Document ZnxkqjojK0xnmD2VJyRMnb8j8

l ead pain t in g es t io n s t v o v *" -l ::* n .- v * FINAL REPORT 14 Sap unbar 1973 Contract No. 6'-tf-62CCSSPC MRI Projact So. 3729-9 For National Paint and Coating* Aaaoeiation 1500 Rhoda Xiland Avanua, S.U\ Vartiington, D.C. 0005 Attn: Mr. Royal A. Srown 0007--SWP--03 6972 ii o PREFACE This report was prepared at Midwest Research Inatitues, All Volker Boulevard. Kinsai City, Miaaouri 64110, under Contract No. 62--62SCU>PC, >2U 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, 3.C. 20005. Dr. John P. Frauley. Chief Toxicologist, Hercules, Inc., Wilmington, Delaware 19B99, 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. Thecas R. Castles, Principal Pharmacologist, was che principal investigator, assisted by Dr. Jaime L. Sanysr, Associate Pathologist, and Mrs. Jane Koch, Biology Research Assistant. Dr. James L. Splgsrelli, Senior Chemist supervised the lead analysts with eha sssistanca of Mrs. Hops Millar, Assistant Chemist. j-tC ' i '** * .to. V V42tj LJvpJK. u.a>a>< Approved for: o MIDWEST RESEARCH IXSTTTTTE S. House, Director Bioiaglet1 Sclancas Division 14 September 1973 ii !r.: *\ *r'. 0007"SUP-036973 i 0007-SWP-000115541 3 I. Introduction II. Methodi . . :a c l s or c o k t ih t s Isa. 1 A. Preparation of Paint Filma Containing Different Caneen* tree ione of Lead..................................................... B. Preparation of Dietr....................................................................... C. Experimental Procedure .................................................................. D. Ana lytee.............................................................. E. Pathology............................................................................................ F. Statietical ....................................................................................... 1 2 2 3 5 6 III. aceulee............................................................................................................ 6 Coneentracione of Laad in Paint Filae ................................. Lead Aoalytet of Diete Containing Different Paint filna. Preliminary Feeding Studiaa .................................................. . Greta Obaervationa, Feed Contuapcion and Body Weigher . Weakly and local Land Coneuapeion .......................................... Hematology of Bata Fed Paine Flint Containing Different Coneentracione of Lead .............................................................. Tlecue and Fluid Cheaiecry of Kata Fed Paint Filme Containing Different Coneentracione of Lead ................. Lead Concent in Tleeuee of Rate Fed Paint Fllma Containing Different Cone antra done of Lead . . . . . Pathology of Rata Fad Paine Filar Containing Oiffarane Cone entree tone of Lead .............................. ............................. 6 6 9 9 14 14 14 36 61 IV. Dieeueaion 68 V . Coneluaion 69 Raferencee 60 Sag m m cAai; gtfifVtV mn J-.'&r.xK k-s. -ri**Z` 3 ill *. Vfc. 0007-SWP-036974 0007-SWP-000115542 i. imooucriai & Few persona will disagree that old leed-baicd point protone* ^1 Hi I......... .......... imlll 'll no~li yimiootn. At t rctult of radorte oy Kehoe.-i' Chitholnand King.-'' nodical raaearchert, paint menu- A j{curort and legislators havo recognized tho seriousness of this problem aid) art acting to cttablith tafe conccneracion* of load in paint, to do chi*, cltoy htvo voluminous report* on elinieol oboarvaeiont and animal rttoarch from wtfich to draw. Unforcunotoly, mote of chit information it bated uoon_ch*_ald painct a r.T^nier. JUU. ace sne name for- liiUl dill doneoneraeion* of lead in paint can be intolligontly ettablithad, wo mutt evaluate the coxlelty of the load ae it oxitet currently in modern point*. l_ua_tho jarao*o_of_thlj ttudyfto ova lute a eh* toxicity ln_rac* prgdused_by_aodorn paint* containing load oetoacejplasd chromate !i .......... in i and^ to^oBoarothomtommold ipainc fomilationtevoieal of \ ehoei^TeeDonaibEr^^rTSjnSKmKM^^IhereeuTtiTrepreeantad in tho following page*. \X |'.U U-< #-r II. MET,HOg /Jr 0 / A. Prooiratiof! of Point Fllme Conttinina Different Concentration* of Load ^v^r*****^ / Paint film*!ij without added lead end^eddedylLeeead compound* ir. tho form of laad octoato. load chromate, or load"carbon*to were supplied by s*m bars of tno National Point and Coatinga Aiaoclacion. Painte which contained load octoato.* Lod chromate (medium yellow) a Mmafeamjeam were prepared using a flat alkyd enamel formula baaed upon ...... Cleee elkyd The tlet alkyd paint without addad laad waa uaed for controls* The percent of lead in each laeipla wet calculated on the betie of cbapdnvoletile mete' rial. Diepertion wee eceompliihed with a Coulee DUsptfor and the pigment - volume eoncencracion wee held at on* level. Xheurffu* were pieced In pena and allowed to nr dry. Tbit vaa followed by^#*reed air drying at 120*F to volatilize any wining eolvent. Paint tdriM were ground^sieved, end ehipe ranging from 0.3 to 1.0 mm in ett* wege^ueed for this study. This chip tisa vat sal acted on tha baeit of a prpKmtnary rat loading atudy which aatablithad :.-.at rate would eeloctivtly eep^round largor tized chipa. . * Aafcmflb*'*-*-* The concaatratwui of lead in the different paint films were deterr.inec independently b/'oeSoto Incorporated, the Shervin-Wil liana Company end Kievest Research Ipdtituce, utiog Atomic Abeorpcion Spcctrophotoeecry. o **-.*** 0007-SWP-036975 0007-SWP-000115543 Sample^ of old lead-based peine. JiC* obtained tram eh* U.S. Depart- mane of Commerce, National Buraau of Standards, Washington, D.C.a Tbit paint had baan pulvatiaad and aiavad through 325 acsh screen (0.06-0.OB i sis*) and contained U.87% lead by weight. This saepia will b* refarld to aa 'TBS lead paint" in this report. B. Preparation of Piets Diets were prepared by mixing paint tel me with Purina Rat Chow cash in eh* eoneancracion of ^J^% (weight/weight). this was aceesvlishsd by first preparing a 101 concentrate (paint fila/foed, w/w) in a high-speed twin shall aixar. This concentric# was ehan added to eh* appropriate asount of Purina Rat Chou mash to give a final concentration of O.lt (paint film/feed, w/w) and aixad for 10 am' in a bulk mixer. lech diet was prepared thras etnas during cha experiment and ssoplte of each of eheto preparation* war* assayed for laid content. C. Experimental Procedure TwgJyiijgred'Waanllng (40-60 ga) Charles River rati (100 n*les and 100 foul* rata) were used for ehii study. Upon arrival rats vers housed individually in air conditioned quarters in polycarbonate cages containing' hardwood bedding and filter topi. After 5-7 days of equilibration, all race war* assigned one of eh* following diets. U> l. ftJC chou plus paint fMa without added lead (control) 2. Rat chow plus paint fA* containing 0.08% laad as lead oetoata 3. Rat chow plus paint fda containing 0.53% laad as lead oc teats P . Rat chow plus paint film containing 2.05% laad a* laad octoate tftf fUC ehov plus paint fila containing 0.42% laad as laad chromic* 6. Xac chow plus paint fila containing 1.95% laad as laad chromae# s 7. Rat chow plus paint fila containing 12.43% laad as laad chroaaee 1. Rat chow plus paint fila containing 6B.05X lead a* laad carbonate S. Rat chow plus X8S lead paint containing 11.92% lead M4 2 ..tj'ij jeu i t 0007-SWP-036976 Twenty Ml* and_20 foul* rats were assigned to eh* control dice, and 10 mala* and 10 few la ret* were assigned to each af :n* t training diets. 2atn diet was fad in a 9-os vidc-mouth glass jar winch was secured to the cage. Tap water was available *ey I lbiturn during the entire experiment feed consumption was measured twice each week and calculated on a daily basis. The animals were weighed once a watx. All rats were observed twice or more each week for toxic signs. After A, 6 and 13 weeks, a selected number of msle and female rata from each group were placed in msteboUsa eages for eh* collection of 2A-hr urina samples. A portion of the collected urine wea used for urinalysis and ch* ramainder frosen for assay of delta-aminolevullnlc acid and coproporphyrin. After urine collection each rat was anesthetized with ether, exsanguinated vu the abdominal aorta end bone marrow ameers were prepared, blood samples ware heparizined, cooled and used inmediately fo^s hamate logy end enzyme assays. Aliquots of ch* reaMinlng blood ware cakan for protoporphyrin and lead assays. After urine and blood samples were obtained, each rsc was nacropsled and tissue taken for lead enslysli microscopic examination. 3. Analysts 1. laid analyses of atUit_films and discs containlnt 9tint U*m-. s. Paint film: tan.isilUgram/ samples of paint fiUme con taining no lead or 0.08% lead octosta were eharrad with 3 ml of concentrated nitric acid end dry-aehed et 500*C for 2 hr. The eah wet dissolved in 1 ml of scua regie, diluted to 3 ml with distilled water end this final dilution measured for lead content by atomic absorption. All other paint Mama (10 mg samples) were dig as ted in 15 cl of a mixture of concentrated nitric acid end 37* perchloric acid (2:1, V/V), evaporated end ch* remaining perchloric acid solution diluted to 25 ml with distilled water. This final dilution was usad for comic absorption spec trophotometry . b. Piets; One-gram samples of fsad war* digested in a eixture of concentre tad nitric acid and 371 perchloric acid (2:1, V/V) and their final dilutions adjusted to make their lead concsntrscions within the oateenon limits of the atomic absorption ttehniqu*. 3 0007-SWP o 7% V 2. Analyses nerfnrred upon each rat: t. Haaatolcev: (l) Hematocrit: Hematocrit was dettntined in capillary tubes using s microcapillary centrifuge (International Equipment Company, Model .'3). oechemoglobin.-' (-) Kemoclobir.: Hemoglobin was measured as cvano- (3) En'throcvce end leukocyte counts: Total erythrocyte and leukocyte wora counted using a Coulter Electronic Particle Counter with 100-u aparature.J^ M Reticulocytes: Reticulocytes were counted by the aacnylena blue method using the Miller dise.i/ (5> Differential leukocyte counts: Wright's stain uaa used to stain the leukocytes /or examination. (6) E_rythtocuta osmotic fragility: Osmotic fragility of erythrocytes vas quantitatively determined by subjecting heparinized whole blood to sodium chloride solutions of different otmolarieiaa.2' The concen tration of sodium chloride vhleh hanalyzed SOI of the a^throcyces was obcainad froa a plot of parcant hamolysis versus (odium chloride concentration. b. Body fluid end cietue chmristrv: JlMiM-arftMin- rleetroohoretic patterns: Total plains protein veae determined using the riyctl Biurac Reagent (Hycel. Inc., Houston, Texas). The quantity of each plasma protein vie dtctrrur.ed electro- pnoretieally on ca'.luloia acetate end expressed as a percentage of the total plains protein. (2) Ervthrocvto-8-emlnolevulinic acid dehydrate fALAP): 6-Aminoievutir.it scid dehydreee activity wae determined by tna method of lizhtsen end Feldman.^ This eroeedure wee started 30 min after each blood sample was taken. (3) Ervthrocvte orotoootohvrin: Protoporphyrin in erythrocytes was sutured by the method of Heller, ee a 1.2/ This analysis was performed the day after blood was wiehdrewo. <-) Urinary ^-aminolevulinic acid fAIJO: Urinary 4-aminolevulinic acid was measured according'"to "the Davie and Andelseftsjy modification of .Vouzerail 's and Granule's Method.These analyses ware periorrec m subdued lignt. 4 0007-SWP-036978 0007-SWP-000115546 (3) Urinary cod^Shnornhvrin: Urinarv cepr0?orphyrin* wars determined by Chs method of Sehlanker and KitenaU.--! Tha* analyse! were performed in subdued light. e. Urinalvs is: (1) Urinary nrocaln (albi:rin>: Urinary protein vas measured with "Uriatix" reagents strips (Anas Company, Elkhart, Indiana). (2) Microscopic examination of urine: Urine samples vert centrifuged, the residues resuspended, and examined microscopically for the presence of erythrocytes end leukocytes under high power field and for easts under leu power field. d. Tissue lead! (1} Blood: One milliliter of blood was mixed with 1 ml of a mixture of 5X trichloroacetic acid-.37% perchloric acid (3:1 v/v). The sample ves centrifuged end the supernatant flltarad through on JJUt 0.8 u Killipore filter. Tho supornata was analysed for lead with an aeomie absorp tion spectrophotometer using standards prapaTad in control rat blood. (2} Other tissues: Bone, liver, kidney, and brain war* digested in concentrated nitric acid, evaporatad to dryneaa ond reconstituted to the lowest possible volume with 2OX nitric acid. These solutions were analysed for lead by atonic absorption spectrophotometry. (3) Atonic absorption analysis': Lead concentrations were measured using e Varlsn-Ttehcron AA-5 atonic absorption ipoesrophotonetor. Sample solutions vtre aspirated into an sir-aeatylana flame and the absorbance was stasured se 2B3.3 nm. Background interference waa determined wieh the use of a hydrogeo concimim lamp at 283.3 nm and subtracted from the absorbance obtained et 283.3 nm with the ?b hollow cathode lamp. . ?atholorv 1. Cross oechcloev: At necropsy, rats were axeeinad for gross abnormalities, chair livers, kidneys, spleens, hearts, gonads, thyroids, orair.s, and adrenal* weighed, end the rslscfv* organ weight* calculated. After weighing, a pertlee f the liver, kidney end brain were taken for sicroseopie examine don end the rut of each organ wee measured for Load content. A faaur we* removed ce be used for tho moaeuromoat of lead in bone. 2. Microscopic pathology: Bo m marrow smears wort prepared for a cycicid/arytnrotd call count. The following tiatuts wert fixed in buffered neutral 12X formalin: brain, liver, spleen, stomach, email Intestine 3 z: '* ' irSSj-t t?rz >5:tj&L I ** !>>,v * `wl / laL+f&li 0007-SWP-036979 0007-SWP-000115547 (duodtnum, jejunum, ileum), colon, pancreas, kidneys, urinary bladder, adrenals, thymus, gonad, tnyroid (with parathyroid attached), mesenteric salivary eland, lymph nod**, heart, lunga, diaphragm, akalatal rustle, and prostata or uterus. Tissues from all 13 week rat* led t.nc control. 2.037, lead occoate, 12.43?. lead entoeate, bb.OS" lead carbonate, and t.nc 11.92" SBS lead paint dices were embedded in paraffin, section to * thickness of 6 u, stained with hematoxylin and eosin and examined for histopathoiogy. F. Statistical Analyses Control and treatment values were compared statistically using Dunnsct's mulcipla-eempariaon tut with P < 0.05 aa tha criteria of significance. III. RESULTS Cl. T*e A. Concentrationa of Laid in Paint Mm The cone antra eiena of lead In tha pome rmwma supplied by tha Members of the Naeienal Paint and Coatings Association were analysed for lead content by Se5oeo Incorporated, eh* Shetvin-Hillias* Company, and Midwest Research InacicutaCiai). The reaulca of the** analyse* are shown in Table 1. The control which did not contain lead octoate or chromate wet found to contain approximately 0.01% laad. There was e reasonable agreement beewe'es the difftreat analyte*, ao an average ppm of Iced wet computed and used for the actual lead concentration of each paint film. Lead concentra tion* of the lead paint obtained from the N'aciona) Bureau of Standards were averaged in a similar manner. 8. laad Analyses of Slats Contelnine Siffarant Paint Films Table 2 shows the lead content of each diet. Purina Rat Chew mesn was found to contain approximately A ug/gm of lead (assuming 0.1 ng/ga cf lead was contributed by the control point). The tig of lead/gm of feed which was attributable to each added peine film wa* reasonably close co its respective theoretical value. Tha sveragt coral ug of laad/gm of feed for eacn disc was used for the calculation of weekly and total lead consumption ft; seen rat. if*. '-r-y; 6 r,-4.1*9 7* 0007-SWP-036980 0007-SWP-000115548 r* t- Paint Pi'.r. Control Paint Lead oetoata '.tad oetoata laid oetoata Lead chromaca Lead chromate Lead chromate lead carbonate MBS lead paint coscEyrnATTo?: or ie a o in p a in t p iim Theoretical Load Concentration w 0 0.06 0.50 2.00 0.30 2.00 15.00 64.12 Measured Lead Cnre<- trjcis?! Paint Cottpamas Mill /".i /pi Average!'' /* 0.01 0.08 0.50 1.90 0.42 1.80 12.3T 63.20 , < o.o: 0.07 0.36 2.20 0.41 2.10 12.30 68.90 11.9B 0.01 0.08 0.53 2.03 0.42 1.95 12.43 66.05 ./ Maaaured by V.S. Department ot Ceauaerte, National Bureau of Standard*. Jahin$ton, O.C. ,6/ Average of paint companle* and MBI value*. / Avarafa of Bureau of Standard! and MRI valuaa. H ... *V.; >38* * .-h/Sife V*vT^iSS5-; ' yV-v*'3. 0007-SWP-036981 t abl e 2 > LEAP AVALY5ES 0? 21ETS CJSTAtVtrX "IfTi-irT PAINT ?'-V.`S 2iet Control paint plus rat chow Lead octDaee (0.082) plua rat chow Lead oetoata (0.337.) plua rat ehow Lead octoate (2.052) piua rat chow Lead chromate (0.422) plua rae chow Lead chromate (1.952) plua rat chew Lead chromate (12.437.) plua rat chow Lead carbonate (66.057.) plua rat ehow NSS lead amomd paint (11.922) plua rat ehow hcoricieil-^ 0.1 o. 3.3 20.5 4.2 19.3 124.3 660.5 119.2 Measured TocalS' ?alnt Leae fos/nn) 4.1 (6) 'ic ~ |g mm mm3.2 1.1 (6) (6) 12.0 (6) 7.9 -- .7 (6) mm27.2 (6) 23.1 (6) mm11.0 (6) 6.9 gjs (6) 23.3 (6) 19.4 (6) 140.2 (6) 516.0 CJ> 124.0 (3) 136.1 (6) III511.9 Jim(5) 119.9 (3) */ Theoretical concentration of load in feed contributed hy indicated paint film, b' ``can ng of laad/gm feed for number of sample* aiiown in paruitcliuaia. ".can ug of lead froa paint fllma/gm feed for nurber of lamplc* shown in parentheai*. 3 ___ _ n- --. 'w 0007-SWP-036982 ... :: :i. . -v- iv. .:? 0007-SWP-000115550 C. Preliminary Foedinc Studies C Before beginning chi* study. a pilot experiment was performed to sae if tne rats were consunins cn lead paintfTT> along with tne:r tied. Three rat* each were, placed on die control diet and the 66.03% lc-J caroor.ace diet. After AS hr^.e.a were collected and lead content r.easurec. feces tram rats on tne control diet contained11.5 ug of leed/gn faces vrvile faces fro* rscs fad the 66.03% lead carbonaecdiet contained 3,683 -j of lead'ga -`aces. Thus, rat* etc constant tna paint trim sJong with tneir fteo. CVaape e - - 0. Cross Observationi Feed Consunoeions and 3odv U'alehts Durineeh^jntiT|^jtudvjn^j!j<^jjj^Jiai^rjjgaj];|y^jgjymjf. One female rat on 2733%4caaooete developed an aocM^^Jje^la^JJJjiitBaf during the ninth treatment week and one fanala on 66.032 leao carbonate developed abcessed hind feet in the_10th week and chmrnag toes off. 1 The feed consumption*of aale and female rata 2* shown la Figures 1 and 2, respectively, Ixcapt far the rata fed the diet containing 11.82% lead as KBS laad paint, all groups consumed feed at the sane rate as tna con* trol group throughout the entire experiment. The raaaon the 11.82% KBS laad paint group ata significantly lass initially, was that they ware started a month after the other rats and ware initially smaller. Sine* this group uat consuming faad at the seme rate ea the control group by 4 weeks, we consider tneir initial food consumption normal also (for thair site). The body weight geint for eeeh group of rets are shown in Figures 3 and 4. At mentioned above, the rats on the 11.92% KBS lead paint diet ware small initially hut bteams similar to centrals by slthar tha 4th (females) or 6th wetx (sales). Throughout the entire period the male rats in tha other treatr.ee: groups gained weight ec e normal rats. The female rats did snow a change m booy weights during tna first 6 weeks of faading which appears related to the concentration of laad oecoate. After 4 weeks of feeding, rata m tna l.C3% lead occoacc group weighed significantly lass than control. Tha patter.-: was the seme at fl weeks. After 12 weeks chair average body weignta still inhibited the same patcem, but the 2.03% Iced occoate group was not significantly different from control. This waa dut to fewer animals and a slight increase in variability. &+<* v'Ob/s'W*e-- Axs( BWi Ut "Wbw.. .4u ix^Vuiff/ *\ 7** . S' i 0007--SWP-036983 0007-SWP-000115551 ---- !---- " WSREBSiifc *" -i H .i " - '*' '> ' - * , j -- i ------ ____**>__li nT"b*Liki i--': ...... ." '**;................. - - - . * . .... a ._ ?r >. .* . , -- v .* Hi'* * --t m '-i ^ .||^L Jiiin : i. '!''~iiin' T ~- ` --.1--.! .' mniH i; 3 j i! i i sbsitcgjp ~-23=fc ijCEaoao c s z e b s s h e i' "l?1 --..................... _----------------------------.,. -....,,. %r >4 !'* .3, .i) .5^/.*!'.-*: \j ; :o V:"v*.; -:rW- iZ&r Vi " Vr'- '. 1 '. 0007-SWP-0369B4 0007-SWP-000115552 n w' ^mgMiK.aiaur'i K*<S- I*4 1 #1 ItJ 'rSf i iUMUi !1 1 1 1 1 1 1 S ad (*/,./-> MgiMJvntNCS Silt 11 4*'/* * U'VV JO':.. 0007-SWP-036985 0007-SWP-000115553 Figure imni I I ? 2 l I >s\ Hi : ;i 21 ?W :caazacu 1 l ~i J tii .-c;t i: -cssssm iitiKafct i imi u mu* '... r'w-V i-v-.-:. - >: /': >. i... T1 0007-SWP-036986 0007-SWP-000115554 5V.V. .. S. Figure 1-5 1 !* '*: * ii "---- H*V" ' "= i *m*& igY".?y i 0007-SWP-000115555 e. ehl and Total CarSu.-->;icn The weekly and total lead cansur.pf.cn caring the experiment were calculated an the basis of ttie tetal luad/dict and amount of each dice eaten The results of the hematology of rat* fed paint films containing different concentrations of lead are shown in fables 5*13. No differences were observed in ttie erythrocyte count, reticulocyte count, hanatocric, leukocyte count, differential leukocyte count, and the erythrocyte osnotie fragility. G. Tissue end Fluid Chemistry of Rats Ted Feint Sours Cc-:a*r.irc 3'.ffcren; Concentrations of lecd t q V i Tables 11-11 show the tissue end fluid chemistry of rets fad point ra-fets containing different concentrations of lead. Ths serum proteins, erytnrocytt protoporphyria, urinary coproporphyrin, and urinsry deUa-ajtmo- ievuiinic acid ware not altered by any* of the diets. The activity of eho erythroeyte enrymo, delta-aminolevulinic acid cenydrije (ALAI)), in the control, lead oetoete (O.OSt, 0.33Z, end 2.03*) end '.sad cnrorate L-1.55V and 12..31.) .groups dm not differ thrpjighautje experiment -Taolei ||). ALADiotiyitv ms ccaroteed in rata fed 66.Q5% lead caraer.sts anc 92-, ::as lead oaint. AC & weeks tnerc we* a depression of ioi and 591. in these two grOuoLiZrggbectAVeiy. weeks the ALAD activity wa na-*nan w Icoj^jaroonar^^jip, but tne AUD activity -r. tr.a :;!S lead paint group we* not significantly different from control. At this time a sex diffstance was osserved in both of these groups. The A1AS activity m the male rats remained depressed at the 6waeklevels while the t'iJO activities of the fssnalo rats rttumed to control level*. This sex difference say be linked to the fact that these female retsVhed matured end wore prrujbiy oeginmns trt. sir nenityfal eyelet. n nv) >1, ssbr Zb*#* *~i.. y*.*. . * `1 0007-SWP-036988 0007-SWP-000115556 '7V7?rr- s i. ~ i i - i| * i+=Z *5 s||= j S *J 2* 5' i *. i :r* ^;- ;: - is 55 - s* * \1 pp *i i *. 5s i* p p P P P p p p*3 i 5 *3 t 5s i* ;* t' ~*/i i * r r ri :* :s i v v Pi5 'j J: *j m 3 it; !!:: ii i 3 if i I3 ii i i1 rS-;' -:- -v i* ?s ;5 ii= l! Is 5s P ;* s s ; * I-5 is p p r r r Is V P 5s i* Is r i 3 il i -1 i l1 !* 51"i ;i 5 mi s* lt : i= S.= \}* i * ; j*. i*i- :;i 2;iS* 5 i** iiii !!3 P l si jl il j! -! it il is - ii ;i ii ;i ii ;t is isJ I, I* * u. -* "r 'i ? ;i 1ii! 13 > if'-: ?*?'*' i> i. - 0007-SWP-036989 0007-SWP-000115557 o Q / > M- "i -i . : . : ; ^ i i t s' r ; r rr !'>i M V- \l *s s 5' J- I *7 *S *1 22 = 3 := :i i r'i r*j i r i1 r* S1 =3 i i! * Ji i5 i5 ur. r* a i ;i k r- V-4i *3 it i* ;i ? ? *s r ; i- =S 2 a s ;i; v J ,=- \1 *. s : *s r s5 V- a t t! I i'! 51 is ?! V i! V < 5- i * Si .. S a * i ii v- r- is *; : s S' %* s- *r: - s i i * ii V i" r * P ; : * i r : I 3! i* Jt i* i* !M* *3I! it i! *f I :* = - -i : :i r s- i: ii j! i 1i ? i* * >a H i* * if l!' a ;s ?- -i3 i |-3 j! ! , r ?! *i %- 'i 5= j- ?i HJ iqt JiH >3 :* H i* is ii i* ;I i- i t * : i - *i: js, , i \ i-Ui iilir 5 j? 5 : * ; -s: i : i ]V 16 \r-* * * W* S'"*' . : ^ r ............... - - ....... .. ., 0007--SWP-036990 s, , ____________ ___ : --------------------------------------- - ' 0007-SWP-000115558 *- TABLE S c o n t a in in g n o i LEAD AnfIvses Treat-cnt Vesk 6 8 13 fS'6i Erythrocyte* (x 106/c3) 6.17 6.33 0.19 6.42 s 9.29 Reticulocyte*, T. 1.5 0.7 0.8 = 0.3 1.4 0.1 Hematocrit, vol.,7. 66.1 0.8 64.6 1.2 44.6 * 0.3 Hemoglobin, ga, S 16.0 = 0.3 16.6 * 0.3 16.0 * 0.1 Leukocytes (x 10^/en*) 5.0 t 0.5 7.6 * 1.3 8.0 a 0.6 Neutrophils, T, 11.8 s 2.9 21.7 * 5.6 13.1 * 1.5 Lymphocytes, T. 86.3 * 3.3 75.4 * 5.3 84.1 s 1.6 Sonde, ?. 0.3 0.2 O 0.1 = 0.1 Eosinophils, 7. 0.9 0.6 1.3 o 0.6 0.5 s 0.2 Btsopniis, " 0 00 Monecyeot, T, l.l s 0.6 1.6 0.6 2.1 0.4 Acypleol, T. 0 0 0 Nucleated SBC, S 0 0 0.1 * 0.1 o Erythrocyte esaocic fragility [SC1^ 0.311 * 0.004 0.391 0.008 0.400 = 0.004 i Nusor of rae* per period, b/ ;:tn = condo rd orror. c,/ Canetncrotian of XoCl Shot hamolyred SOS of the erythrocytes. : * 9 " ; 8 ^* j... ' 7* v*V"v -' * . * ' * K4. ` . <k p* *#.*-*? ,,.*-. -*i"r.a'W7arSb^l''! 0007-SWP-036991 0007-SWP-000115559 TABLE 6 HSHATPIPGY 0? \ATS TKO FAIVI **~T coNTAir.'t::; r.'vy. :;An as '.t ab o c t o at e A.-alvses _________________Trucr-enr 'Vtik A3 (JtmL.) 13 Erythrocytes (x iO^/nsi^) Reticulocytes, % Hematocrit, vol.# 7. Hemoglobin, t~y % leukocytes (x IC^/k o P) Neutrophils, * lymphocytes, " Bond*. 1 Eosinophils, % Sssophils, * Monocytes, X Atypical, " Nucleated UC, * Erythrocyte osmotic fragility CSaClZ--/ 5.14 O.lli1 0.9 = 0.5 41.3 * 3.9 14.1 s 0.4 5.1 a 0.S 24.5 a 1.6 74.3 * l.B 0 0.B a 0.3 0 0.5 = 0.5 0 0 0.331 > 0.005 4.33 = 0.09 1.0 * 0.5 46.0 * 0.7 15.4 * 0.2 5.5 0.7 20.3 * 4.7 76.3 * 4.7 0 l.S * 0.6 0 2.0 a 0.3 0 0 0.410 = 0.007 5.92 * 0.39 1.4 a 0.2 43.0 a 0.5 14.7 * 0.3 7.7 * 0.9 16.3 * 1.7 30.9 s l.S 0.3 i 0.2 0.7 * 0.3 0 1.9 0.5 0 0 0.395 s 0.005 iJ Number of rats par period. Sean = standard error. Concentration of NaCl cim htmolytcd 50*. ef Che erythroeytes. *1 sASSr* _ m>VAf f i 1 s&s r-.-rr i.' ~ _ / ...au. |> Hr - 0007-SWP-036992 0007-SWP-000115560 t abu ; O^r0 HEWAT07 0PY OF HATS TtTl ?A'.::T agfcMg CONTAlXtNC - rSi*'. ISA!) AS MAD OrrOATE Anaivbcr Treatment 'Jack A 8 13 IX41 Erythrocytes (x lofyam3) Reticulocytei, Hematocrit, vol.y * Hemoglobin, 7. Leukocytes (x ioVmm^) Neutrophils, 7. lymphocytes, % Banda, r. Eosinophils, Z 3stophiIs, T Monocytes, Z Atypical, Z Nucleated XBC, * Erythrocyte oanotie fragility [jtaClS/ 6.27 = o.' 0.9 s 0,7 42.3 * 0.7 14.2 a 0.2 5.0 a 1.4 15.8 * 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 * 0.3 7.5 * 2.8 19.3 * 3.8 77.3 * 3.8 0 1.0 s 0.4 0 2.3 s 0.5 0 0 0.388 3 0.012 3.6 x 0.4 1.6 s 0.2 43 s 0.7 15.3 3 0.3 8.3 s 1.0 16.2 s 1.9 81.4 x 1.3 0.1 3 0.1 0.8 s 0.3 0 1.6 3 0.3 0 0 0.401 3 O.OC a/ Number or rata par period, b/ Mean s acandafd arror. c/ Concentre;ion of N'aCl that henolyzed 307. of Che erythroeyraa. 1 > * '. k 19 0007-SWP--036993 0007-SWP-000115561 TABLE 8 HSMATOICCV ?" RATS FED PAINT CONTAINING 2.M* IS ad As"'as!raef3Stt......... AnpIvi^i Treatment Keek 48 13 Eryehrocytes (x l06/ws3) Reticulocyte*, T. HtMCoeric, vol.* X Hemoglobin, ga f. Leukocyte* (x lo^/s**3) Neutrophil*. % Lymphocytes, Sand*, T. Eosinophils, * Basophils, X Monocytes, X Atypical, * Nucleated RBC, 7. Erythrocyte osmotic fragility ^SeCl^ 6.30 * 0.2&7 1.1 x C.6 41.8 0.3 14.4 x 0.2 3.1 1.1 13.3 * 3.0 83.8 * 3.2 0 0.8 * 0.5 0 0 0 0 0.381 x 0.006 6.46 x 0.08 0.7 s 0.1 45.0 x 0.4 15.3 * 0.2 6.7 * 1.7 13.0 * 2.1 85.0 = 2.3 0 0.3 0.3 0 1.8 * 0.9 0 0 6.72 = 0.42 :.o x c.6 43.4 * 0.8 13.6 x 0.3 8,1 * 0.7 14.1 x 2.6 82.8 US 0 0.9 x 0.3 0 2.2 * 0.7 0 0 0.399 x 0.008 0.393 s 0.006 1/ Kumar of race par period, b/ Kaon x standard error. ' Concentration of SaCl that hamolyzed 50* of the erythrocytes. -<v-: - r***,' K&v as Sgg? ;V"?i r&*r.. . B5S&. - S4 Mwir (;*&-fpa. rififcsi 20 s;i 7* ? ; v-t.VviV-TVi sgS 0007-SWP-036994 0007-SWP-000115562 TAKE 9 fcL^r* HEVATO'-OCY 0? ?ATS F** p it ..- f-u. C0StAty^S'~".'.'!T''igAP"''AS''''^Bvn^8<AT5 An lvses Treatment Vick 4* 8 'V41 Erythrocyte* (x 106/im3) Reticulocytes, 7. Kenicoertt, vol.f " Hemoglobin, gm; 7. leukocytes (x lO^/aa^) Saucrophllt, 7. lynpnocytts, 7. Banda, S Eosinophils, * Basophils, * Monocyte*, 7. Atypical, " Nucleated RBC, 7, Erythrocyte osmotic fragility, *SaCl;/ 6.71 s 0.16&' 0.9 X 0.8 A3.3 * 0.6 1A.7 * 0.A S.l * 0.6 8.3 * 1.7 90.3 * 2.0 0 0 0 1.3 * 0.8 0 0 0.383 s 0.003 6.18 x 0.16 0.8 x 0.2 A5.0 it 1.1 13.1 0.3 3.1 x 0.3 19.0 x 2.0 79.5 x 1.3 0 0.8 * 0.8 0 0.8 * 0.8 0 0.3 X 0.3 0.393 * 0.12 13 cy52' 6.6 x 0.3 1.6 x 0.2 A3.6 x 0.7 13.6 x 0.3 6.9 x 0.7 U.3 x 2.1 83.A x 2.0 0 . 1.2 x 0.4 0 1.2 x 0.A 0 0 0.392 = 0.0C wtl* a/. Number of race par parlod, axcape where indicated otherwMr. b/ Mean X (tandard error. c/ Concentration of Sad tiiat heaolyzed SOU of cha erythrocyte*. 4. t * &&.V- ShiT*? ?23k-:i> 5>-s?i C**;^ v'5- Rf .ss-^sr u r^Yf - ` T0f$$(rZ .?7'V - ; v**v*- - ... v-'V ^6' '*' - *'*'-* 0007-SHP-036995 0007-SWP-000115563 Afalvtt* Erythrocyte* (x 10* '=vrs^) Reticulocytes, \ Hanatoeric, vol.y Z Htnoglobin, tv ' Leukocyte* <x lOVsaJ) Sautrophila, ". Lyophoeytti, 7. Sand*, % Eo*inopnili, * basophil*, T. Monoeycaa, Z Atypical, * Nucleated RSC, Erythrocyte oxnotic fragility, CsaCl^ 6.17 s 0.0iSJ 0.S r 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 z 0.007 13 6.21 = 0.13 0.8 z 0.3 44.3 s 0.3 15.3 s 0.3 6.1 * 1.2 18.3 z 6.6 80.5 z 6.6 0 0.3 s 0.3 0 0.8 0.3 0 0 0.385 = 0.007 6.37 z 0.52 1.2 z 0.2 44.1 s 0.7 16.2 s 0.4 8.2 z 1.0 11.4 r 2.3 85.9 z 2.2 0 1.5 s 0.3 0 1.2 z 0.3 0 0 0.404 s J.003 / .`.'unbar of -tit per pariod, S/ Mean s aear.dtrd error. c/ Concentration of I.'aCl edit htaelyted SC7. of cha erythroeytt* ** SggSf $&z ^7 " *% cr`.7&S*. J-".- ' .*- I"y ^ 22 0007-SWP-000115564 TABLE 11 HEY-ATP'-QCY 0? BATS ~P AI3? ***T-3*- '-Tad as is a u rnovATC Analyses Erythrocytes (x ftetieulocyte^jfc hematocrit, vol., % hemoglobin, gm^ 7, Leukocytes (x 103/sm3) Sauerophlla, % Lymphocytes, % Bands, * Eosinophils, % 3ssophils, * Monocytes, % Atypical, 5 Nucleated BBC, % Erythroeyta osmotic fragility [Sadi*7 A 6.32 * Q.IL&/ 1.95 a 1,25 41.3 * 1.1 13.9 a 0.4 4.5 a 0.3 21.5 * 6.3 73.0 s 6.0 0 0.5 * 0.5 0 0 0 0 .354 a 0.007 rcatment "eek 5 />r4i 13 'S-12' 6.23 a 0.15 0.9 a 0.1 43.5 * 0.5 15.2 a 0.1 5.3 a 1.5 15.0 * 2.1 B2.S a 2.0 0 1.0 a o 0 1.3 * 0.3 0 0.3 a 0.3 0.35$ a 0.010 6.61 = 0.33 1.6 a 0.2 44.1 a 0.7 15.3 * 0.2 7.6 a 0.6 12.5 a 1.6 54.5 a 1.7 0.1 a C-.l 1.0 a 0.4 0 1.3 a 0.3 0 0 0.396 a 0.006 a: NuaSer of rats par period. 6/ Mean a standard error. Concentration of XaCl that hemolyzad 3051 of the erythrocytes. R .* . ov'cis 23 0007-SWP-036997 0007-SWP-000115565 'JEJIATW.PSV 5? *A?S ?sn PAl'.T * CONTAINING i-OSVlEAD AS ICAO CAttOSATE Analv*ea a 'S3'4' Traatncnt Veeit 8 !2sl Erychrocytaa (x 10*/TM*) Atciculoeysei, \ Haoatoerit, vol./ 7. Kanoilobin, in/ % Laukoeytta (a l03/an3) Sautrophlls, 7 lyapnoeyta*, % Banda, % Eosinophils, 7. Baaophila, X Monoeytaa, X Atypical, X Nuelaatad RBC, X Erythrocyte oootic frajillty [MaClI-' S.JA * 0.36^/ 0.4 a 0,2 41.3 0.9 13.9 0.3 4.3 s 1.2 14.3 s 4.1 82.0 3 J.8 0 0.7 4 0.7 0 3.0 = 2.0 0 0 0.388 * 0.009 8.13 * 0.06 1.1 * 0.3 44.0 4 1.3 1S.0 * 0.2 5.0 * 0.9 13.0 * 4.1 14.0 * 4.2 0 0.3 s 0.5 0 0.5 * 0.3 0 0 0.394 4 0.012 13 Oa#!2' 6.60 3 0.37 2.9 s 1.3 44.7 3 1.2 15.6 s 0.6 7.7 s 0.5 13.2 3 2.6 82.7 s 2.7 0.2 * 0.1 0.9 s 0.3 0 1.0 s 0.3 0 0.1 * 0.1 0.397 = 0.003 a/ Suobar of rats per paciod. V Mean s standard arrer. c/ Concentration of JfaCl that hamolyxad S07. of cha srythroeytas. l >% rn&g S-*?sS? SW/51*. yet 3588 te v'%--r+ ' : * b ^a t\ . ^ 2. 0007-SWP-036998 TABLE 13 7o l c o y or ?.vrs r*o ::*s LEAD :n t co::rA:*;:No n. : E43 4 I'y.iii'1 Treatnenc 'JmmV 8 13 5.72 s 0.15^ 2.5 * 0.3 39.7 * 0.8 13. * &! 9.7 1.4 9.5 = 2.1 90.2 * 2.3 0.3 * 0.3 0 0 0 0 0 0.389 * 0.006 4.49 0.83 1.0 * 0.4 42.8 1.1 14.7 * 0.3 6.7 * 1.2 7.0 * 1.7 92.0 * 2.1 0 0 0 1.0 0.7 0 0 0.412 * 0.006 5.63 = 0.34 1.3 * 0.1 42.9 s 0.4 14.7 * 0.1 -- 7.0 0.5 14.2 ss 2.1 83.6 = 2.2 0 1.4 = 0.4 0 0.8 0.3 0 0.1 0.1 0.408 s 0.005 :*?lod, exempt wham indicacad ocherviaa. .1 chat hanolyxaa 50% of cha arycltrocytaa. :s ;*> *' 0007-SWP-036999 0007-SWP-000115567 o & TABLE 14 c !mv t is s u e as p r-rin oiE:rsT!w o f s at s t es ?Ai:rr c q n t ai:::n c m, l eap Antivie* Traatr-ertc LVk 48 fS-81 Serum electrophore*!* Albumin, % Alpha 1 globulin, 1 Alpha 2 globulin, 7. Beta globulin, 5 Gamma globulin, X Total protein, 1 Aibuain/giobulin ratio 43 * Si/ 28 a 1 5*1 20 * 1 6* t 5.6 * 0.1 0.76 * 0.03 63 s 2 23 * L 5*1 20 * l 10 * l 6.2 s 0.2 0.77 * 0.07 Srythroeyta ALAD, umol. PBS/ Male*: 23.2 4 2.3&/ 100 ml ABC/hr Females: 16.6 * 3.iir Total: 19.9 2.2 Protoporphyrin, 19.6 1.6 ug/100 ml RBC 15.1 * i.j/ 16.3 * 2.2&' IS.6 * t.S 23.3 * 1.9 13 '>26) 48 * 1 21 s l 5*1 21 * 1 6* 1 6.3 s 0.1 0.93 * 0.04 14.5 * !. >/ 13.4 * 0.7^/ 13.0 * 1.0 29.2 * 2.0 L'rine Copreporphyrir., ug/26 hr ALA, ug/24 hr 2.3 1.2 78.0 * 7.6 3.3 * 7.6 83.3 s 11.0 6.0 * 1.9 63.6 * 3.0 a/ Nusbar of rat* por period, ixeopt where indicated othervia*. s'- Mean = standard error. ./ Four ret*, d/ Twelve rtc*. i.* W. WV" 26 0007-SWP-000115568 o o TABLE 15 eJU*** rzssi-E a::o fir13 CVS'TSTAY OF sATS FS1 ?s t jrr-wt- c o n t a in '::.g :. !.eat as '.EAl* CTTTATf Ara!v*s 4 _^X;4V3.t Treatrent Veck 8 (Vail Sarum olacerophorotls Albumin, ~. Alpha 1 globulin, X Alpha 2 globulin, Bata globulin, % Gamma globulin, % Total protain, jig % Albumin/globulin ratio 40 x W 26 1 58l 21 a 1 8*1 5.9 * 0.1 0.66 * 0.02 44 * 3 20 * 2 4* 1 20 * 1 12 * 1 6.3 * 0.2 0.79 x 0.08 Eryehroeycaa ALAD, uaol. PBG/ .Hala> : 24.8 * 7. 100 ml RBC/hr Famalaa : 19.5 * 4.i' Total:s 22.2 * 3.9 Protoporphyrin, 28.4 x 2.4 ug/100 ml ABC U.9 a 1.3/ 18.7 * 0.3S/ 16.8 * 1.2 24.2 * 1.6 13 48 8 1 20 s 1 58 1 22 s 1 7*1 6.2 s C.l 0.93 s 0.04 12.0 = 2.^./ 16.1 s l.J^ 14.0 s 1.4 22.8 s 2.5 L'rina Coproporphyria, ug/24 hr A!Ui ug/24 hr 1.2 * 0.2 86.4 s 16.6 4.8 * 2.4 107.6 8 34.1 2.9 * 1.4 3S.0 s 6.7 *i Suabor of race par pariod, axeapt wbtre indltacad otharviaa. / Maan s atandard arror. i/ Two rati. i! Slx raea. 0 o 27 ss m i.'v&V sc2x> VipdV"- rv< j-- ;! >v&8&t fry-*-. &Qtr} '7?*, A-'V*-'S 0007-SWP-037001 0007-SWP-000115569 TABU 16 o ; '* coNT-.:'::n o -`.S3'- \no a orroAtr CW^t * Ana !-.** m fr.'.i'.i' Traatmant Maak S 0>41 ;3 Strum alactrophoraiia Albumin, Alpha l globulin, a Alpha 2 globulin, * Bata globulin, % Cana globulin, % Total pretain, i| X Albuain/globulln ratio a i - ' 26 x 1 5* 1 21 x 1 Sx 1 3.7 * 0.1 0.6B 0.04 41 * 4 ' 23 s 3 6*2 20 * 1 11 x 2 6.3 x 0.1 0.70 X 0.13 50 x i 20 x 1 4x l 20 - 1 7x1 6.0 x 0.1 1.00 x 0.07 Erychroeytsa ALAD, uaol. BBC/ Malta: :i .a x t.& 10.3 x 0.8y 12.5 X O.ri'' 100 ol XBC/hr rtnalaa: i5.s x 15.3 x 0.2' 17.0 X Total: 18.4 a 3.3 12.8 x 1.3 14.7 x 0.9 Protoporphyrin, 23.1 x 1.0 21.7 x 1.1 27.7 x 3.3 0 Ug/100 ml X3C Urina Copraporphyrm, Ug/24 hr AU, ug/24 hr -* -* II O Ob 10.7 x 5.9 97.4 x 19.2 101.4 x 30.1 4.9 x 2.4 50.5 x 6.6 a,/ .Nuaoar oi rata parlod, txcapt wnara indicattd ot.lan.-Ua b/ Maan x atandard arror. c,/ Two ratl. / Six r :i. wr <ptv * ?>rX ?. |HK7 iy?- 1-- ?#s? '=3*v>$#r. zvZ.f. r:.V*c !.r**jr*Ti.i*vxi-*-. w*. - * **# . % T '>-/'.** . *, ,y Jr.W"7 j^V^&'*.' rt *C*w 0 V, i ' :s *A''i/y 'jv`. 0007-SWP-037002 0007-SWP-000115570 TABLE IT A !ii lvm t is s it >vo Borv fu`?n cst^sw *r r at s ' A::r c o n t a :vtvc 2. os*, l t a d as og-s;,-? ftix* Trracaant "aak -8 13 Strum alactrophortiis Albuain, T. Alpha l globulin, % Alpha 2 globulin, * Saca globulin, % Sam* globulin, $ Tout protain, ^ ! Albuain/globulin ratio 41 s 3^ 27 X 2 4*1 20 s 1 9x1 5.7 X 0.2 0.70 * 0.07 Erychroeytaa AUD, wool. ?BG/ Malta: 100 ml KBC/hr Fanalaa: Total: Protoporphyrin, ug/100 ml XBC 24.1 18.7 21.4 29.3 X 4.0&' X l.o^ x 2.3 * 2.8 46 s 2 22 * 1 4*1 19 X 1 9*l 8.0 * 0.1 0.85 * 0.07 48 i 2 20 x 1 6x1 21 x 1 6x1 6.2 x o.l 0.96 x 0.07 12.1 X 1.7--^ 11.3 * 0.z; 11.7 x 0.7 30.8 X 2.5 11.4 x O.S^ 16.3 x 3.9^ 13.8 x 2.0 24.0 2.2 i'rina Coproporphyrin, ug/24 hr ALA, ug/24 hr 1.6 X 0.6 80.7 x 17.3 3.0 x 0.8 82,7 X 10.4 7.7 X 2.9 64.5 x s.8 a.- -*b*r of rata par pariod, axccpt vhart indicated otharvii*. b/ V.aan x atandard arror. c/ Tuo rata. 5,1' Six rata. 29 " 0007-SWP--037003 0007-SWP-000115571 TABLE IB A-i'.vsa TISSUE AND BODY rr.L'ID CHEMISTRY ffr RATS ?ES ?A:r*7 JOtif COXTAI.VING O.iy. LEA3 AS LiAD CHSOVATE 0*t 4 r$mu\kt raatmcnt Vcah 8 1*=1 13 QI2) Serum elaetrephortais Albumin, Alpha l globulin, * Alpha 2 globulin, % Bata globulin, X Gama globulin, % Total protein, g t Albumin/globulin ratio 42 * 28 * 1 40 1 19 a l 8*3 5.8 * 0.2 0.71 s 0.06 44 s 2 20 * 2 4*1 19 x 1 12 x 2 6.0 * 0.2 0.80 x 0.08 47 x 1 22 X 1 5*1 21 * 1 6xi 6.1 x o.l 0.90 x 0.03 Erythrocyte ALAD, pool. PBG/ Mala*: 23.4 * 3.5&' 100 xl RIC/hr Famalat: 13.4 * 0.9*' Total: 18.4 * 3.3 Protoporphyrin, 27.3 * 2.8 pg/100 ml TBC 14.1 * 0.6*^ 17,4 * 0.1*' 15.7 * 1.0 28.3 x 0.6 15.7 x i.Sjl' 16.5 * 1,4*' 16.1 x 1.0 27.9 * 2.1 Urine Coproporphyrln, ' ug/24 hr AU, ul/2A hr 8.2 * 6.7 69.3 * 6.6 1.3 * 0.7 65.2 * 17.4 6.3 x 2.2 85.3 x 31.4 */ Kumber of rat* par parted, axcapc whara Indicated otharvlsa. b/ Mean s acandard error. cj Two rat*. ij Six rat*. 30 w . k a. wsr* r-rTf^V ***Y ~ . t.l vj'vi .-i** >J7*** t5; * *. 0007-SWP-0370D4 0007-SWP-000115572 Ane'vses TABU 19 TISSUE AMD BODY Fin* CHEMISTRY OF RATS FrrcD p a :; a. *W CONTAt ':i\*C 1.95", MAD as ma d c h r o mat e CUo Trca:-cn: '..'et>. 4 fX4H/ 6 fS-ll 13 Serum electrophoresis Albumin, % Alpha 1 globulin, TS Alpha 2 globulin, % Baca globulin, ", Caana globulin, 7. Total protein, fg % Albumin/globulin ratio 41 2&/ 25 * 1 4*1 20 * 1 9*1 5.7 * 0.1 0.73 * 0.06 45 * l 19 * L 4* 1 20 * 1 U* l 6.20 * 0.04 0.81 * 0.04 47 s 2 19 * 2 5*1 23 * 1 6*1 6.4 * 0.1 0.94 s 0.08 Erythrocyte ALAD, uaol. SBC/ Males: 29.5 * 3.l' 100 al RBC/hr resales: 20.4 * 4.3i/ Total: 24.9 3.4 Protoporphyrin, 26.3 * 4.7 Ug/100 al BBC 12.3 * 1.0/ 14.7 * 0.6E' 13.3 * 0.8 26.8 * 1.8 13.3 * 1.1& 12.9 * 2.2&> 13.1 * 1.3 27.4 * 1.3 Urine Copropotphyrin, Ug/24 hr AU, Ug/24 hr 0.9 * 0.1 60.8 * 7.2 7.0 * 6.4 67.7 * 6.8 / Number of race par period, except where Indicated otherwise. / Mean * standard error. / Two rat*. Six rata. `AV.V.- : - ip ,** j* e ' ' >.Tj '/.vty: M&'1* 1^"Kv ?iV*s:i;ifV^Svv. STJjWr.. P fees* HJF5V VAffTf !f?SiV 31 >< *Vr !. . tdIVtv _ r v *w* iw.- * * , .' * I -*? "* v 0007-SWP-037005 III. (I. fV l 0007-SWP-000115573 TABLE 20 TISS11 AN5 SOTVY rV25 EijgMIST^f P" -ATS *22 TAUT ocr-TAi^rs ::.-3- i -ad as l e a s Analvsas Treat-ant * >'< 6 13 Strum eleeerophorttis Albumin, X Alpha 1 globulin, 7, Alpha 2 globulin. ?, 3eta globulin, " Gama globulin, 7. Total protain, ^g 7. Albumin/globulin ratio 40 * f 27 a l 3a1 22 a 1 8* 1 S.6 a 0.1 0.68 a 0.03 47 a 3 21 a l 4a 1 18 a l 9 2 6.1 a 0.1 0.90 a 0.12 46 a 2 21 a 1 5a1 22 a 1 6a 1 6.4 a 0.1 0.88 a 0.36 Erythrocyte ALAO, ml. PSG/ lilies: 100 ml RBC/hr Famalat: Protoporphyrin, ug/100 ml BBC Total: 26.4 a 7,4 14.0 * 3.2-7 20.3 a 4.9 2S.S * 1.6 16.5 a l.t^ 13.4 a 3.B5/ 13.0 a 1.9 24.2 a 2.6 if12.6 13.7 a a 1.6 l.t il 13.1 a 1.0 24.6 a 2.3 I'rina Coproporphyrin, ig/24 hr ALA, ug/24 hr 2,6 a 1,8 89.7 a 19.3 0.9 a C.l 78.4 a 6.8 2.6 a 3.3 49.7 3 lM%t Nusbtr of rati par period, axeept uhara indict ted otherwise. / Keen a standard error. ' Two rats. Six rats. i". F-a.v V. t%*.<53' K; ^.V ' ,v t: j. 32 0007-SWP-037006 0007-SVVP-000115574 A:i*vses TABLE 21 TISJL'f : A";r -ow n vio c h e w s m*1 or t t : ?*: 24K rcNTAIK! '0 11.03-. LEAD A S LSA CAETOVAT? Mw-a* Treac-cnr Vc# a* ! 'X4i *2 5erua alaccrophorssia Albumin, T. Alpha 1 globulin, " Alpha 2 globulin, T. Bata globulin, * Coosa globulin, % Total protain, ^g % Albumin/globulin ratio 41 s 6 37 * 1 3= 1 21 * 2 1i2 5.4 z 0.2 0.73 z 0.17 Erythrocyte ALSO, uaol. PBS/ Males: 12.3S/ 100 al UC/hr raaalaa: 9.1 z 2,li/ 10.2 = 1 .tif Protoporphyrin, ug/100 al RBC 23.2 = 0.9 43 z l 21 z 2 19 z 1 10 s 1 5.0 z 0.06 0.62 * 0.04 47 = 2 22 = 1 5s 1 20 * l 7z l 6.3 z 0.1 3.56 z 3.06 4.4 z O.li7 12.4 z 0.4J/ 6.4 z 2.3X' 20.4 z 2.1 3.4 s :.</ 7.4 z 0.S/ 6.4 z o.:il 27.2 = 6.8 Trine IMJ Cooroporphync, ug/24 hr AUA > ug/24 nr 2.0 z 0.7 90.3 = 27.7 0.6 z 0.3 72.3 = 13.4 6.7 z 2.2 42.2 Z s.a , Sumoar or rats par period, except utiara Indie*tad otherwise. Xaan = standard arrer. Ona rat. Two rats. / Six rats. / S'-ini!leantly different fron control (? < 0.0I> as shown by Tur.netr's nultipleeoepari*OR test- following an analysis of variants. E-;w- t *...* . '* * *4 *y_ * 8S& ,vur*. 33 0007-SWP-0370Q7 li |*> lir l* 0007-SWP-000115575 :a s u TTSSfr .AS`n :*' y or 'a t s lAO .>/. i:.~ c.'N7.-,::::r:c ii.r i 'ao -ss Anjlvsis Treatment "eok < 8 13 Serum olactrepnorasis Albumin, " Alpha 1 globulin, " Alpha 2 globulin, * bats globulin, 7. Canoa globulin," Total protein, f.$ \ Albuoin/'globulin ratio 52 = & 21 = 1 4= 1 20 s 1 3=1 5.4 * 0.2 1.1 * 0.03 irythroeyta ALAD, pool. ?SC/ Hal as: 8.5 i.C^ 100 ml UC/hr Females: 10.1 * 0.1^ Total: 9.3 s 2.1*' Protoporphyrin, 14.8 4.3 us/100 ml MC ;; - ; 21 * 2 8* 5 21 * 3 6a1 6.0 a 0.2 0.78 * 0.06 14 s 2 24 s 1 7* 1 21 = 1 3=l 6.1 = 0.1 0.82 = C.26 5.0 * Q.& 13.0 * S.1&' 10.0 a 3.6 24.5 * 1.1 4.9 * 0.& 88 * 6.9 s O.o1' 26.3 = 1.2 Vrine Coproporphyria, ug/24 hr AlA, ug/24 hr 2.0 = 0.8 lie.i * ::.5 6.8 a 4.9 72.0 * 13.6 4.6 a 2.1 31.3 a 3.6 ' Sushar of rats par parted, axeapt unsra indicated otherwise. 1 >:aan = standard arrer. Two ra ts. Six rats. i Significantly diffaranc from control It < 0.05) as indicated by Ouanett's ajlciple-ecsparison east following aa analysis of variants. rs- .' \ r -v. > ***._ i " ", 1* '***V * Id...-- - 34 0007-SWP-037008 I .nl ti|-> i| i i | 0007-SWP-000115576 s o ai. sis < *a m 9 r 0 B * 40 .. . KN *neo -- -- cof* Vi +l VI <*l Vf VI +i V <Ntn9rA49N iVfwc*)) \ ij * Ot 0 * i> s fVi N N N -O" I +* w9 a e KA ^^^ o 4 N ^M V /i rt n ^NNN 0* v^ s *i s v m5 < m --i ! * Ss w <` 9* 9 ^* - f* t. O * -- *M V* *1 +1 c ^ -- ---- in ff m w^* t+a r ^IN^NMNNNN XI 0 V# *f III a0* 8 0* 8 5 to 5 0V U V U______ 11? 15 ?5 40 ^f<vn vMi r*4 >1 v1' 4t** 4 ** * 9 \ <N *v u i V a i S * d m m o H -g *> 29 ;v la a 1f * S" l : s b 0 . *to S s9 9-- <* 0W l*a --v W It 1 a ofc w w-- > * *5S* i. i ' 0 -- * q >* n --Z>aS* u a a^ y vt U ~*va6u a --:>aKi im Z < tfi tl| I a|9| *-> .& ` f--, . *?.*.* .. >; i . -."?r.i- `V*" m:-i f. * .:? ; *-iU '- > v .`r.. ?r.; ... * >.:*.,. ijS* - > *v, ti>v **>* >#/* 'V* *'*rt. < f 'v. 2?.v}>Mr'*-S'v.M*T'.--4^T;,aT.>< t-. " 0007-SWP-037009 0007-SWP-000115577 By 13 weak* the sex diffsrcnc* hsd disapo**red_cnd the ALAD activity in 66.051 ~>co'citaottatc'^cSy^naTa^TTmSr^B^Mcalist trcjo -ttt tesress *d 56oni 53%. rasptccively. Urinalysis of rsts fd paint ^'r containing different concentra tions of lead are shown in Table 24-31. No marked difference* in urinary preeein or sediment (eryenroevee, crystals, eaat*. etc.). Thu*, urinaiysu can be considered within normal ranges for all ereitmene groups. a* H. lead Content in Tissue* of Bees Fed Paint Concentration* of lead , Cor.tainir.t Different Tables 33*37 show the/lead content o^lood, brain, liver, kidney, ( bon* *ro rltl P*lnc *** con Mining different concentrations of lead-. Vpri values for eh* lead concentration* in blMd at 4 and B week* were ooitted^p*^ / from Table 23 because it was discovered that/ehe** sample* had been contaryW Li-*ted by their storage container*. At 13 <A*k the[ blood, lead leveijjg^ the ms fed lead octoate and eh* rats fee D^4K7onr?jsi^i**Tic^S2I2tliiL noccjwgjjmncswwm^tjfglatroi.Tn the rat> ;tajj STcnrcatSasg66.05% lead carbonfcg^jade 11.92%- MBS lead pa HU., blood, level* were found to be elevated ignj/icandy* -"'CSLr^vw^ At 4 weeks, tissue lead* levels in, liver end kidney were belmtf eh* detection liaiiu of our assay.^Tb* result* were the same at 8 week* with eh* exception of the JJB$><il*aj|ainiijrou. AlL_four_tati in thi* group showed detectable lead in their kidneys. ^^ /'Aj^JLSjueekj^eteetabl^evjUljJflej^Jtjjthei/Ao^rarel ir^jjgiijid^onl^ in the ret* fed l^ySyTSiTearBaMTyisl^^^e^^e^eebiyTeVe^jj ' ieadwjge^Jgjyj^ in two r*li_f*d 6a7032 laid carbonate* and nine ret* fed 11.92* ? KBSJLeedaeint, SSoq_lel*a*d vw*ee detected in echhni kiddnneeyye_ocff_ine_soatToJ_ group ef li wttas. in centralt, i**o was detected in the-kidneys of onerevo rats ir. ~ each lead octoate group) and the 0.42% and 1.85% lead chromate group*, lead was detected in the kidneys of thro# rate, sight rata and 12 rats in tha 12.43% lead chromate, 66.05% lead carbonic* and 11.95% XBS lead paint groups, tetpec lively. Th*_ femur* of rat* fed the control diet, lead octoate or lesd_ chronet* axnibite^Wry^c^e^eao at 4 weans. "Itowever. ;ne feaurs of rets fc_o..'4^t_ligj_Egrasnate or 1L^&2%-115S--l*Ad_aiint contained 13.3 and '.-.i -S of laad/gm of bene, reapactivsly. i$a$1 Vi SF fnfclAjg f-S&Sfcs pfej-V. t-;i- 36 0007--SWP-037010 0007-SWP-000115578 TABLE 14 _ u r in al s sis or r a t s ~ta pain t say c o n t ain in g ?, i-iz./a-pJ Treatment '.'c*k 4 1 i2 Protein: Negative 7 6 20 * 100 Dig 1 l2 4 __________ > 100 mg_T.__________ .Microscopic Examination H3Ci/: Normal 7 5 23 Moderate l2 1 _____ _ _ E*isiv,4 _ _ 1 HiCi': Normal *7 Moderate Excessive EpicneliauE': Normal 22 7 a a 4 Moderate _ Excessive Crya ta1*;: Norma1 4 8 23 Moderate 2 * Casts: Negative _ oiirive __ J 8 8 24 Nuaors indicate number of rats at response level. 1/ Normal, 10 oe 1(* cell:.; raoCerjtt, 10*100 cells; excess.ve, > 100 ceila/fleld (x 440). y Normal, 5 or less cells; moderate, J-23 ceils; axes live, > 23 cells/tield (x 100). ct Normal, none; moderate, 1*5 crystals; excessive, > 5 crystals/field (x 100). 37 V *.<* 0007-SWP-037011 0007-SWP-000115579 :a s u :s eW4 L'SISAIYSIS OF RATS PS3 PAINT WW CONTAIN! NS o.oas l e a d a s l e a p s c t o a t e Treicmtnc 'ck i ^3 Protein: Negative 4 < 100 0| 1 > 100 ist 1 Microacopic Examination UCi/; Noraal 4 Moderate ;_________ 3*S.***ivS___________ _ WBCi'; Noraal 3 Moderate 2 38 3 10 12 3 ""To" 12 ----------------s%*Lm - -_________ Epienaliuml': Noraal " - -\" " " " 12" Moderate ------------------- ----------________________ Cryical*': Noraal Modaraca 2 2. 4 1 Cues: Negative 4 4 12 Nuabart indicate nusbar of rata at reaponi* a/ Noraal, 10 or lata calla; eedarata, 10*100 call*; axeaativa, > 100 eellt/field (x 440). b/ Noraal, 5 or laat calla; moderate. $*25 call*; excaasiva, > 25 calla/flald (x 100). zi Notnal, non*; moderate, 1-5 cryatalt; txcaiilva, > 5 etyatala/field (x 100). rV. .. *jv 1 --. M:r 3S " 0007-SWP--037012 0007-SWP-000115580 'ABU 26 i'k ik a iv s is or r a t s f e d p a in t c o n t ain in g 0.53% ISAfl AS tA8 OCTPaTS Proto;::: Nesative < 100 n| TM __________ > U> r.:_%_ Microscopic Examination RBCi^: Normal Moderate _______ ______________ WBCi': Normal Moderate __________ Exeats Wa _ _ _, Epit.ieliuoi': Normal Moderate _________ _Excsivsi Crystals!/: Normal Moderate .Excessive_ _ Caaca: Nafaclva ___ _ Posieive_ _______ Treatment Week. 4 8............ 13, 43 l ___ __ _ _ 10 1 _1,, 2 2 10 1 21 1___________________ 1. *19 33 4 12 34 1 12 4 4 12 Number* indicate nunbar o rata ac response laval. a/ Normal, 10 or laaa eclli; aodoraca, 10-100 calls; axcaasiva, > 100 cells/field (x 440). if Normal, 5 or lass calls; moderate, 5-23 eells; cxcaaelve, > 25 cells/field (x 100). tl Normal, none; moderate, 1*5 crystals; excessive, > 5 crystals//icld (x 100). 39 .1*1 > VJ`* *7 ; * ' w_.- -iOT W .'s*T. ,-TH U* \. *_,v--m *J i#a* ]* -'..>*. * " ya>s *^..*""'"'.'t* jC. ili* 0007-SWP-037013 0007-SWP-000115581 TABU 27 URINALYSIS OF RATS TEO '?A1jgt~ WiAl CONTAINING 2.05II LEAD AS OCTvdiTE Protein: .VfcpaClVf < 100 mg u > 100 t -if H Microscopic Examination RBCi-': Normal Modernla Excassiva UBCS' : Normal Modarata _ Exeaisiva Epitasliuaii': Normal Modarata Excjssiva Crystals^': Normal Modarata __ ... Cases: Najjativa Positive Tre*crc*nc >*k 2 ^2 44 11 1 4 <* 12 33 1 7 4u _ 24 * 4* 8 4 1+ 12 12 NunBtr* indicat* numbar of rat* at rasponsa laval, / Normal, 10 or lass call*; nodarac*. 10*100 culls; exetssiva. > 100 ealU/fisld (x 440). b/ Normal, 3 or lass calls; modarata, 3-23 calls; xcusaiva, > 25 cU*mcld (x 100). c/ Normal, non*; wdarata, 1-5 crystals; axcastlva, > 5 erystalx/iiald {* 100). . : .'-.TV ~f. 40 0007-SWP--037014 0007-SWP-000115582 TABLE 2B TRJSALYSIS OF RATS PtD PAINT e,V)' --r*' CONTAIN>**> 0.42', LEAD AS LIAO CHRO'.-.TI Protein: Negative < 100 mg % > 100 ra T. Microscopic Examination HBCj> : Normal Nodtrees Excessive WBCi': Normal Moderate Excessive Epitneliutai': Normal Moderate ___ _ _ Excessive crystals^'; Noma 1 Moderate Excessive Casts: Negative Positive Treatment Week 48 4 2 :o 2l % 31 13 8 3 8 23 8 21 1 3 1* 4 12 2 4 12 1 4 4 12 Numbers indicate number of rats at response ltvtl. / Norms 1, 10 or less cslli; moderate, 10-100 etUi txcuuvt, > 100 eslli/field (x *40). b/ Normal, S cr lest calls; moderate,. J-2i eslls; excessive, > 25 cells/field (x 100). e< Normal, none; moderate, 1-5 crystals; excessive, > 5 erystals/field (x 100). h K 41 0007--SWP-037015 0007-SWP-000115583 TABLE T9 o (klP* URINALYSIS OF RATS FE3 PAINT "W CC.'.TA l.?sr. LEAP AS LEAP CHROMATE Protein: Negative < 100 mg % i2 2S-\__________ Microscopic Examination MCI': Normal Moderate WBCi': Normal Modsrsee Epithelium!': Normal Moderate _________________Exejsiive Crystals!/; Normal Moderate _Ecjaive_ _ _ Casta: Negative Positive Treaenene Week kS & i 3 3 1 ' 11 1 4 3 9 *9 l 23 2 _____ 1 _ 44 B 3 9 *fc 4 4 12 4 4 12 Numbers indicate number of race at response level. J Normal, 10 or lea* cells; nodersce, 10-100 cells; excessive, > 100 c<.lls/fieid (x 440). b/ Normal, 5 or less cells; noderete, 5>25 cells; excessive, > 25 cells/fielo (x 100). e/ Normal, none; eoderste, 1-5 crystals; excessive, > 5 crystals/field (x 100). ... , '* 9 * * ... * ' * ;.r . * v; i i >, . -'.'-V; -.-..a T *7 .2 tr*. .v-.Wk;;- w-V*<U<r v OO 07-SWP-037016 0007-SWP-000115584 TABU 30 ckiir1 I'RLNALYS IS OF RATS FE2 PAINT r*U C2: TAININC 12.43'. LIAO AS LEAD CHROMATE Treatment week i H ^3 Protein: Negative < 100 mg 2, i1 32 9 3 Microscopic Examination RBCi'1: Normal Moderate Excessive WBCi': Normal Moderate 4 1 3 Epicneiiums' : Normal 4 Moderate _________________ Excessive Crystals^: Normal Moderate Excessive Casts: Negative 2 2 4 -- -- -- -- -- Z0!.1!---B_ ^ __ -- __ W M 1 3 1 1 2 4 4 4 m s *H-- 11 1 1 11 1 12 12 mm erne Numbers indicate number of rats at response level. J Normal, 10 or less cells; modnrate, 10*100 cells; excessive, > 100 cells/field (x 410). b! Norms 1, 3 or less cells; moderate, 5*25 cells; excessive, > 23 cells/field (x 100). i! Normal, none; moderate, 1-3 crystals; excessive, > 3 cryseals/fiuld (x 100). r.iar.* S5 ** 43 :v- j 0007-SBP-037017 0007-SWP-000115585 ?A2L 31 ~ VRIN-Airsis cr .h at s F-3 p a in t ** ............................... --.o .- MXi c ax ;j :;a~s Protein: Negative < 100 e.t 1 Microscopic Examination RiC/: Normal Moderate *BCi': Normal Moderate Excessive Epienaliuat-': Normal Moderate ________ _Exeassiva Crystals^': Normal Moderate Casts: Negative Positive Trotffanc l'ocx a* 13 3 i: 14 1 42 2 2 22 a 4 u * 10 2 12 34 12 4 4 12 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!, nona; moderate, i-5 crystals; excessive, > S erysiais/field lx 100). Sa. '* ** I----- -T k>~? S^SK t ?-"- a. 0007-SMP-037018 0007-SWP-000115586 t a 3:2 :: -r in a iy s is or s at s no ?,u:~ -inti wtjy.?s -a frostsent V -4 12 Protein. Ntgaeiva < 100 nB r. 2 2 > *.00 ag *. Microscopic examination RBCi'; Normal Modaraca 2 2 im______ wad-; Normal 2 Modaraca Ixeass^v* Spittle! uni': Normal * Modaraca _ _ _ .15*4*11**. -- -- -- _ _ _ Crystal si' : Norms 1 4 ModarJta Excessive Cases: Negative * Positive 3 4 1 4 4 4 5 12 11 *1 12 _---- .2 12 Nye^cri incicat* nurbar of rats at response level, a/ Sorisal, 10 or laaa call*; moderate, 10-100 cell*; excessive, > 100 eallsffield (k 440). b/ Normal, 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). h`M 1.-T5 itr `v1* ai.. :&r:P 0007-SWP-0370I9 0007-SWP-000115587 lASLi 33 LEAD CrXTEN" :c d Oirt Control"^ 0.03% Lead Cctosce 0.53% lead Octoace 2.05% Lead Oecoaca 0.42% Lead Chromace 1.95% Lead Chromat 12.43% Lead Chromate 66.05% Lead Carbonate 11.92% KSS Lead Paint ' Tt h fon c Ve<t Si' d(3)-' 13.1 = 0.9 (16)*/ 12.3 5 :.s (3) -6*< z 2.* (?>!' 14.5 - l.S <8) 13.2 : 1.0 (S) 12.0 : 1.4 (8) I 19.4 - 2.2 (3)*' I 24.9 = 2.2 ( ' ^23.9 = 1.3 ciiiiii/ a/ Values deleted due to coneacinacion. b/ Four valuta delated dua to contamination. e/ Number of rat blood* analyzed per group unit** indicated otherwise. it Sixteen blood samples were analyzed. &t Average ug of l**d/100 ml blood - standard trror of number of rat* in parenthesis. it On* blood sample lost. &/ Significantly different from control CP < O.CJ) as shear. by Ou.nnett'$ suitiple-cenparison test following sn analysts sf variance. 2/ Tualva blood samples analyzed. %* / v*t * * .v \.j : -J * ` '?:' rsSJ . - SOS'*.' ...H5P3C *,'... 46 >7.702: ir *r J Ijfi 0007-SWP-000115588 IP- |n In' Ik 3:at Control-^ 0.08Z Load Oetoata 0.33S Laad Oetoata 2.031 Laad Oetoata 0.42J Laad Chroataca 1.95X Laad Chroaata 12.A3* Laad Chronica 66.031 Laad Carbonata 11.92Z NBS Laad Paine :a b l s -i LEAD CONTENT O? liV.TN a* (I)-1' n.d.-^ n.d. n.d. n.d. n.d. n.d. n.d. a.d. n.d. Tr.atr-.nt "en 3 W n.d. n.d. n.d. n.d. n.d. n.d. a.d. n.d. n.d. ii u:> n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. 0.2:0 / Nuooir of braina analyzed par group ualaaa indicated ochezviae. / Eight braina vara analyzad at A and t vaeka. Twanty-four braina wara analyzad at 13 waaki. / Not datactable. Below aenaltlvity of 0.07 ug/g of brain. / Avaraga ug of lead/ga brain - acindard arror nuabar of rata shown in oaranthaiia. N T T i.'i-L'i.i.-.' t*d *J.^aT Jb^bisar S'**.'? i >w** -: ' 0007-SWP-Q37023. 0007-SWP-000115589 rue Control^ 0.08% Lead Oeteaea 0.33% Lead Octoata 2.03% Laad Oecoaea 0.42% Laad Chromate 1.95% Laad Chromate 12.43% Laad Chronaca 66.05% Laad Carbonaea 11.92% XSS Laad fame (i)i' n.d.-S' n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. Treatment Weak ;-) 11 CD n.d. n.d. r..d. r..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_S.0.6/ C) 0.4 s 0.1 (9) a/ Number o liven analyzed par group unlm indicated ptharviae. b/ Eight livarawara analyzad ae 4 and B vaaka. Tvanty-four liverz vara inalyzad ac 13 week*. / Noe detectable. Balov aanaitivity of 0.07 ug/ga Uvtr. / valuet art avtraga ug of lood/gn Uvtr s standard arror of number of rata snovn in parentheai*. ... V .. r^rrr^rv .a 0007--SWP-037022 0007-SWP-000115590 ZABLE 36 LEAD CCNTIST Of VlONSV JJ* j J Control-^ 0.087. Lead Occoats 0.53% Lead Octoat* 2.03% Lead Octoate 0.42V Lead Chromate 1.93% Lead Chromace 12-43% Lead Chromate 64.03% Lead Carbonate 11.92% XBS Lead faint (I)i' n.d.^ n.d. n.d. n.de n.d. n.d. n.d. n.d. n.d. - it o Trcatnenc '->< a (-) n.d. n.d. n.d. n.d. n.d. n.d* n.d. n.d. r..d. :.3l C) 0.31 (1) 0.** -0.1 C) '*.5i (i) 0.4 - 0.0 (3) 2.3 - 0.2 (8) 1^ = 0.1 U2) /* xx*. && ma: a/ Number of kidneys analyzed par group unlaaa otherwise indicated. Eight kidney* vara analyzed at 4 and 8 veeke. Twenty-four kidney* were analyzed at 13 weak*. c7 Not deteetablt. Below tencitivicy of 0.07 ug/gn kidney. d/y Values ar* ug cf lead/gn kidney or average ug of lead/gs kidney : standard error of number of'rats in parenthesis. -I . iV:: N. p* h""iX r\ i:. - & * .. V" - ^r--ov 0007-SWP-037023 0007-SWP-000115591 s - ?; TABLE 37 l e a s c c n t ;:.t o f bo n e 'rs^.-sn W*' rraatsarc Vttk i (i> L-.LfiV >V: Control^ o.oas Load Oetoata 0.53i Laad Oetoata 2.05. Laad Oetoata 0.L2S Laad Chrooata 1.952 Laad Chroaata 12.43" Laad Chroaata 66.05?. Laad Carbonaca SBS Laad ?air.t 3.9 i 0.6 ce) 6.2 - 0.6 W 3.9 s 0.2 (6) 3.0 s 0.7 (4) 2.6 r 0.9 (4} 1-8 r 0.2 (4) E 8y3..ir 1.1 6.0 2.1 l-4 s 0.4 wif 3.2 = 0.2 (24) 3.3 = 0.3 (12) 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 (10)^ 21.7 (12)^ 10.9 r C.9 C12)/ a; .'.ueear si bonaa analyzad par group unlati indieatad ocharvi.it. b/ Sight bonaa wara analyzad tc 6 and 8 uaaka. ?uanty>four bonaa wara analyzad at 13 uaaka. cl Valuaa ara ug of laad/ga bona or avaraga ug at laad/gn bona - atandard arror of nuabar of rata in paranchazia. i! Net datactabla. Balow aanaicivitv of 0.4 ug/ga of bona, a ^ Significantly difftranc iron control (9 < 0.5) aa ahown by Cunnatt'a aultipit ccrparuon caat following an anajyaia of varianct. If* a*. *aA . r*. .. t -BW X S3 0007--SWP-037024 0007-SWP-000115592 At 8 weeks, the femurs ef rat* id tne control, lead oetoatc or lead chromate diets contained similar quantities of lead, ranging from l.4.2 ug lesd/gs bone. Leadcrr^n^nih^femu^^nf^jeJi^JJJft^Iead^aroo^^e 3otn*:cr.it;cart.y corve ujr.trbl. 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 m tne control group. The-6.6.03* lug carbonate group exploited nigner .evtisotieTPin their femurs i21.7 ug ieia.ga eons; tsar, at 8 weeks. ThallJJ2jgJ<BS lead paint group had less lead in their femurs (10,9 ug lead/ga bona> man at 6 weeks. Ihj_laid_ly^ls in both of these groups wore s--WsignificantW irsater than choss obsaryea tn tnt c55tryr*TnTT {Mr1 1. Pstholotv of Rats Fed Paint P*lm Containing different Concentrations sLiss* The relative organ weights of the thyroid, spleen, heart, kidneys, Uver, adrenals, brain and gonada are shewn in Table 38. The 3unnect`s siultiple-conpsnson test showed that none of the relative organ weights dif fered from control veluee. 0 Ihe pathology of rats fad the control, 2.032 lead occoata, 12.432 lead chromate, 66.OK lead carbonate or 11.922 BBS lead paint diets for 13 weeks is shown in Tables 39-43.^Tj^e control rats had a few naturally occurring lesions. Fourteen of these ra1 this! mild to moderate lymphoid hyperplasia m the lungs, characteristic of early minute p^ntumonia. One of these rats had a coderate pneumonia and another one a miId esphystms., ^4Other lesions included en unspecific myocardieds in no rats; foci ef subacute inflacaetion in the liver of two rata; a cross-see cion of a parasite (roundworm) in the colon ef ese rat; and foci of mononuclear cell infiltration in the kidneys ef two rats. Tha bona marrnr myeloid/crytnroid tell ratios (M/I) of all rata war* within normal limits. Tha lesions observed in rats fed the 2.3iT lead oetoate diet were, similar so th* lesions assn in the eantrol rite. Seven out of the 12 racsir*'*"hae slid to eoderata lymphoid hvserplaaia in she lungs and one had mild pneumonia. Four rats had mild a^E^bf subacute infiarmatipn in tne liver characterlied by e leas of aavarsl hepatic cord calls which wars replaced by aecropoagea and lymphocytes. Another rat bed a foea^&f inflance cion in the interstitial tissue of tha panereas. Tha K/E ratios of tntst rsts were normal. <1 0007-SWP-037025 0007-SWP-000115593 jiilM f i_t a W lin o ta w ia 'S * |i.irJ m SB# 0 ** $5 E *. - fiwas- g& WK&: sifijiaiir "rji*** ' r?r^tiT*.' c-fv-: lilM TTSiinil ijM -iiiam ii&i Vm A il ilii j iiiriim mv imi fc lm ij & 2 0007--SWP--037026 0007-SWP-000115594 0007-SWP-037027 0007-SWP-000115595 0007-SWP-000115596 <m 5 KS *51 is 1to * 8 :i o *-8 ** = w X o *i a> *h U M> MN4a1 (OA ? *1 Y>V;V7 v\ i '.Va-,`n* ^ *?i2i SK H5?2& ass - *w.:i.r ;**. * ^SgC rvi 0007-SWP-000115597 0007-SWP-037030 0007-SWP-000115598 0007-SWP--O37031 0007-SWP-000115599 S' aa leeie tha ^iSOLJUSSL^L^S^SSaiS^ilii- s,v*n of U rscsi rvd eilc to rod tract 4 ilymphoid hyperplasia, and one had mild pneumonia. Onl ' one rat had a moderate unspecific myocarditis. The M/E ratios wars normal. ThtUsionjii|jttn-inehsraHj|4l>thall;92Ri2SMis*dll>oaintdiec wars 2tSfilSSLJESSSi Bight of 12 rata had different degrees of lymphoid hyperplasia in the lungs. In one rat ^ pneumonia was evident. Cnspecific myocarditis occurred in 2 rats while foei of Inflammation in tha liver war* observed in only ena rac. The M/E ratios ware normal. ,\ **/ In conclusion^ tha pathol o|y_sean_in tha control^and^traatsdrats wars characteristic! of these in a notaaT'ooDuTstigir'oilTtr^, No evidence of lcid-injucod retton^^BtouHr"*"**'*"^^^^^ iv. yagyiam The levale of load fad to rata in the itudy ware sufficient to pro duce significant elevation of body lead burdan if tha lead ia in tha form of load nittata.il'' if va equate the amount of lead eonawed by tha average rac fad the 2.051 Laad 'oc coats, 12.431 laad'chroawce, 66.051 lead eerboneta, and 11.921 SIS laad paint diaea to a 14.6 kg (3-year old) child, wa find that tha laad conaumpcion would equal 28.6, 123.7, 503.0 and 113.9 mg of lesd/dey, respectively. On the basis of 21 laad peine, the total gaint intaka would j ? be 1.5, 6.2, 25.0, and 5.7 ga/day. Daily Ingestion of thaaa quantities of / old paint have baan reported to produce laad poitoning.il/ Thera wart no changes In hamatology, urine lysis or serum procaine throughout the entire study. However, chare was one effect on porphyrin metabolism. As early as 4 weeks, rata fad either tha 66.05% lead carbonate diet or tha 11.92% K1S laad paint diet exhibited 501 depression of erythrocyte A1AD activity. This affect appeared to ba directly related to the concentra tion of blood laad as has baan reported by othera.iiuii/ However, chars was one exception, blood laad la the 12.431 laad chromate group became elevated by tne 13th week, but there was no depression of AUD activity. This dis crepancy could ba related to tha duration of alavatad blood lead or tne age of tne animal when blood lead becomes elevated. Evan though erythrocyte ALAD was depressed 501, the overall effect on porphyrin metabolism muse be considered mild, since there was no concoainant change in the levels of protoporphyria (erythrocyte), delta-aminolevulinic acid and coproperphyrin. These observations are consistent with reports that blood load levels greater than 40 ug/100 ml UC are needed before serious charges in porphyrin metabolism oecur.l/ 58 hdS>'4. r'f&V %-'*, Vi S'-.'-.ll v'^la.V QMf* V 0007-SWP-037032 0007-SWP-0001 The bod* Usd butdsn of ract--fcd-saist--sggtaigjns -*sd occosce ar lead chromatewere ne_differtr.t err- coccrsl ujctt-l ' * .. *. At this sire lead was found in the kidney* of one to three rota on eaen of the Usd accosts ind lead chromate diet*, and the blood lead levels ,,r. tn '.:.A32 lead chromate group were significantly eltvaced. Since rats fed Usd nitrate show elevated lead levels in blood, Kidneys, bone and liver within 2 weeks,.12/ ij^aooears fr that the lead octaete end leed chromate wege_nag_cithgs_rgaiIv_abosgtd_gr avsi.aoi* tor sosorption. ..... Pathology wae performed only on the rats chit were fed the eontrol, 2.05* Usd oceoete, 12.432 lead chromate, and 11.922 XHS lead paint diets for 13 weeks. All of these rats wera relatively free of lesions, and those lesions which were observed, oecur naturally in ret colonies. v. cojEisisji?bv^ atijtjjdvjhowsjhe^UMfedO^J^^iCjjESiSS^JJJii-SfJiiJSiScontaining 11.922 Usd and^maw'psincconttlnlng 66.J)j2 >*<L as*Tsc carPonaea txjuil t*o aarly signs of laad poiionin|. Ihasa signs conaistad of trythrocyta ALAS depression (502) and significant increases in blood, kldnsy, bone, liver and/or brain lead, hats fd_g_ diet consisting of 0.12 new paint _gilg_contalninf 12.132 _1 sad ei lead chrawate exhiPttsp only elivsTadblooPisajL-tltarTT"" weeks, of leading. A nailsr_aiet_gx_ new paint"cont*Tn?ng'7TtSSr leed at isT~ occosts produced no signs of lead poisoning. The lack of any marksd changt m the body leed 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 Usd in ssveral forms, ona must conclude chat the lead in the paint used in this enperisient was r.ot readily available for absorption. In conclusion. th results reported nersm support tbs contention thdt lead is not rsadilv spsoreed from cna newer osints. ......... 59 r rr.v.rw-PT/"" ' -`"A.V. ' 0007-SWP-037033 0007-SWP-000115601 o REFERENCES 1. Kchoe, R. A., "The Xatobolism of Lead m Man in Health and Disease," Laccurs II, J. Rov. lose, Public Haalch Hvg. . 24:101 (1961). 2. Chisholm, J. J.. fed. Clin. Sorch Alter.. 17:591 (19*0). 3. King. 8. G.. Amur. J. Pis. Child.. 122:337 (1971). 4. Selagson. D., Standard Methods of Clinical Chemistry. Academic Press, Inc., Sew Yorls, Vol. 2, p. 52 (1958). 5. Brother, G., M. Schneiderman, and C. Z. William, Am. J. Clin. Path.. 26.: 1639 (1956). 6. Brasher, S., and H. Schneiderman, Ami__^_Ciin_1_Fj^hi, 20:1079 (1950). 7. Faulkner, W. R., and J. W. King, Ede., .Manual of Clinical Laboratory Procedures. 2nd Ed., p. 178 (1970). 6. Liehtman, H. C., and F. Feldman, J. Clin. Invest.. 4:830 (1963). 0 9. Heller, 5. R., R. F. Labbe, and J. Sutter, Clin. Cham.. 7:525 (1971). 10. Pavia, F. R., and S. 1. Andelman, Arch. Envir. Health. 13:53 (1967). 11. Xauzerall, P,, and S. Cranick, J. Biol. Cham.. 219:435 (1956). 12. Schlenker, T. S-, and C. L. Kicehell. Am. J. Clin. Pathcl.. 29:593 (1958). 13. Gate, J. C., and K. H. Litchfield, J. Oil. Col. Cham. Assoc.. 5:236 (1969). 14. Goldwaear, L. J., Induct. Med.. 41:13 (1972). 15. Killar, 3. A., V. Bettis tint, R. 1- C. Cuaaaing, F. Caravail, and A. Goldberg 4--at. 3_:695, October 1970. /Un+' . r 60 0007-SWP--037034 0007-SWP-000115602 Older paint fomul ation* contained soluble lend salts. Two samples of paint chips representative or these formulations were fed to rats at 0.1% of theiT diets. One was supplied by the National Bureau of Standards and was removed from interior walls o: alder hares. It contained 11.92% lead. The other was a formulation csr.tair.in; 66.0S'. lead carbonate. Both of these samples produced classical signs of lead poisoning in the rats. These signs consisted of erythrocyte ALAD 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. The only evidence ef toxicity or effect on body burden appear in the Tats fed the paint containing lead chromate. This consisted of an increase in blood lead, withouc ether sips of poisoning. Paint containing 1.95% lead ehrcaata and pair.t containing 2.05'. lead octoate produced no detectable changes in the rats. 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 lsck'of toxieity with paints containing 2.0% lead chromate or lead octoate undoubtedly it due to the lack of sipiflcant absorption from the gastrointestinal-tract. 0007-SWP--037035 4 r \ I1 4 5 . mo d if t c v t io v c"rrscvssro* The levels of lead fed to rats in the study were sufficient to pro* duce significance'evacion of body lead burden if the lead is in tr.e fern of lead mtrate.ii/ If we equate the amount of lead consumed oy the average rat fed paint chips containing 3.05% lead ectoate, 13.45* lead chromate, 66.05% lead carbonate, and 11.93% MBS lead name diets to a 14.6 kg (3-year old) child, we find that the lead esnsuration 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 5.3 gm/day^tssgi Daily ingestion of these Quantities of old paint.have been reported to produce lead poisoning.il' A $$:{In c -v ^'C' There were no changes in hematology, urinalysis or serum proteins throughout the entire study. Hqapyy, porphyrin metabolism was defective w as early as 4 weeks in rats fed either the 66.05% lead carbonate diet or the 11.92% X8S lead paint diet as evidenced in a 50% depression of erythrocyte ALAD activity. This effeet appeared te be directly related to the coneentra* tion of blood lead as has been reported by others.liiil' ii<--ri there was ona exception; blood lead in the 12.45% lead ehrqgate group became elevated by the 13th week, but there was no depression of ALAD activity. This dis* crepancy could be related to the duration of elevated blood lead or the age of the animal when blood lead beeasw elevated....... ,tV*i _.l? r *{-* Even though erythrocyte ALAD was depressed 50%, the overall effect on porphyrin metabolism oust be considered gild, since there was no concomitant change in the levels of protoporphyrin (erythrocyte), delta-Mtmoievulir.ie acid and ccpropoiphyrin. These observations are consistent with reports that blood lead levels greater than 40 ug/100 al RBC are needed before semi* us changes in porphyrin metabolism occur.ii/ I-.. ` . The body lead burden of rats fed paint containing lead octoate or lead chroaatt SSJTno different from control until 13 weeks, a; this time lead was found in the kidneys of one to three rats on each of the lead octoate and lead chromate dicta, and the bleed lead levels in the 12.45% lead ehrsrate group were significantly elevated. Since rats fed lead nitrate show elevatcetdoj lead levels in blood, kidneys, bone and liver within 2 veus.li.' it appekatrsi yF that t*h'e letoarnd* VoWctloiVaBtke* and laad chromate were ms cithtr'rtadily absorbed\ orV* 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 chromate, and 11.92% N8S 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 eolonies. <4 *+*C*' sr - , .....-* S' ",*..>....*....*.... ... **.f. %i ,, W% 1* % >-:.- sir*** tsj ***4*i*tr~* 4* ,..i . ; 1 I 0007--SWP-037036 PREFACE This resort was prepared at Midwest Research Institute, 425 Volker Boulevard, Komos City, Missouri 64110 under Contract No. 62-W-62GC & NFC, MRl Project No. 3729-B, "Lead Paint Ingestion Study.* The Research was sponsored by the National Point ond Coatings Association, 1500 Rhode Island Avenue, N, W., Washington, D, C. 20005. Royal A. Brown, Teehnieoi Director, Notional 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 Sanyar, Associate Pathologist, and Mrs. Jane Hoch, BToIbgy Research Assistant. Dr. James L. Spigorelli, Senior Chemist supervised the lead analysis with the etiistonee of Mrs. Hope M. Millar, Assistant Chemist. Members of the National Paint and Coatings Association Industrial Metals Task Fores prepared the paint chips aid consulted with Dri>Cattla, Sanya, Spigorelli and House during the course of the study. The personnel of the NPCA Industrial Metals Task Force is os follows; Richard A. Moore Royol A. Brown Dr. John P.Frowley Charles M. Joekson Joseph G. Kingston S*. Sidney Lauren William W. Ringle Edwin . Swain Jean P. Teas Domcnic J. Tenor! The Sherwin Williams Company Chairman The National Paint & Coatings Association Hercules, Inc. Celanese Coatings Co. GISddenDurkoe Division of SCM Corporation Coatings Research Group, Ine. Pratt and Lambert, Inc. E. I. duPontde Nemours & Co., Inc. The Flood Company De Soto, Inc. Aooroved for; MIDWEST RESEARCH INSTITUTE W. B. House, Director Biological Sciences Division 14 September, 1973 ii'dr * . .L .. pss X:-'' 0007-SWP-037037 n 'I SLIGHT MODIFICATION OF CASTES' COSCLVSIOS'S This study shows that Tats fad 0.1\ diet (paint/feed) of old paint containing 32.92% lead and new paint containing 66.05% lead as lead earoonate 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. Kata fed a diet con* sisting of 0.2% 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 octoate produced no signs of lead poisoning. N The lack of any narked change in the body load burden in these leed chronate and lead octoate rat* fad paint containing ehronato and octoate suggests that very little lead was absorbed iron the gastro intestinal trset. Since there is considerable evidence that the rat can absorb lead in several forms, One oust conclude that the lead in the om of lead chromate and land octoate and incorporated into paint was not readily available for absorption. In conclusion, the results resorted herein sunoort the contention that lead is not readily -worced rrc- "ne newer ;a:r.:s. ...... " ................. . *> :*V /Ktfr 2*V', s' uv$r 1*?*i."K'SiZ" % * i .Sr?**? 0007-SWP-000115606