Document rB6dOm55GLLzO6ggk57Nv3xDJ

CC: E. I. d u Po n t d e , Ne mo u r s 51 Co mp an y lNtO.*?r*OHAYF.D Wil min g t o n , De l a w a r e w s s o ENGj NEERI MG DEPARTMENT G J , Stopps - Haskell Laboratory R. S4 Waritz - 11 " A. Azar "" P. E. Smith - .. 11 *' D, R. Diggs/J, J..Mikita - Wilm. W. E, Bettoney/ A. J. Pahnke - Pet, Lab E, N. Cantwell/K. Habibi -" r J, E. Riegel/W, E, Morris - 11 11 J. H Whelan - Louviers P. J. Peltier- " R. W, Kennard- " D, W, Marquardt- M D. H. Doehlert " C. M. Drummond " RC-13 - Math, and Stat, Dev, December 9, 1970 P. D. GRIFFITH HASKELL LABORATORY EFFECTS OF LEAD AND CARBON MONOXIDE ON BLOOD ALA DEHYDRASE This letter summarizes our discussion on the results of your lead-carbon monoxide study. The experiment was conducted pri marily to determine the effects of lead and carbon monoxide on blood ALA dehydrase in rats. Five other measurements were also' taken; DALA, osmolality, hematocrit, hemoglobin, and carboxy hemoglobin. \ The analysis indicated that.ALA dehydrase decreases in the pres ence of lead and increases in the presence of carbon monoxide. It further showed that each of the other quantities measured is affected by lead, carbon monoxide, or both, A description of these effects is contained in the discussion of the analysis, which is attached. ENGINEERING SERVICE DIVISION Mary E. Finan Mathematics and Statistics Development Ronald D, Snee Applied Statistics MEF/RDS:kk Attach. DEC HASKil N40749 K, *1, E$XPERIMENTAL DESIGN . Four groups of ten rats each were included in the experiment. The first group served as a control. The other three groups were exposed to lead and carbon monoxide as follows: group two: - lead alone; group three: carbon monoxide alone; group four: lead and carbon monoxide. The test was conducted over a four-week period. Five complete sets of measurements were taken: one prior to treatment, and the other four at weekly intervals.. The quantities measured, in addition to ALA dehydrase, were: DAIA , osmolality, hematocrit, hemoglobin, and carboxy hemoglobin. In addition, the weight of each rat was recorded at regular intervals throughout the four weeks. 2. ANALYSIS The data constituted a partially nested and crossed factorial design and were analyzed via analysis of variance. In the . case of ALA dehydrase, data were transformed to logarithms prior to analysis to ensure that the variability would be independent of the response. These analysis of variance tables for each response are included in Appendix B. In all cases, pretreatment data were' analyzed separately, as a one variable design, with each lead-carbon monoxide com bination considered one treatment. 3. RESULTS A. Pretreatment Data For ALA dehydrase, DALA, osmolality, and hematocrit, there is no difference among average response for the four treatment groups. Differences are apparent, however, in the other two responses: the average hemoglobin measure ment for the control group is lower than for the other; the average carboxy hemoglobin measurement for the same group is higher than for the others. See Table A for a listing of the mean values and the least significant differences between those mean values. B. Growth Data W Neither lead nor carbon monoxide had any consistent ^ effect on the weights of the rats over the period of experimenta tion. On day 17, the lead/carbon monoxide group showed a slightly higher average weight than the others: on day i 19, the control group was slightly lower than the others. The major effect seen was a significant weight gain over time. Table gives average weights for each treatment ' group on the days when these measurements were taken; Table B2 gives the average weight" of all forty rats on the days In question* C. Treatment Data Effects of lead and carbon monoxide upon each of the six responses are described below. Tables of means and graphs of significant interactions appear in Appendix A.. N40749.01 DUP050058506 Page 2 December 9, AM 1970 1. ALA-DEHYDRASE shows a significant decrease In the presence of lead, and a significant increase in the presence of carbon monoxide,- There is also a time effect. The weekly responses, averaged'over all groups, are different; however, here, as in the'cases to follow, there is no. trend apparent. The largest ' value occurs at week 1; the smallest, at week 3, The lead x week interaction is significant: the non-lead groups show a fairly sharp drop in ALA dehydrase over the first three weeks, and. an increase in the fourth week. The lead group measurements remain fairly stable throughout the period. (See Figure 1.) 2. HEMATOCRIT increases with carbon monoxide; there is no effect due'to lead. There is a difference among weekly measurements: week 2 is significantly higher than the others. There is also a CO x week interaction: in the fourth week, the non-CO groups show a decrease in hematocrit, while the CO groups show .an. in crease, (See Figure 2.) 3. DALA increases in the presence of lead and shows no change in the presence of carbon monoxide. Again, there is.a time effect: the-average measurement in the second week is higher than the others 4 OSMOLALITY decreases with carbon' monoxide; there is no effect due to lead. 5 HEMOGLOBIN increases in the presence of carbon monoxide and shows no change In the presence of lead. There is a time effect: Y the measurements in the second and fourth weeks are higher than the others. No significant interactions are evident in the data collected over the four-week period. How ever, the hemoglobin measurements taken prior to the.experiment indicate that the control group averaged lower than the other three. A comparison of the pretreatment averages for the four groups with the fourth week averages shows that the hemoglobin inersase for the lead group is substantially smaller than that for the other three, (See Figure 3.) GROUP PRETREATMENT FOURTH WEEK d if f er s : Control Lead CO Lead/GO 14.07 15,04 14.96 14.62 ' 15.51 15.39 16.33 16.06 1,44 .35 1.37 1,24 The average difference for the lead group is significantly different from that for the control; those for the CO and lead/CO groups are not different. This indicates that an interaction is present after all: lead alone supresses hemoglobin; lead in combination with carbon monoxide does not, . 6. CARBOXY HEMOGLOBIN decreases in the presence of lead and increases in the presence of carbon monoxide. There is an effect due to DUP050058507 par:e 3 pocember 9, 1970 .time. Average measurements in waeks 2 and 3.are lower than those in weeks 1 and 4* The lead'x CO interaction is significant: lead decreases carboxy hemoglobin in the presence of carbon monoxide. Diff erences in the initial values'of carboxy hemoglobin (high values in the control group) do not affect the interaction appreciably. (See Figure 4*) A CO x week interaction is also present, but small. (See-Figure 5.) DUP050058508 APPENDIX A 1. TABLES OF MEANS 2, GRAPHS OF SIGNIFICANT INTERACTIONS DUP050058509 TABLE A : AVERAGE PRETREATMENT RESPONSES BY GROUP # 4 o n O O ON On Pi CO IS O -4 NO ON <N Pi o n O CvJ to O O ..4 on ' . d CD > H bO 0) P O -4 O O Os* O 0 O HI O H to ON 0 . u\ O H UN 4 O pT on -4* O rH < * to IS Pi rH CO X5 CD 0) O H 0 P rH rH rH CD *> CD H O 4 O O \o O UN C^ r~i vO vO QN 4 O 0 . * 0 0 0 H O cv vO 4 O o n 4- 4 rH to > rl CD & P P Ctf O 4 O P 4' O ON H UN H O rH PI o.r 0 4 1--I O CN2 vO UN rH 3 ON Pi 4 B rH H O 4O O O to H rH ON O O O * . .. : 04 O O rH UN 4* ON E-* ON 4 4 rH to O O rH d H HO 0) Pi CO 'CO cd 5 d O xf b0 O & P *>> d 0) H P rH Pi Pi CO ,d d H ,0 0 rH O W w CD 04 p toO <d rH P rH 0 < <d O Q & 4 Pi d & d d Pi 4 CD CO 0) <d Q p : O p-i p rH c\* ON 4* UN vO cod dbHO CO CO H CO r i <H O d9 rl CD G$ O P*d CD d<D d0)<CPh CD *H P CD 43 ^d 0O O *rH d<H CD rrl d0 R bO p *H <H CO H X* P CO Q> <d ,d CD IH rH * /01 6/z x DUP050058510 TABLE Bl: AVERAGE WEIGHTS BY TREATMENT GROUP MEASUREMENT DAY 4 7 9 12 14 17 19 CONTROL 252.30 290.00 306.50 316.SO 329.10 349.60 362.70 LEAD 250.90 291.50 310.00 317.10 32S.40 353.20 370.SO CO 251.10. 291.00 305.90 31S.90 329.00 353.20 374.30 LEAD/CO 253.30 294.50 310.70 323.20 335.60 360.60 3SO.50 LSD 6.66 6.66 6.66 6.66 6.66 6,66 6.66 TABLE B2: AVERAGE WEIGHT BY DAYS SUREMENT DAY AVERAGE WEIGHT 4 251.90 7 291.75 9 306.2S 12 319.00 14 330.53 17 354.15 19 372.0S LSD 3.33 12/9/70 rr* DUP050058511 RESPONSE ALA Dehydrase Hematocrit BALA ' Osmolality Hemoglobin Carboxy Hemoglobin TABLE Cl: LEAD MEANS* WITH LEAD [ A. 6l** ' 46.09 \ .20** 1,345.35 1 15.26 j 3.59** WITHOUT LEAD 31.93 45.91 .15 1,371-53 15.49 4.51 LSD 1.35 1.09 .02 139.32 .32 .32 . TABLE C2: CARBON MONOXIDE' MEANS* . RESPONSE ALA Dehydrase Hematrcrit DALA Osmolality Hemoglobin Carboxy Hemoglobin WITH CO > / 21.56*4 / 47.03** ' / .13 ' (1,750.3 3** ! 15.71** 5.20-r* WITHOUT CO 14. >:' 44 'so .16 1,967.00 15.05 2.90 LSD 1.35 1.09 .02 139.32 l2 .32 * Each figure is the average measurement for twenty rats over the four-week period. * ** Significant difference between means at the .05 level 12/9/70 DUP050058512 TABLE C3: WEEK MEANS* RESPONSE ALA Dehydrase Hematocrit DALA Osmolality Hemoglobin Carboxy Hemoglobin WEEK 1 2 25.76** 16.53 4496** 47.20 .17** .20 1,671.06 1,669.35 . 15.20** 15.69 4.50** 3.70 3 12.90 45.76 .17 1,626.95 14.60 3,35 k -15.67 46.05 .16 1,645.36 15.62 ' 4.65 LSD 1,44 .69 .02 119.99 .22 ,75. * Each figure is the average measurement over all forty rats. ** Significant difference among means at the ,05 level. 12/9/70 DUP050058513 DUP050058516 APPENDIX B ANALYSIS OF VARIANCE TABLES: 1. ALA Dehydrase 2. DALA 3. Hematocrit 4. Osmolality 5. 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