Document EdN3d9KDY33VKd4Ywg2bvmj44

I THE EFFECT OF PROLONGED FEEDING OF LOW LEVELS OF LFftD TO LABORATORY KiWM S He n r y J, Tr o c h imo w icz. Sc,D,, Jo h n R. Ba r n e s , Ph ,D,, He n r y Sh e r ma n , Ph .D,, Ed w in F, St u l a , D.V.M,, a n d No r ma n W, He n r y , III Pr e s e n t e d b y J,He n r y Tr o c h imo w icz AT THE Ame r ic a n In d u s t r ia l Hy g ie n e As s o c ia t io n Me e t in g Sa n Fr a n c is c o , Ca l if o r n ia Ma y 16-19, 1972 N40755 I. INTRODUCTION Lit t l e e x p e r ime n t a l w o r k h a s b e e n p o n e o n t h e BIOLOGICAL EFFECTS OF LONG-TERM EXPOSURE TO LOW CONCENTRATIONS OF LEAD, RECENTLY/ A COMPREHENSIVE STUDY ON THE CHRONIC TOXICITY OF ORALLY-ADMINISTEREO LEAD IN LABORATORY ANIMALS WAS RELEASED BY THE 1971)Ame r ic a n Pe t r o l e u m In s t it u t e (Ap r il / , Th is s t u d y SHOWED THAT THE ADMINISTRATION OF LEAD ACETATE TO RATS/ DOGS/ AND MONKEYS FOR 22 MONTHS AT A DIETARY LEVEL OF 10 PPM AS LEAD PRODUCED NO EFFECTS THAT COULD BE DETECTED BY ANY OF THE TECHNIQUES THAT WERE EMPLOYED; 50 PPM PRODUCED MINIMAL EQUIVOCAL EFFECTS/ AND 100 PPM PRODUCED HISTOLOGIC CHANGE WITHOUT FUNCTIONAL CHANGE IN THE KIDNEYS OF RATS, THE PRESENT STUDY WAS UNDERTAKEN TO FURTHER CLARIFY THE POSSIBLE BIOLOGIC MANIFESTATIONS OF LONG-TERM/ LOW-LEVEL/ DIETARY ADMINISTRATION OF LEAD IN LABORATORY ANIMALS, II. METHODOLOGY A, Ex p e r ime n t a l De s ig n (SLIDE #1) Th e b a s ic e x p e r ime n t a l d e s ig n c o n s is t e d o f t w o Is t u d ie s . n St u d y l, r a t s a n d d o g s w e r e f e d d ie t s o f . Gr o u n d Pu r in a La b Ch o w t o w h ic h w a s a d d ed 0/ 10/ 50/ 100/ PAGE TWO AND 500 PPM LEAD ADMINISTERED AS LEAD ACETATE Pb (C H 0 ) 2322 3H 0 , Th e a mo u n t o f l e a d a d d e d t o t h e 2 DIET WAS CALCULATED ON THE BASIS OF ACTUAL LEAD, THIS BEING 54,6% OF THE WEIGHT OF THE ACETATE. Bo t h ma l e s a n d f e ma l e s o f e a c h s p e c ie ? w e r e u s e d , Th e v a r io u s b io l o g ic in d ic e s u n d e r o b s e r v a t io n w e r e MEASURED AT 10 DIFFERENT INTERVALS THROUGHOUT THE TWO YEARS OF FEEDING. (SLIDE #2) IAf t e r St u d y h a d b e e n in p r o g r e s s f o r s e v e r a l mo n t h s , IT APPEARED THAT A DIETARY LEVEL OF 500 PPM LEAD WAS NOT AFFECTING THE CLINICAL, NUTRITIONAL, OR REPRODUCTIVE INDICES OF RATS, THEREFORE, A SECOND TWO-YEAR FEEDING STUDY WAS INITIATED IN RATS TO PROVIDE DIETARY LEAD LEVELS OF 1000 AND 2000 PPM, B, Pa r a me t e r s Un d e r Ob s e r v a t io n Th e n e x t t h r e e s l id e s w il l s h o w t h e b io l o g ic r e s p o n s e s WHICH WERE OBSERVED OR MEASURED PERIODICALLY THROUGHOUT THE TWO YEARS OF DIETARY LEAD ADMINISTRATION, DUP050058635 PAGE THREE (SLIDE #3) 1, Cl in ic a l Ob s e r v a t io n s Th e a n ima l s w e r e o b s e r v e d f o r v a r io u s g r o s s a s p e c t s OF APPEARANCE AND BEHAVIOR, DOGS WERE GIVEN A NEURO LOGICAL EXAMINATION INITIALLY AND AT ONE AND TWO YEARS OF f e e d in g , We ig h t g a in , f o o d c o n s u mp t io n , a n d in c id e n c e o f MORTALITY WERE ALSO RECORDED FOR ALL TEST GROUPS, 2. Le a d An a l y s e s Bl o o d , u r in e , f e c e s , a n d v a r io u s t is s u e s w e r e . .. ... ... .. _ PERIODICALLY ANALYZED FOR LEAD IN ALL DOGS AND IN FOUR TO " SIX RATS SELECTED AT RANDOM AT EACH DIETARY LEAD LEVEL, Al s o , t e s t d ie t s f o r a l l g r o u p s w e r e a n a l y z e d p e r io d ic a l l y FOR THEIR ACTUAL LEAD CONTENT, 3, - He ma t o l o g y (s l id e my He ma t o l o g ic e x a min a t io n o f b o t h r a t s a n d d o g s in c l u d e d A COMPLETE BLOOD COUNT, HEMATOCRIT, HEMOGLOBIN, AND NUMBER OF BASOPHILIC STIPPLED CELLS, 4. Bio c h e mis t r y Bio c h e mic a l a n a l y s e s in v o l v e d a n e v a l u a t io n o f l iv e r FUNCTION, A COMPLETE URINALYSIS, AND SPECIAL TESTS TO DUP050058636 PAGE FOUR DETERMINE DELTA-AMINOLEVULINIC ACID (DALA) IN URINE AND DELTA-AMINOLEVULINIC ACID DEHYDRASE <ALAD) IN BLOOD IN BOTH RATS AND' DOGS, (SLIDE m 5. Pa t h o l o g y Al l a n ima l s t h a t w e r e s a c r if ic e d / d ie d , o r s u r v iv e d TWO YEARS OF DIETARY LEAD RECEIVED A THOROUGH NECROPSY, ALL MAJOR ORGAN SYSTEMS WERE EXAMINED/ BOTH GROSSLY AND MICROSCOPICALLY, 6. Th r e e -Ge n e r a t io n Re p r o d u c t io n St u d y in Ra t s A THREE-GENERATION/ SIX-LITTER REPRODUCTION STUDY WAS CARRIED OUT ON RATS RECEIVING 0/ 10/ 50/ 100/ 1000/ AND 2000 PPM (P )DIETARY LEAD b , CRITERIA EVALUATED INCLUDED VARIOUS INDICES OF REPRODUCTION AND LACTATION/ THE AVERAGE BODY WEIGHT OF WEANLINGS/ AND A HISTOLOGICAL EVALUATION OF 21-DAY OLD WEANLINGS, C, St a t is t ic a l An a l y s is Th e DATA FROM EACH RAT STUDY AND FROM THE DOG STUDY WERE SUBJECTED/ RESPECTIVELY/ TO AN ANALYSIS OF VARIANCE TO DETERMINE THE SIGNIFICANT EFFECT OF THREE VARIABLES (DIETARY LEAD/ DUP050058637 | PAGE FIVE |. ! .. _ . _ ....................... . SEX/ AND TIME) ON THE VARIOUS QUANTITATIVE RESPONSES WHICH WERE MEASURED DURING THE TWO YEARS OF DIETARY LEAD ADMINISTRATION, i I;II, i RESETS, Fo r t h e p u r p o s e o f t h is p r e s e n t a t io n / I w il l d e a l MAINLY WITH THE SIGNIFICANT EFFECTS OF ONE VARIABLE DIETARY LEAD - ON THE VARIOUS BIOLOGIC RESPONSES THAT WERE MEASURED. (SLIDE 6) A, Cl in ic a l Ob s e r v a t io n s Th e r e w a s n o s ig n if ic a n t e f f e c t o n a p p e a r a n c e / BEHAVIOR/ NEUROLOGICAL EXAMINATION/ BODY WEIGHT/ OR MORTALITY IN DOGS RECEIVING AS MUCH AS 500 PPM Pb IN THEIR DIETS. IN RATS RECEIVING AS MUCH AS 2000 PPM DIETARY LEAD/ INDICES OF APPEARANCE AND BEHAVIOR WERE NORMAL. HOWEVER/ THE PRESENCE OF 1000 AND 2000 PPM LEAD IN THE DIET APPEARED TO DEPRESS THE RATE OF WEIGHT GAIN IN BOTH MALE AND FEMALE RATS. ALSO/ AN INCREASE IN MORTALITY WAS SEEN IN MALE RATS FED 500 PPM LEAD AND ABOVE. DUP050058638 PAGE SIX B. Le a d An a l y s e s (SLIDE m Th is t ime g r a p h s h o w s t h e p a t t e r n s o f b l o o d l e a d CONCENTRATIONS IN DOGS AT DIFFERENT DIETARY LEAD LEVELS. Th e r e w a s a s ig n if ic a n t t r e n d t o w a r d h ig h e r b l o o d l e a d VALUES OVER THE TWO YEARS OF LEAD FEEDING, ALSO/ A HIGHLY SIGNIFICANT RELATIONSHIP (P < 0,01) WAS FOUND BETWEEN BLOOD LEAD AND DIETARY LEAD, THE BLOOD AND URINE LEAD LEVELS OF DOGS RECEIVING 10 PPM OR MORE LEAD (Pb ) IN THE DIET WERE SIGNIFICANTLY GREATER THAN THE CONTROL VALUES, SIMILAR RESULTS WERE FOUND IN THE RAT DATA. ' (SLIDE. #8) On THIS SLIDE WE SEE THAT THE LEAD CONTENT OF BONE. BLOOD, LIVER, KIDNEY, AND BRAIN IN THE DOG INCREASED AS THE AMOUNT OF LEAD IN THE DIET INCREASED. THE RELATIVE RATE OF INCREASE WAS GREATEST FOR LIVER LEAD AND LEAST FOR BONE LEAD. In RATS, SIMILAR RESULTS WERE FOUND EXCEPT THAT THE KIDNEY, RATHER THAN THE LIVER, SHOWED THE SHARPEST RATE OF INCREASE. DUP050058639 PAGE SEVEN (SLIDE #9) Th e r e w e r e s ig n if ic a n t c o r r e l a t io n s b e t w e e n t is s u e , LEAD LEVELS AND BLOOD LEAD LEVELS, FOR EXAMPLE/ THIS SLIDE SHOWS THE CORRELATION BETWEEN KIDNEY LEAD CONCEN TRATION AND BLOOD LEAD LEVELS IN DOGS. A COEFFICIENT OF CORRELATION <R> OF 0,88 INDICATES THAT >77 0% OF THE VARIATION IN KIDNEY LEAD CAN BE ACCOUNTED FOR BY BLOOD LEAD, Th e LEAD CONTENT OF BONE/ LIVER/ BRAIN/ AND KIDNEY WAS SIGNIFICANTLY CORRELATED WITH BLOOD LEAD IN ADULT DOGS AND ALSO IN RATS FED OVER A LIFETIME, THIS INDICATES THAT THE BLOOD LEAD IS A GOOD INDEX OF TISSUE DEPOSITION AS WELL AS EXPOSURE, (SLIDE #10) Th e r e w e r e n o s ig n if ic a n t h e ma t o l o g ic e f f e c t s in d o g s EXCEPT FOR AN INCREASE IN THE NUMBER OF STIPPLED CELLS SEEN AT 500 PPM, IN RATS, A STATISTICALLY SIGNIFICANT INCREASE IN STIPPLED CELLS WAS SEEN AT 10 PPM AND ABOVE AND A DECREASE IN HEMOGLOBIN CONCENTRATION AND HEMATOCRIT WAS NOTED ONLY AT DIETARY LEAD LEVELS OF 1000 PPM AND ABOVE, Th e LIVER FUNCTION TESTS PERFORMED AND THE RESULTS OF URINALYSIS WERE WITHIN NORMAL LIMITS IN DOGS RECEIVING - DUP050058640 PAGE EIGHT AS MUCH AS 500 PPM LEAD OR IN RATS RECEIVING AS MUCH AS 2000 PPM LEAD IN THE DIET. (SLIDE #11) ON THIS SLIDE WE SEE THAT LEAD DID INHIBIT ALA-D ACTIVITY AT A CONCENTRATION OF 50 PPM IN RATS AND 100 PPM IN DOGS. HOWEVER/ THIS INHIBITION WAS NOT ASSOCIATED WITH ANY INCREASED URINARY EXCRETION OF ITS SUBSTRATE BALA UNTIL A CONCENTRATION OF 500 PPM LEAD (Pb ) WAS ADMINISTERED IN THE DIET. THESE FINDINGS ARE CONSISTENT WITH THE FUNCTIONAL RESERVE EXHIBITED IN MANY BIOLOGICAL SYSTEMS, Sig n if ic a n t c o r r e l a t io n s w e r e a l s o f o u n d b e t w e e n ALA-D ACTIVITY AND BLOOD LEAD/ AND BETWEEN DALA AND BLOOD LEAD IN BOTH RATS AND DOGS, THESE RELATIONSHIPS WERE VERY SIMILAR TO THOSE DESCRIBED BY HERNBERG FOR HUMANS, (SLIDE #12) Th e r e w e r e n o p a t h o l o g ic e f f e c t s o f d ie t a r y l e a d o n ANY ORGAN SYSTEM IN FEMALE DOGS FED 500 PPM LEAD OR LESS, PAGE NINE In ma l e d o g s , h is t o l o g y w a s n e g a t iv e e x c e p t f o r a s l ig h t DEGREE OF CYTOMEGALY' FOUND IN THE PROXIMAL CONVOLUTED TUBULES OF THE KIDNEY IN TWO OUT OF FOUR DOGS FED 500 PPM LEAD, In RATS, THERE WAS NO PATHOLOGIC EFFECT OF DIETARY LEAD ON ANY ORGAN SYSTEM AT LEAD LEVELS UP TO AND INCLUDING 100 PPM. At 500 PPM LEAD AND GREATER IN THE DIET, BOTH MALE AND FEMALE RATS SHOWED HISTOLOGIC LESIONS ATTRIBUTABLE TO LEAD IN THE KIDNEY, At THREE MONTHS OF FEEDING AT 500 PPM, RATS SHOWED CYTOMEGALY, INTRANUCLEAR INCLUSIONS, AND CYTOPLASMIC PIGMENTATION IN THE PROXIMAL CONVOLUTED TUBULE OF THE KIDNEY, At 12 MONTHS OF FEEDING, ADDITIONAL CHANGES OCCURRED SUCH AS METAPLASIA AND HYPERPLASIA. Kid n e y t u mo r s w e r e a l s o f o u n d in ma l e r a t s f e d 500 PPM OR MORE LEAD (Pb ). THESE TUMORS WERE FIRST OBSERVED AT 14 MONTHS WHEN A RAT RECEIVING 2000 PPM LEAD DIED. TUMORS AT LOWER LEVELS WERE DISCOVERED AT TIME OF SACRIFICE (24 MOS.), (SLIDE #15) Th e INCIDENCE OF SUCH KIDNEY TUMORS IN RATS AT 500 PPM LEAD AND ABOVE IN THEIR DIETS IS SHOWN IN THIS TABLE, DUP050058642 PAGE TEN It is s e e n t h a t t h e in c id e n c e o f k id n e y t u mo r s in c r e a s e s IN MALE RATS FROM 10% AT 500 PPM DIETARY LEAD/ TO 50% AT 1000 PPM/ t o 80% a t 2000 p p m, Fe ma l e r a t s s h o w e d n o KIDNEY TUMORS UNTIL A DIET CONTAINING 2000 PPM LEAD WAS ADMINISTERED, MOST OF THE TUMORS WERE CONSIDERED TO BE ADENOMAS DERIVED FROM THE TUBULAR EPITHELIUM, THE GREATER SUSCEPTIBILITY OF THE MALE RAT TO KIDNEY TUMORS FROM LEAD MAY BE RELATED TO THE HIGHER INCIDENCE OF SPONTANEOUS CHRONIC NEPHRITIS IN MALE RATS OF THIS STRAIN AS COMPARED TO FEMALES, (SLIDE m) Th e RESULTS OF THE THREE-GENERATION/ s ix -l it t e r REPRODUCTION STUDY ON RATS ARE SHOWN ON THIS SLIDE, THERE WERE NO MAJOR'DIFFERENCES BETWEEN CONTROL AND TEST GROUPS WITH RESPECT TO REPRODUCTION AND LACTATION INDICES IN RATS FED UP TO AND INCLUDING 2000 PPM LEAD (Pb ), 1000 2000: . At and p p m d ie t a r y .l e a d / h o w e v e r / t h e ... AVERAGE WEIGHT OF WEANLING RATS WAS SLIGHTLY DECREASED, ALSO/ HISTOLOGIC EXAMINATION OF ALL ORGAN SYSTEMS IN THE 21-DAY OLD WEANLING RATS (F B) REVEALED LESIONS IN THE 3 DUP050058643 PAGE ELEVEN KIDNEY ATTRIBUTABLE TO LEAD AND COMPARABLE TO THOSE PREVIOUSLY FOUND IN ADULT RATS RECEIVING 500 PPM LEAD OR MORE IN THEIR DIETS, SlNCE WEANLING RATS DERIVE MOST OF THEIR NOURISHMENT FROM SUCKLING IN THE FIRST THREE TO FOUR WEEKS AFTER BIRTH/ IT IS ASSUMED THAT LEAD WAS TRANSMITTED TO THE NEWBORN THROUGH THE MILK/ RESULTING IN HISTOLOGIC CHANGES IN THE KIDNEY, SUMMARY (SLIDE #15) IN SUMMARY/ THIS STUDY SHOWED THAT THE -INGESTION OF 10 PPM LEAD (Pb ) BY RATS AND DOGS RESULTED IN A DETECTABLE INCREASE IN BLOOD/ URINE/ AND KIDNEY LEAD LEVELS, HOWEVER/ NO EFFECT ON CLINICAL OR BIOCHEMICAL PARAMETERS WAS SEEN AT THIS DIETARY LEVEL, At 50 PPM LEAD (Pb ) IN THE DIET 100OF RATS/ AND PPM IN DOGS/ A SIGNIFICANT DECREASE IN ALA-D ACTIVITY OCCURRED, THIS DECREASE WAS NOT ASSOCIATED WITH A CORRESPONDING INCREASE IN THE EXCRETION OF BALA/ THE SUBSTRATE FOR ALA-D, IN OTHER WORDS/ ALTHOUGH ALA-D ACTIVITY WAS INHIBITED/ SUFFICIENT ENZYME RESERVE REMAINED TO HANDLE THE BODY'S REQUIREMENTS, DUP050058644 PAGE TWELVE (SLIDE #16) I 500t w a s n o t u n t il t h e d ie t c o n t a in e d ppm l ead (P )b t h a t a n ima l s d id s h o w a s ig n if ic a n t in c r e a s e in DALA 500e x c r e t io n , Al s o , a t ppm d iet ar y l ead an d abo v e, KIDNEY TUMORS AND AN INCREASE IN MORTALITY WERE SEEN IN RATS, Ev id e n c e o f d e p r e s s e d h e mo g l o b in , h e ma t o c r it , a n d WEIGHT GAIN WERE NOT FOUND UNTIL RATS WERE FED DIETS CON TAINING 1000 PPM LEAD (Pb ) OR MORE. INDICES OF REPRODUCTION AND LACTATION WERE NORMAL AT ALL DIETARY LEVELS TESTED, Ho w e v e r , a d e p r e s s e d w e ig h t g a in a n d r e n a l l e s io n s w e r e 1000FOUND IN THE WEANLING RATS OF PARENTS INGESTING PPM l e a d (Pb ) o r mo r e in t h e d ie t . Re s u l t s f r o m t h is s t u d y c a n b e in t e r p r e t e d in t w o w a y s , Fir s t , o n e c o u l d s a y t h a t s u c h in f o r ma t io n c o u l d n o t b e e x t r a p o l a t e d t o t h e h u ma n s it u a t io n b e c a u s e t h e d o s e s OF LEAD FED TO BOTH RATS AND DOGS WERE MUCH HIGHER THAN THE DAILY DIETARY LEAD INTAKE OF HUMANS, FOR EXAMPLE/ A 10 70LEVEL OF PPM ?B FED TO RATS AND DOGS IS TIMES HIGHER THAN DAILY HUMAN DIETARY INTAKE, IF DOSE IS CALCULATED ON THE BASIS OF /JG Pb /k G BODY WEIGHT. (SLIDE #17) PAGE THIRTEEN j HOWEVER/ THE RESULTS SHOWN ON THIS TABLE SUGGEST ANOTHER POSSIBLE INTERPRETATION. HERE/ WE SEE THE BLOOD/ KIDNEY/ AND BONE LEAD LEVELS IN DOGS AND RATS AFTER 24 MONTHS OF FEEDING. Th e NORMAL RANGE OF LEAD CONCENTRATION IN THE SAME TISSUES IS ALSO SHOWN FOR HUMANS, ITj IS INTERESTING THAT AT 100 PPM DIETARY LEAD AND BELOW/ BLOOD LEAD LEVELS AS WELL AS KIDNEY AND BONE LEAD LEVELS/ IN RATS AND DOGS ARE COMPARABLE TO HUMAN TISSUE LEAD LEVELS. In THIS NORMAL RANGE OF BLOOD LEAD FOR HUMANS (11-40)/ INVESTIGATORS HAVE REPORTED A SIGNIFICANT DECREASE IN ALA-D ACTIVITY,* IN THIS STUDY, WE SAW A SIG NIFICANT DECREASE IN ALA-D ACTIVITY AT A BLOOD LEAD OF 31,5 IN THE DOG AND AT 35,9 IN THE RAT, ALSO/ IN HUMANS/ A BLOOD LEAD OF 80^6/100 ML IS THE LEVEL AT WHICH CLINICAL SIGNS OF LEAD POISONING BEGIN ITO APPEAR IN HUMANS. n OUR STUDY/ 500 PPM DIETARY LEAD IN RATS AND DOGS PRODUCED A SIGNIFICANT INCREASE IN DALA EXCRETION IN THE URINE; THE BLOOD LEAD CONCENTRATION IN BOTH SPECIES WAS ABOUT 80/jG/100 ML. ALSO., AT 500 PPM 80 JUG/100 ML-BLOOD LEAD)/ RATS BEGAN TO SHOW HISTO LOGICAL CHANGES IN THE KIDNEY ATTRIBUTABLE TO LEAD/ AND AN INCREASE IN MORTALITY. KlDNEY LEAD LEVELS AND BONE LEAD LEVELS IN RATS FED 500 PPM LEAD ARE SEEN TO BE WELL ABOVE DUP050058646 PAGE FOURTEEN THE LEVELS OF LEAD REPORTED IN THE KIDNEY AND BONE OF HUMANS, i I Th u s , ANOTHER INTERPRETATION f r o m t h e p r e s e n t ANIMAL STUDY IS THAT RESULTS DO SUPPORT SOME ASPECTS OF LEAD TOXICITY WHICH HAVE ALREADY BEEN REPORTED ON HUMANS, HJT/u t d 5/5/72 DUP050058647 Sl id e #1 EXPERIMENTAL DESIGN to to CM CM o>% E co ro o * o CM tO f* o o SI* CM I O II >- o ~ CO o o z < to w o =5 H* co ' * OW cc LJ CL CJ * CO 0 tr E Q. CL O o to e o 2 _i w UJ o> *1 H 2 tr 0 * CO CO to m CM *? CM IO CO Q <ZZ O * CC UJ CL CM * 00 1 UJ CL co -J Ll h< 111 >' cr S o UJ -J 2 < CO GO O CL 35 to sr O 1 co -j o to 5 UJ > Ul --I o z <c tr UJ co co jo m o o to to C/> tf) O t~ O O (X jI I DUP050058648  -%r- Sl id e #2 CONTINUED Q Z I ly H ,,> >" 50 0 {-" CO & a. o. O o o CM TMJ Ul o > Ul o O OS ft Ul 0 L 60 - CM 1L1 ro CM 1i Q <sf CM os Ul O. 1 U. Z O CO CM J~ & -J US > Ul o z < >? OS UJ Li CO <23 CL ^ O ro c m CP . m l-- '< cc I V I i7 > DUP050058649 PARAMETERS UNDER OBSERVATION o Sl id e #3 o I-- UI < X co X 2 .g 5Q X-- UJ co jg ^< oX x UJ >* t -j <Z h" x o CO UJ UJ 2 o. Q UuJ. SC X UJ < UJ UJ 1 UJ <r o o __ 2 X co <S 2 < X o -J X 2o X 00 => ^ * 3 <2ft Jo <s O uj x a- CL' UJ H s o x L Ju & og O < .2 o CD f--: **4 3 3 Mj vj N '2 c --I: DUP050058650 CONTINUED SPECIAL TESTS ( DALA, A L A -0 ) Sl id e #4 $ 3 o $ ffc 1 R >4 S a CO -J -J yj O O UJ o o -J . CL q , yj o o co -J CD o3 < yj i-- o LU $ X ~J 3L X a. <>r oj~ o os o x a. x s a I 03 .u Q. O O C/3 a: O G 03 U o s to- X CX a. x ( yj a. co < a. O IS < CO o #* 35' -J o h- > 22 CO y. CO >* -J cs < tu > 22 OS 23 DUP050058651 C O N T IN U E D Sl id e #5 X H* U, ' o cr. fc **S LJU S3* ca O $ JSi UJ Cl. > > o J OX 1P v> < 51 a. * Q. '22 <b CL <S} 2 UJ ~ H* S to <s >X to <o o oH oS eg CO Q Z < UJ -J < u z < o 02 CP O I *s5 UJ a <9 z < n O iz ' CO .o lo co O d cs s oJ o J o UJ h* I-- ^ *0 to _ o Uu o Ift Xo Q z IK U. o &OK u- * o& oo 1s35 ft* 5t Cb $5 CO CO UI CO o CK CO CO OH Htbi to co o o CO UJ < d o Z a z < X $ &?> o IK iff I5 K H S 9N H N V 3M r '* DUP050058652 EFFECT OF DIETARY LEAD ON CLINICAL OBSERVATIONS go. tr> Q. O HO <0 cc CM 1 O ts o. a. O E O 10 cx cx <w O O WQ O UJ CD < z. 1 a ! < o Z 5' Sl id e #6 oa. , CO to o O 0in z z z oi O < X UJ -J UJ .< oo z ec ac o o o X -j > o H < UJ d *- x CL uj GC ID o uj ac cc o Q 3 o s Q UJ .s . ec Ho U to. UJ QJ u- UJ to. rt UJ t-- OO Z" z It It zl DUP050058653 Sl id e #7 TWO YEAR LEAD FEEDING STUDY IN DOGS 500ppm lOOppm 50 ppm 10 ppm Oppm DUP050058654 Sl id e #8 TWO YEAR LEAD FEEDING STUDY (DOGS) TISSUE Pb vs. Pb IN FEED BONE LIVER BLOOD KIDNEY BRAIN WMwj 10 100 LEAD IN FEED (^g/g) DUP050058655 TWO YEAH LEAD FEEDING STUDY {DOGS) REGRESSION : KIDNEY Pb on BLOOD Pb BLOOD LEAD (M g/iO O DUP050058656 EFFECTS OF DIETARY Pb ON HEMATOLOGY a. Q. O CO O 0 UJ > L'J <c oc CM o1 O UJ Lj u U. CO g a. a UJ o o O 0 a to 1 O U4 CO 22 o a, co UJ /V* cl Q. o Q. CL o O a. OL O O OO a. CL UJ O .2 OO to 2 UJ 2 o 2 co UJ o 2. o o CQ O UJ o _J -J f-- O CL CL < s O s h~ UJ UJ CO X X 5~ lO 2o <o o Ev 2a 2 i~ CO < CO i UJ > UJ < x; o ESC UJ > 2 O UJ o -J > <r 2 o cuc. UJ o h2 H CO UJ X UJ UJ u. o u> -J a *St' i* DUP050058657 SIGNIFICANTLY DIFFERENT FROM CONTROL LEVEL AT p < 0 .0 5 Sl id e #11 O JQ 0 #* i 1 ^ (O 00 (> ** < TO F*1 0'*` fO vj CM CM -- ~ CO o o o lx. c*5 <C --J TV ro o CO <CJin tv. CO fv CM CM- CM CM in.# Q d Od o 5 co . LU 3 <-J .> z CM j Sc s < Q O CM ^ FO CD LU I CM CM ^ <^` FO <S CM CM 5 co < an # IS c 2 sj- <3" ro to CM hto M* 00 o> 6 o o d - .4 ' * 55: F.O 3** C O fv CM ~~ CM t *' fv M* o Tv to o CM CM CO H 2 3 f* Q O O _J CO u o O > > o Ih- EO < UJ Ui CO S' < HE T Q > X UJ o Q o < o< cj K <0 "i>3ii -3 > UJ q e 33 0 Z` Oz Us Ou >0C I <c --` P UJ UJ > OO o o -- m , o* o m 0.0 o o o CM S < s < 1i << b S yj --J UJ Ui o CM DUP050058658 . Sl id e #12 PATHOLOGIC EFFECTS OF DIETARY LEAD DOGS AT MALES o. Q. o o m VI LJU1 < S UJ U- - z: UJ ZL O JZ (/> O O Q > h~ UJ XS OO _J cn < LU O o LU o o o. X CO 5 <t o LU oS' N h- CM o>- * E LU cl O Qu O LU O LU LO DC O LU <C- O CM eox. tOOo <hr 4- o Q. >LZU o i<. . zr-~ >oo-J o LU X < O> CD Q- O < <o O Z M .X <C C0 h" < * cc L>U. oCO <C O Z < CEexL O too h<-t s *- CO ODC sCD h U> J z o . *5, * ci -- i .1 i. , r? < :* DUP050058659 INCIDENCE OF KIDNEY TUMORS IN RATS FED LEAD 2 CO CO LU q : -J O< 2E ZD LU h~ U. o O O 10 ro LU 2 CO o LU --1 < fsf*g oooo -- 10 oo Sl id e #13 oooo OO lO w E L>U o. CL Ui JQ Q- O o . .6 , E u. o. o. Cl Q. Q. O >QZ IO O m tO O o o O J2 Q LU ` _o o a: LU O o s DUP050058660 scmm J2 k. ^ SV 6 m &> $g ^ & K f: k to to to to b* ^ as* C fe |g * M e o. o. o O OCVJ ** E o. o. O oO o o O (M VI o o JZi # a. X X UJ LU X UJ X UJ QO Qo Z z -- z z *MW >* z >- z j~ o I-- o J H zf H* h- 2 CO 01 UJ u_ UCOJ o 2 > < -J c6x Co& o o o CVJ o z < o o o o lo UJ > X) js UJ 0Q 1 U- CO O UJ > UJ a -J LU IX. ja UJ Gu. o >* Z CUOJ z o z. < esc o 03 z oUJ CCOL K => CL CL lx. u Oo z CK O CO CO CL Z> CL U- O OO O s: . z z CL > ir J2 co z o UJ CO UJ --I ll o >* Ho* UJ z u. Qyj HK9? lx, id CC <1 h- LU < CO Q B O D Y W T. OF F -8 W E A N LIN G S t i ) S' k / i; '! DUP050058661 >U*J zQ * O zsj Sl id e #1! \ Z' -- o z MIM <fiC 9 < Q3 * 0 0 Ift <6 o CO UCOJ < UJ > C < H UJ h<? fK lJ z z UouuJ r UJ : G=> CO UoJJ UJJ UJ CzoO 0 O OJ GO ' O Uo- z\ a.tmni a,' aX.3 {C-O* z-- UeeJ XQJ S J 1 < -J < ~- CO H* :<^rww>in o IS! UJ u. u. UJ i,. t- <o UJ 22 o UJ O U UJ >* Z o CO C> CO o Io $ ***iS* K Z UJ VC z o - Sf* o oO o sg :W*J <g%*. 25 ^ UJ T X C3^ z o z UJ axa. > z < K m z o,, 0 1 < ~J < -J o> UJ > UJ J UJ o .- m m o DUP050058662 Jo I 1 <0 SO o 13J fa 2 2 oo <2s -J esc 2? i X Qc 5 Sl id e #16 ***** CO CO Cf> ss o<sX-53 2 H* <?. UJ O S5 WT. 0 KIDNEY TUMORS o u<j - 2 < in <H *** <n 'b UJ iD X > G > <X to CO tHDi O < 5 X2 oCO Uo 2 Zj C2O 2O < UJ 55 CO U~JJ UJ O li- O <,,J? >* UJ G <OsgSr? 2cOunt* < Sg O a <2- r o .S3 X 35 U. > UJ 2 O s*: Hco* o uj uy, Ly m_-jj UoJ 2 < o Q UJ *Umnu* 2 a. L <D tn in o O o {" u j co *|5*i*P* o o ca. <' -J .O < .a, o 2Q0O UJ > mJ o l&J if) o sn O o o oo DUP050058663 PB LEVELS IN BLOOD, KIDNEY, AND BONE (M E A N VALUES A T 2 4 NOS.) DUP050058664