l e a d p a ;:.t :::c s s t t o s s t v o y '*' W ' - w V I FINAL REPORT 14 Sopuobor 1973 *-.■ > i* Contract So. 62-tf-62GOkSPC MR! Projocr So. 3729-B For National Paint and Coating* Aatoelatien 1500 Rhode Xiland Avanua, S.W. Waihingcon, D.C. 20005 -f Aten: Mr. Royal A. Brown •3 0007—SWP—03 6972 i i N21656 o ?xsra c e This report was prepared at Midwest Research Institute, tli Volker Boulevard. Kansas City, Missouri 64110, under Contract No. 62-W-62CCANPC, J3U Project No. 3729*8, "Lead Paint Ingestion Study." The research was sponsored by che National Paint and Coatings Association, 1500 Rhode Island Avenue, N.U., Washington, O.C. 20005. Dr. John P. Frawley, Chief Toxicologist, Hereules, Ine., Wilmington, Delaware 19899, was the project monitor. The research was conducted in the Biological Sciences Division, under the direction of Dr, W. 8. House free 1 December 1972 through 31 July 1973. Dr. Thomas X. Castles, Principal Pharmacologist, was the principal investigator, assisted by Dr. Jsios L. Senyer, Aesoeiatt Pathologist, and Mrs. Jane Koch, Biology Research Assiecant. Dr. Jamas L. Splgsralli, Senior Chemist supervised the lead analysis with the assistance of Mrs. Hope >!. Millar, Assistant Chemist. m Approved for: o MIDWEST RESEARCH IXSTTTLTE igg; w, S. House, Director Biological Sciences Division eStete 14 September 1973 6Hg Wik iv ‘A ’ » '"X • • ■ -------0007-SWP-036973 0007-SWP-000115541 :ac ix or c o k t s h t s •'3 ’ Psee I. Introduction II. Methods A. III. Sfi? . . i 2 2 3 j 6 Results........................................................................................................... 6 A. B. c. D. E. r. © 1 Preparation of Paint Files Containing Different Cancan* traciona of Lead............................................. Preparation of Diets ....................................................................... Experimental Procadura .................................................................. Analyaaa.............................................................. Pathology............................................................................................ Statistical ........................................................................................ B. C. D. E. T. H. I. 'mil Concentrations of Laad in Paint Filaa ................................. Laad Analyaaa of Olata Containing Different Paint Filaa. Fraliainary Feeding Studies .................................................. . Croat Observations, FeedConauaiption and Body Weights . Weakly and total Land Consuaption .......................................... Kasaoology of Hats Fed Paint Filaa Containing Different Concentrations of Lead............................................................ Tissue and Fluid Chaaistry of Kata Fed Paint Files Containing Different Concentrations of Lead ................. Lead Content in tissues of Rats Fed Paint Flint Containing Different Coneancraciona of Lead ................. Pathology of Rats Fed Paine Filaa Containing Different Coneaneraciena of Lead ............................... e 6 9 9 14 14 r&y§ 36 51 :v. Discussion 58 V. Conclusion 59 References E.’feair 14 m 60 k-s. 3 iii 0007-SWP-036974 0007-SWP-000115542 I. INTNODUCTIOH ftSeJut Few parsons will disagree that old laad-basad paint praaenta a^ awg imiaicn. At a result of »d ................... iiwlrh hJ’.il ,JL forts ay Kehos, i> Chisholm,!*1 and King,-'' aiadical raaaarchars, paint menu- « icurars and legislators hava recognized the aariouanaaa of tbit problem j » acting co iscabliah aafe concantraciona of lead in paint, to do this, ly have voluminous reports on clinical observations and aniaial research from ich to draw. Unfortunately, moat of chia information ia based upon eha aid i*'r"i* °» tn;i Jf ti.-o.a.-aad u.:s^ mien not tMUuUc: cn« name rar' mu TETST on toaav ' t aia 2? V. Tslifl ill! loneantraclone of load in paint can ba intalligantly established, we must evaluate the cexlelty of the lead as it exists currently in modern paints. l£_uat_tht puroosj of thla ttudyj to evaluate eha toxicity in_raea produjed__,by modern paints containing lead octoacajplaad chromata_?r» iwod' eaweawsesjindj^o^ottparsthai^ommol^painc^jotgilationjibyfc^e^f N chosi^ras^^^i^T^^^^^S^^^^ST^p^'lbrTe'tuTTs ar* presented in cue following pages. ' |sU t—* #■ iz. Q A. MEimpg **. + ^•fW A/- / Preairatiof! of Paint Films Containlnt Different Concentrations ef Laad Paint filmsij without added lead and^addodXai [aid eompounda ir. the form of lead octostt. laad chromate, or lead"carbonate ware supplied by mam* bars of tna National Paint and Coatings Association. Paints which contained laad octoate.*lead chromate (medium yellow) am*«^rti>m*aibmmjgwe were Pr*Par,<* using e flat elkvd enamel formula baaed upon M'TttTTwTjtwil Class I alkyd. The flat elkyd paint without added laad was used for control^ The percent of lead ia aaeh aampla was calculated on tha beats of tbepsfnvoletila matsrial. Dispersion waa accoapUahad with a Cowlea DlaspJdar and eha pigment volume concentration was held at one level. TheinjKfita ware placed In pana and allowed to air dry. Tbit was fallowed by^Mfrcsd air drying at 120*r to volatilize any remaining solvent. Peine ££Qm were ground^sieved, end chips ranging from 0.5 to 1.0 mm in alas wKr^usad for this study. Ihlc chip sire was saltotad on tha basis of a preliminary rat feeding study which aatablisbad :r.at rata would selectively eap/£round larger aisad chip*. , lr*Jr i»> Misled 'Whd fcWO Tha concent ratStfns of lead in the different paint fllma wars datarr.inae indapasdantly b//DaScto Incorporated, the Shetvin-Wiliians Company and Kicvest Research Ipdcituca, using Atomic Absorption Spectrophotometry. jfe fb TZ**' '3 ■ ■ ■ .H-jpmw, ».iiu 0007-SWP-036975 0007-SWP-000115543 Sample^ of old lead-based psinc.iS^ fron eh* uU.S. Dsparc• obtained from none of Commerce, Sac ion* 1 Bureau of Standards, Washington, O.C.» Thi* paint had bean pulvtntad and aiavad through 323 M*h seratn (0.06-0.OB i» sis*J and contained 11.872 lead by vitight. Thi* sample will b* rafarld to aa "J'BS lead pain:" in chit report. B- 'Sliw. f-*J«ra_e-.on_ofi.O:ecs , CW*» . Oieta u*r* prepared by mixing paint tela* with Purina Rat Chow each in the concentration of 0ilt (weight/weight). thi* wa* aceompllined by first preparing a 102 conctncraca (paint film/feed, w/w) in a high-speed twin ahall aixar. This concentrate was ehan added to the appropriate amount of Purina Rat Chou mash to give a final concentration of 0.11 (paint flla/fnnd, w/w) and mixed for 10 mm'in a bulk mixer. lech diet was prepared three times during thn experiment and samples of tech of these preparations were assayed for lead content. C. Experimental Procedure ■ ^g_tmgdred weanling (40-60 fa) Charles River rati (100 mslas and 100 female ret*) were used far thia a tody. Upon arrival rets were housed individually in air conditioned quarter* in polycarbonate cage* containing' hardwood bedding and filter topi. After 3-7 days of equilibration, all rats were assigned ons of the following diets. 1. Rat chow plus paint f*4m without added lead (control) 2. Rat chow plus paint fA* eeneainlng 0.082 lead AS lead ACCOAtt 3. Rat chow plus paint concainlng 0.331 laad AS laad OCCOACS * containing 2.052 laad AS laad octeAti «. Rac chow plus paint 3. 6. Rat ehow plus paint £ila containing 0-422 laad AS laad ehrooACA ♦ i Xac chow plus paint fila containing 1.952 laad AS laad e.SroesiCA 7. Rat chow plus paint film containing 12.432 lead as lead chrosace I. Rat chow plus paint film containing 66.051 lead as laad csrbenata 9. Rat chow plu* KBS lead paint containing 11.922 lead stedte 2 ■ >'» • .r'*'•« . -vV..., s ■ • ”r r '*........-5 '• '•'v.* '' 0007-SWP-036976 0007-SWP-000115544 i\>v* Twenty ih U end 20 female rats were assigned to the control diet, and 10 males aha 10 fcsiala rats were assigned to each ef :ne remaining discs. Seen diet was fad in a 9-os widc-mouth glass jar which was secured to the cage. Tap water wee available acv libitum during the entire experiment feed consumption was measured twice each week and calculated on a daily basis. The animals were weighed once a weex. All rats were observed twice or more each week for toxic signs. Afeer A, 6 and 13 weeks, a selected number of male and feaiale rats from each group were placed in mseabolisai eages for ehe collection of 2A-hr urine samples. A portion of the collected urine wee used for urinelysis and che remainder frosen for assay of delta.aminolevulinic acid and coproporphyrin. After urine collection each rat was anasehetixad with ether, exsanguinated vis the abdominal aorta and bone marrow smears were prepared. Blood samples ware heparixined, cooled and ueed imaedlacely fogp hametelogy and enzyme sxsayz. Aliquot* of the resistning blood were cakan for protoporphyrin and laid assays. Afcar urine and blood sample# were obtained, each rat was necropsied and tissua taken for lead analysis ft microscopic examination. 3. Analyses 1. (WiM laid analyse* of osinc films sad diets containlns saint Me: a. Paint film: T«n.ailligrsmf samples ef paint fUae con­ taining no lead or O.OflZ lead oetoate were eharrtd with 3 ml of concentrated nitric acid and dry-ashdd at 500*C for 2 hr. The ash was dissolved in 1 ml of ecus regie, diluted to 5 ml with dlseillad water and this final dilution mtasurad fer lead content by atomic absorption. All other peine M4me (10 mg samples] wars digested in 15 el of a mixture of concentrated nitric acid and 37& perchloric acid (2:1, V/'V], evaporated end ehe remaining perchloric acid solution diluted eo 25 ml with distilled water. This final dilution wae used for atomic absorption spec­ trophotometry. b. Diets: One-gram samples of feed were digested in a eixture of concentrated nitric acid and 371 perchloric acid (2:1, V/V) end their final dilutions adjusted eo make their lead concentre cions within the detec­ tion limits of che atomic absorption technique. u i -1.0-f 3 0007-SWP-036977 Z. Ars«Ivi»> r>»rfsrrtd uoor. each rat: t. Haaatolcev: (1) Hematocrit: Hematocrit was deticninad in capillary cubes using * microcapillary centrifuge (International Equipment Company, Model MS). (2) Ksmoclobin: Hemoglobin was measured as evano- nechenogisbin.i' (3) Smhrocvce end leukocyte counts: Total erychroeys# and leukocyte were counted using e Coulter Electronic Particle Counter with 100-u spareture.l^ (4) Reeiculocvtee; Reticulocytes were counted by the aetnylene blue method using the Miller disc.-^ (5) Differential leukocyte counts; used to stain the leukocytes for examination. ! i i o Wright's stain use (6) Ervthrocvte osmotic fragility; Osmotic fragility of erythrocytes was quantitatively determined by subjecting heparlmted whole blood to sodium chloride solutions of different Osmelerieioe.2^ The concen­ tration of sodium chloride vhieh hemolyted 301 of the o$chroeytes wee obtained from a plot of percent hemolysis versus sodium chloride eoneentretion. b. Body fluid end tisaue chgriitrv: tl) E,;emjr^|^p,,t}(|p^OBj}lffX«ii^ rotil plasma proeain w h dace rained using che dyes l Biuret Reagent (Hycal, Inc., Houston, Texas). The quantity of each plasms protein vas dtcernir.ed aieetropnoretically on ca'.luloia aettaca end expressed ea a percentage of tho total plains protein. (2) Irvthrocvtc-6-amlnolevulinic acid dehydrate fAlAD): b-Amicolevulir.ic acid dehydraae activity waa dateimmad by tna method of — ch tsan and .reismao.i/’ This eroeaduta vas a carted 30 min aftar each blood sampit was taken. (3) Ervchrocvta orotooorohvrin; Protoporphyrin in erythrocytes was ossturad by the method of Heller, ee el.2/ This analysis was performed the day after blood wee wiehdrewn. -J- (!) Urinary i-tminolevulinic acid fAIJll: Urinary 4-aminolevulinic acid was measured eccerding to the Davis end «ndelaar.2£/ modification of Maui trail't and CranicU'e Method.22/ Thcet ana lye ea were perforret :n subdued lignt. «r V >SS?: ! • 'Vjr I » /n’’ 3 0007-SWP-036979 0007-SWP-000115547 (duodenum, jejunum, ileum), colon, pancreas, kidney*, urinary bladder, adrenal*, Chymu*, gonad, enyroid (with parathyroid atcacned), reseneerlc salivary (land, lymph nod**, heart, lung*, diaphragm, skeletal ru*ele, and pro*cate or uterus. r _ ST v*oRw ~ Tissue* from all 13 week rat* led tna control. 2.057. lead octoat*, IZ.uj?. lead entoeate, 66.03” lead carbonate, and t.nc 11.92* SBS lead pain; dices were embedded in paraffin, section te * thickness of 6 u, stained with hematoxylin and eosin and examined far histopathoiogy. •• " F. _ . Statistical Analysts Control and craatment value* were compered statistically using Dunnect's multiple-comparison cast with P < 0.05 ss tht criteria of signifiesnes. III. A. ttSCLCT Cl. im Ccncencrstions of Lead in Psir.c Prta* The eonetntroeiens of lead in the paint two* supplied by th* Members of tho National Paint and Coating* Association wort analysed for lead content by S*5oto Incorporated, the Shatvin-Uilliaae Company, and Midwest Research ln*ti tutaO*l). Tha result# of the** analyst* or* shown in Table 1. Th* control f*Hri which did noc contain l*ad octoat* or chromate was found to contain approximately 0.01% lead. Thor* wo* a reasonable agrees*.-.; beewt'es the different analyses, so an average ppm of lead wa* computed and used for tho actual lead concentration of oach paint film. Load concentra­ tion* of the lead paint obtained from the National Bureau of Standards wort averaged in a similar manner. B. ?. i • load Ar.elvs«j of Siets Contelnine Siffarent Paint Film* table 2 shows the lead contone of tach diet. Purina Rat Chew -as-, was found to contain approxiaiataly A ug/gm of load (assuming 0.1 ug/ga cf lead was contributed by th* control paint). 7h* ug of lead/gm of food which was attributable to each added paint filsi was reasonably elos* to its respective theoretical value, the average total ug of laad/gm of feed for each dice wis used for th* calculation of weekly end total load consumption fir seen rat. 6 : *• 0007-SWP-036980 0007-SWP-000115548 TABLE l COSCENT’LVHP: OF LEAP IN PAINT PILM Paint Tils Control Paint Lead oetoat* Lead oetoaee Lead oeeotce Ltad chranaca Lead chromate Laad chromate Lead carbonate SIS laad paint */ bf £/ Theoretical Load Concentration f”.'i 0 0.06 O.SO 2.00 0.30 2.00 15.00 64.12 • Veaeured lead Cercentratinn Paint Companies “HI Average,*/ /•> r-,\ /pi 0.01 0.08 O.SO 1.90 0.42 l.BO 12.37 62.20 , 11.87-' < 0.32 0.07 0.36 2.20 0.41 2.10 12.30 68.90 11.98 • ; 0.01 0.38 0.53 2.05 0.42 1.95 12.43 66.05 11.92&' Maaaurad by V.S. Departnant of Ceauaarea, National Bureau of Standard*, Vaihlngcon, a.C. Av«ri|* of paint coapante* and MB I value*. Average of Bureau at Standard* and MRI value*. 0007-SWP-036981 0007-SWP-000115549 -£a d a s a l y s s s or r:rrs c o s t a tvryn n:rr;--T ?a ::.t Theorottcali' Diet Control paint plus rat chow 0.1 4.1 (6) Lead octoate (0.081) plua rat chow o.a 5.2 (6) i.i (6) Lead oetoata (0.537.) plua rat chow 5.3 12.0 (6) 7.9 (6) Lead octoate (2.05.) plua rat chow 20.5 27.2 (6) 23.1 11.0 (6) 6.9 23.5 (6) 19.4 140.2 (6) 136.1 Lead chromate (0.62L) plua rae chow 4.2 * o Measured TocalS' ?elnt laaei f-js/sm) -.Carr.? - Lead chromate (1.95L) plua rat chew 19.5 Lead chTsrata (22.43-) plua rat chow 224.3 (6) (6) (6) (6) leae carbonate (66.057.) plua rae ehow 660.5 516.0 (5) 511.9 (5) NBS ; cad ammmd paint (.11-92%) plua rat chow 119.2 124.0 d> 119.9 (3) */ b' c/ 7'ii-orecieal concentration of lead in Iced contributed by indicated paint film. waan sig of load/pm feed for number of «am|ilo* aiiown in iiarunchuaia. ".can ug of lead froa paint fllms/gm feed for number of sample* shown in parenthetis. i 3 a ■ ■ ■ jdw'ii ■ " ■ .j"! t V-.. 0007-SWP-036982 0007-SWP-000115550 ... .■ -V- r. iv.-? .:■ v. w C. Preliminary Foedinc Studies i •:*.“•** Cl>T» Before beginning this study, a pilot experiment was perfersee to see if tne rite were consuming tax iced peine fTT> along with tne:r toed. Three rets each were, pieced an (lie concrol diet end the 66.03% lc-J ceroor.ece diet. After AS hr^*tece» were collected and lead content measures. ?cces tram rets on tne control diet concerned 11.5 eg of lesd/gn faces vnile (sees from rate ted the 66.05% Iced cerhoneee diet concerned 5,683 eg of leadiga fsees. Thus, rats etc esnsume tne peiot «■ *• along with tneir feeo. 0. Cross Observations ■ 4: « Feed Consunoeione and Sodv U'alehts Duri^eth^jntir^jtudvjj^jjagji^Jie^jgjejl^ig.siymm. One female ret on 2.05% iceo oeeoete developed an aoysT'w^isrjj^jjjygg^^j during the ninth ereaemenc week end one female on 66.0& leeo carbonate developed abcessed hind feet in the 10th veek end chewier toes off. ............ ■ ' 1 0 The feed eonsumpeientof male and female rats m* shown la Figures 1 end 2, respectively. Ixcept for the rats fed the diet containing 11.82% lead as KSS lead peine, all groupe conaumad feed at the tame race as tne con* tool group throughout the entire experiment. The reaaon che 11.82% KBS lead pair: group ate aignificantly lass initially, wa* that they ware started a month after the other rats and war* initially smaller. Since this group css consuming faad ae tha saaa rata at eha control group by 4 weeks, we consider tneir initial food consumption normal also (for their site). The body weight gains for each group of rets are ahown in Figures 3 snd i. a s mentioned above, the rata on che 11.92% KBS lead peine diet were small initially but bteamt similar to controls by either tha 4th (females) or Sen cask (males). Throughout she entire period the male rats in the other treatment groups gained weight at e normal rate. The female rats did snow a enanga m booy weights during tns first S weeks of fseding which appssrs related to the concentration of laad oeeoete. After 4 weeks of fseding, rats in tne T.C5% lead oeeoete group weighed significantly lass then concrol. Tha pattern was the same at 8 weeks. After 12 weeks chair average body veignss sttll exhibited the seme peccern, but the 2.05% lead oeeoete group vet not significantly different from control. This was due to fewer animals end a slight increase in variability. *0 *»—■ «-» mth 1 U-t. %. mG i*i t ^ fir-' ir v’v -: . > >- .* e»*' ” w h ;«-i 'i Fwir'i.'+***- T *• ^ . , xr 0007—SWP-036983 0007-SWP-000115551 I - vwif. J ......... . -. v^V*.'»<*** '>• . ■ «>* •-**■ ■ •* < • ■ .^.v k-. ; cv- v. .. •’*%*:-. r. , • ."V .* .: _"' :. '=\'..-*li^aL;;.. v-s.r/sr.K'.f ...*..'*. w -< ? ii Q i 3 ■ s^ !tr ^Igtfjag^^ytv:'gaafiMsaggl au J*>i_L. mini] ] f s ____ □ ] 1 1 ] I ] ffg.*w?»aa»i.a' -g- u l m s-v-e-^Jl^X^a aa ■ **$'• ' ' • • . .»: ?[- i -u--------------------------------- i—; sites?: ~ • “ 2 3 = «. []£'§□ □ ^ JJ ' frrz^=pL V ;-y □ -.1 .3. «) •.“IJJ/.V. •.; : ’.0 fM*. M - C-'^-r.;■ ;--=- 5: -. . 0007-SWP-0369B4 0007-SWP-000115552 5 ’•Otjtf.'tf'WCS :}3> n j '»Tr; ^ ^ ... p„ I 0007-SWP-036985 I 0007-SWP-000115553 Figure ■ =! I«*1 ' -• Cl* ’■ J ~nr 0007-SWP-036986 0007-SWP-000115554 I ______ _________________ * 7"! .". ’-** ' • ~A7» "n’‘ IEs " *_! »T3 ’ ^ •-* i—J ^WA\ = ta-t .......... ’.••*•. “t pj • , . . • _____ *—1 .. .......................................... ■ • c ■ «. : v ; k_\. •■ • .,•. - -a ■ •■ / ■ ■ a vE ' — ..... . ... ...... . . • W-9*.: *>: •' Ji “ ■ 32 n___ ___ -n o . . * . . . ■ 1 .... ............ • . ' • «• . . • kv ^;r-v~ ■ ■ •?•> •• ■ # Ilf !! rv‘ I l <5 7 I n i ] 1 ids 1 7 1 io *io %N■ s CCG ID 1 w 1 j; 5 •SSESE3 s i a s 3 r 1------H CIj O I*-5 1 n 0007-SWP-036987 0007-SWP-000115555 '-'ttk jv and Tata i a Corsur^ticn The weekly and total, leas consumption djring th« experiment were eaieuiaced sn the basis of the tetel iuad/diet and amount of each diet eaten by each rec. These calculations are snsun in Tables 3 and a. During the 13th week period, the control group consumed approximately 1C3 ug of lead/da while rats eating 2.:sr. lead octoate, 12.4311 lead chromate, 66.03!; lead careonace, and 11.92". MS lead paint eonsunad an average of 786.5, 3,391.1, 13,435.8. and 3,184.3 ug of lead/dav, reapeeeively. I'aing the average body weights of rats surviving 13 days, the rats m the control, 2.035 lead octoate, IT-431 lead chromate, 66.051; and 11.927. KBS lead paint groupl coneumad 245.2; 1,957.5; 8,469.6; 34,430.8! and 7,804.7 ug of lead/kg/day, raapeceivaly. F• Hegatoloay of Bata Fed Palr.c Swims Containing Oiffsrent Concentrations The results of the hematology of rats fad paint films containing different concentrations of lead are shown in Tablas 3*13. No differences were observed in the erythrocyte count, reticulocyte count, hasatocric, leukocyte count, differential leukeeyte count, and the erythrocyte oamotie fragility. o C. Tt«n.« and Fluid Chemistry Concentrations of letd • lea • it Rats Ted Psint 6 cx-r Tables 14-22 show the tissue end fluid chemistry of rats fed paint ms* containing dlfferant concantraclona of laad. The serum proteins, crytnrocytt protoporphyrin, urinary coproporphyrin, and urinary dalta-asmoiavulin;: acid ware not altered by any of the diets. . •- '*i \ ,!TS The activity of ehe arythroeyta aasyma, dalta-aainolevulinie acid cenydraja fAlAD). is thi control, laad octoata (0.085, 0.335, and 2.035) and '.sad enrsraea /l.-111. 1.537. and 12.«35) irouoa am not diifdr tnrouchout tna axnar'.r.snt -Taolas 13). a IAP scttvicv was eoBresaad in rats fed 66.03% laad carO£ngy,_gjg>ii^^2%K^ At 4 weeks thara was a depression Of 36% and 39% in thaaa two erouos, ratesdttvsiv. At 8 wteka tna ALABactivicy was -^ffSnara hug in* A LAD activity in tna MS laad paint group was not significantly different from control. At this time a sex diffaranca waa observed in both of theso groups. The AIAS activity m the male rats remained depressed it the 4-waek>laveIs while the r.L/O activities of the fssnale rets returned to control levdls. This sex difference say be liniied to Che fact that these female retsNttad matured and wore prruaoly Beginning chair mensArul cycles. (TsUglinv) 1P*sk ;fc- r'W; 7> %C revi>t 7#0 «»p* 0007-SWP-000115556 t'iiv u *> z = i. = « * ; ; ; \i ;: ti r i’ ;" m ? X •• A i £ il m t t a* i£ i= X • • * ; i * .*2 *• 7 i4 i*: - zX •s£ * * .2 :• - >* >• •- y~- - >»• «.... x. * «5 V- 8* V is X x X X il Asl l s* i* v> X i r Is si5 Is i s = i* !* is it i* i* i£ -• x i* i£ 35 i.1 i5 i;: <* i! j* j/: < z : A H Is i5 m Is i5 i‘ s* * : z ii Is V V i5 i5 *• £ ii si n * I3 X V z i= H li A r * ii i* £ :» s« ii :i *- -- n u * s i ii H i' j;i xii 7£ £li £ * ii !3 ii ii •*:i n ll ii x x ■4 r 5 A 7 s i? ii i ■ ;] ! 2 r * < i 13 n ?* 5= ;ii ' i ; i, -■ ?zzrrr?r. 0007-SWP-036989 0007-SWP-000115557 o - £ '* i t L£k- •t -5 •5 ^ s I s ; £ • s •I *7 =i P P p *s r s- r :■ 'i Zi i i • • A-.-.' 'i * :■> - :* : :* ; 'i r r ! f i* !* P P P j* r s* 5* ;£ ;i * J ; j I ■ * s= =• «! p i5 . *' “; *; 5= i- S- a SI 71 t! U*-'' »• s^C*. 5* 5* 51 ■ 5* : I s *. s * s p 5­ S5 *• -p s* s * 5* : ^7 Zi i: ;s i! i* 1! P Is *7 * 7 -3 ji =5 p j* p :■ :* Is 5s .* -• s s •­ r is i5 I *2 I§ i*- i.*- i* P i5 5 i j I S‘s -5 »ia !^ Q 1 j-* i! ^ P i5 «* siIs - - * I1- II P r 3‘ i! -3 if j! i! P P P a M - * a i •a '5 5 = il is |'3 i- ?i \l j* j* ?= V £ rt\* 55 il P a it il 1n »;* ‘-J i? 5? s* • i » .r> T 5 I ! !, I: j? 5 V * 2 r { E:^ ‘■ r.. * i* i •• , « , !M 1i ? i* * & if l iU & s <71 Ji;4 T=*: -*i: ■ iili 16 r . ‘**V ?• 0007-SWP-036990 0007-S WP-000115558 *- TABLE 5 HEMATPTOCY ™ »ATS TE" ?A?*:T coxTA::.ir:c n o l e a d Treat-tnt Veck 6 s 13 ; i9 ) . \ o irythroeyte* (x 106/r «3) Reticulocytes, " Hematocrit, vol.^T. Hemoglobin, ga, L Leukocytes (x 103/os3) Seu crop bill, 7, Lyaphocvtei, ' Benda, 7. Eosinophils, " Basopnils, * Monoeyres, ?, Atypical, T. Nucleated BBC, " irymrocytt o ie io c ic fragility [SaCl]£> i b/ SJ 6.17 ± O.iiS^ 1.5 ± 0.7 66.1 ± 0.6 14.0 s 0.3 5.0 * 0.5 11.a * 2.9 86.3 x 3.3 0.3 i 0.2 0.9 ± 0.4 0 1.1 s 0.6 0 0 0*311 * 0*004 6.33 ± 0.19 0.8 = 0.3 44.6 « 1.2 14.6 a 0.3 7.6 * 1.3 21.7 a 5.6 75.4 * 5.3 0 1.3 a 0.4 0 1.6 ± 0.6 0 0 0.391 * 0.008 6.42 x 9.29 1.4 s 0.1 44.6 * 0.3 16.0 * 0.1 8.0 x 0.6 13.1 x 1.5 84.1 s 1.6 0.1 = 0.1 0.3 = 0.2 0 2.1 * 0.4 0 0.1 x 0.1 0.400 x 0.004 Nusoar of race par parted, Mean s icandard error. Concentration of XaCl that hetnolyred J0& of tho erythrocytes. 3 . ■ •« •’ '■ X:' * ’■ " ./V -»'■ 1* H:5: m ? *.*• 0007-SWP-000115559 TABLE 6 0 HEMATPIOGY C? 3-ATS 7E0 PAINT ■ ?*•— coNTAir.'t::; r.'vy. ’.t a p a s '.t a b o c t o a t e 4 Trucr-enf V««k S 5.14 ± 0.32-1 0.9 = 0.5 41.3 * 0.9 14.1 s 0.4 5.1 a 0.S 24.5 a 1.6 74.3 * 1.8 0 0.8 a 0.3 0 0.5 = 0.5 0 0 0.381 a 0.005 6.33 = 0.09 1.0 a 0.5 46.0 a 0.7 15.4 a 0.2 5.5 a 0.7 20.3 a 4.7 76.3 a 6.7 0 1.3 a 0.6 0 2.0 a 0.8 0 0 0.410 = 0.007 A.-s Iv s b s o Ervihroeyt** (x iO®/B»^) Baclculoeytts, % Kraatotrit, vol.j T. Hemoglobin, gry % laukoeyta* (x 103/(0x3) Neutrophil*, *. Lvnphoeyti*, * Bond*. ~ Eoiinophils, \ Saaophils, * Manoeyt**, X Atypical, * Nucleated %BC, * Eryehrocytt osnocic fragility [SaClZ-/ S**X-‘ >V V-y-VA"' 13 r;;.12> 5.92 a 0.39 1.4 a 0.2 43.0 a 0.5 14.7 a 0.8 7.7 a 0.9 16.3 a 1.7 SO. 9 a 1.8 0.3 a 0.2 0.7 * 0.3 0 1.9 a 0.5 0 0 0.395 a 0.005 ;v **%*... £/ b/ c,/ Numbtr of :s:s par period* Mean = ie*ndird arror. Concentration of Nad chic htmolyred 50" of Che tryth?oeyc«£» .VAf * l ' r-.-rir Ir? ,. / ...du. 0 8 0007-SWP-036992 0007-SWP-000115560 * “ABLE 7 tJU*HEMATOJWY OF HATS FFO ?a :n t s«-9 CONTAINING -IS}''. LEAD AS LEAD OCTOa TS A reatment ’Jack 8 fS.il 13 6.27 = 0,±7 0.9 = 0,7 42.3 * 0.7 14.2 * 0.2 5.0 a 1.4 15.8 a 3.8 83.0 * 4.4 0 0.8 * 0.3 0 0.5 ± 0.5 0 0 0.383 a 0.006 6.04 s 0.31 0.9 * 0.4 43.8 a 0.5 14.9 a 0.3 7.5 * 2.8 19.3 « 3.8 77.3 * 3.8 0 1.0 « 0.4 0 2.3 a 0.5 0 0 0.388 a 0.012 3.6 s 0.4 1.6 a 0.2 43 s 0.7 15.3 a 0.3 8.3 s 1.0 16.2 s 1.9 81.4 a 1.3 0.1 a 0.1 0.8 a 0.3 0 1.6 a 0.3 0 0 0.401 8 0.006 Analyses Erythrocytes (x 106fsm3l Reticulocytes, Z Hematocrit, vol.y % Hemoglobin, gm^ Z Leukocyte* (x ioVam^) Neutrophils, Z Lymphocytes, % Banda, Eosinophils, Z Sisophils, Z Monocytes, Z Atypical, * Nucleated BBC, 7. Erythrocyte osmotie fragility [NaCli^ a/ Jb/ t/ I v; y , > Nursbar or rata par period. Mean a acandafd error. Concentration of SaCl that heaolyaed 507. of Che erythrocytes. ■ • 1 !- k -y-;- 19 0007-SWP-036993 0007-SWP-000115561 TABLE 8 o HEMATOLOGY g? RATS F~0 PAINT g;*" l e a p Ai aaraefsstt c o x t a in in c ! Treatment Votk 8 CN-41 4 Ay Ivifi Erythrocytaa (st X06/«r *) Retlculocytaa, « Heaaeocrit, vol.a X Heaojlobin, ga» .. leukocyte* (x lO^/W) Neutrophil*. % lynpheeytea, « Band*, T. Sotinophils, * Basophil*, X Monocyte*, S Atypical, ” Nucleated RBC, 7. Erythrocyte otaocic fragility 'SaCl'^ a/ b/ £■ ' 13 n:»i2' 6.30 * O.li7 1.1 = C.6 AX.8 * 0.3 14.4 a 0.2 3.1 * l.X 13.3 a 3.0 83.8 * 3.2 0 0.8 * 0.5 0 0 0 0 6.46 s 0.08 0.7 a 0.1 43.0 * 0.4 15.3 *■ 0.2 6.7 * X.7 13.0 * 2.1 83.0 = 2.3 0 0.3 ± 0.3 0 1.8 * 0.9 0 0 43.4 * 0.8 13.6 a 0.3 8.1 ± 0.7 14.1 a 2.6 82.8 a 2,3 0 0.9 a 0.3 0 2.2 a 0.7 0 0 0.381 a 0.006 0.399 s 0.008 0.393 a 0.006 6.72 = 0.42 :.o = c.6 :.'uso*r of rat* par period. Mean a standard error. Concentration of SaCl that hemolyied 50* of the erychr ocytu. f*; SftI'dcfc- • *'« Ki* V-* * P5, &srf>Arf! 1 L**fi*^!W* ” -U’. #?&*!? 4&&S {Sss^ W0$- yj/i“ ;5":V-} -?h\ ■ «s 20 **£ 0007-SWP-036994 0007-SWP-000115562 ms 9 HEVATOIOCY 7? EATS b a t -?- &L*r* r-u. &&>:• 7 reatment Vick An lvsas Erythrocytes (x 10®/m*31 Reticulocytes, % Hematocrit, vol./ % Hemoglobin, gm^ 7. leukocytes (x lO^/ax^) Seucrophlls, % lyupnocytts, 7. Banda, % Eosinophils, % Basophils, % .Monocytes, % Atypical, 7. Nucleated RBC, 7, Erythrocyte osmotic fragility, CsaClji' a/ b/ c/ a 13 f*ei >' 6.16 x 0.16 0.8 x 0.2 65.0 X 1.1 15.1 ± 0.3 5.1 x 0.5 19.0 x 2.0 79.5 x l.J 0 0.8 * 0.8 0 0.0 X 0.8 0 0.3 * 0.3 0.395 X 0.12 6.6 s 0.3 1.6 x 0.2 43.6 x 0.7 15.6 x 0.3 6.9 x 0.7 16.3 x 2.1 83.6 x 2.0 0 . 1.2 x 0.4 0 1.2 x 0.4 0 0 0.392 = 0.007 fSW.i' 6.71 x 9.16^ 0.9 ± 0.8 63.3 * 0.6 14.7 ± 0.6 5.1 x 0.6 8.5 * 1.7 90.3 X 2.0 0 0 0 1.3 x 0.8 0 0 0.383 x 0.003 *•/*»* Number of rata per period, except where indicated otherwMr. Mean X standard arror. Concentration of NaCl that heaolyied 50% of :ha erythrocytes. m * * •i/a.-* tt.*****^ -p’-c».• n 4 ■ r^'.r *■ '***:-.••*4 0007-SKP-036995 0007-SWP-000115563 I 7AS1Z 10 •3 >cr~ l**Y.VT0Lf *v HATS r-t) u' CflT.TA "C :.-5* -Jha' a s mm2 crsnv.vr- •/ V c/ 6.37 s 0.52 1.2 3 0.2 44.1 s 0.7 16.2 s 0.4 8.2 z 1.0 II 6.21 = 0.13 0.8 * 0.3 44.3 s 0.3 15.3 s 0.3 6.1 3 1.2 18.3 s 6.6 80.3 s 6.6 0 0.3 s 0.3 0 0.8 0.3 0 0 0.383 * 0.007 £-i\r* ' UrS^v.j* «.» 6.17 s 0.0j£' C.S s 0.4 40.8 z 0.6 13.9 z 0.2 4.9 s 0.8 10.0 3 2.4 92.0 z 1.8 0 0.5 * 0.3 0 0 0 0 0.383 = 0.007 13 f' Erythrocytes (x 106,rp) Reticulocytes, \ Hcnatoeric, vol.jZ Hanot lob in, tv ' Leukocyte* T- 22 0007-SWP-000115564 t abl e o u HEJWTe’.ocv or b a t s ~o **~r CONTAIVINC: i;.-3*T LEAD AS LEAD S!3eVATS Erythrocytes (x 10°/ni«^ ftetieuioeyte^jfc Heaatoerie, vol./ 5 Hcaeglobin, jiy 7. '.aukoeytei (x lOVsm^) Seutrophils, 7. Lymphocytes, % Bands, 7. Eosinophils, T, 3asophils, * Monocytes, % Atypical, 5 Nucleated BBC, T. Erythroeyee osmotic fragility L-VaClI-7 a: 6/ 0/ . • L"!*-^4a r- A Treatment ”«ck 6 /?r»4i 13 'S-12' 6.32 = 0.14^ 1.95 a 1,28 41.3 * 1.1 13.9 = 0.4 4.6 a 0.3 21.3 a 6.3 73.0 a 6.0 0 0.5 * 0.3 0 0 0 0 0.384 a 0.007 6.23 s 0.18 0.9 a 0.1 43.8 * 0.5 15.2 a 0.1 5.3 a 1.8 15.0 * 2.1 B2.8 a 2.0 0 1.0 a 0 0 1.3 * 0.3 0 0.3 a 0.3 0.385 a 0.010 6.61 a 0.33 1.6 a 0.2 44.1 a 0.7 15.8 a 0.2 7.6 a 0.6 12.8 a 1.6 34.8 a 1.7 0.1 a 0.1 1.0 a 0.4 0 1.3 a 0.3 0 0 0.396 = 0.006 AnaK'sea o SS£ , • • i*' f ’ . * • «. W'.*r:i£ q*£nMt’ !*rf; *v*vS NueSer pi rats par period. Mean a icandard error. Coneantratlon of NaCl chat hemolyred 305 oi the erythrocytes. o 23 I® g!§ 0007-SWP-000115565 TABLE 12 3 . , nEMAToiosy gy r a t s rsn ?A?vr •r^T. CONTAIS'rS’G LEAD AS LEAS CARBONATE Analyses 13 t"..\ v 6.60 3 0.37 2.9 s 1.3 44.7 m 1.2 15.6 s 0.6 7.7 s 0.3 13.2 3 2.6 82.7 s 2.7 0.2 s 0.1 i-e .•*! ?:2£?C 1* n o S.13 3 0.06 1.1 s 0.3 44.0 * 1.3 13.0 * 0.2 3.0 * 0.9 13.0 3 4.1 14.0 * 4.2 0 0.3 s 0.3 0 0.3 3 0.3 0 0 0.394 * 0.012 o */ V c/ 3.94 * 0.36^ 0.4 s 0.2 41.3 ± 0.9 13.9 * 0.3 4.3 s 1.2 14.3 s 4.1 82.0 x 3.8 0 0.7 4 0.7 0 3.0 s 2.0 0 0 0.388 3 0.009 psag TreitncnC Ve*'« 8 Oh © Erythrocytes (x 10*/™*) Reticulocytes, % Hcautocrit, vol.y 7. Kaaoglobin, gay % Leukocytal (x I03/soi3) Seutrophill, r. Lyapnoeycei, % Banda, X Eoi inop hill, 7. Baaophila, X Monocytes, X Atypical, 2 Nucleated RBC, X Erythrocyte osmotic fragility [NaCl’- A 'S»3'l/ 0 1.0 s 3.3 0 0.1 3 0.1 0.397 = 0.003 sS8 Xuobtr of rats per period. Mean s standard error. Concentration of NaCI chat hamolysed $0% of the arythrocytet. *3r£ v-—»«* • 1 4» *• b Me 2. TajyaB'f 0007-SWP-036998 0007-SWP-000115566 TAIC.E 13 ••roLC'r or r«\t s r*o ::ss l s /o :sr •ctwiwjfsss K.«r :e a d 6 {Smi.'li1 3.72 s 0.15^ 2.5 * 0.3 39.7 x 0.8 13.6 * if 9.7 ± 1.4 9.5 s 2.X 90.2 * 2.3 0.3 * 0.3 0 0 0 0 0 0.389 ± 0.006 Treatnenc Waat" a rv»4i 4.49 £ 0.83 1.0 e 0.6 42.8 * l.l 16.7 * 0.3 6.7 * 1.2 7.0 * X.7 92.0 * 2.1 0 0 0 1.0 £ 0.7 0 0 0.412 * 0.006 U 5.63 = 0.36 1.3 s 0.1 62.9 s 0.4 14.7 * 0.1 — 4* 7.0 X 0.5 14.2 * 2.1 83.6 = 2.2 0 1.4 s 0.4 0 0.8 ± 0.3 0 0.1 X 0.1 0.408 x 0.005 :iriod, excapc vhara indicaced oehcrvise. chat hamolyia* 50% of eha arychroeytaa. :s ^TV^rT,; ■ > '‘ 0007-SWP-036999 0007-SWP-000115567 TABLE 14 c !mv t is s u e a s p r-vin oitirrsisy o f mt s TE3 ?Ai:rr c Sn t a i :::^ k >, l e a p ................... Traarr-artc 4>«k 8 r:.g> 4 Analyses Serum *l«c:rophor«*i* Albumin, % Alpha l globulin, % Alpha 2 globulin, T, Bara globulin, 5 Gamma globulin, % Total prorain, fa X Albuain/globulin ratio '.3 <>24) 43 28 s 1 J a 1 20 * 1 6 s l 5.4 3 0.1 0.76 £ 0.03 10 x 1 6.2 s 0.2 21 3 A 6 3 1 6.3 s 0.1 0.77 * 0.07 0.93 s 0.94 Erythroeyte ALAO, umol. PBS/ Hale*: 100 ml ABC/hr Female*: Total: Protoporphyrin, ug/100 ml RBC 23.2* 2.3S7 16.6 x 3.1*' 19.9 x 2.2 19.6 * 1.6 15.1 * 16.3 s 2.2&' 15.6* 1.5 23.5 * 1.9 14.5 * l.i£f 13.4 x o.-i' .. 15.0 * 1.0 29.2 x 2.0 L'rina Copreporphyrir., ug/24 hr 2.5 * 1.2 3.3 * 7.4 6.0 * 1.9 78.0 s 7.4 83.3 * 11.0 43.4 * 5.0 ALA, 43 * 2 23 * L 5 * 1 20 * l 48 x 1 21 3 l 5 s 1 ug/2A hr a/ 3/ it Cushat oi ran par period, except where indicated oeharviaa. .‘lean = standard arror. Four rasa. Twelve rac*. 26 0007-SWP-000115568 ■ .................TABLE 15 .............. ..............>......... t is s u e a :.'p firu cvr.v:sTBY o f s at s rsi COKTAWta i. 25". tljiAi AS '-ini' OCTOATE Treacrent :.*eek 4 Artlv*»s Sarua electrophoresis Albumin. *. Alpha 1 globulin, X Alpha 2 globulin, 1. Baca globulin, X Camma globulin, X Total pretain, ft X Albumin/globulin ratio Erythroeyces AUO, uaol. PBC/ .Ha lac 100 ml R»C/hr Famalaa: Total: Protoporphyrin, ug/100 ml BBC Jrint Coproporphyrin, ug/24 hr AtaAg ug/24 hr 4/ £/ :/ i! 13 a iiil 40 ■ ii' 26 * 1 5 3 1 21 3 1 8*1 5.9 * 0.1 0.66 * 0.02 44 — 3 20 3 2 4 * 1 20 * 1 12 * 1 6.3 * 0.2 0.79 3 0.08 48 = 1 20 3 1 ..... S 3 1 22 3 1 7 * 1 6.2 3 0.1 0.93 s 0.04 26.8 * 7.7i' 19.5 * 4.iSJ 22.2 * 3.9 28.4 x 2.4 14.9 s 1.3£' 18.7 * 0.3&' 16.8 3 1.2 24.2 3 1.6....... 12.0 = o.ci/ 16.1 3 1.3^ 14.0 s 1.4 22.8 x 2.5 1.2 * 0.2 4.8 * 2.4 2.9 = 1.4 86.4 * 16.6 107.6 3 34.1 35.0 3 6.7 • » c •» * . .•’ :sv-Jfr »p' ©aKi®r: feSs.- Musbcr of rata par period, except where indicttad othtruiae. Maan * acaadard error. Two rata. S ix ra ca. n h TL_> : "TiX, 0007-SWP-037001 0007-SWP-000115569 as^ n?j: TAS1* 16 23&i- T.ISSCE ACT K03V• =*s :<:s t »Y OP PATS *?D PAT' •COSTA':*:2N0 \53*- MV0 A< errOATT ......... ' ........... CW^e Traacnanr Weak 6 ('^41 « Ana!-.-*** Serum alectrophoreiia Albumin, r; Alpha l globulin, a Alpha 2 globulin, T. lata globulin, * Cana globulin, X Total protein, dtg X Albuain/globulin ratio Erythroeyta* A LAD, uaol. PBC/ lUlaa: 100 nl RBC/hr rasalei. Total; Protoporphyrin, Hg/100 ml X3C Urine Coproporphyrm, Pg/24 hr AU, ug/24 hr V b/ SJ e/ rsai'i' 13 '•■ VIA.- •«•:-£■ ... 41 * •£' 26 * 1 5 * 1 21 s 1 6=1 3.7 = 0.1 0.66 * 0.04 41 * 4 ’ 23 x 3 6 x 2 20 x 1 11 x 2 6.3 x 0.1 0.70 ± 0.13 21.4 x 6.& 15.5 x 2.T&1 18.4 s 3.3 23.1 = 1.0 10.3 x 0.8£/ 15.3 x 0.2£> 12.8 x 1.5 21.7 * X.l 12.5 * 0.7&' 17.0 x 1.2$/ 14,7 X 0.9 27.7 x 3.3 10.7 x 5.9 1.1 x 0.6 4.9 x 2.4 97.4 s 19.2 101.6 x 30.1 50.5 x 6.6 50 = l 20 x 1 4 x l 20 X 1 7xi 6.0 s 0.1 1.00 X 0.07 {£& £*?&■ m ?**>;!■ e***V%*^.— £* V^C 4*1 .^unoer oi rata period, axeape wnert indicated otharviie. V.aan r *tandard error. • **> Two rat*. Six rat*. 4- •.- r - :s • 1 •■ •' ■ nnna.03D.naanaa 0007-SWP-000115570 TABLE IT "" ' TTSSl’S AND BORY fU'Tn c v e v t s w f'T SATS rr*> ».»• S*T* C3.\Ta :v T-.'G 2.05’. LEAD AS LU.‘.0 0CT3ATE ftk* • i* rracsent "ask 8 .\Va4> 13 41 * 3^ 27 * 2 4 a 1 20 a 1 9 x 1 5.7 a 0.2 0.70 x 0.07 46 a 2 22 a l 4 a l 19 a i 9 a i 6.0 a o.l 0.85 a 0.07 48 a 2 20 a 1 6xi 21 a l 6 x 1 6.2 x o.l 0.96 x 0.07 24.1 a 4.0*' 18.7 a l.o£> 21.4 a 2.3 29.5 a 2.8 12.1 a 1.7S' 11.3 a o.z4; 11.7 a 0.7 30.8 a 2.5 11.4 a 0.8^ 16.3 a 3.9^ 13.8 a 2.0 24.0 ± 2.2 1.6 a 0.6 3.0 a 0.8 7.7 a 2.9 80.7 * 17.3 82.7 a 10.4 64.5 x s.8 AM lviai Sana electrophoresis Albumin, T. Alpha l globulin, X Alpha 2 globulin, X Seta globulin, X Casons globulin, X Total precaln, tl X Albumin/globulin ratio Erythrocytes AiAD, unol. ?BC/ Malta: 100 ml XBC/hr Females: Total: Protoporphyrin, ug/100 ml BBC "fine Coproporphyrin, ug/24 hr ALA, ug/ 24 hr a.b/ c/ »•' " : -xbar of rot* ptr poriod, except vhart indicated otherviee. Moan a standard arror. Tuo rat*. fix rats. 29 w ” 0007-SWP-037003 0007-SWP-000115571 TABLE IB TISSUE AND BODY fr.CIP CHEMISTRY ffr RATS rso ?a :?.t XW* COmiKISS 0.;r, LEA3 AS LEAP CHROMATE i'l: V 'rtatncnt Vcak 4 An !'■ »«« 8 13 <»41 OM2) Albumin, X Alpha l globulin, * Alpha 2 globulin, X Beta (lobulln, X Cima (lobulln, % Total protein, £g X Albuain/globulln ratio 42 * & 28 * l 4 * i 19 * i 8 * 3 3.8 X 0.2 0.71 * 0.06 44 s 2 2Q * 2 4 X 1 19 X 1 12 X 2 6.0 X 0. 2 0.80 X 0. 08 47 X 1 22 X 1 3 X 1 21 X 1 6 X 1 6.1 X 0..1 0.90 A 0..03 ".... ' Erythrocyte AUO, pool. PBS/ Male*: 100 ml MC/hr Pi■ ulet: Total: Protoporphyria, uj/100 al TBC 23.4 * 3.5fJ 13.4 A 0.9^ 18.4 A 3.3 27.3 A 2.8 14.1 X 0. 6*/ 17.6 X 0. & 15.7 X 1. 0 28.3 X 0. 6 15.7 X i .,s*i 16.3 X i. 16.1 X i. 0 27.9 X 2.,1 ^ .Tj * '?• 25 a 1 4 S l 20 * 1 9 * 1 5.7 * 0.1 0.73 ± 0.06 o.si a 0.04 47 a 2 19 a 2 5 * 1 23 a l 6 * l 6.4 a o.l 0.94 a 0.08 29.5 * 3.& 20.4 a 4.3S/ 24.9 * 3.4 26.5 * 4.7 12.3 a 1.0£; 14.7 a 0.6£/ 13.5 * 0.8 26.8 a 1.8 13.3 A l.id/ 12.9 a 2.34' 13.1 * 1.3 27.4 a'1.5 0.9 * 0.1 7.0 = 6.4 4.4 a 1.9 S0.B a 7.2 67.7 = 6.8 66.5 * 17.5 20 * 1 11 * 1 6.20 * 0.04 fev p; v'y-v I •***;•*< *•**< « -vvr*» Suober at rare par period, except when indicated otherwise, Mean * icmdard error. Two race. Six rat*. 3 •V .. . 31 0007-SWP-037005 0007-SWP-000115573 TABLE 20 TISS11 AN5 BOW r VtS C^SMISTB? 0" -ATS *ST'PAT.'.T 1 ccs?Arnsg "::.^r LEa c *^'L£X5". :^t:aTTreat-ant *•>«’< 6 ....... iS»l) Anaivss* Strum tlettrophortsis Albumin, X Alpha 1 globulin, * Alpha 2 globulin, * Seta globulin, X Gama globulin, 7. Total protein, ^g 5 Albumin/globulin ratio Erythrocyte ALAO, uaol. PSG/ >!alea: 100 ml RBC/hr Famalat: Total: Protoporphyrin, ug/100 ml BBC t«M« l«* l*» Irina Coproporphyrin, ;s|/24 hr ALA, ug/24 hr / ' ■’ i&f*. :3 AO * £/ 27 a l 3 a 1 22 a 1 8 * 1 5.6 a 0.1 0.68 a 0.03 47 * 3 21 a t A a 1 18 a 1 * * 2 S.l a 0.1 0.90 a 0.12 46 = 2 21 a 1 5 a 7 ”2 a 9A. 6 a X 6,4 a 0. X 0.88 a 0*36 26.4 a 7'4=£/ 14.2 a 3.2-; 20.3 a 4.9 25.3 * 1.6 16.5 a 1.4*' 13.4 a 3.8—^ 15.0 * 1.9 24. 2 a 2.6 12.6 a 13.7 a l.i13.1 = l.G 24.6 a 2.3 2,6 * 1.6 0.9 a C.l 2.6 a 3.3 as.7 a 19.3 78.4 a 6.8 49.7 a Uii j-fc'*;. ‘3$. rfafcrfA Vv t: LV'-? Suaber of ra:a par period, exeepc vhara indicated otherwise. Mean a standard error. Two rats. Six rats. • **• 4 V rt-.Buwipr 32 u• r".i .... »................. .. >7 n^aplBp 0007-SWP-037006 0007-SWP-000115574 .e^r- TABLE 21 A‘;r r.mv rt.:*T9 CKEWSM*■ or a a -v t t o : ?*:■ SU rc.NTAIS: •:s -j0.051 LEAD A S ISAS CAKOVATt t is s u e UMf* Tresc-cnr ’.t** ! ^y»4i am A:i*vses 5erua alaccrophorasia Albumin, T. Alpha 1 globulin, " Alpha 2 globulin, % Bata globulin, * Cacma globulin, % Total protain, fg % Albumin/globulin ratio 41 s 6 27 * 1 3*1 21 * 2 a 12 5.4 a 0.2 0.73 * 0.17 Erythroeyta A1A0, uaol. PBS/ Miles: 12. & 100 ml ABC/hr females: 9.1 * 2.11/ 10.2 = 1 .blf Protoporphyrin, 23.2 = 0.9 ug/100 ml ABC Trine 4HJA Cooroporphyrtc, ug/2A hr AUA > ug/ 24 nt 45 a l 21 a 2 19 a 1 10 s 1 6.0 a 0.06 0.12 a 0.04..... ■ s 47 z 2 22 = 1 5 * 1 20 * l 7 z l 6.3 r 0.1 3.56 s 0.06 4.4 * O.li7 12.4 a 0.4J/ 6.4 z 2.3X' 20.4 s 2.1 27.2 z 6.8 2.0 * 0.7 0.S z 0.3 6.7 s 2.2 90.3 * 27.7 72.3 = 13.4 42.2 *5.8 t *...' 3.4 a l.Ofi/ 7.4 a 0.5*/ 6.4 z O.rP to* t«> |«i •* * , •’ ! / / Sumoar of rats par period, except utiara indicated otherwise. >’.«an a standard error. Ona rat. Two rats. Six rats. Significantly different iron control (? < 0.03> as shown by Synnett's nulciple«eoeparisoB test- following an analysis of variants. v. if sags At*** 23 0007-SWP-037007 0007-SWP-000115575 rs- :a »u TtSSrE vry ' 1£AD ?. • fV^rT!6??::? ST EATS ??2 ' SS .»*» ii.®r is.\o....... a Treatment •■ .'oek 8 '.W> <• Anjlvsis Sarum alectropnoresis Albumin, ” Alpha 1 globulin, * Alpha 2 globulin, * beta globulin, 7. Caiaoa globulin,' Total protein, f.g % Albumin/'globulin ratio . 13 /w::- 52 = :i = i i = i 20 * 1 3 = 1 5.4 * 0.2 1.1 * 0.03 44 = : 21 si 2 8 * 5 21 * 3 6 a 1 6.0 a 0.2 0.78 * 0.06 44 s : 24 s 1 7 * 1 21 * 1 5 = 1 6.1 = 0.1 0.82 = C.26 Erythroeyta ALAD, nmol. ?SC/ Males: 8.5 si 5.0i/ 100 ml UC/hr Females: 10.1 * o.i—/ Total: 9.3 s 2.1*/ Protoporphyrin, 14.8 a 4.3 ug/100 ml sac 5.0 * 0.3*/ 15.0 * 5.1—/ 10.0 * 3.6 24.5 * 1.8 4.9 a 0.61/ 8.8 a l.C^/ 6.9 s O.S—' 26.3 a 1.2 2.0 = 0.8 6.8 * 4.9 4.6 a 2.1 nc.i * :i.5 72.0 * 13.6 51.3 a 3.6 Trine Cop roporphy rir., ug/24 hr a ia . ug/24 hr I * *' '** ;v*‘’ * r'-. N'usber of rats per period, axeapt unsrs indieatad otherwise. Mean = standard arror. Two rats. Six rats. Significantly diffaranc from control C? < 0.05) as ir.dicacad by Ouanttt's ajlsiple»cot?ariaon test following aa analysis of variants. Si.-.— - i 34 0007-SWP-037Q08 0007-SWP-000115576 0 o '•■ v! TABI.E 2 I . e*>r~ SIIHHAHY OK rilKrilYtlH HKIAMUI.ISH 1H BATS FEU FAINI.FOB >1 WEEKS CONTAIN INK DIME RENT CONCENTBATIONS OF TJAI) m M yllirocvte__________________ ___________ III unify *7 AIAI)S Diet ?iv •<;:.'ia :51? # £J (■ •mil. l'BC/100 oil KMC/lir) lBfi/100 ■ 1 BMC) *¥ Control 0.U8Z l.end Oct oatc II.VJl l.cml Octoate i/S.QYL U'ol Octoate 0.421 leal Chromate 1.951 l.ead Chromate I2.4U CliriMate 66.U5Z la‘»d Chromate 11.721 IMS l.ead Faint 24 12 12 12 12 12 12 12 12 15.0 4 1.0^ 14.0 4 1.4 (ft. 2 4 0.9 u. a t 2.0 16.1 ♦ 1.0 11.1 f t.l 11.1 * 1.0 - 6.4 t 0./£/ - 0.9 4 0.8“/ 29.2 f 2.0 22.8 4 2.5 21.2 i 1.3 24.0 t 2.2 22.9 i 2.1 22.4 * 1.5 24.6 1 2.0 22.2 4 0.8 20.1 1 1.2 6.0 4 2.9 ± 4.9 4 2.2 4 1.9 1.4 2.4 2.9 4.3 i 2.2 4,4 4 1.9 2.6 t 0.8 0.2 4 1.2 4.6 4 2.1 (1‘g 41.4 15.0 50.5 64.5 85.1 66.5 49.2 42.2 51. :V1 ,. ** ?•» •• ,).• •« *• * **•». *V1 |i iM ■ ;M 1 2 o o 0 -j CO $ "0 1 o o o tn on -d -vl n Vi af b/ c,/ d/ e/ 6-Anliioleviitlitic acid ddiyilMsc. 6-Aminolevulinic acid, Ntiwb.r u( rnta |>ar diet. Avi-race i standnrd error of nuaber of rats indicated. Si|>ol(lcant ly dlfteren) Iron control (F < 0.05) as shown by Ounnett** Hull Ipte-comparison tes on analysis at vatlaorc. By 13 vaoks the tax difference had disappeared and the ALAD activity ;TT. ;.'3S -«Jg oai-.t ;::j » depressed in 66.05* -caa caroonacc trevp a.-.c 5b :. and 53». respectively. l U> Urinalysis of rats f«d paint Mb * containing different concentra­ tions of lead are shown in labia 24-31. No marked difference* in urinary protein or sediment (erytnroevee, crystals. tuts. etc.). Thu*. urinaiysia can be considered within normal rangea for all traatmant groups. tXtf H. load Contant in Tlaauaa of !Uti Fed Paint Concantrationa of laad ^er.tdinjr.g.piffaran^ i^|. '•/ft » Tab lea 33-37 show the/iiad contant of^lood, brain, livar, kidney, ( and bona from rata fad paint mu containing /iffaranc concaatrationa of laad. \Zlfea valuaa for tha laad eoncantratlona in bl^d at 4 and 8 weak* vara omxttad^P^ I from Tab la 23 bacauaa it waa diaeovarad that/cheat templet had baan eontaiy/’ LiaateC by thair atsraga containara. Atl3^«aki_thj^bloodleadleve3j^£ thaj£tafadlaa^o«toataand^h^r«tafaa^i^Kaodl^5^1aa?|c^S*S*JM£t» hot ceewenanc control.Tntht rata^eaTTjj^iaae cnroaatifrjb 6.0 521 «a d ’ 11.92* MBS load pmCfft, blood. 1 avals waraTound to bo ilavacao ------- ----------------------------------------f'2...... ' " ..— ■ »ncty . i«y & £SpA«b ?"*U *»<%}* St. ky-et. K? b!&. At 4 waaka, tiaaua laad lav*la livar and kldnay vara balm* tha dataetioa limit* of our assay.^Tbo rasults vara Che aamt at 8 waaks with tha exception of chtJjBS<>leedj«lncjreu£. All four rats in this group showed dataetabla lead in chair kldnaya^ tCs^e A^-Wjjaok*jdataec*blj^iavjl|^MjM^Binilliehj^bralrMt£g|j£jd_on^ injjherac^ednjj^&jjU^E^et^m'IjtSCiSOy^SSSiSii.iSiSiS-im lead vara found •.nTwora^r'TedbijaTl laid csrbonatV and nlni rics fed 11.52* SBSlaadsaint, So load waa dataecad in tha kidneya of tn«_so8trol group at 13 wtaaa. in centralt, load waa ootaecod in tha-kidneys of onartvo rats in ~ tseh laad aecoata group) and thd 0.421 and 1.9JZ laad ehroaata groups, laid was dsttecsd in tha kldnay* of three rats, eight rats and 12 rats in tha 12.431 laad chromate, 66.051 laad earbonaca and 11.95* NBS laad paint group*, rtspeetivsly. Ths_ femurs of rat* fed the control diet, load oeto*e*_or_lft*d_ chromate exnibi ttj^!eryTrcsIe*?Bp'T?y weans ■ However. tne iaauts of rata [*aj__£trBonate or l.L^&21_HSS_liAd-ai.,--nr eon gained 13.0 and 1».$ -S of laad/gu of bene, raapactively. ? r-:i- 36 *: ' 0007-SWP-037010 TABU 24 L’aiNALv 100 mg r. Microscopic Examination HiCi' : Normal Moderate Excaniva _ _ UBCi': Normal Modaraea Excessive Epiensliaaic': Normal Nod a rat a Excessive Crya ta1si': Norma 1 Modaraea Caat*: 4 1 i2 7 1 6 2 20 4 7 l 5 2 l 7 23 1 * 2 ______2_ . a a 4 2 -i. 8 Negative _ £oiiriva 2 7 •4 a 23 * a 24 • "»rS -* Kuao«rs andicaca number of rats at response level. £/ Normal, 10 or lass calU; moderate, 10*100 call*; excess:ve, >100 cails/fiald (x 440). b/ Normal, £ or laaa call*; modaraea, J-23 ceils; excessive, > 23 cells/fiald (x 100). c/ Normal, non a; modaraea, 1*3 crystals; excessive, > 3 crystals/field (x 100). ■ v 37 0007-SWP-037011 0007-SWP-000115579 TABLE IS isimvsis o f s a t s f t p f a in t -aw con ?a :n :;;o 0.0HL LSAO AS ITA.0 OCTOAIE ___ L:,gU3« -Cjak 1 ± A Proeain: N'acaeiva < 100 mi H _ > 100 >51 Microscopic Examinecion RBCi/: Normal Medaraca -------- ; . W»Ci': 4 3 8 4 3 1 10 2 .. •/, y.vv.. 1 ____________ 3 3 “IT” Normal 2 1 2 Modsrace __________ Exeat s^vj_____________ — — _ '■ ■ "■ a" EpieneliucE^: Normal ■» 4 12 Moderate _ _______ ________ txcfiiivi_____ __ _ _ _ _ ________ _ _ Cryscals£/: Normal 2 4 11 Hodarata 2. 1 -------------------------- _______________ u 4 Caaca: Negative 12 Numbers indicate number of race at responds level. ■ / Normal, 10 or Its* call*; eedsrste, 10*100 cells; excessive, > 100 eells/field (x 440>. b/ Normal, 5 or lass calls; moderate. 5*25 calls; excessive, > 25 calla/field (x 1001......... £/ Normal, none; moderate, 1-5 crystal*; excessive, > 5 cryatala/fiald (x 100}. 3B 0007-SWP-000115580 'ABU 26 VKiSALYSIS OF RATS FE3 PAINT ?4W CONTAINING 0. j 3* UAO AS IlAO OCTOATS ....... 4 Preccr.: Najativa < 100 r ib r. __________ > i0£ r.:_%_ _ _ , Microscopic Examination RBCi^: Normal Moderate _ _£*£*l*iv« WBCi': Normal Moderate __________ Exeats Wa _ _ _ ^ £picnelius&/: Normal Moderate ________ _Exc£»siv« Cry seal si'': Normal Moderate _Exe£ssiye_ _ Caaca: Na(aciva ___ ?os:c1va ___ 4 Treatment Weak, 8................ 13 3 l 10 1 ___ __ _1. 2 10 2 1 2 1 1___________________ l. 4 19 3 3 4 12 3 1 4 12 4 4 12 Numbers indicate nunbar of rata ac response laval. a/ Normal, 10 or laa* eetla; aoderata, 10-100 calls; axcaasiva, > 100 cella/field (x 440). ....... ......... £/ Normal, 5 or lass calls; moderate, 5-23 calls; excessive, > 23 cells/field (x 100). c/ Normal, none; moderate, 1-3 crystals; axcaasiva, > 3 erystaia/field (x 100). r j-saii itr-rt i sKw 39 »v- 0007-SWP-037013 0007-SWP-000115581 TABU 27 t>’vT4 ■ jRiXALvsts o f Ba t s rso p a in t »w ai 2.05*; LEAP AS UA3 QCTvATE * Frotum: Negative < 100 mg u > 100 t..K X Microscopic Examination RdCi': Norasl Moderate Excessive UBCS': Norms l Moderate £*« Epitasliusti': Crystal*^; Casts: Trcscrcnt Week 8 ;3 4 4 11 l 4 <* 12 3 1 3 8 4 7 ___________________ 4 U Sternal Modarsco llCiliiVO ____________ _ _ 2 4 Normal Mod*roc* * 4 Negative Positive * r+ ______ 12 12 Kunbera indicate number of me * at rospons* level. */ Normal, 10 or l*i* call*; moderate, 10*100 culls; excossivo, > 100 eolU/fiold (x 440). .................................. b/ Normal, 3 or 1ms colls; moderate, 3-23 colls; excessive, > 25 eells/field (x 100). ................. e.‘ Nora* l, none; u.-dorno, 1-5 cry* to Is; excessive, > 5 crystels/iield (x 100)................................................... -' 4a -:: • ' • -i.'1’ \ l-.i ~v-'' • 40 0007-SWP-037014 0007-SWP-000115582 TABLE 2B CONTAIN eom or r a t s p t d PAINT *.iw • 0.42', LSAO AS '•_TAD CMM5KA T t r in a l y s is Protein: Negative < 100 mg X > 100 ra X Microscopic Examination HBC£j> ; Normal Moderate Excessive VBCi'; Normal Moderate cpitneluc—': Crystal si.': Casts: Normal Moderate Normal Moderate Negative Positive 4 Treatment Week 8 4 2 :o 2 l % 3 1 1 3 8 3 1 2 2 3 1 8 1 1* 4 12 2 1 4 12 4 4 12 Numbers incicace number of rats at response level. »/ Normal. 10 or less cells; moderate, 10*100 cells excessive, > 100 calli/field (x *40). ■ >/ Norm*!, S cr less calls; moderate.. 5-25 cells; excessive, > 25 celle/field (x 100). £/ Normal, none; moderate, 1*5 crystals; excessive, 5 5 erystala/field t* 100). r k £t :.r. 41 < 0007—SWP-037015 0007-S WP-000115583 TABLE T9 o (klP* l 'r is a l y s is ?a :?.t HtM c c n t a :n :n s l.?sr. L£A3' AS LEAP CHROMATE .............. of r at s f ed Treacnenc Week Procter.: Negative < 100 sig 1 i°2 SSj^__________ Microscopic Examination RBC1': Normal Moderate WBCi': Normal Moderate kS £ 3 1 3 __ A 2 2 o Normal Moderate _________________Exejsiive Cryittlsi'; Normal Moderate ------- _E£tJsiive_ _ _ Casts: Negative Positive A . *. 1• ' 1 • • ... , , *9 * * ... '■ 1 3 l 9 *9 3 B 3 9 •• *fc _____ 1 Epithelium-'; •i * _ A ;.r . * • v 4 A 12 A A 12 ; i■ i >, Numbers indicate number of race at response level. •J Normal, 10 or lea* cells; moderate, 10-100 cells; excessive, > 100 etlls/fieid (x 4A0). b/ Normal, 5 or less cells; noderace, 5>25 cells; excessive, > 25 eells/fielo (x 100). e/ Normal, none; eoderste, 1-5 crystals; excessive, > 5 erystals/field (x 100). . -'.'-V; -.-..a "ffr}-' ............................ i'.'f'':'’ »■ ••»£ *7 .2 irt .v-.wi;;- . '. • ' : rC— -- ■ -* v-v--.—.'. 0007—SWP-037016 0007-SWP-000115584 TABU 30 tktr1 CR1NAIYSIS Of RATS FE3 PAINT rUE CONTAINING 12.-3'. LEAD AS LEAS CHROMATE r.0st>v- To eatment week Protein: Negative < 100 mg 2, Microscopic Examination RBCi'': Normal ......... Moderate Excessive WBCi': Norma 1 .Moderate Epitneiiums' : Normal Moderate ___ ______Excessive Crystals^: Normal Moderate Excessive Casts; Negative i H ^3 i 3 1 2 9 3 4 1 11 1 1 3 3 1 1 2 4 4 1 11 1 2 2 4 12 4 4 12 11 Posicive m Numbers indicate number of rats it rttponii level, a/ Normal, 10 or lass cells; moderate, 10*100 calls; excessive, > 100 cclls/field (x 440). ........ b! Norms 1, 3 or lest c«Us; moderate, 5*25 cells; excessive, > 23 cells/field (x 100). i! Normal, none; moderate, 1-3 crystals; excessive, > 3 cryseals/fiuld (x 100). ■ cs i*. • : r»* j 43 0007-SWP-037017 0007-SWP-000115585 spar- :a 2L£ 31 ~ . c*"* ■ **« VRIN-Airsis cr .h a t s F-3 p a in t ............................... —.o .-£ MXi c a x ;j :;a ~s I*KXV. It* '* •* Trotffanc l»'ocx 13 a* Protein: Negative < 100 e.t 1 3 Microscopic Examination RiC±/: Normal Moderate ^ Normal Moderate Excessive Epienaliuat-': Normal Moderate ________ _Exeassiva Crystals^': Normal Moderate I----'T «• u 2 * k>~? S^SK 2 2 2 10 2 t ?-"- ■ a 4 12 3 4 12 4 4 12 •» Negative Positive i: 1 2 4 *BCi': Casts: ' 1 4 Numbers incicste number of test at response level. *,! Normal, 10 or lass calli; moderate. 10*100 calla; excessive, > 100 cslls/fULd (x 440). b/ .Normal, 5 or laai call*; moderate, 5*23 calls; excessive, > 25 cells/fleld (x 100). e/ Nonas!, non a; moderate, i-5 crystals; excessive, > S erysiais/field lx 100). a. 0007-SMP-037018 0007-SWP-000115586 :a b i 2 * % "RINA .vs is or s .ms 7zo ?,u: — N«gaciv« < 100 nB r. > *.00 ag *. .Microscopic examination RBCi'; Normal Moderate Protein. Trticnep.c V«.u k 3 13 2 2 3 2 2 4 ■ - .. 'v *-.. 7 5 - ** v • / > V i *^. L.* i 12 i««m________ ”3 2 11 Normal 1 *1 Moderate 2 Excessive * 4 12 Spittle! uni': Normal Modaraca _ — _ _1x c «i j i »i _ _____ _ _ ______ _ _ 4 4 .2 Crystal si' : Norms 1 ModarJta Excessive Cases: Negative *» 4 12 Positive - • waci-: . ii-r-js-. ; .* . ; i*,’.. * 1 .* H ;*• P _ k .».»*• ~ ,.v: • . . N‘.a ■ », t *\.c. ■ •' :•». *, 1.-T5 itr Nye^cri ineicar* number of rats at response laval. a/ Sorisal, 10 or laaa call*; moderate, 10-100 cell*; excessive, > 100 eallsffield (k 440). b/ Kansal, 5 a; last calls; moderate, 3*25 calls; excessive. > Zi ealli/ileld (x 100). £•' Normal, non*; moderate, 1-5 crystals; excessive, > 5 cry*tai*/fi*ld (x 100). ‘v1* ai.. :&r:P -'V* 0007-SWP-0370I9 0007-SWP-000115587 \i3Li 33 LEAD CrXTEN" :c d Tru fon c V»e!' 14.5 - 1.5 (8) 13.2 : 1.0 (S) 12.0 : 1.4 (8) I 19.4 - 2.2 (S)i' I 24.9 = 2.2 Wi/ ^£3.9 = 1.3 ciiiiii/ Values dtlatad due to contaminecion. Four value* dilated du* eo contamination. Number of rat blood* analysed par group unit** indicated otbarvis*. Sixteen blood samples war* analysed. Average ug of ltad/100 ml blood - standard trror of number of rat* in parenthesis. On* blood sample lost. Significantly diffarar.t from control CP < O.CJ) as shear. by Ou.nnett’s eulcipls-cempariion cast following in analysts of variance, Twelve blood sampla* analvzed. ......................... ........................... .•v \.j . *"5 *. -J * • -'yy ■ ‘ '£?:'■ rfsJ . - ■ vlSJ SOS'*.' H5P3C 46 ir *r» J 0007-SWP-000115588 TABLE li LEAD CONTENT C? liV.TN • "e«k 2;*= Control^ 0.08Z Lead Oetsata 0.53* Lead Oetoata 1.051 Lead Oetoata 0.42Z Lead Chromate 1.95X Lead Chromate 12. A3* Lead Chromate 66.051 Lead Carbonate 11.92X N8S Lead Paint a* (A)-1’' 9 (A) n.d.£/ n.d. n.d. n.d. n.d. &ad. 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 r.:> ' n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. 0.2 g Q.li/ (7) - * * • f.r .i ’• ' ?. • '• at a/ b/ e/ it Nuaoer of braina analyzed par group unlaaa indieaced otherwise. Eight brain# ware analyzed at A and B week#. Twenty-four brain# war# analyzed at 13 weak*. Not detectable. Below sensitivity of 0.07 ug/gs of brain. Average ug of laad/ga brain - atandard error number of rat# shown in oarenthaiis. ........................... •N m»i :• T T :i ru. i.'i-L'i.i.'.' >a..'^a* ‘■ ■ (ir'JS. t*< * J.^aT Jyibear •S’**.'? c*-®^ - : \ . * **'»•• • o4»**y ': 0007-SWP-037021 0007-SWP-000115589 TABLE 35 i cr:m:.T c f Treatment Keek 9 ' ;-) Ttet (4)*'' Control^ 0.091 Lead Oetcate 0.531 Lud Octett* 2.051 Lead Oetoate 0.421 Lud Chromate I, 951 Lud Chromate LI.431 Lud Chromate 66.051 Lud Carbonate II. 921 KBS Lud faint n.d.-S' n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. ,. .. n.d. n.d. Red* r..d. n.d. n.d. n.d. n.d. n.d. ........... B& 13 (12) n.d. n.d. n.d. n.d. n.d. n.d. n.d. 1.1 g_0.6j/ (1) 0.4 sjl.l (9) r>»/ v5v V »» .'r» life gfe ISSr? . 30saa>* a/ b/ «/ £/ Kumber ot 11vara analyzed per group unleaa indicated otherwise, Cighc livari were analyzed at 4 and B waaka. Twenty-four liven were analyzed at 13 waaka. Not dacaetabla. Below sensitivity ef 0.07 ug/ga liver. valuer are average ug of lead/gn liver s standard error ef number of rati anown in parentherii. i *y* - » •• “* v\* .9 0007—SWP-03 7022 0007-SWP-000115590 ZABLE 36 LEAD CCNTIST Of VlONSV n.d.^ 0.087. Lead Octeats 0.53% Lead Octoats 2.03% Lead Ottoace 0.42% Lead Chromate 1.93% Laad Chromate 12-43% Lead Chromate 64.03% Lead Carbonate 11.92% XBS Laad faint 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. o Control-^ it (I)i' Trcatnent '->«< a (-) »-» Jit* a/ .b/ r..d. :.3l C) 0.31 (1) 0.** -0.1 C) '*.5i (i) 0.4 = 0.0 (3) ............ 2.3 - 0.2 (8) ‘ ' CSrr*- &&■ ma: 1^ = 0.1 U2) /•* Number of kidneys analyzed par group unlaaa otherwise indicated. Eight kidneys vara analyzed at 4 and 8 weeks. Twenty-four kidneys vara analyzed at 13 vaaka. e/ Not dataecablt. Balov sensitivity of 0.07 ug/ga kldnay. d/y Values ar* ug of lead/gn kidney or average of lead/gs kidney : standard error ef number of'rata in parenthesis. N. xx*. m* - I . fM ? *&.:• iV: , p* h""iX r • •• \ i:». •- & * - ^r—o»v •.. V" *’ 0007-SWP-037023 0007-SWP-000115591 s - ■ ?; Z-7yii\ ‘ABLE 37 >V- t-£A3 ecxTsrr or s q n e ^7:3) Us.1 rraatsarc yaak £ W*' Concrol-/ O.OB» Laad Oetoact o.jji Laad Occoaec 2.05. Laad Oetoaca 0.42S Lead Chrooace I.95S Lead Chraaac* 12.437. Lead Chresata 66.057. laad Carbonaca II.5:7. N3S laad Paine ■ 3 (4> 1.6S/ (1) 3.1 (1) n.d.i/ n.d. 2-9 (D 0.9 (1) 3.3 (1) U.O - 4.1 (4) 14.9 i 2.5 (4) 3.9 * 0.4 ce) 4-2 > 0.6 (4) 3.9 s 0.2 (4) 3.0 s 0.7 (4) 2.6 r 0.9 (4} 1.8 r 0.2 (4) 5 1.1 8* E»‘l 6.0 • 2.1 1-4 S 0.4 w£f 3.2 = 0.2 C4) 3.3 = 0.3 (.3) 3.1 r 0.2 (12) 3.9 s 0.5 (12) 3.8 = 0.6 (12) 4.1 r 0.5 (12) 3.5 s 0.5 (12)^ 21.7 20.9 r C.9 C22>£/ £.>r-. If* a*. *aA . r*.. • a; b/ .'.ueetr si bonts analyzad par group unlati indicated otherwise, Sight bonaa wara analyzad ac 4 and 8 uaaks. Twanty.four bonai ward analyzad ac 13 uaaka. cl Valuta ara ug of taad/ga bona or avaragt ug at lead/gw bona - standard arrot of number of rati in paranchasia. i! Net datactabla. Balow sensitivity of 0.4 ug/ga of bona, a ^ Significantly difftranc iron control (9 < 0.5) aa shown by Ounnett's sail c ip la ccrpiruon cast following an analysis of variance. t -1Wi%4a^»ta->. ■ X 9 S3 0007—SWP-037024 0007-SWP-000115592 At 8 weeks, the femurs ef rat* i«d tne control, lead oetoste or lead chromate diets contained similar quantities of lead, ranging from l.»4.2 ug lesd/gs bone. L«adccr«n^nth^femu^^rcneJj9J?!iZiiJ1e*d>^aroo^^e andJ11ij^>2»J>35jCTOuo^>uer«^h^janel^^£jjyl-j^Ci-jmi_£^ll£j£ig— 3otn*:cr.it ;car;.y coh-Js control. , ‘‘’ After 13 weeks, the rats fed lead Detract or lead chromate had r.o more lead in their femurs than at 6 weeks ana then tnose in tne control group. TheJo.P5% lug csrhonatt grous exmaited ntgntr .svtisotTaUPin their ftmurs i21.7 ug ieia.ga eons; tr.ar. at 8 weeks. ThelLJJ2Zj t: icA. Si r*:*. i it ««. *1 » •* 2 *1 i'-£s & r *» r?#5S5 1VAJ. 1 ii s a raSEb. SsSr i =1 o X&X-' £ •* r *t » m S a -a-vi; i&Ts* gS&ft i .. s; " P £e *;t -V-vV •• 53p *£{N .»—’-T*. •, l i fc a ii *1 .i A . t ; 1 >4 » l> ii j ?!| ms t11; * I ii in l d • a : 3 # -i i! ? i, -11 i1 »*-* A S3 T.''UV? srts. >«'JiCCj J^u w iu j u . J5HJ «*• * ». * -•’.»* fc«’rt“yr 0007—SWP-037027 0007-SWP-000115595 0007-SWP-037028 0007-SWP-000115596 -3 TM > » «* 4T w « » ftl ■ 0 m < $ v \ _“T*s.C 1 '.Va- ,‘n* K i fm. #m IN *51 IN 1to ■ 8 : i « o * -8 *» « «* ms .A#:vy.; - S 8 O a> •i J f mi ll ll ll * h» U M > 41 O a (A !- 4 ".-w*. Av-^2y>' ■ • .... .. . 0007-SWP-037032 0007-SWP-000115600 Thi bad* Usd burden at rscs ftd rr.rt ewtuaiM leed octoite or lcad_chra»ate_tfer« no-different :r~~ cartrcL .umJ '.3 At trul tire lead was found in the kidneys of one to three rota on asen of tha load octoace ind lead chromate ditts. and chc blood lead levels ir. tnc '.2,417. lead chromate ' group wart significantly elavatad. Sines rata fed lead nitrate show elevated lead levels in blood, kidneys, bone and liver within 2 weeks.i27 it^eooears that the lead octaate and lead chroma ee_we£e_n£i_£l£i££_£lg^ill^b»g££il_°r euaTTSSTa tor sosorption/^^^................................... ... ...... ... . • ,, .V Pathology was performed only on the rats that ware fed the control, 2.OS* lead oceoate, 12.43% lead chromate, and 11.92% KBS lead paint diets for 13 weeks. 411 of these rata were relatively free of lesions, and those lesions which were observed, oecur naturally in rat colonies. V. conclusions 'CWsjjtjjdviijhoj»!i^ha^Mitafed^iJ^^|t|iJjjaint/feed2o£c^dj^int containing 11.92% lead andjS^paTnt^ontalnlni bfeToi^Taad'^NrTearTsrPonrca amuaitaa early si|ns of lead poiaonin|. these nans consisted at arvthrocvte A LAO depression (30%) and significant increases in blood, kidney, bona, liver and/or brain land. katf_Jf4_t_>iat consisting of 0,1% new paint Ilia contalnint I2.i32_lcad as lead chrawate exhibited only elevated blooa lead_elter 13 ~ weeks of leadini. 4 nailar^aiet oi. new oaijtconteum?g"r^%la»daa*TeeT~ oceoete produced no signs of lead poisoning. Tha lack of any marked change in the body lead burden in these rats suggests that very little lead was absorbed from the gastrointestinal tract. Since there is considerable evidence that the rat can absorb lead in several forms, one muse conclude chat the lead in the paint used in this anparimene was r.ot readily available for ebaorption. In conclusion, tha raaults reported nereis support the contention that 1 tad is not readily aosoroed from tha hewer otir.it. ..........................'... ■ ..................................... 39 n.i >l"%» »•:>/ fi- 60 -r. s ew cc\zv:s:y: Older paint formulations contained sol uble lend salts. Two samples of paint chips representative or these fomulstisr.s were fed to rats at 0.1% of their diets. One was supplied by the Sational Bureau of Standards and was removed from interior walls o: alder hares. It contained 11.92% lead. The other was a cejAa formulation containing 66.OS', lead carbonate. Both of these samples produced classical signs of lead poisoning in the rats. These signs consisted of erythrocyte A LAD depression (£0%) and significant increases in blood, kidney, bone, liver and/or brain lead. Newer paint formulations contain insoluble lead pigments and low levels of lead driers. Several samples of paint chips containing us to 12.43% lead chromate pigment and up to 2.0% lead octoate drier were also fed to rats at 0.1% of their diets. Tht only evidence of toxicity or effect or. body burden appear in the rats fed the paint esntiinir.g 22.13% lead chromate. This consisted of an increase in blood lead, withouc ether sips of poisoning. Paint containing 1.95% laad ehrsaata and pair.t containing 2.05% lead octoate produced no detectable changes in the rats. j. *3 > Y> him - These studies show a marked difference in the toxicity of lead in paint formulations depending on the nature of the salt form and its solubility. The lack’of toxielty with paints containing 2.0% lead chromate or lead octoate undoubtedly it due to the lack of sipiflcant absorption from the gastrointestinal-tract. W •' . IBJ % s Tj i Ee ?? v\V-V' ! •* - •%> r. V**i tv. -■ f >.:-1 • V mfc r«< 4 :W- ... 0007-SWP-037035 0007-SWP-000115603 •1 • 4 ■ r MODincvr:ov CrCisrirs* :rs:’-'ss:ov •• ?*-' The levels of lead fed to rats in the study were sufficient to pro* duce significant^e’evation of body lead burden if the lead is in tr.e fern of lead nitrate.ii' If we equate the amount of lead consumed oy the average rat fed paint chips containing 3.05% lead cotsate, 13.43* lead chromate, 66.05% lead carbonate, and 11.93% XBS lead ratal ditts to a Id.6 kg ;3-ytar old) child, we find that the lead consultation would equal 3S.6, 135.3, 503.0 and 113.9 ng of lead/day, respectively. On the basis of a 3*. lead paint, the total paint intake would be 1.5, 6.2, 25.0, and 5.? gm/day Daily ingestion of these Quantities of old patnt.hsvs been reported to produce lead poisoning.!" ir L+J■ « 1 3 PJtS£S.V There were no changes in hematology, urinalysis or serum proteins throughout the entire study. Hqa/iyy, porphyrin metabolism was affective w as early as J weeks in rats fed either the 66.05% lead carbonate diet or the 11.93% XBS lead paint diet as evidenced in a 50% depression of erythrocyte ALAD activity. This effeet appeared to be direstlv related to the concentre* tion of blood lead as has been reported by others.iitii/ !ie» eeeri there was one exception; blood lead in the 12.45% lead ehrpaate group became elevated by the 13th week, but there was no depression of AUD activity. This dis* crepancy could be related to the durttion of elevated blood lead or the age of the aniaal when blood lead became elevated,..... .....'...... .................... - ■ ■ ■ ■ f.®- *• ‘ *"SL Ner r . V •*• . %»«■ ,. «*' * ♦ Even though erythrocyte ALAD was depressed 50%, the overall effeet on porphyrin metabolism must be considered mild, since there was no concomitant change m the levels of protoporphyrin (erythrocyte}, delta*asmoievulir.ie........ acid and cepropoiphyrin. These observations are consistent with reports that blood lead levels greeter than 40 ug/100 al ABC are needed before seeteus changes in porphyrin metabolism occur.!!/ The body lead burden of rats fed paint containing lead octoate or lead chroaate Seam no different from control until 13 weeks. At this time lead was found in the kidneys of one to three Tats on each of the lead octoate and lead chroaate diets, and the blood lead levels in the 12.45% lead ehronate group were significantly elevated. Since rats fed lead nitrate show elevated w lead levels in blood, kidneys, bone and liver within 2 weafre.ii/ it appears Jf] that the lead octoate and lead chromate were m« eitherTfeadily absorbed ex’ available for absorption. .......................... * Complete gross and microscopic pathology was performed on the rats that were fed the control, 2.05% lead octoate, 12.45% lead ehrosate, and 11.92% SIS lead paint diets for 13 weeks. All of these rats were relatively free of lesions, and these lesions which were observed, occur naturally m rat eolonies. . . • ’ . .. ..""~T7.............. * ' ",............ / r~ **^ .a w* ‘Wk . i •. - . 64C u 0007—SWP-03703 6 0007-SWP-000115604 PREFACE This resort was prepared at Midwest Research Institute, 425 Volksr Boulevard, Kama City, Missouri 64110 under Contract No, 62-W-42GC & NFC, MRI Project No, 3729-B, “Lead Paint Ingestion Study.* The Research was sponsored by the National Point and Coatings Association, 1500 Rhode Island Avenue, N. W., Washington, 0. C. 20005. Royal A, Brown, 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 I 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, BToIbgy Research Assistant. Dr. James L. Spigarelli, Senior Chemist supervised the lead "analysis with the assistance of Mrs. Hope M. Miller, Assistant Chemist. ft:-' i-. > ' Members of the National Paint and Coatings Association Industrial Metals Task Force prepared the paint chips aid consulted with Dri>Castle, 5anyer, 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 Dr. John P.Frowley Charles M. Joekson Joseph G. Kingston S*. S idney Lauren Wiiliom W. Ringle Edwin £. Swain Jeon P. Teos Domenic J. Tessari The Sherwin Williams Company Chairman The National Point 6 Coatings Association Hercules, Inc. Celanese Coatings Co. Glidden-Durkpe Division of SCM Corporation Coatings Research Group, Ine. Pratt and Lambert, Inc. E. I. duPontde Nemours & Co., Inc. The Flood Company De Sato, Inc. AoDroved for: MIDWEST RESEARCH INSTITUTE W. B. House, Director Biological Sciences Division 14 September, 1973 ■ W .* » 0007-SWP-037037 0007-SWP-000115605 SLIGHT MODIFICATION OF CASTES' COXCU’SIQSS This study shows that Tats fad 0.1% diet (peint/feedl of old paint containing 11.92% lead and new paint containing 66.05% lead as lead carbonate exhibited early signs of lead poisoning. These signs consisted of erythrocyte ALAS depression (S0%) and significant increases in blood, kidney, bone, liver and/or brain lead. Rats fed a diet eon* sisting of 0.1% new paint fila containing 12.45% lead as lead chromate exhibited only elevated blood lead after 15 weeks of feeding. A similar diet of new paint containing 2.05% lead as lead octoatc produced no signs of lead poisoning. The lack of any narked change in the body lead burden in these lead chromate and lead oetoate rats fad paint containing chromate and octoate suggests that very little lead was absorbad frost tht gastro­ intestinal traet. Since there it considerable evidence that ths rat can absorb lead in several forms, one must conclude that the lead in the forts of lead chromate and lead ectoste and incorporated Into paint was not readily available for absorption. In conclusion, the results retorted herein sunaort the contention that lead is not readily snsoroed rrcnewer ;a:r.:s. ........ ■ ..................... m'-\+ \\££ SStt fS if %*S»-5 n.r