1 2 0 1 0 0.0 50.0 0. I 0 0 0 0.0 0.0 0. Appendix 7 individualSkeletalFindingsfor Each Litter Pilot TeratologyStudy of T-3551 in Rats Animal Number: Number of fetuses examined Numberof fetuseswithinnormal limits Number of fetuseshaving one or more skeletal variants Percent of fetuseshavingone or more skeletal variants (%) Number of fetuseswith: L!VpiUg ossification skuI I - incompleteclosure (lessthan 75%) - lntraparietalIncompleteossification - supraoccipital Incompleteossification nonfused nonossifled - hyoid- nonossifled - parietalsIrregularlyossifled - zygoma incomplete Rib cage - sternebraebipartite Vertebralcolumn- thoraciccentra- Incomplete - nonfused - sacral arch- Incomplete - nonfused - caudals < 3 Pelvic girdle- lschitm Incomplete - pubis nonasslfled '%A' rnel&Maeent chectered Angulated ribs Extra ribs Number of fetuses having one or more skeletal mmal Ies Percentof fetuses havingone ormore skeletal anomalies (%) 0 7 3 7 3 2 1 33 1 0 o.0 G7roup I i 0 Mg/ki 0 0 0 7 7 7 3 3 4 8 9 0 3 4 4 0 0 1 3 4 3 100 100 is 2 3 2 2 1 I 3 I I 1 2 I 1 2 0 0 0 0.0 0.0 0.0 z 0 7 4 1 4 2 2 50 1 1 1 0 0.0 Gro!l 2 z z 0 0 0 7 7 7 4 4 4 3 4 5 4 4 4 2 3 0 2 1 4 so 25 100 2 2 4 2 1 3 I 1 4 1 1 4 2 2 0 0 0 0.0 0.0 0. Appendix 7 - Continued individualSkeletalFindingsfor Each Litter Pilot TeratologyStudy of T-3551 In Rats G7roup 3 5 mg/k 2 z i i z z z 0 0 0 0 0 0 0 0 7 7 7 7 7 7 7 7 Animal 5 5 5 5 5 5 5 5 Ntnber: 0 1 2 3 4 5 6 7 Nmber of fetusesexmined 4 4 4 4 4 4 4 4 Numberof fetuseswithin normallimits 2 1 0 0 0 3 0 2 Nmber of fetuseshaving one or more skeletalvariants 2 3 4 4 4 4 2 Percent of fetuseshlving one or more skeletalvariants %) so 75 100 im too 25 100 so Number of fetuseswith: Lasping ossification skuil- incompleteclosure(less than 75%) - lnterparietalIncomplete ossification 2 1 4 2 4 1 - supraoccipital- Incomplete ossification 2 2 4 2 4 1 - nonfused nonossified - hyold- Incompl-ete 2 2 - nonossified - parietalsIrregularly ossified Rib cage - sternebraebipartite ribs Irregularloyssified Vertebral-column - thoraciccentra- Incomplete - Imbar arch Irregularly ossi(IW 2 - sacral arch Incomplete Pelvicgirdle- lschim and pubis Irregularly ossified 2 Group 4 2 2 0 0 7 7 5 5 8 9 4 4 1 3 3 75 25 2 1 Appendix 7 - Continued IndividualSkeletalFindings for Each Lltter Pilot TerstologyStudy Of T-3551 In Rats 2 0 7 Animal 5 Nmber: 0 one Ar ent ' St-e n"e!:!ae cneckerea Fused ribs;galaligned thoracic centra and arches Angulated ribs 2 Extra ribs Femur short Nmber of fetuses having one or owe skeletal anomalies 0 Percent of fetuses having one or more skeletal anomalies 0.0 Nmber of fetuses with: Bone AlIpment tusea iscnis ano cojoinea hind limbs; pubis small Group 3 i 5 m,,kl z ---r 0. 0 0 0 1 7 7 7 5 5 5 5 1 2 3 4 3 4 2 2 0 0 0 1 0.0 0.0 0.0 2S I z z 0 0 7 7 5 5 s 6 4 0 0 0.0 0.0 Group 4 - z z z 0 0 0 7 7 7 s 5 5 7 8 9 2 0 0 0 0.0 0.0 0.0 42 Appendix 8 ReTerences Pilot TeratologyStudy of T-3551 in Rats STATISTICAL METHODS/LABORATORY PROCEDURES 154-158 Bartlett, M. S., "Some Examples of Statistical Methods of Research in Agriculture and Applied Biology." J. Royal Statist. Soc. Suppi.., IV:137-170, 1937. Draper, N. R., and Hunter, W. G., ntransfamations: Some Examples Revisited." Technometrics, 11:23-40, 1969. Dunnett, C. W., "A Multiple CornoarisonProcedure for Compari-ngSeveral Treatments with a Control." J. Am. Stat. Assoc., 50:1096-1121, 1955. Dunnett, C. W., "New Tables for Multiple Comparisons with a Control." Biometrics,,20:482-491, 1964. Games, P. A., and Howell, J. F., "Pairwise Multiple Comparison Procedures with Unequal N's and/or Variances: A Monte Carlo Study." J. Ed. Statist., 1:113-125,.1976. Fisher, R. A., StatisticalMethods for Research Workers Oliver & Boyd, Edinburgh, 1950. Levene, H., "Robust Tests for Equality of Variances,nin I. Olkin edited, Contributionsto Probabilityand Statistics,Stanford University Press, Palo Alto, 196U. Staples, R. E. and Schnell, V. L., "Refinementsin Rapid Clearing Technic in the KOH-AlizarinRed S Method for Fetal Bone." Stain Technology, 39:61-63, 1964. Wilson, J. G. and Warkany, J., editors. Teratology: Principles and Techniques, The Univ. of Chicago Press, Chicag-o,TW, pp. z5i-zii. Winer, B. J., Statistical Principles in Experimental Design, 2nd Ed., McGraw-Hill,N.Y., Chs. 3-10, 1971.