Document kaLN1baV5RvKV6NkX8bJmOjd0

s9qf = Suae 3 yey ARQ&_6 1133 UNITED STATES EANVSINROGNMTEONTNA5L..PRSOToECTION AGENCY eva eanrnSn ceeesio 21 October 2002 MEMORANDUM: ) SUBJECT: Focussed statistical analysesof APFO datma en wd oveinglos FROM: Efabeh i Margesches PhD, Stasis 5 , { oe S o RExiissktiAnsgseCshsemmeinctalDsivAisssieosns(m4en0t3BMr)anch OY ; S2338 0: Katherine Asitole, Ph.D. Toxiologis // Existing Chemicals Assessment Branch 23 BAE25 ~ Risk Ascossment Division (7403M) BpgA s0t0u1d3y0bAyugYuosrtk2(0200022s)eavnerdalsommeemboethresrosfudiindcussttrhyamietgwhithhiEvePbAeasrtinagtfoonditshceums,s rTetswulatss asgereeeidnthe that certain additonal analyses would be caried out sgYirotorsukpps(e2ri0nd0en2l)uimvebeperoerrdstolefdtdt,ahmegrseeswtdaeetlriieovnenroiinnsdgteaxit,itsentriuc,matlblehyrssidguonfrdiaftaiicmoannstowdfgiieftsfhtealrteinlocens,raavmeporuanpgsge,stdhaoenrzsciamwtpialtnahennattlaltion epiutphserdtyhiengF,OloirveFbIorpnaraenndtasltglebneormatpiuopnss,avnidabitleitiy linidteexs,, pAucpcosrexdirnagtioos pa.nd45boofdythweeeigphotrs,ftorhese hcanooammlpoyagsreeansbeliwete,yr.ethcearKrriuesdkaolu-tWuaslilnigs atneaslty(sniosnopfarvaamreitanrciec,oro,f,ifftorhcgoruonutpdavta,RAaRCchEiS-Wsqeuraerenofsor J"Tihveebaodrdni)t/ioinmapllaanntaaltoynsse)raenldatoedntoo fanprienwdeeaxnoifnpgremnoartaallimtoyrttaalkiteyn (2tak(evneabo(rpnlmainntuastitohnossemsiinvues asonanamdlpyalsyees2p2rc)oalpniovrbetebiocoransri)r.ierdeTlhoaeuttsedeutsqoiutnahgnetdiarttiuaeestparraeonpspofrrotoirpomoner.dtiioWnnhaseSwnittathnhidenardedeanfcaohsmhiiineoanrt,oaurnssdinatgrheeAnrveaacrrsilayinneceq+uo(f,e the Gaylor, 1978). Analysisof variance of the numbers of mplanations, and those nombers - properly transformed for being counts ((iy2+(i+l)) assured that this condition was pretty well fulfilled. To illustrate the effect of the transformation of the proportions, plots and analyses are provided both with and without the transformation, although the untransformed statistics should not be used. All analyses were implemented using the computer program Statistix (Statistix 7, Analytical Software, 2000). In light ofthe other tests already carried out by York (2002), the high dose prenatal mortality index was statistically compared only to the control. Plots ofthe index for control and four positive doses (p. 1 of the 36 analysis pages) showed that, in both FO (red squares) and Fl(blue circles), the treatments did not appear to differ. The comparison was carried out using Student's t-test. No difference was seen between high dose and control (p. 3). The preweaning mortality index was examined across all dose groups, within generation. Here again a plot ofthe index across all treatments (p. 12)suggested no differences. This comparison was carried out using Analysis of Variance (p. 20 for P or FO, p. 26 for Fl), followed by a Scheffe comparison of the individual treatment means (p. 21 and p. 27, respectively). Using the Scheffe basis permitted all contrasts of means to be considered in follow-up comparisons while controlling for an overall error rate. While the order of treatment-group means appeared to differ between FO and Fl, in neither case were there any significant pairwise differences between the dose groups and the responses did not have any trend in dose. Gaylor, D.W. 1978. Methods and Concepts of Biometrics Applied to Teratology. In: J.G. Wilson and F.C. Fraser, ed., Handbook of Teratology. Vol. 4. New York: Plenum Press. pp. 429-444. Statistix Version 7.0 [computer software]. Analytical Software. 1985, 2000. cc: J. Seed [ o- 8: a EE oo mmouwems oi 8 8 | 2 i: y | Ha | 1 . Ha if y :E 3Fea 5 8 33 oo we ommam |& oo 58 [ . . owesamo owe cmmme coscmocm 2 : 33 ((Guerduwian-| Jubs)uisoe)gFuduL. an FE . u 28 2 x ag k] 55a 8 Ea 28 oe omomwo|8 | 8 | Ind |lw g Cees r| e -- | coo ocmwmm | conmom 0 | (ueiduwian-L HLT 2 STATISTIX 7.0 TWO-SAMPLE T TESTS FOR TRPREB BY DOSE CONT--HIGHONLY, 10/15/02, 4:03:39 PM p DOSE MEAN SAMPLE SIZE S.D. S.E. 0 0.2020 28 30 0.2825 29 DIFFERENCE -0.0805 0.1941 0.2945 0.0367 0.0547 NULL HYPOTHESIS: DIFFERENCE = 0 ALTERNATIVE HYP: DIFFERENCE <> 0 ASSUMPTION EQUAL VARIANCES UNEQUAL VARIANCES T --1.21 --1.22 DF 55 48.6 P 0.2299 0.2273 95% CI FOR DIFFERENCE (--0.2134, 0.0524) (--0.2129, 0.0518) TESTS FOR EQUALITY OF VARIANCES F 2.30 NUM DF 28 DEN DF 27 P 0. 0166 CASES INCLUDED 57 MISSING CASES 3 TRPREB = transformed (loss prior to birth) In parents (p. 3) and Fl (p. 8), control and high dose groups not significantly different under assumption of unequal variances (or equal ones). The test for variance equality is sensitive to normality assumptions, and these data are not normally distributed. The indication the variances are unequal may not be correct for P (see the box and whiskers plot p. 4) although it's likely for Fl (see p. 9) --3-- Box and Whisker Plot P 1.5. * 1.0~ 0.5 0.0. 0 30 DOSE 57 cases 3 missing cases --4-- The Box and Whisker Plot procedure computes box plots that graphically present measurements of central tendency and variability. A series of box plots can be displayed side by side, which can dramatically illustrate differences between groups. Bach box plot is composed of a box and two whiskers. The box encloses the middle half ofthe data. The box is bisected by a line at the value for the median. The vertical lines at the top and the bottom of the box are called the whiskers, and they indicate the range of "typical" data values. Whiskers always end at the value of an actual data point and can't be longer than 1 ~/2 times the size ofthe box. Bxtrerne values are displayed as "s" for possible outliers and "0" for probable outliers. Possible outliers are values that are outside the box boundaries by more than 1times the size of the box. Probable outliers are values that are outside the box boundaries by more than 3 times the size of the box. Copyright 2000 Analytical Software --5-- STATISTIX 7.0 TWO-SAMPLE T TESTS FOR PREBIRTH BY DOSE 10/15/02, 3:34:53 PM P DOSE MEAN SAMPLE SIZE S.D. S.E. 0 0.0727 28 30 0.1274 29 DIFFERENCE -0.0547 0.0826 0.1942 0.0156 0. 0361 NULL HYPOTHESIS: DIFFERENCE = 0 ALTERNATIVE HYP: DIFFERENCE C> 0 ASSUMPTION EQUAL VARIANCES UNEQUAL VARIANCES T --1.38 --1.39 DF 55 38.1 P 0.1747 0.1719 95% CI FOR DIFFERENCE (--0.1344, 0.0250) (--0.1342, 0.0248) TESTS FOR EQUALITY OF VARIANCES CASES INCLUDED 57 F NUN DF 5.52 28 MISSING CASES 3 DEN DF 27 P 0.0000 PREBIRTH = loss prior to birth --6-- Box and Whisker Plot P 1.0 0 0.8 0.6 * 0.4 0.2 0.0 0 30 DOSE 57 cases 3 missing cases --7-- STATISTIX 7.0 TWO--SAMPLE T TESTS FOR TRPREB BY DOSE DOSE MEAN SAMPLE SIZE S.D. CONT--HIGHONLY, 10/15/02, 4:05:52 PM F: S.E. 0 0.2497 28 30 0.2521 29 DIFFERENCE -2.41E--03 0.1488 0.2223 0.0281 0.0413 NULL HYPOTHESIS: DIFFERENCE = 0 ALTERNATIVE HYP: DIFFERENCE <> 0 ASSUMPTION EQUAL VARIANCES UNEQUAL VARIANCES T --0.05 --0.05 DF 55 49.1 P 0.9619 0.9616 95% CI FOR DIFFERENCE (--0.1032, 0.0984) (--0.1028, 0.0980) TESTS FOR EQUALITY OF VARIANCES F 2.23 NUM DF 28 DEN DF 27 P 0.0201 CASES INCLUDED 57 MISSING CASES 3 --8-- Box and Whisker Plot 1.0 0.8 0.6 0.4 0.2 0.0 0 30 DOSE 57 cases 3 missing cases --9-- STATISTIX 7.0 TWO--SAMPLE T TESTS FOR PREBIRTH BY DOSE 10/15/02, 3:27:53 PM Ft DOSE MEAN SAMPLE SIZE S.D. S.E. 0 0.0801 28 30 0.0998 29 DIFFERENCE -0.0197 0.0600 0.1258 0.0113 0.0234 NULL HYPOTHESIS: DIFFERENCE = 0 ALTERNATIVE HYP: DIFFERENCE <> 0 ASSUMPTION EQUAL VARIANCES UNEQUAL VARIANCES TESTS FOR EQUALITY OF VARIANCES T --0.75 --0.76 F 4.39 DF P 95% CI FOR DIFFERENCE 55 43.4 0.4563 0.4524 (--0.0723, 0.0329) (--0.0722, 0.0328) NUM DF DEN DF P 28 27 0.0001 CASES INCLUDED 57 MISSING CASES 3 --10-- Box and Whisker Plot 0.8 0.6 0.4 0.2 0.0 0 30 DOSE 57 cases 3 missing cases --11-- -- [= 8El 2 3 2 HLi:2 go8w El $ for H:PRT5 g td 5 2 g s2 Eo. gz a esoo eee com eB | y| | `3 | | || lIe | |la oe | | coo mm oe come 9 gE: 2 ((ann/zzAep-| ubs)ursoe) MIL 12 1.6 1.2 08 0.4 0.0 0 Box and Whisker Plot 1 3 10 30 DOSE 283 cases 16 missing cases p. 12 and pp. 13--15 illustrate two ways of looking at the transformed preweaning loss, to clarify the conclusions of the analysis of variance on pp. 20 and 26. --13-- 1.6 1.2 0.8 0.4 0.0 0 Box and Whisker Plot 1 3 10 30 DOSE 142 cases Parent(F0) 8 missing cases --14-- 0.8 0.6 0.4 0 0.2 0.0 0 Box and Whisker Plot * * 0 0 1 3 10 30 DOSE 141 casesFl8 missing cases --15-- | oo o fo. on comme 8 w 8 5 3 coms . N Fl ows cme Be cms gg (onirzzAep-LINVIMI Box and Whisker Plot 1.2- 0 0 0 0.8 - 0.4 _ 0.0- I 0 * _ II 0 6 * II I 0 _ II 1 3 10 DOSE 283 cases F0&F1 16 missing cases 0 0 * _ ____ 30 --17-- Box and Whisker Plot 1.2 0 0 0 z 0.8 cC w w a. 0.4~ _ _ 0.0 - I I I 0 * 0 0 _ 0 _ II I 0 1 3 10 30 DOSE 142 cases Parent(F0) 8 missing cases --18-- 0.35 0.28 0.21 0.14 0 0.07 0.00 0 Box and Whisker Plot 0 0 0 0 0 1 3 10 30 DOSE 141 casesFi8 missing cases --19-- STATISTIX 7.0 ONE-WAY AOV FOR TRPREW BY DOSE SOURCE DF SS MS F P BETWEEN 4 WITHIN 137 TOTAL 141 0.31524 9.85073 10.1660 0.07881 0.07190 1.10 0.3611 CHI--SQ DF P BARTLETT'S TEST OF EQUAL VARIANCES 32.85 4 0.0000 COCHRAN'S Q LARGEST VAR I SMALLEST VAR 0.3314 6.0991 COMPONENT OF VARIANCE FOR BETWEEN GROUPS 2.433E-04 EFFECTIVE CELL SIZE 28.4 DOSE MEAN SAMPLE SIZE GROUP STD DEV 0 1 3 10 30 TOTAL 0.1378 28 0.1532 27 0.1162 29 0.0661 29 0.2089 29 0.1362 142 0.3110 0.3150 0.1632 0.1403 0.3465 0.2681 CASES INCLUDED 142 MISSING CASES 8 10/16/02, 3:23:20 PM P --20-- STATISTIX 7.0 I SCHEFFE COMPARISON OF MEANS OF TRPREW BY DOSE 10/16/02, 3:21:29 PM P DOSE MEAN HOMOGENEOUS GROUPS 30 0.2089 I 1 0.1532 I 0 0.1378 I 3 0.1162 I 10 0.0661 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --21-- One-Way AOV Residual Plot 1.5- + + + 1.0- + 0.5- + t + + + * 0.0 - + + + -0.5- -1.0- -1.5- o.be o.bg 0.12 0.15 0.18 0.21 Fitted values:arcsinsqrt(1 -day22/Iive) parent generation Using dose alone somewhat underestimates the pattern of preweaning loss. --22-- STATISTIX 7.0 ONE-WAY AOV FOR PREWEAN BY DOSE ALLDOSESD22, 10/17/02, 8:51:04 AN 12 SOURCE DF SS MS F P BETWEEN 4 WITHIN 137 TOTAL 141 0.10272 3.38109 3.48380 0.02568 0. 024 68 1.04 0.3887 BARTLETT'S TEST OF EQUAL VARIANCES CHI-SQ 70.93 COCHRAN'S Q LARGEST VAR I SMALLEST VAR DF P 4 0.0000 0.3627 15.017 COMPONENT OF VARIANCE FOR BETWEEN GROUPS EFFECTIVE CELL SIZE 3.520E-05 28.4 DOSE MEAN SAMPLE SIZE GROUP STD DEV 0 1 3 10 30 TOTAL 0.0591 28 0.0638 27 0.0372 29 0.0221 29 0.1005 29 0.0564 142 0.1889 0.1914 0.0641 0.0549 0.2126 0.1571 CASES INCLUDED 142 MISSING CASES 8 --23-- STATISTIX 7.0 SCHEFFE COMPARISON ALLDOSESD22, OF MEANS OF PREWEAN BY DOSE 10/17/02, F 8:52:43 AN DOSE MEAN HOMOGENEOUS GROUPS 30 0.1005 I 1 0.0638 I 0 0.0591 I 3 0.0372 I 10 0.0221 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --24-- One-Way AOV Residual Plot 1.0 ++ + 0.5 + + + + + 0.0 + -0.5 -1.0 I I I I 0.01 0.03 0.05 0.07 0.09 0.11 Fitted values:1-day22live/Iiveborn parent generation --25-- STATISTIX 7.0 ALLDOSESD22, 10/17/02, 9:01:05 AM ONE-WAY AOV FOR TRPREW BY DOSE SOURCE DF BETWEEN 4 WITHIN 136 TOTAL 140 SS 0.04543 2.65791 2.70334 MS 0.01136 0.01954 F 0.58 P 0.6768 BARTLETT'S TEST OF EQUAL VARIANCES CHI--SQ 6.38 COCHRAN'S Q LARGEST VAR I SMALLEST VAR DF P 4 0.1726 0.3339 2.3935 COMPONENT OF VARIANCE FOR BETWEEN GROUPS --2.903E-04 EFFECTIVE CELL SIZE 28.2 DOSE MEAN SAMPLE SIZE GROUP STD DEV 0 1 3 10 30 TOTAL 0.0593 28 0.0710 28 0.1114 28 0.0878 28 0.0721 29 0.0803 141 0.1168 0.1299 0.1807 0.1308 0.1323 0.1398 CASES INCLUDED 141 MISSING CASES 8 --26-- STATISTIX 7.0 SCHEFFE COMPARISON ALLDOSESD22, OF MEANS OF TRPREW BY DOSE 10/17/02, F' 9:02:27 AM DOSE MEAN HOMOGENEOUS GROUPS 3 0.1114 I 10 0.0878 I 30 0.0721 I 1 0.0710 I 0 0.0593 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --27-- One-Way AOV Residual Plot 0.6 - + + + 0.3. + + i 0.0 - + + -0.3 - -0.6 o.d58 0.067 0.076 0.685 0.094 0.1103 Fitted values:arcsin(sqrt(1 -day22/Iive)) Fl dams; F2 pups 0.112 --28-- STATISTIX 7.0 ONE-WAY AOV FOR PREWEAN BY DOSE ALLDOSESD22, 10/17/02, 8:56:04 AN Ft SOURCE DF SS MS F P BETWEEN 4 WITHIN 136 TOTAL 140 0.01010 0.33704 0. 347 14 0.00253 0.00248 1.02 0.3999 CHI-SQ DF P BARTLETT'S TEST OF EQUAL VARIANCES 32.45 4 0.0000 COCHRAN'S Q LARGEST VAR I SMALLEST VAR 0.5270 6.0289 COMPONENT OF VARIANCE FOR BETWEEN GROUPS EFFECTIVE CELL SIZE 1.665E-06 28.2 DOSE MEAN SAMPLE SIZE GROUP STD DEV 0 1 3 10 30 TOTAL 0.0162 28 0.0205 28 0.0408 28 0.0236 28 0.0214 29 0.0245 141 0.0329 0.0425 0.0809 0.0358 0.0413 0.0498 CASES INCLUDED 141 MISSING CASES 8 --29-- STATISTIX 7.0 ALLDOSESD22, SCHEFFE COMPARISON OF MEANS OF PREWEAN BY DOSE 10/17/02, F' 8:57:42 AN DOSE MEAN HOMOGENEOUS GROUPS 3 0.0408 I 10 0.0236 I 30 0.0214 I 1 0.0205 I 0 0.0162 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --30-- One-Way AOV Residual Plot 0.3- + 0.2 - + + + 0.1 - + ~t+ *+ * 0.0 - + + -0.1 - -0.2 - -0.3 - I ---- I 1.6 2.1 I 2.6 3.1 3.6 4.1 Fitted va!uesXl OE-2: I -day22/Iive Fl generation dams; F2 pups --31-- Statistix offers a number of procedures to test hypotheses about the central values of the population distributions from which the samples are drawn. These procedures are often referred to as tests of location. Several of these tests are parametric and require the assumption that the data are normally distributed. Nonparametric tests are provided for situations where the assumption of normality is not appropriate. When their assumptions are appropriate, parametric tests are generally more powerful than theirnonparametric equivalents, although nonparametric tests often compare quite well in performance. The parametric versions test hypotheses concerning the group means. The nonparametric procedures test central value hypotheses based on measures other than the mean. Copyright 2000 Analytical Software --32-- STATISTIX 7.0 ALLDOSESD22, 10/17/02, 2:28:50 PM SCHEFFE COMPARISON OF MEANS OF TRIMPLANT BY DOSE F, DOSE MEAN HOMOGENEOUS GROUPS 10 8.0142 I 1 8.0054 I 30 7.9451 I 0 7.9304 I 3 7.9130 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. pp. 33 and 35 show the numbers of implantations (transformed because they are counts) are essentially the same across treatment groups. --33-- STATISTIX 7.0 ALLDOSESD22, SCHEFFE COMPARISON OF MEANS OF IMPLANT BY DOSE 10/17/02, P 2:27:40 PM DOSE MEAN HOMOGENEOUS GROUPS 10 15.621 I 1 15.593 I 30 15.345 I 0 15.286 I 3 15.241 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.438 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --34-- STATISTIX 7.0 ALLDOSESD22, 10/17/02, 4:03:39 PM SCHEFFE COMPARISON OF MEANS OF TRIMPLANT BY DOSE DOSE MEAN HOMOGENEOUS GROUPS 1 8.0345 I 10 7.9177 I 30 7.8713 I 0 7.8501 I 3 7.8387 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.404 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --35-- STATISTIX 7.0 ALLDOSESD22, SCHEFFE COMPARISON OF MEANS OF IMPLANT BY DOSE 10/17/02, F' 4:05:19 PM DOSE MEAN HOMOGENEOUS GROUPS 1 15.709 I 10 15. 2 98 I 30 15.086 I 0 15.036 I 3 14.982 I THERE ARE NO SIGNIFICANT PAIRWISE DIFFERENCES AMONG THE MEANS. CRITICAL F VALUE 2.404 REJECTION LEVEL 0.050 STANDARD ERRORS AND CRITICAL VALUES OF DIFFERENCES VARY BETWEEN COMPARISONS BECAUSE OF UNEQUAL SAMPLE SIZES. --36--