Document zo99Qjg2Qj3q6Rn6kJKbpjZg6

LEAD PAINT INGESTION STUDY FINAL REPORT 25 February 1974 Contract No. 62-W-62GC&NPC MR! Project No. 3729-6 For National Paint and Coatings Association 1500 Rhode Island Avenue, NW. Washington, D. C. .20005 DUP040008876 PREFACE This report was prepared at Midwest Research institute, 425 Volker Boulevard, Kansas City, Missouri 64110 under Contract No. 62-W-62GC & NPC, MR1 Project No. 3729-B, "Lead Paint Ingestion Study." The Research was sponsored by the National Paint and Coatings Association, 1500 Rhode Island Avenue, N.W., Washington, D. C. 20005. Royal A. Brov/n, Technical Director, National Paint and Coatings Association was the project monitor. The research was conducted in the Biological Sciences Division, under the direction of Dr, W. B. House from 1 December 1972 through 31 July 1973. Dr, Thomas R Castles, Principal Pharmacologist, was the principal investigator, assisted by Dr, Jaime Sanyer, Associate Pathologist, and Mrs. Jane Hoch, Biology Research Assistant, Dr. James L, Spigarelli, Senior Chemist supervised the lead analysis with the assistance of Mrs. Hope M. Miller, Assistant Chemist, Members of the National Paint and Coatings Association Industrial Metals Task Force prepared the paint chips and consulted with Drs. Castle, Sanyer, Spigarelli and House during the course of the study. The personnel of the NPCA Industrial Metals Task Force is as follows: Richard A, Moore Royal A, Brown John P, Ffawley Charles M. Jackson Joseph G. Kingston Sidney Lauren William W. Ringle Edwin E. Swain Jean P. Teas Domenic J, Tessari The Sherwin Williams Company - Chairman The National Paint & Coatings Association Hercules, Inc. Celanese Coatings Co, Glidden-Durkee Division of SCM Corporation Coatings Research Group, Inc. Pratt and Lambert, Inc. E, I. duPont de Nemours & Co,, Inc. The Flood Company De Soto, Inc. Approved for: MIDWEST RESEARCH INSTITUTE W. B. House, Director Biological Sciences Division 25 February, 1974 11 DUP040008877 TABLE OF CONTENTS Page Introduction . , . , e e a .- 9 .. ' .. 1 II Methods .9 o eop e o A Preparation of Paint Films Containing Different Concen trations of Lscid .0 .0 0 P 9 * . .9 . * o p ; o O Bp Preparation of Diets . . * C Experimental Procedure ** . . D Ana lyses *..*.**#.# E 0 Pathology * . . o * > . * * F,u Statistical ...*> . # ..* 1 2 2 3 5 6 111 Results ,..* p .o60 .90V 6 A, Concentrations of Lead in Paint Films B, Lead Analyses of Diets Containing Different Paint Films , Co Preliminary Feeding Studies . . ,, . , . .. , Da Gross Observations, Feed Consumption and Body Weights. E. Weekly and Total Lead Consumption F,, Hematology of Rats Fed Paint Films Containing Different Concentrations pf Lead Go Tissue and Fluid Chemistry of Rats Fed Paint Films Containing Different Concentrations of Lead...... H. Lead Content in Tissues of Rats Fed Paint Films Containing Different Concentrations of Lead ..... . I. Pathology of Rats Fed Paint Films Containing Different Concentrations of Lead 6 <5 9 9 14 14 14 36 51 1V. D is c u s s i on ....... ... .... . . ....... . . .. 50 V o Conclusion * 59 References . . 60 Hi DUP040008878 I, INTRODUCTION Few persons will disagree that the ingestion of old lead-based paint presents a , potential health hazard to young children. As a result of reports by Kehoe,-/ Chisholm/--' and King,:/ medical researchers, paint manufacturers and legislators have recognized the seriousness of this problem and are acting to establish safe concentrations of lead in paipf. To do this, they have voluminous reports on clinical observations and animal research from which to draw. Unfortunately, most of this information is based upon the old white lead paints or soluble lead salts, which are not representative of the paint formulations on today's market. Some studies have been reported ^ ~ which indicate that lead compounds are not as readily leached from modern paint films. Before safe concentrations of lead in paint can be intelligently established, we must evaluate the toxicity of the lead as it exists currently In modem paints. It was the purpose of this study to evaluate the toxicity in rats produced by different concentrations of modern paints containing lead octoate and lead chromate at varying concentrations compared to an aid paint formulation containing white lead. The results are presented in the following pages. II, METHODS A. Preparation of Paint Films Containing Different Concentrations qf Lead Paint films without added lead and with added lead compounds in the form of lead octoate, lead chromate, or lead carbonate were supplied by members of the National Paint and Coatings Association, Paints which contained lead octoate, and lead chromate (medium yellow) were prepared using a flat alkyd paing formula based' upon a Federal Specification TT-R-266D Type ! Class B alkyd resin. A white lead in oil paint was prepared from a formula typical of this type paint as it was used in the 1920's, This contained a high percentage of lead carbonate. The flat alkyd paint without added lead was used for control. The percent of lead in each sample was calculated on the basis of the nonvolatile materia!. Dispersion was accomplished with a Cowles Dissolver and the pigment volume concentration was held at one level. The paints were placed in pans and allowed to air dry. This was followed by forced air drying at 120F to volatilize any remaining solvent. Paint films were ground and sieved, and chips ranging from 0.5 to 1.0 mm In size were used for this study. This chip size was selected on the basis of a preliminary rat feeding study which established that rats would selectively eat around larger sized chips but would eat chips in the selected size range. The concentrations of lead in the different paint films were determined independently by De'Sota Incorporated, the Sherwin-Williams Company and Midwest Research Institute, using Atomic Absorption Spectrophotometry , 1 DUP040008879 Twenty male and 20 female rats were assigned to the control diet, and 10 males and 10 female rats were assigned to each of the remaining diets. Each diet was fed in a 9-az wide-mourh glass far which was secured to the cage. Tap water was available ad libitum during the entire experiment. Feed consumption was measured twice each week and calculated on a daily basis. The animals were weighed once a week. All rats were observed twice or more each week for toxic signs. After 4, 8 and 13 weeks,r a selected number of male and female rats from each group were placed in metabolism cages for the collection of 24"hr urine samples, A portion of the collected urine was used for urinalysis and the remainder frozen for assay of delta-aminolevulinic acid and coproporphyrin, After urine collection each rat was anesthesized with ether, exsanguinated via the abdominal aorta and bone marrow smears were prepared. Blood samples were heparizined, coded and used immediately for hematology and enzyme assays. Aliquots of the remaining blood were taken for protoporphyrin and lead assays. After urine and blood samples were obtained * each ret was neeropssed and tissue taken for lead analysis or microscopic examination. D 1 , Lead analyses of paint ehips and diets containing paint chips; a. Paint film: Ten-mill igram samples of paint chips containing no lead or 0.08% lead octoate were charred with 3 ml of concentrated nitric acid and dry-ashed at 500 C for 2 hr. The ash was dissolved in 1 ml of aqua regia, diluted to 5 ml with distilled water and this final dilution measured for lead content by atomic absorption. All other paint chips (10 mg samples) were digested In 15 ml of a mixture of concentrated nitric acid and 37% perchloric acid (2;'l, V/V), evaporated and the remaining perchloric acid solution diluted to 25 mi with distilled water. This final dilution was used for atomic absorption spectrophotometry, b. Diets; One-gram samples of feed were digested in a mixture of concentrated nitric acid and 37% perchloric acid (2:1, V/V) and their final dilutions adjusted to make their lead concentrations within the detection limits of the atomic absorption technique. 3 DUP040008880 A sample of old lead-based paint collected from walls of old dwellings was obtained from the U. S. Department of Commerce/ National Bureau of Standards, Washington, D.C. This paint was intended as an analytical standard and had been pulverized and sieved through 325 mesh screen (0.06-0.08 mm size) and contained 11.92% lead by weight as carbonate. This sample will be referred to as "NBS lead paint" in this report. B. Preparation of Diets y Diets were prepared by mixing paint chips with Purina Rat Chow mash In the concentration of 0.1% (weight/weighf). This was aecornplSshed by first preparing a 10% concentrate (paint film/feed, w/w) in a high-speed twin shell mixer. This concentrate was then added to the appropriate amount of Purina Rat Chow mash to give a final concen tration of 0,1% (paint film/feed, w/w) and mixed for 10 minutes in a bulk mixer. Each diet was prepared three times during the experiment and samples of each of these preparations were assayed for lead content, C, Experimental Procedure Two hundred weanling (40-60 gm) Charles River rats (100 males and 100 female rats) were used far this study. Upon arrival rats were housed individually in air conditioned quarters ip polycarbonate cages containing hardwood bedding and filter tops. After 5-7 days of equilibration, all rats were assigned one of the following diets. 1. Rat chow plus paint chips without added lead (control) 2. Rat chow plus paintchips containing 0,08% lead as lead octoate 3. Rat chovv plus paint chips containing 0.53% lead as lead octoate 4. Rat chow plus paint chips containing 2.05% lead as lead octoate 5. Rat chow plus paint chips containing 0.42% lead as lead chromate 6. Rat chow plus paint chips containing 1.95% lead as lead chromate 7. Rat chow plus paint chips containing 12.43% lead as lead chromate 8. Rat chow plus paint chips containing 66.05% lead as lead carbonate 9. Rat chow plus NBS lead paint containing 11,92% lead as carbonate 2 DUP040008881 I3 (5) Urinary coproporphyrirr, Urinary coproporphyrms were determined by the method of Schienker and Kitchell These analyses were performed in subdued light. Cm Urinalysis' (1) Urinary protein (albumin): Urinary protein was measured with "Uristix" reagents strips (Ames Company, Elkhart, Indiana). (2) Microscopic examination of urine; Urine samples were centrifuged, the residues resuspended, and examined microscopically for the presence of erythrocytes and leukocytes under high power field and for Casts under low power field. d. Tissue lead; (1) Blood: One milliliter of blood was mixed with 1 ml of a mixture of 5% trichloroacetic acid: 37% perchloric acid (3:1 v/v). The sample was centrifuged and the supernatant filtered through an AAWP 0.8ju Millipore filter. The supernote Was analyzed for lead with an atomic absorption spectrophotometer using standards prepared in control rat blood;, (2) Other tissues: Bone, liver, kidney, and brain were digested in concentrated nitric acid, evaporated to dryness and reconstituted to the lowest possible volume with 20% nitric acid. These solutions were analyzed for lead by atomic absorption spectrophotometry. (3) Atomic absorption analysis: Lead concentrations were measured using a Varian-Techtron AA-5 atomic absorption spectrophotometer. Sample solutions were aspirated into an aif-aCetylene flame and the absorbance was measured at 283.3 nm. Background interference was determined with the use of a hydrogen continuum lamp at 283.3 nm and subtracted from the absorbance obtained at 283.3 nm with the Pb hollow cathode lamp, ^ E. Pathology 1. Gross pathology: At necropsy, rats were examined for gross abnormalities, their livers, kidneys, spleens, hearts, gonads, thyroids, brains, and adrenals weighed, and the relative organ weights calculated. After weighing, a portion of the liver, kidney and brain were taken for microscopic examination and the rest of each organ was measured for lead content. A femur was removed to be used for the measurement of lead in bone. 2. Microscopic pathology: Bone marrow smears were prepared for myeloid/erythroid cell count. The following tissues were fixed in buffered neutral 10% formalin: brain, liver, spleen, stomach, small intestine (duodenum, jejunum, ileum), 5 DUP040008882 20 Analyses performed upon each rat; a. Hematology: (1) Hematocrit: Hematocrit was determined in capillary tubes using a microcdpillary centrifuge (International Equipment Company, Bode! MB). (2) Hemoglobin: Hemoglobin was measured as cyanomethemoglobin.-13/ (3) Erythrocyte and leukocyte counts: Total erythrocyte and leukocyte were counted using a Coulter Electronic Particle Counter with IGO-j j aperature (4) Reticulocytes: Reticulocytes were counted by the methylene blue method using the Miller disc;-- (5) Differential leukocyte counts: Wright's stain was used to stain the leukocytes for examination. (6) Erythrocyte osmotic fragility; Osmotic fragility of erythrocytes was quantitatively determined by subjecting heparinized whole blood to sodium chloride solutions of different osmolarities,--' The concentration of sodium chloride which hemolyzed 50% of the erythrocytes was obtained from a plot of percent hemolysis versus sodium chloride concentration. b,, Body fluid and tissue chemistry; (1) Plasma protein electrophoretic patterns: Total plasma protein was determined using the Hycel Biuret Reagent (Hycel, Inc., Houston, Texas), The quantity of each plasma protein was determined electrophoretically On cellulose acetate and expressed as a percentage of the total plasma protein. (2) Er/throcyte-6-aminolevulmic acid dehydrase (ALAD): 6-Aminolevulinic acid dehydrase activity was determined by the method of Lichtmah and Feldman.-- This procedure was started 30 min. after each blood sample was taken. (3) Erythrocyte protoporphyrin: Protoporphyrin in erythrocytes was measured by the method of Heller, et alju This analysis was performed the day after blood was withdrawn, (4) Urinary 6-aminolevulmic acid (ALA^/ Urinary &-arnioo)evulinic acid was measured according to the Davis and Andelman-^= modification of MauzeraU's and Granick's Method.---^ These analyses were performed in subdued light. 4 DUP040008883 Paint . Film Control Paint Lead octoate Lead octoate Lead octoate Lead chromate Lead chromate Lead chromate Lead carbonate NBS lead paint TABLE 1 CONCENTRATION OF LEAD IN PAINT FILM Theoretical Lead Concentration m 0 0,06 0.50 2,00 0.50 2.00 15,00 64,12 - Measured Lead Concentration Paint Companies MRI Average**/ ............ m ____ . 0.01 0.08 0.50 1,90 0,42 1.80 12.57 63.20 . 11.87--^ < 0,02 0.07 0.56 2.20 0.41 2.10 12.30 68,90 11.98 0.01 0.08 0,53 2.05 0.42 1.95 12,43 66.05 11.92& a/ Measured by tj.S. Department of Commerce, National Bureau of Standards, Washington, D.C. b/ Average of paint companies and MRI values. cj Average of Bureau of Standards and MRI values. 1 DUP04C008884 colon, pancreas, kidneys, urinary bladder, adrenals, thymus, gonad, thyroid (with parathyroid attached), mesenteric salivary gland, lymph nodes, heart, lungs, diaphragm, skeletal muscle, and prostate or uterus. Tissues from all 13 week rats fed the control, 2a05% lead octoate, 12.43% lead chromate, 66,05% lead carbonate, and the 11.92% NB5 lead paint diets were embedded in paraffin, section to a thickness of 6 /j , stained with hematoxylin and eosin and examined for histopatho!ogy. F. .Statistical; Analyses Control and treatment values were compared statistically using Dunnett's multiple-comparison test with P<0,,Q5 as the criteria of significance. Ill, RESULTS A. Concentrations of Lead in Paint Chips The concentrations of lead in the paint chips supplied by the Members of the National Point and Coatings Association were analyzed for lead content by DeSotp Incorporated, the Sherwfn-Williams Company, and Midwest Research Institute (MRI), The results of these analyses are shown in Table 1. The control paint which did not contain lead octoate or chromate was found to contain approximately 0.01% lead. There was a reasonable agreement between the different analyses, so on average ppm of lead was computed and used for the actual lead concentration of each paint film. Lead con centrations of the lead paint obtained from the National Bureau of Standards were averaged in a similar manner. B. Lead Analyses of Diets Containing Different Paint Films Table 2 shows the lead content of each diet. Purina Rat Chow mash was found to contain approximately 4jug/gm of lead (assuming 0.1 jug/gm of lead was contributed by the control paint), thepg of lead/gm of feed which was attributable to each added paint film was reasonably close to its respective theoretical value. The average total jug of lead/gm of feed for each diet was used for the calculation of weekly and total lead consumption for each rat. 6 DUP040008885 C. Preliminary Feeding Studies Before beginning this study, a pilot experiment was performed to see if the rats were consuming the lead paint chips along with their feed Three rats each were placed on the control diet and the 66.05% lead carbonate diet. After 48 hr^. their feces were collected and the lead content measured. Feces from rats on the control diet contained 11.5 jjg of lead/gm Feces while feces from rats fed the 66.05% lead carbonate diet contained 3,683 jug of lead/gm feces. Thus, rats did consume the paint film along with their feed. D> Gross Observations, Feed Consumptions and Body Weights During the entire study only two rats had remarkable changes. One female rat on 2.05% lead octoafe developed an abcess On her left shoulder during the ninth treatment week and one female on 66,05% lead carbonate developed csbcessed hind feet in the 10th week and chewed her foes off. The feed consumptions of male and female rats are shown in Figures 1 and 2, respectively . Each bar represents the mean standard error of six to twenty rats. Except for the fats fed the diet containing 11.92% lead as NBS lead paint, all groups consumed feed at the some rate as the control group throughout the entire experi ment. The reason thd 11.92% NBS lead paint group ate significantly less initially, was that they were started a month after the other rats and were initially smaller. Since this group was consuming feed at the same rate as the control group by 4 weeks, we consider their initial food consumption normal also (for their size). The body weight gains for each group of rats are shown in Figures 3 and 4. Each bar represents the mean t standard error of six to twenty rats. As mentioned above, the rats on the 11.92% NBS lead paint diet were small Initially but became similar to controls by either the 4th (females) or 8th week (males). Throughout the entire period the male rats in the other treatment groups gained weight at a normal rate. The female rats did show a change in body weights during the first 8 weeks of feeding which appears related to the concentration of lead actuate. After 4 weeks of feeding, rats In the 2,05% lead octoafe group weighed significantly Jess than control . The pattern was the sCme at 8 weeks. After 12 weeks their overage body weights still exhibited the same pattern, but the 2,05% lead octoafe group was not significantly different from control!. This was due to fewer animals and a slight Increase In variability. 9 DUP040008886 TABLE 2 LEAD ANALYSES OF DIETS CONTAINING DIFFERENT PAINT FILMS \ Diet Control paint plus rat chow Theoretical--^ ....lEg/gilL___ _ 0.1 Measured Totai/ Paint Lead--^ (hg/sm) fog/gm) 4.1 (6) Lead oetoate (0.087.) plus rat chow 0.8 5.2 yl.i (6) (6) Lead oetoate (0.53%) plus rat chow 5.3 12,0 7.9 (6) 0) Lead oetoate (2,057.) plus rat chow 20.5 27.2 (6) 23.1 (6) Lead chromate (0.42%) plus rat Chow Lead chromate (1.95%) plus rat chow 4.2 19.5 11.0 (6) 23.5 (6) 6.9 (6) 19,4 (6) liead ehrbmate (12.43%) plus rat chow t. : ri ' > : Lead carbonate (66.05%.) plus rat chow NBS lead paint (11.92%) plus rat chow 124.3 660.5 119,2 140.2 (6) 516,0 (5) 124.0 (3) 136.1 (6) 511.9 (5) 119.9 (3) a/ Theoretical concentration of lead in feed contributed by indicated paint film. |>/ Mean pg of lead/gm feed for number of samples shown in parenthesis. cj Mean ,ug of lead from paint films/gm feed for number of samples shown in parenthesis 8 -- DUP040008887 (Xop/)Dj/uiS) MOlidWnSNOD Q33J n TREATMENT WEEK DUP040008888 TREATMENT WEEK DUP0400038S9 Control' Bodyweights of Female Rats Fed Different Concentrations o f Lead in Paint Chips TREATMENT WEEK Zl Rodyyiieights of Male Rats Fed Different Concentrations of Lead' Irv Paint Chips. TREATMENT WEEK WEEKLY AMD TOTAL LEAD. CONSUMPTION BY HALE RATS FED P A IM CHIPS DUP040008892 E. Weekly and Total Lead Consumption The weekly aMd (ofaI lead consumption during the experiment were calculated on the basis of the total lead/diet and amount of each diet eaten by each rat. These calculations are shown in Tables 3 and 4. During the 13th week period, the control group consumed approximately 105pg (males) or 79 pg (females) of iead/day, while rats eating 2,05% lead octoate, 12,43% lead chromate, 66,05% lead carbonate, or 11,92% NBS lead paint consumed an average of 787pg (males) or 708 pg (females); 3,591 pg (males) or 2,689 pg (females); 13,436 pg (males) or 9,185 pg (females); and 3,184 pg (males) or 2,363pg(females) of lead/day, respectively. Using the average body weights of rats surviving 13 days, the rats in the control, 2,05% lead octoate, 12,43% lead, chromate, 66,05% lead carbonate and the 11.92% NBS lead paint groups consumed 288pg (males) or 335 pg (females) of lead/kg/day; 2,133pg (males) or 3,278 pg (females) of lead/kg/day; 9,785pg (males) or li,590pg (females) of lead/kg/day; 37,014pg (males) or 42,133pg (females) of iead/kg/day; and 9,888 pg (males) or 10,364 pg (females) of lead/kg/day, respectively. F. Hematology of Rats Fed Paint Chips Containing Different Concentrations of Lead The results of the hematology of rats fed pain films containing different concentrations of lead are shown in Tables 5-13. No differences were observed in the erythrocyte count, reticulocyte count, hematocrit, leukocyte count, differential leukocyte count, and the erythrocyte osmotic fragility.. At 13 weeks, the hematocrits of rats fed the NBS lead paint v/ere significantly lower than control, while the hematocrits of the rats Fed the other diets were normal. G. Tissue and Fluid Chemistry of Rats Fed Paint Chips Containing Different Concentrations of Lead Tables 14"22 show the tissue and fluid chemistry of rats fed paint chips containing different concentrations of lead.. The serum proteins, erythrocyte protoporphyrin:, urinary coproporphynn, and urinary deito-amInolevu11nic acid were not altered by any of the diets. The activity of the erythrocyte enzyme, delta-aminolevulinic add dehydrase (ALAD), In the control, lead octoate (0.08%, 0,53%, and 2.05%) and lead chromate (0.42%, 1,95% and 12.43%) groups did not differ throughout the experiment (Tabies'23), ALAD activity was depressed in rats fed 66.05% lead carbonate and 11,92% NBS lead paint. At 4 weeks: (Table 21, 22) there was a depression of 56% and 59% in these twb groups, respectively. At 8 weeks the ALAD activity was depressed 44% in the 66.05% lead carbonate group, but the ALAD activity in the NBS lead paint group was not significantly different from control. At this time a sex difference was observed In both of these groups. The ALAD activity in the male rats remained depressed at the 4-week levels while the ALAD activities of the female rats returned to control levels. This sex difference may be linked to the fact that these female rats had matured and were probably beginning their menstruai cycles. 14. DUP040008893 TABLE 5 HEMATOLOGY OF RATS. FED PAINT CHIPS CONTAINING NO LEAD Analv:ses 4 ,ftt*8W Treatment Seek 8 MI Erythrocytes (x 10/mm-') Reticulocytes, %> Hematocrit, y:ol % Hemoglobin, gm % Leukocytes (x LO^/mm^) Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility [NaCl]./ 6,17 * 0.14^ 1,5 0.7 46.1 * 0,8 14.0 0,3 5.0 * 0.5 11,8 2,9 86,3 3.3 0,3 0,2 0,9 0.4 0 1.1 0.6 0 0 0,381 0.004 6.33 0.19 0,8 0.3 44.6 1.2 14,6 0.3 7.6 1.3 21.7 5.6 75.4 5.3 0 1.3 0.4 0 1,6 0.6 0 0 0,391 0.0,08 13 (N=24) 6.42 0.29 1.4 0,1 44,6 0,3 16.0 0,1 8.0 0,6 13,1 1,5 84,1 1.6 0.1 0,1 0.5 0.2 0 2.1 0,4 0 0.1 0.1 0.400 0.004 a/ Number of rats per period* b/ Mean standard error, c:/ . Concent rat ion of NaCl that hemolyzed 50% of the erythrocytes. 17 DUP040008894 "(9 O U a Pa tO uO qs3. ap am mV i1-1 Oa u 4> '>S '0 Ho 16 DUP040008895 TABLE 7 HEMATOLOGY OF PATS FED PAINT CHIPS CONTAINING 0.53% LEAD AS LEAD (3CTOATE Analyses fN=4)^ Erythrocytes {x IQ^/mm-^) Reticulocytes, % Hematocrit, vol. % Hemoglobin, gw % Leukocytes (x 10 Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility [NaCl]/ 6,27 * 0. 1 0,9 0,7 42.3 0.7 14,2 0,2 5.0 1-4 . 15.8 3.8 83.0 4,4 0 0.8 0.3 0 0.5 0.5 0 0 0.383 0,006 Treatment Week 8 (N=^a 6, 04 i 0.31 0.9 0,4 43.8 i 0.5 14,9 * 0.3 , 7.5 2.8 19.3 3.8 77.5.4 3,8 0 1.0 * 0,4 0 2.3 0.5 0 0 0.388 0.012 13 (N-12) 5.6 * 0.4 1.6 X 0,2 43 0.7 15,3 0,3 - 8,3 i: 1,0 16.2 1,9 81,4 i 1.8 0,1 0.1 0.8 0,3 0 1,6 0.3 0 0 0.401 0.006 a/ Number of rats per period, Jb/ Mean standard error, cj Concentration of NaCl that hemolyzed 50% of the erythrocytes. 19 DUP040008896 TABLE 6 HEMATOLOGY OF RATS FED PAINT CHIPS CONTAINING 0.08% LEAD AS LEAP QCTOATE Analyses 4 (N=4'J--' Treatment Week 8 (N-4) 13 (N=12) Erythrocytes (x lO^/mm^) Reticulocytes, % Hematocrit, vol. % Hemoglobin, gm % Leukocytes (x lO-^/mm^) Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated BBC, % Erythrocyte osmotic 6.14 4 O.32V 0,9 4 0,5 41.3 4 0,9 14.1 4 0,4 5,1 4: 0.8 24.5 4 1,6 74,3 * 1,8 0 0.8 * 0.3 0 0.5 4 0, 5 0 0 0.381 * 0,005 6.33 4 0.09 1.0 * 0.5 46.0 4 0.7 15,4 4 0.2 5.5 4 0.7 20,3 4 4,7 76.3 * 4.7 0 1.5 4 0 ,6 0 2,Q 4 0.8 0 0 0,410 4 0.007 5.92 0.39 1.4 4 0,2 43.0 4 0.5 14.7 4 0.8 7,7 4 0.9 16.3 4 1,7 80.9 4 1.8 0.3 4 0.2 0,7 4 0.3 0 1,9 4 0.5 0 0 0,395 4 0.005 a/ Number of rats per period, b/ Mean 4 standard error. c/ Concentration of NaCl that hemolyzed 50% of the erythrocytes. 18 DUP040008897 TABLE 9 HEMATOLOGY OF BATS FED PAINT CHIPS CONTAINING 0.42% LEAD AS LEAD CHROMATE Analyses 4 (N=4V- Treatment Week. .8 OL-4) 13 (N=12) Erythrocytes (x lO^/inxn^) Reticulocytes, % Hematocrit, vol, % Hemoglobin, gm % Leukocytes (x Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility, [NaCl]^ 6.71 0,16^ 0,9 0,8 43,3 0.6 14.7 0.4 5,1 0.6 8,5 1,7 90,3 -db 2,0 0 0 0 1.3 i: 0.8 0 0 0.383 0,003 6.18 0.16 0,8 0.2 45,0 1,1 15.1 0,3 5,1 0.5 1.9,0 2,0 79.5 1.3 0 0.8 0.8 0 0.8 0.8 0 0,3 0.3 0.395 0.12 6.6 0,3 ,1.6 0,2 43.6 0.7 15.6 0.3 6.9 0.7 14,3 2.1 83.4 2.0 0 1.2 0.4 0 1.2 0,4 0 0 0.392 0.007 3/ Number of rats per period, except where indicated otherwise b/ Mean standard error* c/ Concentration of NaC1 that hemolyzed 50% of the erythrocytes. 21 DUP040008898 TABLE 8 HEMATOLOGY OF RATS FED PAINT CHIPS CONTAINING 2.05% LEAD AS LEAD OCTQ&CE Analyses c. 'O Erythrocytes (x IQ /mnv ) Reticulocytes, % Hematocrit, vol 7. Hemoglobin, gits % Leukocytes (x 10^/mm^) Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility [NaCl]-- 4 xm*)&/ 6.30 i Q.2^ 1,1 dr 0.6 41.8 0,5 14.4 0.2 5,1 rb 1,1 13.5 dr 3,0 85,8 3. 2 0 0.8 j. 0.5 0 0 0 0 0.381 0.006 Treatment Week 8 (N=4) 6.46 0.08 0.7 0,1 45.0 dr 0,4 15,5 0,2 6.7 1,7 13,0 dr 2,1 85.0 dr 2.3 0 0,3 0.3 0 1.8 0.9 0 0 0.399 dfc 0.008 13 ( N= 12) 6,72 * 0,4.2 2,0 0.6 43,4 0,8 15,6 *0.3 8,1 * 0.7 14,1 * 2.6 82.8 2,8 0 0,9 0,3 0 2,2 * 0 ,7 0 0 0.393 * 0,006 aj Number of rats per period, b/ Mean! * standard error. cj Concentration of NaCl that hemolyzed 50% of the erythrocytes. 20 DUP040008899 TABLE 11 HEMATOLOGY OF RATS FED PAINT CHIPS CONTAINING 12.43% LEAP AS LEAD CHROMATE Analyses Treatment Week 4 8 13 (N= 12) Erythrocytes (x lQ^/mnP) Reticulocytes, % Hematocrit, vol, % Hemoglobin, gm % Leukocytes (x lO^/min^) Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated SBC, % Erythrocyte osmotic fragility ffxNt aoCl la]^c'/ 6.32 .0.14^/ 1,95 * 1,28 41.3 1.1 13.9 0.4 4.8 0.5 21.5 6.3 78.0 6.0 0 0.5 0,5 0 0 0 0 0.384 0.007 6.23 0,18 0.9 0.1 43,8 0.5 15,2 0,1 5,3 1,8 15,0 2.1 82.8 2.0 0 1.0 0 0 1.3 0.3 0 0.3 0.3 .0.385 0.010 6.61 0.38 1.6 0,2 44,1 0.7 15.8 0.2 7.6 0,6 12.8 1,6 84.8 1.7 0,1 0.1 1.0 0.4 0 1.3 0,3 0 0 0.396 0.006 a/ Number of rats per period, b/ Mean standard error. cj Concentration of NaCl that hemolyzed 50% of the erythrocytes. 23 DUP040008900 TABLE 10 HEMATOLOGY OF RATS BED PAINT CHIPS CONTAINING 1,95% LEAD AS LEAD CHROMATE Analyses Treatment Week 4 8 13 (N=A) (N=i2) Erythrocytes (x 10^/mnr*) Reticulocytes, % Hematocrit, vol, % Hemoglobin, gm % Leukocytes (x lO^/mm^) Neutrophils, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility, [NaCl]^ 6,17 0.05^/ 0.8 0,4 40,8 0.6 13,9 0,2 4.9 db 0.8 10.6 2,4 92,0 db 1.8 0 0.5 . 0,3 0 0 0 0 0.383 d: 0.007 6.21 0.13 0.8 0,3 44,3 0,3 15.3 0,3 6.1 1.2 18.5 : 6,6 80,5 6,6 0 0.3 * 0.3 0 0,8 0.3 0 0 0,385 0.007 6.37 0.52 1.2 0.2 44,1 0.7 16.2 0,4 8.2 1,0 11,4 2.3 85,9 2.2 0 1,5 0.5 0 1. 2 0.3 0 0 0.404 0.005 a/ Number of rats per period, bj Mean standard error. c/ Concentration of NaCl that hemolyzed 50% of the erythrocytes. 22 DUP040008901 TABLE 13 IiEHA'i'01 OGY 6E RATS FED KBS I.LAI) PAINT CONTAINING 11.92: LEAD Anal vsos ErYthrqcy.tes (x 10, . r.;n3 ) Reticulocytes, /, Hematocri t. vbi, % Hemoglobin, gm % Leukocytes (: : l-0J/nmJI Neutrophils, % Lymphocytes, 1 Bands, % Eosinophils, % Basophils, l. Monocytes, " Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility [NaCl.il 4 (W)2J Treatment: Week_____ _______ ,8 13 (N=4I ' Cl=12; 5.72 X 0,1.5-/ 2.5 X 0.8 39,7 i 0,8 13,6 * / 9.7 4- 1.4 9.5 2,1 90,2 dt 2.3 0.3 0,3 0 0 0 0 Q 1.389 0.006 4,49 0.83 1,0 n: 0.4 42.8 :b 1,1 14,7 0,3 6.7 JU 1.2 7,0 X 1.7 92,0 X 2, 1 0 0 0 1,0 0.7 0 0 0.412 X 0,006 5.63 0,34 1.3 - 0.1 42,9 - 0.4 14,7 = 0,1 7,0 =c 0.5 14,2 2,1 33.6 = 2,2 0. 1.4 0,4 0 0.8 0.3 0 0,1 i 0.1 0.408 = 0.005 aj Numbef of rats per period, except where indicated otherwise, b/ Mean standard error. cl Two rats. a/ Concentration of NaCl that hemolyses 304 of the erythrocytes. e/ Significantly different from control (P<0,05) as hown by Dunnett's multiple-comparison test following an analysis of variance. 41 i t > .j 25 V' 1 DUP040008902 Ta 8LE 12 HEMATOLOGY OF RATS FED PAINT CHIPS CONTAINING 66.05% LEAD AS LEAD CARBONATE Treatment, Week 4 ............... 8 Erythrocytes (x 106D/mn3r) Reticulocytes, % Hematocrit, vpl., % Hemoglobin;, gin, % Leukocytes (x 10-7mm ) Neutrophils,, % Lymphocytes, % Bands, % Eosinophils, % Basophils, % Monocytes, % Atypical, % Nucleated RBC, % Erythrocyte osmotic fragility [NaCl]-- 5.94 * D.56--^ 0,4 0,2 41.3 0.9 13.9 0.5 4,3 1.2 14.3 4,1 82.0 5,6 0 0,7 * 0.7 0 3,0 2,0 0 0 0.388 0.009 6.13 0.06 1.1 0.3 44.0 1.5 15.0 * 0,2 5.0 0.9 15.0 i 4.1 84.0 * 4.2 0 0.5 0.5 0 0.5 I 0.3 0 0 0,394 0.012 13 (N= 12) 6.60 0,37 2,9 * 1,3 44,7 1.2 15.6 0.6 7.7 0.5 15,2 2.6 82.7 2.7 0.2 0,1 0.9 0,3 0 1.0 0,3 0 0.1 0.1 0.397 0.005 aj Number of rats per period, b/ Mean; standard error. c./ Concent rat ion of NaCl that liemolyzed 50% of the erythrocytes. 24 DUP040008903 TABLE 15 TISSUE AMD FLUID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING 0.08% LEAD AS LEAD OCTOATE Analyses 4 (mmhW Treatment: Week 3 Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 40 4 IE/ 26 4 1 54 1 21 4 1 841 5.9 4 0,1 0,66 * 0.02 44 4 3 20 4 2 44 1 20 4 1 12 * 1 6.3 0.2 0,79 4 0.08 13 im 48 4 l 20 4 1 541 22 * 1 71 6.2 4 0,1 0.93 0.04 Erythrocytes ALAD, pmol. PBG/ Males: 100 ml BBC/hr Females: Total: Protoporphyrin, ng/ioo ml BBC 24.8 4 7.72/ 19.5 4 4,4/ 22.2 4 3.9 28.4 4 2,4 14,9 4 i.3/ 18.7 * 0,3^ 16.8 4 1.2 24,2 * 1.6 12.0 4 2.22/ 16.1 4 l.sJ/ 14.0 * 1,4 22.8 4 2.5 Urine Cqpropprphyrin, Ug/24 hr ALA, . jig/ 24 hr 1,2 * 0,2 86.4 4 16.6 4.8 4 2,4 107.6 4 34.1 2.9 4 1.4 35.0 4 6.7 a/ Number of rats per period, except where indicated otherwise, b/ Mean 4 standard error, c/ Two rats, d/ Six rats. 27 DUP040008904 TABLE 14 TISSUE AMD FLUID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING NO :LEAD Analyses 4 =8W Treatment Keek 8 13 01*241 Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 43 1^ 28 : i 5 .dfc i 20 i 6*i 5,4 0,1 0.76 0.03 43 2 23 : 1 5 1 20 1 10 1 . 6.2 0.2 0.77 0,07 48 db 1 21 1 Ml 21 1 6i 1 6.3 0,1 0.93 0.04 Erythrocyte ALAD, jimol. PBG/ Males: 23. 2 2.3S/ 100 ml BBC/hr Females: 16.6 * 3,1-/ Total: 19.9 2.2 Protoporphyrin, 19,6 1.6 Ug/IOQ ml RBC 15.1 2.3^/ 16.3 2. 2--/ 15.8 1.5 23.5 1.9 14.5 i 1,45/ 15,4 0.7--/ 15,0 1.0 29.2 2.0 Urine Cop roporphy rxn, Hg/24 hr ALA, Ug/24 hr 00 o H- 2.5 1.2 7,4 3.3 7,4 83.3 11.0 6.0 .1.9 43,4 5.0 aj Number of rats per period, except where indicated otherwise, b/ Mean standard error. cj Four rats. d/ Twelve rats. 26 DUP040008905 TABLfi 17 TISSUE AND BODY FI,HID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING 2.05% LEAD AS LEAD OCTOATE Analyses '4 ' fN~4)M.( Treatment Week S" (N?4) 13 (N=12) Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 41 * 3b/ 27 * 2 4 1 20 4 1 94 l 5.7 0,2 0,70 4 0.07 46 2 22 1 4& 1 19 l 9i 1 6,0 db 0.1 0,85 0.07 48 4 2 20 .4 1 641 21 4 1 641 6.2 4 0.1 0.96 4 0.07 Erythrocytes A1AD, pmol. PBG/ Males 100 ml RBC/hr Females Total P rotopo rphyrin, ng/100 ml RBC 24.1 4 4,#/ 18,7 1.0SJ 21,4 4 2,3 29.5 4 2.8 12.1 \.0 11.3 0. 2--^ 11.7 & 0,7 30. S d; 2.5 11.4 4 0.8-/ 16.3 4 3.0 13.8 4 2,0 24,0 4 2.2 Urine Cop roporp hyrin, ng/24 hr ALA, ng/24 hr 1.6 4 0.6 80.7 4 17,3 3.0 4 0,8 82.7 10.4 7.7 4 2.9 64.5 8.8 a/ Number of rats per period, except where indicated otherwise bj Mean 4 standard error, cj Two rats, d/ Six rats. 29 DUP040008906 TAiBLE 16 TISSUE AND BODY FLUID CHEMISTRY OF BATS FED CHIPS CONTAINING 0.537, LEAD AS LEAD QCTOATE Analyses 4 * IN=4')S./ Treatment Week 8 13 fN=12) Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, go % Albumin/globulin ratio 41 2-/ 26 1 5i 21 1 8 1 5.7 0.1 0,68 0.04 41 4 23 3 6 2 20 1 11 2 6,3 0,1 0.70 0,13 50 1 20 1 4 1 20 1 7 1 6.0 0,1 1,00 0.07 Erythrocytes ALAD, nmol. PBG/ Males: 100 ml EBC/hr Females: Total: Protoporphyrin, ng/100 ml RBC 21.4 6.3^' 15.5 i.l--t 18.4 3.3 23.1 1.0 10.3 0.8^ 15.3 0 , 2--^ 12, 8 1,5 21,7 1.1 12,5 Q.l-- 17.0 l.W 14,7 0.9 27.7 3.3 Urine Coproporphyrin, Pg/24 hr ALA, Pg/24 hr 10.7 5.9 97.4 19.2 1.1 0,6 101.4 30.1 4,9 2.4 50.5 6.6 a/ Number of rats period, except where indicated otherwise, b/ Mean standard error.. cj Two rats, d/ Six rats. 28 DUP040008907 TABLE 19 TISSUE AND BODY FLUID CHEMISTRY OF RATS FEED FAINT CHIPS CONTAINING 1.95% LEAD AS LEAD CHROMATE Analyses 4 (tM Serum electrophoresis Albumin, 7. Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 41 &/ 25 .db i 4 i 20 i 9& i 5.7 d: 0.1 0.73 0.06 Treatment Week 8 (N=4) 45 1 19 1 4 1 20 1 11 1 6 .20 0.04 0,81 0.04 13 47 * 2 19 2 5*1 23 * 1 61 6.4 * 0.1 0.94 0.08 Erythrocyte ALAD, nmol. PEG/ Males: 100 ml RBC/hr Females: Total: Protoporphyrin, Ug/100 ml RBC 29.5 3. 20.4 4.39./ 24.9 db 3.4 26.5 db 4.7 12.3 1..0E/ 14.7 0.6S/ 13.5 0,8 26.8 1.8 13.3 l.iJ 12.9 Z.3&f 13.1 1.3 27.4 1.5 Urine Coproporphyrin, Ug/24 hr ALA., ng/24 hr -H o r-* 0.9 0.1 80.8 db 7.2 6.4 67.7 6,8 4.4 1.9 66.5 17.5 a/ Number of rats per period, except where indicated otherwise, b/ Mean standard error. cj Two rats, d/ Six rats. 31 DUP040008908 TABLE 18 TISSUE AMD BODY FLUID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING 0.42% LEAD AS LEAD CHROMATE Analyses 4 niM Treatment Week 8 0fe4) 13 (N=12) -H o00 o '4' CO r*4 Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 42 .tfe 2--/ 28 db 1 41 19 1 8 3 5.8 * 0, 2 0.71 -L 0.06 44 d: 2 20 db 2 4 db 1 19 db 1 12 2 6,0 X .0,2 0,80 : 1 22 1 " 5 l 21 1 6 1 6.X * 0,1 0.90 0,03 Erythrocyte ALAD, nmol. PBG/ Males; 23.4 i 3.5^' 100 ml RBC/hr Females: 13,4 db 0.9 Total: d: 3,3 Protoporphyrin, 27.3 2.8 Ug/IOO ml TBC 14.1 , 0.6&' 17,4 0.1-- 15.7 1.0 28,3 d: 0,6 15.7 1.5^ 16.5 .= 1.4--7 16.1 1.0 27.9 = 2.1 Urine Coproporphyrin, ng/24 hr ALA, ng/24 hr 8,2 & 6.7 .69.3 6.6 1.5 0,7 65.2 db 17.4 6.3 2.2 85.3 * 31,4 a/ Number of rats per period., except where indicated otherwise, b/ Mean standard error, c/ Two rats, a/ Six rats. 30 DUP040008909 TABLE 21 TISSUE AND BODY FLUID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING 66.05% LEAD AS LEAn CARBONATE Analyses . _____ Treatment Week 4 .8................... (N=3)-- fN~4) , 13 (S12) Serum electrophoresis Albumin, % Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio H- 6 27 1 3 1 21 * 2 8 2 5.4 0,2 0.73 0.17 45 : 1 21 2 4 1 19 1 10 dt 1 6,0 0.06 0.82 sfe 0.04 47 2 22 1 5 dt 1 20 1 7 1 6.3 , 0.1 0.86 * 0.06 Erythrocyte ALAD, pmol. PBG/ Males: 100 ml RBC/tir Females: Protopstphyrin, Ug/100 ml EBC 12, 3SJ 9.1 2. ll/ 10.2 1, 23.2 0.9 4.4 , 0.1^ 12,4 0.4d/ 8.4 d: 2.3& 20.4 d: 2,1 5.4 1, o^/ 7.4 0.8./ 6,4 * 0.7--^ 27,2 6.8 Urine Coproporphyrin, Ug/24 hr ALA, Ug/24 hr 2. 0 * 0.7 90.3 27.7 0.8 0,3 72.3 13,4 6.7 * 3,2 42, 2 8.8 a/ Number of rats per period, except where indicated otherwise, b/ Mean standard error. cj One rat, d/ Two rats, e/ Six rats. Sj Significantly different from control (P < 0.05) as shown by Dunnett's multiple^comparison test following an analysis of variance. 33 DUP040008910 4^ o> H|Cu |cu TABLE .20 TISSUE AMI) BODY FLUID CHEMISTRY OF RATS FED PAINT CHIPS CONTAINING 12.43%'LEAP AS LEAD CHROMTE Analyses Treatment Week A .......... 8 (N=4) 13 (N=12~) / Serum electrophoresis Albumin, L Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, % Gamma globulin, % Total protein, gm % Albumin/globulin ratio 40 2--' 27 1 3 1 22 l 8 l 5.6 0.1 0.68 0,05 Erythrocyte ALAD, nmol. PBG/ Males: IdO ml RBC/hr Females: Total: Protoporphyrin, Iig/100 ml RBC 26.4 7.4 -/ 14.3 3.2S/ 20.3 4,9 25.5 1.6 47 3 21 , 1 4 1 18 1 9 2 6.1 0.1 0.90 0.12 2 21 1 5 1 22 1 6 1 6.4 0. 0.88 0. 16,5 1.4^ 13.4 3, 8--/ 15.0 1,9 24.2 * 2.6 12.6 1. 13.7 1. 13.1 .1, 24.6 2. Urine Coproporphyrin, ng/24 hr ALA, Ug/24 hr 2.6 1.8 89,7 19.3 0,9 0.1 78.4 6.8 2,6 dt 0.8 49.7 =b 12.5 aj Number of rats per period, except where indicated otherwise., b/ Mean standard error. c/ Two rats, d/ Six rats. 32 DUP04Q008911 SUMMARY OF PORPHYRIN METABOLISM IN; RATS FED FAINT C H IPS FOR 13 WEEKS CONTAINING DIFFERENT CONCENTRATIONS OF LEAD in \ri O N O CO 00 VO i0\0'M* 3OO '<h0. *r**<*i-csifiOQ <n +! +1 +{ +1 +! +1 +1 -H <sf'O10nr,'ii/'i(''-cMeo EOCL 4(fl iniri' <<f uV *M3 O'* ^CM *ir-l' H r-J N CSI N H o n N +1 -H -H *H' -H +1 -H +1 -ft o Gh Ov N fO s \o * : V CM <( i> < <) CM \D Mt d X5U>DQ24 O0. i0j 44, 0u01 OC i>O) 54 xu: 4J ** UP05 W <45 CMrinNHiriOcOd ***:*** CM CM C*") CM CM r* CM rM' 41 41+1 41 41 +1 41 41 41 CM CO l'- <jv *4* v c m C"> * a * OiNN4NP`4'f'MI CM Si CM CM CM CM CM CM CM 4S| <BI <U| O -4 Oi o O CO O r- co <-4 rM CM i-^ p-t W O 41 41 41 41 41 41 41 41 41 OOC^COcHrMr-IMfCS id <$ <f oi vo oi. oi sD ud % CM CM CSI CM CM CM. i-4 r-4 r-4 -4 t-4 4J 0 a) 0 0 0 44 44 r4 <SS 0 Gi 4J 44 0 0 0 4-> 44 44 0 0 B B P4 0 0' 0 B B o o a o O O o 54 54 T5 44 44 44 54 M JS -0 0 o O U XJ X2 o U 0 4J o o a O a 0 t3; x> i4 -0' XS t? xs X) 0 0- CO 05 0 0 0 0 0 0 0 P 000 00 f-q 25' HpPPpP o A* 54 4J 0 B-S co o tTi 10 in o ErS b-s str*': in CM CM 10 4t O CTv o\ O * m CM \D --1 O p o rs? O H H vP r4 35 500 *rf 4o4 4mJas dOto *a'u0rs-i. e0 V01 r*4 *0H4 a. r4 0 x 44 4004) 00 P P>*v a i *0- 43 CO 40-J CO 0o CB *'ri U*4 tn o uro uCO fc 44 O Vt 0 ou P0 0 6O 03 as 00 O u XS U4 oB au xJ 44 0 xCsD 54 a 4J o 4J 0; 0 54 0 0 XS *0 *f4 U 0 r4 r4 "i4 -0 0 54 54 o 0 0U 0 M *0 44 0 > oO H Hi 00 0 0* *4 0 T5 44 >4 xs 44 O 00 4 r4 44 0 P*i- 0 '**t r4 0 44 r-4 r4` 3 54 0 44 CO > > d r0 0 44 +1 0 r--1' 4 --4 d a0 od 0 *r4' d d d; 54 b0 M4 0 *4. H 0 0 i4 EB 54 d d < <ij B 0 00 0 i d > **4. 6 o < CO * Pl^3| Ol -oj 0} DUP040008912 TABLE 22 TISSUE AND BODY FLUID CHEMISTRY OF RATS FED NBS LEAD PAINT CONTAINING 11.927., LEAD Analysis S erum el ec tropho resis Albumin, X Alpha 1 globulin, % Alpha 2 globulin, % Beta globulin, 7, Gamma globulin,': Total protein, gm % Aibtimin/globulin ratio _____- Treatment Week 4$ (H=4)^ (N=4) h-* H- 52 & 21 1 4 1 20 1 31 5.4 0.2 0.03 44 2 21 2 8 5 21 3 6 tfc 1 6.0 & 0.2 0.78 J- 0.06 13 mai 44 4^ 2 24 d: 1 7 1. 21 = 1 5 JL 1 6.1 ' 0.1 0.82 2:- 0.06 Erythrocyte ALAD, pmo1. PBG/ Males: 100'ml SBC/hr Females; Total: P rotoporphyrin,, pg/100 ml RBC 8.5 5. 10.1 0.1^ 9.3 2.1--/ 14.8 4.3 5.0 it 0.3^/ 15.0 Urn- 5.1--/ 10.Q 3.6 24.5 a* 1.8 4.9 0.6^/ 8.8 6.9 -Jr 0.8-/ 26.3 tz 1,2 Urine Coproporphyrin, Ug/24 hr ALA, ng/24 hr 2.0 0.8 110.1 i 21.5 6.8 i 4.9 ' 4.6 i 2.1 72.0 dt 13.6 .51.3 A. 3.6 a] Number of rats per period, except where indicated otherwise. bj Mean standard error. cf Two rats, d/ Six rats, ej Significantly different from control (P < 0.05) as indicated by Dunnett1s multiple--comparison test following, an analysis of variance. 34 DUP040008913 TABLE 24 URINALYSIS OF RATS FED PAINT CHXP.S CONTAINING NO LEAD Treatment Week 4 j} 13 Profein: Negative < LOO mg % ^ .LOO mg_%_ __ _ _ Microscopic Examination REGS/: Normal Moderate _____ ____ Excessive____ __ mC*7i Normal Modera te _ _ _____ Excessive __ _ Epithelium!-/: Normal _ Moderate _ _ _________ _____ _Excessive_ Crystals^/; Normal Modera te __________ ____ _ExGj2ss_ive_ _ Casts: Negative ___ ____ __ Poijtive_ ___ 76 12 20 4 75 23 1 2! 1 1 6 " ~ " ' t ~ ' ' Z\ 2 . _2 8" 8~ 2 1 24~ 48 2 23 1 2_ ___________________________ 8 8 24* Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440), b/ Normal, 3 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). c,t Normal, none; moderate, 1-5 crystals; excessive, >5 crystals/field (x 100), 37 DUP040008914/ By 13 weeks the sex difference had disappeared and the ALAD activity in 66.05% lead carbonate group and the 11.92% MBS lead paint group were depressed 56% and 53%, respectively* Urinalysis of rats fed paint chips containing different concentrations of lead are shown in Table 24-32. No marked differences in urinary protein or sediment (erythrocyte, crystals, casts, etc.) were Seen in the control, lead octoate, lead chromate and lead carbonate groups. In the NBS lead paint group 5 of 12 rats (42%) had a moderate proteinuria compared with only 4 of 24 rats (17%) in the control group. Otherwise, the urinalysis of this group was similar to that of the control group* H Lead Content in Tissues of Rats Fed Paint Chips Containing Different Concentrations of Lead Tables 33-37 show the lead content of blood, brain, liver, kidney, and bone from rats fed paint chips containing different concentrations of lead. AH values for the lead concentrations in blood at 4 and 8 weeks, and 4 values in all but the NBS lead paint group at 13 weeks were Omitted from Table 33 because it was discovered that these samples had been contaminated by their storage containers. The blood lead levels of rats fed paint chips containing lead octoate (0.08%, 0.53%, 2.05%) or lead chromate (0.42%, 1.95%) were no higher than control. In the rats fed paint chips containing 12.43% lead chromate or 66.05% lead carbonate, or the 11.92% NBS lead paint (lead carbonate) blood lead levels were found to be elevated significantly. At 4 weeks, tissue lead levels in brain, liver and kidney were below the detection limits of our assay (0*7 pg per gm of tissue). The results were the same at 8 weeks with the exception of the NBS lead paint group* All four rats in this group showed detectable lead in their kidneys* At 13 weeks, detectable levels of lead in the brain appeared only in the rats fed 11.92% NBS lead paint 0 In the liver, detectable levels of lead were found In two rats fed 66 .05% lead carbonate paint chips and nine rats fed 11.92% NBS lead paint. No lead was detected in the kidneys of the control group at 13 weeks* In contrast, lead was detected in the kidneys of one to two rats in each lead octoate group and the 0.42% and 1 *95% lead chromate groups* Lead was detected in the kidneys of three rats, eight rats and 12 rats in the 12,43% lead chromate paint, 66.05% lead carbonate paint, and 11.95% NBS lead paint groups, respectively. The femurs of rats fed the control diet, lead octoate or lead chromate paints exhibited very little lead at 4 weeks. However, the femurs of rats fed 66.05% lead carbonate or 11.92% NBS lead paint contained 13,0 and 14.9 pg of lead/gm of bone, respectively. 36 DUP04000891S TABLE 26 URINALYSIS OF RATS FEB PAINT CHIPS CONTAINING 0.53% LEAD AS LEAD OCTOATS Protein: Negative < 100 mg % _ ,, ^______ Microscopic Examination RBCS/ : Normal Moderate _ Ejxes^sj-V^e ______ _ megJ: Normal Moderate ___________ Excessive _ ______ Epithelium^/: Normal Moderate __________________ _Exces_sive_ Crystals/: Normal Casts: Moderate _ __ _,Excjessive Negative Positiva_ ___ _ _ Treatment Week 1iM 43 1 ^ _ *. - _ 10 l J-, 22 12 4 ~ "Y' 3 4 `4 10 l 9 3 12* Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 celIs/field (x 440). b/ Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells./le.ld (x 100) . c/ Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/field (x 100).. 39 DUP040008916 TABLE 25 u r in al y s is o f r at s f ed pain t c h ips c o n t ain in g 0.08% LEAD AS LEAD OCTOATE Treatment Week 48 13 Protein: Negative < 100 mg % _ _________> 100 Microscopic Examination RBC~/: Normal Moderate ______ _ Excessive_______ WBClJ: Normal. Moderate ____._______Excessivji _ _ . Epitheliuml^: Normal Moderate ____ .__________ __ _Exeess ive Crystal s/: Normal Moderate ___ t ___________ _Excessive__ _ Casts: Negative ________ __ Poiive_________ 43 _________ 1_^ ^ 8 4- _ 43 l 10 2 ~2 ~ ~ ~ ~3~ 21 To" 2 4 ~ 4~" ~ ~ " If 2' 2 4~~ '4' 4_ ~ If 1 If Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440).. b/ Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). c{ Normal, none; moderate, 1-5 crystals; excessive, > 5 Crystals/field (x 100). 38 DUP040008917 TABLE 28 URINALYSIS OF RATS FED PAINT CHIPS CONTAINING 0.42% LEAD AS LEAD CHROMATE Treatment Week 4 TS-T 13 Protein: Negative 4 2 10 < 100 mg % 21 __________ > .100 mgjL ______ ,_____ 1_ Microscopic Examination RBC^L': Normal 31 8 Moderate 13 3 _______ Excessive___;____ __ mC?J Normal 2 _ _ JL_ 38 Moderate 21 X _ ._____ _ Excessive^ _ ____ _ _ ____ _3__ EpitheliunJI/: Normal 4 4 12 Moderate Excessive Crystals.S/7 Normal 2 4 12 Moderate 1 _ ___ _____ _______Excessive_ _ _ - _ i ____ Casts: Negative 44 12 Positive^ Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 celIs/fieId (x 440). b/ Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100), c/ Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/field (x 100). 41 DUP040008918 TABLE 27 URINALYSIS OF RATS FED PAINT CHIPS CONTAINING 2.05% LEAD AS LEAD GCTOATE Treatment Week 4 8 13 Protein: Negative < 100 mg % ________ > 100 Microscopic Examination RBC^/: Normal Moderate ___ __ _ Excessive ______ _ \i&CS7: Norma i Moderate ___ _____ Excessive ______ Epithelium^: Normal Moderate ___________ _________ Exces_sive_ Crystals/: Normal Moderate _ _ _ ______ __ _Excjasj3ive_ _ Casts: Negative ____ _____ ___________________ 44 11 1 4 4 12 1~ ~3~ ^ ~s" 14 _ _ _ ^ _1_ ^ _____ 4~ ~ "4" " ~2 _ - ~2' _2_ 4 . " "4 ~ 12' Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 ceils/field (x 440). b/ Normal, 5 or less ceils; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). cj Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/fIeld (x 100). 40 DUPQ40008919 TABLE 30 URINALYSIS OF RATS FED PAINT CHIPS CONTAINING 12.43% LEAD AS LEAD CHROMATE Treatment Week 4 8 13 Protein: Negative < 100 mg 1 _ ________ > 100 mg % ^ ^ . Microscopic Examination RBCf/; Normal Moderate -- Excessive _____ _ WBCi7: Normal Moderate ___ _ __ _ Excessive________ Epithelium!^: Normal Moderate __ _______ _____ __ --Excessive. Crystals^/: Normal Moderate ________. _ _ExCjesive_ _ Casts: Negative ___ ____ __ J?os_itive_ _____ _ 11 9 32 3 ___________1_ ^ _______ 4 1 11 1 _ __ J3_ _ _. 1 1 11 31 _ 2________1 4 " " -4" if 1 ~2~~ 2 eT ~ ~ ~ S' 4 ~ ~ ~4~. " ~ " 12 Numbers indicate number of rats at response level. / Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440), b/ Normal, 5 or less cells; moderate, .5-25 cells; excessive, > 25 cells/field (x 100). cV Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/fieId (x 100). 43 DUP040008920 TABLE 29 URINALYSIS OF RATS FED PAINT CHIPS CONTAINING 1.95% LEAD AS LEAD CHROMATE Treatment Meek 4 8 13 Protein; Negative < 100 mg % ___________ > 1.00 __ Microscopic Examina t.ion RBC?J: Normal Moderate ,__________ Excessive _______ WBC&7: Normal Moderate _ ._________Excessive______ EpitheliumSA: Normal Moderate ________ ___ ,___ ...Excessive, Crystal s--1^: Normal Moderate i___,___, _ ,___, _Jxcesjs ive_ _ Casts: Negative Positive 3 3 11 11 1 4 3 12 1 2 ~ "3" ~ ~8 ' 23 _ _ _ ^ _1_ __ ______ L 4 4 ~ 12 4 4 12 44 12 Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 colIs/field (x 440). hj Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). ci Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/field (x 100). 42 DUP040008921 TABUS 32 URINALYSIS OF RATS FSB PAINT, CHIPS CONTAINING 11,92% LEAP fNBS LEAD PAINT) Treatment Week 8 13 Protein: Negative < 100 mg % 23 2 7 3 _____ ____ > 100 mg_7u _ ^ ^ Microscopic Examination RBC~/: Normal 2 4 12 Moderate 2 Exc_e_ssive __ __________ _ Normal 2 3 11 Modera te 11 Excessive ^ ______ j_ Epithelium^T: Normal 4 12 Moderate __ _Excess_ive_ ;_____ _ Crystals--/ : Normal A 12 Moderate _Excessiye_ _/ _ ^ Casts: Negative 4 12 Positive Numbers indicate number of rats at response level, a/ Normal, 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440), b/ Normal, S or less cells:; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). cj Normal, none; moderate, 1-5 crystals; excessive, > 5 crystals/field (x 100) . 45 DUP040D08922 TABLE 31 URINALYSIS OF RATS FED PAINT CHIPS CONTAINING 66.05% LEAD AS LEAD CARBONATE Protein: Negative < 100 mg % ______ _ _ > 100 mg_\_ Microscopic Examination RBC^/Normal Moderate __ _ _ Exej>s iye _ ____ "" WBCSTY Normal Moderate __ ._________ Excesive _ ^ _ Epithelium^/: Normal Moderate _____ _ ___ _____ Excessive Crystals^/; Normal Moderate __________ __ _Exc_es_sive_ _ Ca sts: Negative _____ Positive_______ _ Treatment Week 4 8 13 3 4 11 11 4. 2 2 22 22 44 11 1 10 2 12 _ ___ _ _ __ , 1________________ ,, _ . 4' 4 12 Numbers indicate number of rats at response level. a_l Normal., 10 or less cells; moderate, 10-100 cells; excessive, > 100 cells/field (x 440) . b/ Normal, 5 or less cells; moderate, 5-25 cells; excessive, > 25 cells/field (x 100). jc/ Normal, none; moderate, 1-5 crystals; excessive, > 5 crysi:als/f ield (x 100).. 44 DUP040008923 Diet Control!?/ 0.087. Lead Qctoate 0.53% Lead Octoate 2,05%: Lead Gctoate 0.42% Lead Chromate 1.95% Lead Chromate 12.43% Lead Chromate 66.05% Lead Carbonate 11.92% NBS Lead Paint TABLE 34 LEAD CONTENT OF BRAIN 4 (4)1/ Treatment Week 8 (4) n.d.V Did* n.d. n.d, n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n, d, n.d. n.d. n.d. n.d. 13 (12) n.d. n.d. n.d. n.d. n. d, n.d. n.d. q , d . 0,2 0 4/ Number of brains analyzed per group unless indicated otherwise, b/ Eight brains were analyzed at 4 and 8 weeks. Twenty-four brains were analyzed at 13 weeks. c/ Not detectable. Below sensitivity of 0,07 pg/gm of brain, 4/ Average pg of lead/gm brain standard error number of rats shown in parenthesis. 47 DUF04000S924 TABLE 33 LEAD CONTENT OF WHOLE BLOOD Diet rf/ Control-- 0.08% Lead Octoate 0.537. Lead Octoate 2.05% Lead Octoate 0.42% Lead Chromate I. 95% Lead Chromate 12.43% Lead Chromate 66.05% Lead Carbonate II. 92% MBS Lead Paint AS./ Treatment Week / 13pj (8)/ 13.1 + 0.9 (16)e/ 12.8 0,8 (8) 16.4 + 2.4 (T)!/ 14.5 1.5 (8) 13.2 +1.0 (8) 12.0 1.4 (8), 19.4 2.2 (3)S/ 24.9 + 2.2 (&)/ 23.9 + 1,3 (12)/ a/ Values deleted due to contamination, b/ Four samples were omitted due to contamination. 0/ Humber of rat bloods analyzed per group unless indicated otherwise, d/ Sixteen blood samples were analyzed. e/ Average jig of lead/lQQ ml blood + standard error of number of rats in parenthesis, / Qne blood sample lost* Significantly different from control (P < 0,05) as shown by Dennett's multiple-comparison test following an analysis of variance, h/ Twelve blood samples analyzed. DUP040008925 let Control^ 0,08% Lead Octoate 0.53% Lead Octoate 2.05% Lead Octoate 0.42% Lead Chromate 1,95% Lead Chromate 12.43% Lead Chromate 66.05% Lead Carbonate Tl.92% NBS Lead Paint TABLE 36 LEAD CONTENT OF KIDNEY co |SF _________ Treatment Week (V n.d,-- n.d. n.d.. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n,d. n.d. n.d. n.d. j 1,40J - (4) 13 - 0?) n.d 0,3 0) 0,3 (D 0.4 i 0.1 (2) 0.5 dP} 4.2f(1) 0*4 + 0.0 (3) 2.3 + 0.2 (8) 1,9 + 0.1 02) a/ Number of kidneys analyzed per group unless otherwise indicated, b/ Eight kidneys were analyzed at 4 and 8 weeks, Twenty-four kidneys were analyzed at 13 weeks, c/ Not detectable. Below sensitivity of 0.07pg/gm kidney, d/ No explanation for unusually high value, e/ Values are jug of lead/grn kidney or average jug of lead/gm kidney + standard error of number of rats in parenthesis. 49 DUP040008926 Diet Control]*/ 0.087. Lead Octoate 0.537. Lead Octoate 2.057. Lead Octoate 0.427. Lead Chromate 1.95% Lead Chromate 12.43% Lead Chromate 66.05% Lead Carbonate 11.92% NBS Lead Paint LEAD CONTENT OF LIVER 4 (4)--/ n.d,^ n d n.d. n.d. ci* d * n.d. n.d. n.dn.d. Treatment Week S (4) 13 (12) n.d. nd, n.d. n.d. n.d. n.d. n.d. n. d, n.d. n. d. n.d. n.d. n.d. n.d. n.d. n.d. 1.1 0.6& 0.4 +0,1 Q. a/ Number of livers analyzed per group unless indicated otherwise, b/ Eight livers were analyzed at 4 and 8 weeks. Twenty-four livers were analyzed at 13 weeks. cj Not detectable. Below sensitivity of 0.07 pg/gm liver, d/ Values are average pg of lead/gm liver standard error of number of rats shown in parenthesis. 48 DUP040008927 At 8 weeks, J-he femurs of rats fed the control, lead ocfoqfe or lead chromate diets contained similar quantities of lead, ranging from 1.8 - 4,2 pg lead/gm bone. Lead content in the femurs in the 66.05% lead carbonate and 11,92% NBS groups were 16.0 and 16.4 pg of lead/gm of bone, respectively . Both of these values were significantly above control* After 13 weeks, the rats fed lead octoate or lead chromate had no more lead in their femurs than at 8 weeks and these levels were no higher than those in the control group. The 66,05% lead carbonate group exhibited higher levels of lead in their femurs (21.7jjg lead/gm bone) than at 8 weeks. The 11,92% NBS lead paint group had less lead in their femurs (10.9 jjg lead/gm bone) than af 8 weeks. The lead levels . in both of these groups were significantly greater than those observed in the control rats. I, Pathology of Rats Fed Paint Chips Containing Different Concentrations of Lead The relative organ weights of the thyroid, spleen, heart, kidneys, liver, adrenals, brain and gonads are shown in Table 38. The Dunnett's multiple-comparison test showed that none of the relative organ weights differed from control values. The pathology of rats fed the control, 2.05% lead octoate, 12,43% lead chromate, 66.05% lead carbonate or 11.92% NBS lead paint diets for 13 weeks is shown in Tables 39-43. The control rats had a few naturally occurring lesions. Fourteen of the twentyfour (58,390) rats had mild to moderate lymphoid hyperplasia in the lungs, characteristic of early minute pneumonia. One of these rats had a moderate pneumonia and another One a mild emphysema. Other lesions included an unspecific myocarditis in two rats; foci of subacute inflammation in the liver of two rats; a cross-section of a parasite (roundworm) in the colon of one rat; and foci of mononuclear cell infiltration in the kidneys of two rats. The bone marrow myeloid/erythroid cell ratios (M/E) of all rats were within normal limits. The lesions observed in rats fed the 2.05% lead octoate diet were similar to the lesions seen in the control rats. Seven out of the 12 rats (58,3%) had mild to moderate lymphoid hyperplasia in the lungs and one had mild pneumonia. Four rats had mild foci of subacute inflammation in the liver characterized by a loss of several hepatic cord cells which were replaced by macrophages and lymphocytes. Another rat had a focus of inflammation in the interstitial tissue of the pancreas. The M/E ratios of these rats were normal, 51 DUP040008928 TABLE 37 LEAD CONTENT OF BONE (FEMUR) Diet Control-- 0.08% Lead Octoate 0,53% Lead Octoate 2,05% Lead Octoate 0,42% Lead Chromate 1,95% Lead Chromate 12.43% Lead Chromate 66.05% Lead Carbonate 11.92% NBS Lead Paint Treatment Week 8 ~~ (4) 13.0# 4.1 (4) 14.9 +1.5 (4) 3.9 + 0.4 (8) 4.2 0.6 (4) ^ 3.9 t 0.2 (4) 3.0 0.7 (4) 2.6 0.9 (4) 1.8 + 0.2 (4) 3.1 1.1 (4) , 16.0 2.1 (4W/ 16.4 0.4 (4p 13 (f2) 3.2 t 0.2 (24) 3.3 0.3 (12) 3.1 0.2(12) 3.9 0,5 (12) 3.8 0.6 (12) 4.1+0,5(12) 3,5 + 0.5 (10) st. 21.7 2,4 (12) U. 10.9 + 0.9 (12) 1/ q/ Number of bones analyzed per group unless Indicated otherwise. / Eight bones were analyzed at 4 and 8 weeks. Twenty-four bones were analyzed at 13 weeks* c/ Values are/jg of lead/gm bone or average jjg of lead/gm bone standard error of number of rats In parenthesis* d/ Not detectable. Below sensitivity of 0.4 jug/gm of bone, e/ Two samples lost f/ Significantly different from control (P < 0.05) as shown by Dunnett's multiple comparison test following an analysis of variance. 50 53 DUP040008930 PATHOLOGY OF BATS FED FOR 13 WEEKS WITH PAIHT CHIPS CONTAINING HO LEAD Tissues not lis te d were norm al. aj S e ve rity o f le s io n s ; l-m inim a 1; 2~moderate; 3-severe;. 4-very severe; questionable RELATIVE ORGAK WEIGHTS OF RATS 'FED DIETS CONTAINING DIFFERENT GOKCENTRATlONS OF LEAD IN PAIKT CHIPS FOR 13 WEEKS 52 DUP040008931 _ _Q O<3D* ./ 55 DUP040008932 "PATHOLOGY OF RATS FED FOR 13 WEEKS WITH PAINT CHIPS C O N TAIN IN G * 12.43% LEAD AS LEAD CHROMATE CON'I CM CoPII Mo oCM| oCP SI CM 0C0M C00P I IS I CO M00O I C00P I Pc3) JO CL<U-L D>s- -C ^ 3o oc ng E E a; Xc _J Cl . oc oa EE E j u. tfl O 0 o LL. PATHOLOGY OF RATS FED FOR 13 WEEKS WITH PAINT CHIPS C O N TA I 2,05% LEAD AS LEAD OCTOATE iTissues not listed were normal, a / Severity of lesions; 1-minimal; 2-moderqte; 3-severe; 4-very severe; questionqbl O <D c _0 _Eo u o- U <U CO a0. u0_ 54 Co CO CO cm CM CM DUP040008933 TABLE 43 8 CM CO gr[ a. F-- O CoOs < QU O 0? < in no cOo' l CO UJ UJ o OS CO Di 0 & u_ O 8 (5 o M00f <a. C00O!I CM CM CM CM CM CM CM CO CM CO to CM* CM CM CO CM 0 _Bo Zt & "o-l t: if 1 Oo x <>D 23 E JW 57 DUP040008934 ts <N r! CO o SO CO so rsvo ssoo I CO so X 6e z 2o co O u> -G P ai JO a 3 D a. S_ CL EX c --S CL. cs tn *5 0 u X o2 PATHOLOGY OF RATS FED FOR 13 WEEKS WITH PAINT CHIPS C O N TA IN IN G 66.05% LEAD AS LEAD CARBONATE Heart Liver M iositis Tissues not listed were normal, ia / Severity of lesions: 1-minimal; 2-moderate; 3-severe; 4-very severe; questionable co E to Eo 2 U-. . u- O _ 'o o1J_ to m 3- E "5 56 ' o s_ 0 5 <D C O CQ . CO cs CO cs cs CS o 0 V \ 2 DUP040008935 The body lead burden of rats fed paint containing lead octoate or lead chromate was no different from control until 13 weeks. At this time lead was found in the kidneys of one to three rats on each of the lead octoate and lead chromate diets. The rats with measurable kidney lead in the lead octoate, 0,42% lead chromate or 1,95% lead chromate groups did not have concominant increases in blood, brain, liver or bone lead. Thus, these rats did not have a significant increase in body lead-burden. On the other hand, the rats Fed the 12,43% lead chromate diet had elevated blood lead and thus, a significant elevation of body lead-burden. Since rats Fed leacj nitrate show elevated lead levels in blood, kidneys, bone and liver within 2 weeks,-^ it appears that the lead octoate and lead chromate were either not readily absorbed or not otherwise available for absorption. Complete gross and microscopic pathology was performed on the rats that were fed the control, 2.05% lead octoate, 12.43% lead chromate, and 11,92% NBS lead paint diets for 13 weeks. All of these rats were relatively free of lesions, and those lesions which were observed, occur naturally in rat colonies. CONCLUSIONS Older paint formulations contained soluble lead salts,. Two samples of paint chips representative of these formulations were fed to rats at 0.1% of their diets. One was supplied by the National Bureau of Standards and was removed from interior walls of older homes. It contained 11,92% lead. The other was a recent formulation containing 66,05% lead carbonate. Both of these samples produced classical signs of lead poisoning in the rats. These signs consisted of erythrocyte ALAD depression (50%) and significant increases in blood, kidney, bone, liver and/or brain lead. Newer pairit formulations contain insoluble lead pigments and low levels of lead driers. Several samples of paint chips containing,up to 12,43% lead chromate pigment and up to 2.0% lead octoate drier were also fed to rats as 0.1% of their diets. The only evidence of toxicity or effect on body burden appear in the rats fed the paint containing 12,43% lead chromate. This consisted of an increase in blood lead, without other signs of poisoning, Pairjt Containing 1,95% lead chromate and paint containing 2.05% lead octoate produced rio detectable changes in the rats. The quantity of lead fed as paint chips containing either 2.05% lead octoate or 1,95% lead chromate would have approximated an intake of 40 mg/day in a 3 year old child weighing 14.6 kilograms. This daily intake has been reported to produce lead poisoning in children if the lead was available for absorption. Since the body lead-burdens of the rats fed 2.05% lead octoate or 1.95% lead chromate were not increased, it can be assumed that the lead in these paint chips was not absorbed from the gastrointestinal tract. In addition, lead in paint scraped from old dwellings and not the newly formulated paint, deposited in the brain, even though the blood-lead concentrations of both were similar. 59 DUP040008936 The rats fed the 12.43% lead chromate diet had lesions similar to the lesions seen in the control rats. Seven of 12 rats (58,3%) had mild to moderate lymphoid hyperplasia, and one had mild pneumonia. Only one rat had a moderate unspecific myocarditis. The M/E ratios were normal, The lesions seen in the tats fed the 11.92% NB5 lead paint diet were similar to those lesions seen in the control rats. Eight of 12 rats {66.7%) had different degrees ^ of lymphoid hyperplasia in the lungs. In one rat pneumonia was evident. Unspecific ' myocarditis occurred in 2 rats while foci of infibrnmatian in the liver Wfere observed in only one rat. The M/E ratios Were normal. In conclusion, the pathology seen in the control and treated rats were characteristic of those in a normal population of rats. No evidence of lead-induced lesions was found. IV, DISCUSSION The levels of lead fed to rats in the study were sufficient to produce significant elevation of body lead burden if the lead is in the form of lead nitrate--' If we equate the amount of lead consumed by the average rat fed paint chips containing 2.05% lead octoate, 12.43% lead chromate, 66.05% lead carbonate, and 11.92%*NBS lead paint diets to a 14.6 kg [3-year old) child, we find that the lead consumption would equal 39.4, 156.0, 577.8, and 147,8 mg of lead/day, respectively. On the basis of a 2% lead paint, the total paint intake would be 2,0, 7.8, 28,9, and7.4 gm/day respectively. Daily ingestion of these quantities of old paint containing white lead have been reported to produce lead poisoning.--^' There were no changes in hematology, urinalysis or serum proteins in the rats fed lead octoate, lead chromate Or lead carbonate. The rats fed the NBS lead paint did exhibit a significant reduction In hemoglobin and a mild proteinuria at 13 weeks. Otherwise, the hematology, urlnalysijs and serum proteins in the rats fed NB5 lead paint were normal. Porphrin metabolism wqs affected as early as 4 weeks in rats fed either the 66.05% lead carbonate diet or the 11.92% NBS lead paint diet as evidenced in a 50% depression of erythrocyte ALAD activity. This effect appeared to be directly related to the concentration of blood lead as has been reported by others,--^-- There was one exception: blood lead in the 12,43% lead chromate group became elevated by the 13th week, but there was no depression of ALAD activity. This discrepancy could be related to the duration of elevated blood lead or the age of the animal when blood lead became elevated. Even though erythrocyte ALAD was depressed 50%, the overall effect on porphyrin metabolism must be considered mild, since there was no concomitant change in the levels of protoporphyrin (erythrocyte), delta-aminolevulinic acid and coproporphyrih. These observations are consistent with reports that blood lead levels greater than 4Q/jg/100 ml RBC are needed before serious changes in porphyrin metabolism occur,TM/ 58 DUP040008937 REFERENCES Kehoe, R. A., "The Metabolism of Lead in Man in Health and Disease," Lecture II, J. Roy, Inst. Public Health Hyg., 24:101 (1961), Chisholm, J. J., Fed, Clin, North Amer, 17:591 (1970). King, B. G,, Amer. J. Pis. Child., 122:337 (1971), Gage, J. C, and M. H, Litchfield, J, Oil, Col, Chem, Assoc,, 52:236 (1969), Culp, R. A,, and A. B. Rawitch, J, Paint Tech,, 45:580 May, 1973, Brecher, G, and M, Schneiderman, Am, j, Clin, Path,, 20:1079 (1950), Faulkner, W. 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