Document 5bMz0Ze3w6Lvew75Ob2vLRX78
1 IN THE CIRCUIT COURT TWENTIETH JUDICIAL CIRCUIT OF ILLINOIS
2 ST. CLAIR COUNTY
3 FRANCES E. KEMNER, e t a l . , 4 P lain tiffs, 5 vs. 6 MONSANTO COMPANY, 7 Defendant.
) ) > ) ) NO. 80-L-970
) ) ) )
8 B efore th e HON. RICHARD P . GOLDENHERSH, Judge
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10
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12 TESTIMONY OF DR. FRANK DOST 13 November 5, 1985
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15
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17 APPEARANCES:
18 MR. REX CARR, A tto rn e y a t Law, and MR. JEROME SEIGFRE1D, A tto rn e y a t Lav
19 A ppeared on B e h a lf o f th e P l a i n t i f f s
20 MR. KENNETH HEINEMAN, A tto rn e y a t Law, and MR. JOSEPH NASSIF, A tto rn e y a t Law
21 A ppeared on B e h alf o f th e D efendant
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PATRICIA GANDY, CSR, RPR 23 O f f i c i a l C ourt R e p o rte r
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i
1 INDEX
2 DR. FRANK DOST 3 D ir e c t E xam ination by Mr. Heineman
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5 EXHIBITS
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MARKED
OFFERED
7 D efen d an t's 1281 . . . . . . 8 D efen d an t's 1282 . . . . . . 9 D efendant' s 1283 . . . . . . 10 Defendant* s 1284 . . . . . . 11 Defendant* s 1285 . . . . . . 12 D e fe n d a n t's 1286-89 . . . . 13 D efen d an t' s 1290 . . . . . .
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27 32 33 70 81 93
5 31 82 82 82 82 103
14 D e fe n d a n t's 1291 . . . . . . 15 D e fe n d a n t's 1292-95 . . . .
104 107
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18
19
20
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ADMITTED 5 31
83 83 83 83 104
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1 BE IT REMEMBERED AND CERTIFIED t h a t h e r e t o f o r e , on 2 to - w it: Tuesday, November 5, 1985, b ein g one o f th e re g u la r 3 ju d ic ia l days of th is Court, the m atter as h erein b efo re s e t 4 f o r t h came on f o r h e a rin g b e f o r e th e HON. RICHARD P . GOLDEN5 HERSH, C ir c u it Judge in and f o r th e T w en tieth J u d i c i a l C i r c u it, 6 S ta te o f I l l i n o i s , S t. C la ir County B u ild in g , B e lle v ille , S t. 7 C la ir County, I l l i n o i s , and th e fo llo w in g was had o f re c o rd , 8 to -w it:
9
10 THE COURT: Good m orning. 11 CONTINUED DIRECT EXAMINATION 12 BY MR. HEINEMAN: 13 Q Dr. D ost, I 'd l i k e to ta k e you b r i e f l y back f o r a 14 m inute to th e d is c u s s io n s we had y e s te rd a y on p o rp h y ria and 15 th e no o b s e rv a b le e f f e c t l e v e l , th e NOEL. Have you cau sed 16 a c h a r t to be p re p a re d which would a s s i s t you in e x p la in in g 17 t h i s s i t u a t i o n ? 18 A Yes. 19 (At t h i s tim e D e fe n d a n t's E x h ib it 1281 was m arked 20 f o r i d e n t i f i c a t i o n . ) 21 Q A ll r i g h t , s i r , I ' l l show you w h a t's been marked 22 as D efen d a n t'8 E x h ib it 1281 and ask you to examine t h a t and 23 i d e n t i f y i t f o r me, p le a s e . 24 A T his i s a c h a r t p re p a re d from t a b u l a r d a ta t h a t was
3
1 published by Smith, e t a l . , in the jo u rn a l Biochem ical Pharma 2 c o lo g y , L ine 30 in 1981, and t h i s i s a d e s c r ip tio n o f th e
e f f e c t o f an a c u te o r o f a c u te d o se s, s in g le doses o f TCDD
3
as d istin g u ish e d from the experim ents th a t you have m entioned,
4
I b e lie v e , y esterd ay in which lower doses were given on a
5
6 weekly b a s is over an extended p erio d o f tim e.
Q Now, would you be so k in d as to s te p down h e re and
7
8 explain th a t to the ju ry please? A This is another dose response curve. I t looks a
9
l i t t l e d i f f e r e n t from th e ones t h a t we ta lk e d ab o u t th e o th e r
10
11 day. You r e c a l l t h a t in th e ones t h a t w e'v e seen e a r l i e r
12 th e re i s k in d o f an S shaped arran g em en t. T his i s r e a l l y
13 th e same, th e o n ly d if f e r e n c e t h a t th e , th e change i s v e ry ,
14 v e ry r a p id . The slo p e h e re i s ex tre m ely s te e p , and w hat i t shows
15 i s t h a t
a t a dose, and th is d if f e r e n tia te s between males
16 and fem ales b ecau se th e r e i s some d if f e r e n c e i n th e re sp o n se
17 o f m ales and fem ales to m ost o f th e e f f e c t s o f TCDD* I t i s n ' t
18 much, b u t i t ' s m e asu rab le . And a g a in t h i s i s a lo g a rith m ic
19 s c a le and t h i s would be a d o se, I b e l ie v e , o f ab o u t 6 m ic ro
20 grams p e r k ilo g ra m . And you can see t h a t th e diamond and th e
21 c i r c l e a re superim posed h e r e , th ey b o th have g o t th e same
22 d o se s, t h i s I th in k would be 12 m icrogram s p e r k ilo g ra m and
23 t h i s would be 16. And w hat i t shows h e re i s t h a t th e s e
24 r a t h e r s u b s t a n t i a l s in g l e d o se s, now t h i s i s j u s t a s in g l e
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1 dose, not a chronic exposure but a sin g le dose, th a t i t requires 2 a very su b sta n tia l sin g le dose before th is kind of response 3 em erges. The m easurem ent t h a t i s b ein g made i s th e ap p earan ce 4 o f p o rp h y rin a c t u a l l y d e p o s itin g in th e l i v e r . You r e c a l l 5 yesterday I mentioned th a t they can appear in the u rin e and/or 6 in th e l i v e r , and h e re we a re ta lk in g ab o u t h e p a tic p o rp h y rin , 7 t h a t i s th e s e in te r m e d ia te p ro d u c t in th e s y n th e s is o f heme 8 accum ulating in liv e r c e lls and in the response the anim als 9 manage a degree o f im pairm ent in heme s y n th e s is v e ry , v ery 10 w e ll u n t i l th ey g e t up to some s u b s t a n t i a l dosage, and th en
t
11 th e accu m u latio n s in th e t i s s u e s becomes v e ry , v e ry r a p id b e 12 cause th e -- presum ably b ecau se th e im pact a t th e enzyme 13 le v e l becomes so s u b s t a n t i a l t h a t th e m a te r ia l i s n o t b e in g 14 d isp o se d o f v ery r e a d i l y . 15 Q Now, Dr. D ost, y e s te r d a y you spoke ab o u t th e c h ro n ic 16 le v e l o r th e no e f f e c t le v e l f o r c h ro n ic dosage b e in g a t w hat 17 le v e l? 18 A W ell, i t i s a t a l e v e l down on th e o r d e r o f a te n th 19 o f a m icrogram p e r k ilo g ra m p e r week o r l e s s . S t a t i s t i c a l l y 20 i f you ru n , i f you run th e dose re sp o n se curve a l l th e way 21 down, th e no e f f e c t le v e l app ro ach es a h u n d red th o f a m icro 22 gram p e r k ilo g ra m p e r week. S t a t i s t i c a l l y th e no e f f e c t le v e l 23 i s on th e o rd e r o f a te n th o f a m icrogram , and by t h a t I mean 24 t h a t i t comes up a l i t t l e b u t i t i s n o t enough t h a t a s t a t i s -
7
1 we s t a r t d e a lin g w ith a d is e a s e s t a t e , a g a in , in c r e a s e d b lo o d 2 lip id s a l l by them selves could no t provide a d iag n o sis. A 3 d ia b etic w ill often have elevated blood lip id s , b u t th a t 4 w ouldn't diagnosis d ia b ete s. I t depends on o th e r kinds of 5 e f f e c t s . I t I s a s s o c ia te d w ith d ia b e te s . Smokers may have 6 e le v a te d b lo o d l i p i d s , b u t th e y may n o t, to o . B ut i t seems 7 to be -- th e r e seems to be some a s s o c ia tio n . P eo p le w ith 8 hypertension o ften have elev ated blood lip id s , b u t here w e're 9 dealin g w ith a chicken and egg s itu a tio n in which th e co n d itio n 10 t h a t le d to e le v a te d b lo o d l i p i d s may have cau sed o v e r a lo n g , 11 long p e r io d o f tim e a change i n th e a r t e r i e s t h a t we w ould 12 c a l l a r t e r i o s c l e r o s i s t h a t r e s u l t e d i n th e h y p e r te n s io n , so 13 w hat I am sa y in g h e r e i s t h a t a lo n g term , a lo n g s u s ta in e d 14 e l e v a t io n i n b lo o d l i p i d s and b lo o d c h o l e s t e r o l c o u ld v ery 15 w e ll le a d to and I n f a c t th e s ta te m e n t i s som etim es made t h a t 16 .p ro lo n g ed e le v a tio n o f b lo o d l i p i d s cau ses c a r d io v a s c u la r 17 d is e a s e . O th er p e o p le a r e more c o n s e r v a tiv e and say t h a t i t 18 i s a s s o c ia te d w ith I t . W hether t h a t 's a w ea sel w ord, I d o n 't 19 know. In any c a s e , th e r e i s a t l e a s t an a s s o c i a t i o n w ith 20 c a r d io v a s c u la r d is e a s e , and t h a t 's why w e 'r e t a l k i n g ab o u t 21 i t p erh ap s i n th e c o n te x t o f c a r d io v a s c u la r d is e a s e . But 22 th e r e a re a v a r i e t y o f o th e r c o n d itio n s t h a t w i l l cau se e i t h e r 23 a p ro lo n g e d o r a r e l a t i v e l y f o re s h o rte n e d p e r io d i n w hich b lo o d 24 l i p i d s a r e e le v a te d .
8
Q And a r e th e r e s u l t s in , th e an im al d a ta dose dependent?
1
A Yes, th e y a re , dose dependent. The d i f f i c u l t y i s th a t
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in th e c a se o f l i p i d a b n o r m a litie s w ith r e s p e c t to TCDD a t l e a s t ,
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th e doses have n o t been c a r r ie d downward a l l th e way to c l e a r
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5 no e f f e c t l e v e l s . The dose re s p o n s e i s v e ry c l e a r , b u t we d o n 't id e n tify a s p e c ific no e f f e c t le v e l below which ho
.6
ab n o rm a lities^ i n l i p i d m etab o lism a p p e a r. Many o f .the s tu d i e s
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8 use only a sin g le dose because they are try in g to understand the mechanism by which th is event o c c u rs. But i t would appear
'9
10* t h a t th e no e f f e c t l e v e l i s s u b s t a n t i a l upon th e o r d e r o f a microgram p er kilogram*
11
12 Q Now, w ould you say i t would ap p e a r t h a t i t i s sub s ta n tia l?
13
14 A By t h a t I am j u s t making a judgm ent on w hat th e dose 15 re sp o n se in fo rm a tio n is* You r e c a l l t h a t th e o th e r day I t o l d 16- you t h a t i f you d o n 't have enough p o in ts on a dose re sp o n se 17 cu rv e to c a r r y a l l th e way to a no e f f e c t l e v e l , i t i s p o s s i b le 18 to p r o j e c t w ith in l i m i t s from w hat d a ta th e r e i s w here th e dose 19 re s p o n s e , w here th e no e f f e c t l e v e l m ight be e x p e c te d . However, 20 one c a n n o t sim p ly say h e r e i s w here th e d o se, h e re i s w here th e 21 no e f f e c t le v e l i s . X w ould ju d g e on th e b a s is o f th e e x i s t i n g 22 d a ta t h a t i t i s on th e o r d e r ,o f a m icrogram p e r k ilo g ra m . 23- Q Now, b a se d upon a judgm ent o f a m icrogram p e r k ilo g ra m
t/
24 and th e dose t h a t you c a lc u l a te d w ith r e s p e c t to th e S tu rg eo n
fi \
. -j'
1 ex p o su re o f th e 10 grams o f s o i l , w ould t h a t S tu rg eo n dose be
2 s u ffic ie n t to generate elev ated blood lip id s?
3 A W ell, we c a lc u la te d t h a t a dose o f .001 m icrogram s
4 p e r k ilp g ra m p e r day w ould r e p r e s e n t a 222n d , i f you w i l l
;
5 I beg y o u r p ard o n , t h a t i s 222 tim es h ig h e r th a n t h a t ex p o su re
6 t h a t we p o s tu l a te d , and s in c e we a r e ta lk in g about, 1 "m icrogram ,
7 then th a t number would be expanded by a thousand.
8 Q So th e S tu rg eo n dose w hich you c a lc u l a te d w ould be
9 222,000 tim es 2 low?
1 A I t would be t h a t much l e s s th a n th e 1 m icrogram p e r '
11- k ilo g ra m t h a t I am p o s t u l a t i n g .
12 Q Now, I 'd l i k e t o t a l k to you t h i s m o rn in g .a b o u t
13 n e u r o lo g ic a l e f f e c t s o f TCDD. Now, b r i e f l y d e s c r ib e th e
14 n e u r o lo g ic a l system t h a t we a re t a l k i n g a b o u t.
15 A W ell, th e n e u r o lo g ic a l system i s a l l o f th e - -
16 i t i s th e c e n t r a l n erv o u s sy stem , th e b r a i n and th e s p i n a l
17 c o rd , and a l l o f th e p e r i p h e r a l n e rv e s . So th e n e u r o lo g ic a l
18 system i s o u r system f o r tr a n s m i t t i n g r a p id m essages from
19 one p a r t o f th e body and f o r i n t e g r a t i n g them and a c tin g
20 upon them i n th e c e n t r a l n erv o u s sy stem . I t i s a l l o f th e
21- nerves o f the body.
22 Q Now, had th e r e b een n e u r o lo g ic a l e f f e c t s o b serv ed
23 i n humans exposed to d io x in ?
24 A To my r e c o l l e c t i o n , n o . I am n o t e n t i r e l y f a m i l i a r
''r 10
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1 w ith a l l o f th e ep id em io lo g y , b u t I do n o t r e c a l l any in
2 humans,
.*
3 Q A ll rig h t, are you fa m ilia r w ith the exam ination
4 by Dr, S uskind o f w orkers a t th e N itro P la n t who were exposed
5 to in th e cleanup o f an a c c id e n t th e re in 1949?
6 A I remember some o f t h a t in fo rm a tio n , y e s , I remember
7 one in d iv id u a l th a t from which a nerve bio p sy was taken th a t
8 was found to have s u f f e r e d some d m y lin is a tio n .
9 Q Now, do you r e c a l l w h eth er o r n o t th e r e w ere r e p o r te d
IQ n e u r o lo g ic a l e f f e c t s i n th o s e in d iv id u a ls who w ere exam ined by
11 Dr. Suskind?
12 A I 'm s o r r y , I d o n 't remember.
13 Q A ll r i g h t , l e t ' s d e a l w ith th e anim al d a ta . What d id
14 th e s tu d i e s show w ith r e s p e c t to an im al d a ta and th e e f f e c t o f
15 TCDD i n c a u sin g n e u r o lo g ic a l e f f e c t s ?
16 A I t shows t h a t TCDD i s e s s e n t i a l l y i n a c t i v e as f a r a s ~
17 e i t h e r th e c e n t r a l o r th e p e r i p h e r a l n erv o u s system . T here
18 a re r e p o r ts o f an im als a t h ig h doses showing l i s t l e s s n e s s and
19 e f f e c t s l i k e t h a t , b u t t h a t can h a r d ly be d e s c r ib e d as an
20 e f f e c t on th e n erv o u s sy stem . These a r e v e ry , v e ry s ic k 21 an im als and any s ic k anim al I s going to be l i s t l e s s , and so
22 I d o n 't th in k t h a t d a ta l i k e t h a t o r s ta te m e n ts l i k e t h a t can
23 be i n t e r p r e t e d as r e f l e c t i n g n e u r o lo g ic a l change i n th e 24 e x p e rim e n ta l a n im a ls. What we have to r e l y on a r e th e --
* -W.\
1 p rim a rily the p ath o lo g ic a l exam ination of anim als in which the 2 c e n tra l and the .p e rip h e ra l nervous system a re examined m icro-
i
3. s c o p ic a lly i n d e t a i l to d eterm in e w h eth er th e r e hah been any 4 change in the a rc h ite c tu re o f the nerve c e lls . 5 Q Now, when you say t h a t someone r e p o r ts t h a t th e 6 anim als look l i s t l e s s , a t what doses have those rep o rts been 7 made? 8 A W ell, those have been a t high doses th a t are p ro 9 ducing, they are on the verge o f a le th a l e ffe c t. These are 10 an im als t h a t a r e c l e a r l y i n t o x i c a t e d by th e TCDD, th e y have 11 been b a d ly i n ju r e d . 12 Q A ll r i g h t . And so th e l i s t -- to w hat do you 13 ' a t t r i b u t e th e l i s t l e s s n e s s ? 14 - A A v e ry s ic k a n im al, and I would n o t t r y to a s s ig n 15 any s p e c i f i c -- any s ic k an im al i s going to be l i s t l e s s . 16 These an im als don' t c o n v u lse , th ey don't show s ig n s t h a t w ould 17" s u g g e s t t h a t t h e i r c e n t r a l n erv o u s sy stem has b een s tim u la te d 18 as f a r as d e p re s s io n i s co n cern ed . There i s no way o f 19 a s s ig n in g t h a t k in d o f e f f e c t . However, th o s e an im als have 20 s u f f e r e d a g r e a t d e a l o f o th e r k in d s o f damage, and a s ic k 21 anim al i s l i s t l e s s . 22 Q A ll r i g h t . Now, w hat do th e s tu d i e s d em o n strate 23 w ith r e s p e c t to n e u r o lo g ic a l e f f e c t s on an im als due to TCDD?
i
24 A R e a lly , n o th in g . The o n ly e f f e c t t h a t , f o r exam ple,
*v-
.12`V 4
1 th a t Kociba re p o rte d in h is life tim e stu d ie s on r a ts was, and 2 th is i s a t a dose range t h a t was a c tu a lly in c re a s in g th e 3 m o r ta lity o f th e an im als, th e re was a good d e a l o f p ath o lo g y 4 g e n e r a lly i n th o se a n im a ls. There w as, and I b e lie v e I men 5 tio n e d som ething ab o u t t h i s y e s te r d a y , some c a p i l l a r y b r e a k 6 down i n th e c e n t r a l n erv o u s sy stem so t h a t th e r e was b le e d in g 7 around nerve c e lls , around nerves, b u t th a t's not a function 8 of e ff e c t on the nervous tis s u e . T h a t's a fu n ctio n o f an 9 e ffe c t on the c irc u la to ry system . 10 Q A ll r i g h t , now, K ociba, he c a r r i e d on two d i f f e r e n t 11 s e t s o f s tu d ie s ? 12 A. T his i s th e two y e a r s tu d ie s * In th e t h i r t e e n week 13 s tu d i e s i n w hich th e y ra n s t i l l h ig h e r doses o f m icrogram p e r 14 k ilo g ra m p e r day,, w hich a g a in cau sed a good d e a l o f o th e r 15 k in d s o f p a th o lo g y , th e r e was no m e n tio n , th ey s p e c i f i c a l l y 16 d e s c r ib e th e p a r t s o f th e b r a i n t h a t th e y exam ined p a t h o lo g ic a lly 17 a n d 'th e y r e p o r te d no in fo rm a tio n ab o u t any e f f e c t s on th e b r a i n . 18 T he-custom i s i n a p a th o lo g ic a l stu d y to i d e n t i f y e v e ry t i s s u e 19 t h a t i s exam ined, and th e n to r e p o r t o n ly on th e t i s s u e s i n 20 w hich some k in d o f a b n o rm a lity h as been found, a n d 5f o r t h i s 21 re a so n th e r e i s o f t e n 'a s ta te m e n t in th e methods o f th e p a p e r 22 d e s c r ib in g th e methods o f ex a m in a tio n and th e t i s s u e s exam i 23 n a tio n and no f u r t h e r d is c u s s io n o f c e r t a i n o rg a n s, and t h a t 24 means t h a t th e r e h as been n o th in g i d e n t i f i e d as b e in g abnorm al.
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1 Q Have th e r e been any s tu d ie s done? You m entioned 2 y e s te rd a y a Dr. C a ld e r h av in g done some s tu d ie s ? 3 A Yes, w e ll, he was concerned about e f f e c ts on the 4 p e rip h e ra l in n e rv a tio n , th a t i s o u t in the le g s and arms and 5 so fo rth , o f course he is studying r a ts , so they have four 6 le g s, and I looked s p e c if ic a lly fo r evidence, p a th o lo g ic a l
7 evidence of in ju ry to p erip h e ra l nerves a fte r su b sta n tia l 8 doses o f XCDD and found n o n e .
9 Q Now, w ith r e s p e c t to m y e lin iz a tio n , we ta lk e d b r i e f l y 10 y e s te r d a y ab o u t m y e lin , w hat had th e s tu d ie s d em o n stra te d w ith 11 r e s p e c t to th e e f f e c t on th e m y elin su rro u n d in g th e n e r v e s 12 a f t e r ex p o su re to TCDD?
13 A D e m y e lin isa tio n i s one o f th e e a s i e r m o rp h o lo g ic al
14 -- by t h a t I mean a n a to m ic a l changes to d e t e c t i n n e r v e s ,
15 i n n erv o u s t i s s u e , and th e r e have been no r e p o r t s w h atso ev er
16 o f changes i n th e m y elin s h e a th o f n e rv e s in an im als i n t o x i 17 c a te d w ith TGDD.
18 Q T his i s an e f f e c t t h a t can be o b se rv e d upon exam i
19 n a tio n ?
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As i t happened 1 am n o t a p a t h o l o g i s t , you m ust
21 u n d e rs ta n d , b u t I 'v e done r e s e a r c h on h e x a c h lo ro p h e n e , w hich i s
22 dem yelin a g e n t, and w h ile I had a p a t h o l o g i s t d i r e c t and
23 a n a ly z e p a th o lo g ic a l e f f e c t s on th o se a n im a ls, I was a b le to
24 I d e n tif y w ith o u t any i n s t r u c t i o n s a t a l l j u s t w ith th e p a th o lo g y
i.14
1 t h a t I le a rn e d as a v e te r in a r y s tu d e n t t h i r t y y e a rs ago o r more* 2 I was a b le to id e n tif y th e area s o f i d eray e lin iz atio n . I t i s a
3 very easy thing to see, 4 Q Now, Dr, S ilb e r g e ld has t e s t i f i e d to t h i s ju r y th a t 5 t h e r e 's b een work to d em o n strate t h a t 2 , 3 , 7 ,8 TCDD ca u ses 6 p e rip h e ra l neuropathy and she c ite s th e work o f anim al stu d ie s
7 done b y , among -- she t a l k s ab o u t th e S t r i k e Croup i n th e 8 N eth e rlan d s, E lo v a a ra 's Group in F in la n d , and w orkers a t
9 NEIHS, a r e you f a m i l i a r w ith Dr. E lo v a a r a 's work?
10
A E lo v a a ra w as, I b e l ie v e , a p o s td o c to r a l
in any
11 c a s e , I was a w orker i n th e la b o r a to r y o f H arry Vanio i n F in la n d
12 who h as done a good d e a l o f work on TCDD and phenoxy h e r b ic id e s .
13 I t h ad n o th in g to do w ith m yelin* They w ere w orking w ith a
14 s t r a i n o f r a t t h a t h as a g e n e tic d e f e c t t h a t - - l e t me see
15 i f t can e x p la in t h i s . When hem oglobin o r heme p r o t e i n s a r e
16 u sed u p , when th e y b re a k down, th e heme has to b e , has to be
17 a ls o d e s tro y e d . And one o f th e p ro d u c ts o f th e heme breakdown
18 i s a pigm ent c a l l e d b i l i r u b i n , and i t i s e x c re te d In th e b i l e ,
19 by and la rg e * T hat h as to be done a c t u a l l y i n th e c e l l s j u s t
20 as heme i s made i n each c e l l t h a t u se s i t , t h a t breakdown h as
21 to ta k e p la c e i n th e c e l l when f i n a l l y t h a t enzyme becomes
22 i n e f f e c t i v e and h a s . t o be d isp o s e d o f . But you d o n 't r e p a i r ; {
23 p r o t e i n s , you d o n 't r e p a i r enzymes, d is c a r d them a n d 's t a r t
;
24 o y er a g a in . , The s t r a i n o f r a t t h a t E lo v a a ra was u s in g , and
. J/
r. 15
1 he d id t h i s d e lib e r a te ly ., was a s t r a i n t h a t does n o t d isp o se 2 o f b iliru b in very w ell, 1 th in k I mentioned e a r lie r th a t in 3 m ice, why, th e r e a r e s t r a i n s and s t r a i n s an,d s t r a i n s o f r a t s . 4 There are pro b ab ly a hundred d if f e r e n t s tr a in s o f mouse used 5 in re se a rc h , and they are alm ost designed fo r c e rta in kinds?:of 6 i n v e s t i g a t i o n , and th e Gun r a t , w hich i s w hat E lo v a a ra was 7 using, is a r a t w ith a g en etic d efect th a t d o esn 't perm it i t S to dispose of b ilir u b in p ro p e rly , and what they found was in 9 th o se r a t s th e r e was i n e f f e c t , in r a t s t r e a t e d w ith TCDD 10 w ith t h a t g e n e tic d e f e c t1, th e y w ere a b le to f in d a somewhat 11 a m p lif ie d t o x i c i t y , some b io c h e m ic a l changes i n th e n erv o u s 12 system o f th e r a t s w ith t h a t g e n e tic d e f e c t. The doses a g a in 13 w ere f a i r l y s u b s t a n t i a l , we a re n o t ta lk in g ab o u t f r a c t i o n s 14 o f m icrogram as we have b ee n . 15 Q M d th o se s tu d i e s d e m o n strate p e r i p h e r a l n eu ro p ath y ? 16 A No, th ey had n o th in g to do w ith th e p e r i p h e r a l n erv o u s 17 sy stem . They w ere w orking w ith th e b r a i n . 18 Q Now, I ' d l i k e to d e a l n e x t w ith th e immune sy stem . 19 I w onder i f you would d e s c r ib e th e immune sy stem i n an im als 20 and hum ans. 21 A W ell, th e Immune sy stem i s i n an im als and humans i s 22 in e s se n c e th e same. There a r e d if f e r e n c e s among s p e c ie s b u t 23 th e f u n c tio n and th e g e n e ra l co n cep t a re th e sam. The immune 24 system f i r s t i s th e p r o t e c t i o n system t h a t th e body has f o r '
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1 dealing w ith in fe c tio n , th a t is ex tern al organisms th a t can * 2 cause damage to th e organism . I t i s a ls o In te n d e d to d eal 3 w ith f o r e ig n s u b s ta n c e s , f o r e ig n p r o t e i n s . One o f th e re a s o n s 4 th a t oftentim es an in s e c t b ite can cause such a ferocious*response 5 is because of foreign p ro tein s th a t the in se c t in je c ts in to the 6 skin and the body im m ediately sends c e lls in to try to deal 7 w ith t h a t fo re ig n m a te r ia l. The mechanism th a t r e j e c t s organ 8 g r a f ts , and X th in k everyone i s aware o f the p re se n t m edical 9 advances i i n g r a f tin g o rgans, k id n ey s, h e a r t, etc* .And one 10 o f th e problem s t h a t th e y have i s p re v e n tin g r e j e c t i o n . T h a t's 11 an immune re sp o n se to th e f o r e ig n p r o t e i n o f th e new p a r t . The
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12 immune system a p p a re n tly a ls o resp o n d s to c e l l s w hich r e b e - * 13 coining c a n c e r c e l l s b ec au se th e y have become new and s tr a n g e .
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14 So w hat we have h e re i s a p r o t e c t i v e system t h a t i s 'p r i m a r i l y 15 c i r c u l a t e d in th e b lo o d t h a t resp o n d s to e x t e r n a l a s s a u l t , 16 p r im a r ily i n f e c t i o u s a t t a c k . I t has two g e n e ra l d i v i s i o n s , 17 i t i s an e x c e e d in g ly complex system , and I w ould n e v e r c la im 18 to be im m unologist, b u t i t i s a v e ry complex sy stem t h a t can 19 be d iv id e d I n to two p a r t s . One p a r t i s known as th e c e l l 20 m ed iated immune re s p o n s e , w hich means t h a t f o r th e m ost p a r t 21 i t c o n s is ts o f c e l l s w hich a t ta c k f o r e ig n o rg an ism s, w hich 22 a t ta c k f o r e ig n c e l l s . I f we have c a n c e r c e l l s s t a r t i n g to 23 d ev elo p , i t may a t t a c k th o s e c e l l s and i t i n g e n e ra l resp o n d s 24- to , i f you w i l l , th in g s th a t a re p a r t i c u l a t e in n a tu re by
* 4 't ' j
1 o fte n tim e s f o r e ig n i n e r t m a t e r i a l , some components o f th e c e l l . ' 2 m e d iated immune response, may a c tu a lly , e n g u lf them, may a c t u a l l y 3 su rro u n d them and remove them . The o th e r p a r t o f th e immune 4 sy stem i s c a l l e d th e .hum oral immune sy stem . Humoral a r i s e s 5 from th e same ro o t', as hormone, and w hat i t means i s th a t th is 6 is re la te d to m aterials th a t are a c tu a lly in so lu tio n in the 7 blood, p rim arily antibody,, p rim arily antibodies,, and antibodies 8 a re p r o te in s t h a t have -7 t h a t a r i s e from e x p e rie n c e . In th e 9 immune system th e r e a re some p r o t e i n s t h a t a r e .v e ry n o n s p e c if ic , 1 th e gamma g lo b u lin i s a n o n s p e c if ic immune re sp o n se t h a t w i l l 11 t e n d .t o i n t e r a c t w ith f o r e ig n o rg an ism s, and i t ' s t h e r e , a l l th e 12 tim e . I f you have b een v a c c in a te d a g a in s t so m e .d ise a se o r i f 13 you have h ad ex p o su re to some d is e a s e , you w i l l d evelop a n t i 14 b o d ie s an d .y o u a ls o develop a.memory in th e immune sy stem , so 15 t h a t i f ' you a r e exposed a t a n o th e r, tim e , the. body, im m ed iately ` 16 makes more a n tib o d ie s to resp o n d to th e c h a lle n g e , in o th e r 17 w ords, i f you have had sm allp o x o r ch ickenpox, alm o st everyone 18 my g e a t l e a s t had ch ick en p o x a t sometime o r a n o th e r * You 19 d o n 't g e t i t a second tim e,, and th e . re a s o n you d o n 't g e t i t 20 a second tim e , i t happens,' to be a v i r a l d is e a s e , b u t th e re a s o n 21 you d o n 't g e t ch ickenpox a second time i s b e c a u se th e body has 22 le a r n e d ab o u t ch ickenpox and i t i s s i t t i n g th e r e p o is e d to 23 resp o n d to th e n e x t i n f e c t i o n . S o .th e. hum oral immune sy stem 24 has a memory t h a t resp o n d s to th e seco n d a s s a u l t , l e t ' s s a y ,
>**
19
1 th e re is probably a g en etic change in these c e lls as a r e s u lt 2 o f some i n t e r a c t i o n w ith th e e p ith e liu m o f th e thymus and t h a t 3 i s n o t c le a r ly understood, the mechanism. However, th e se c e lls 4 are given the c a p a b ility of responding to the kind of challenge 5 th a t the c e ll m ediated system has to meet. That i s , they w ill 6 go and a t ta c k any organism s t h a t a r e f o r e ig n , t h a t d o n 't b e lo n g . 7 I t w ill a tta c k c e lls th a t d o n 't belong, and th ese lymphocytes 8 leav e th e thymus w ith t h is change in t h e i r g e n e tic in s t r u c tio n s . 9 They go to th e thymus and th e y form c lo n e s . Now c lo n e s a r e 10 j u s t c o l le c tio n s o f c e l l s . A c e l l h as d iv id e d , made a n o th e r 11 c e l l j u s t l i k e i t , th e y d iv id e and make c e l l s j u s t l i k e them, 12 and so we have a co lony o f c e l l s in each lymph node and th ey 13 a r e c o n tin u a lly shed i n t o th e c i r c u l a t i o n . There a r e a l s o 14 c e l l s i n r e s e r v e , and when some k in d o f i n f e c t i o n t h a t r e q u ir e s 15 t h e i r s e r v ic e s a r i s e s , th e n th e y move i n t o th e -- som etim es 16 r i g h t o u t o f th e b lo o d v e s s e ls i n t o th e t i s s u e s to resp o n d to 17 th e c h a lle n g e . These a re th e k in d s o f - - I would im agine 18 t h a t when you have an i n f e c t i o n on a s k in wound and i t g e ts 19 p u sey , t h a t p r im a r il y i s lym pocytes o f t h i s s o r t , as w e ll as 20 c e r t a i n kindscof c e l l s c a lle d m acrophages t h a t a c t u a l l y e n g u lf 21 f o r e ig n m a te r ia l and th e y move i n to th e a r e a to c le a n up th e 22 m ess. 23 Q Now, w hat i s th e r e l a t i o n s h i p between^ th e thymus
gland and the age of a person, w hether he be in infancy or
20
1 adulthood? 2 A The thymus h as i t s g r e a t e s t im p o rtan ce in e a r l y l i f e , 3 v e ry e a r l y . And in th e human i t b e g in s to f u n c tio n , and as a 4 m atter o f fa c t has accom plished a major p a rt of i t s a c tiv ity 5 probably b e fo re b ir th . I t begins to fu n c tio n s e v e ra l months 6 before b ir th and is going through th is p ro cess, because th e 7 f e tu s has i t s own c i r c u l a t i o n , i t i s p ro c e s s in g lym phocytes 8 f o r a lo n g tim e p r i o r to a c tu a l b i r t h . So t h a t th e human i n f a n t 9 i s b o m w ith a f a i r l y e f f e c t i v e immune sy stem , i t i s n ' t p e r f e c t , 10' b u t i t i s v e ry n e a r ly i n p la c e . And th e thymus p ro b a b ly h a s , 11 w i l l have d isa p p e a re d i n a c h i ld o f p erh ap s te n y e a rs . In th e 12 r a t , and m ost o th e r s p e c ie s i n w hich we do e x p e rim e n ta l w ork, 13 th e an im als a r e b o m v e ry v e ry e a r l y , I th in k I m entioned 14 y e s te rd a y t h a t th e r a t pup i s b o m a t ab o u t th e same s ta g e o f 15 developm ent as a two and a h a l f month human f e t u s , so t h e t r a t 16 h as g o t a long way to go i n term s o f i t s developm ent b e fo re i t 17 a c t u a l l y i s co m p eten t. And t h i s f u n c tio n o f th e thymus i n th e 18 r a t i s n o t r e a l l y f u l l y d ev elo p ed , and th e thymus as a con 19 sequence i s r e a l l y v e ry s e n s i t i v e i n a v e ry young r a t . 20 i Q Now, a r e T c e l l s th o s e c e l l s t h a t a r e coded o r p r o 21 grammed in some way in th e thymus gland? 22 A Yes. 23 Q Humans a re b o m w ith c i r c u l a t i o n T c e l l s ?
A A pparently, yes.
*t
21
1 Q How ab o u t r a t s ? 2 A No, th e y d o n 't seem' t o .b e , a lth o u g h a t th e tim e o f 3 b ir th i t is a l i t t l e d i f f i c u l t to examine them. But they appear 4 n ot to have s ta rte d the process of T c e lls u n til a f te r b irth . 5 Q Now, w hat does t h a t mean th e n i n term s o f w h eth e r 6 a chem ical w ould have an immune e f f e c t on a r a t as opposed to 17 a human? Does t h i s d if f e r e n c e i n developm ent betw een th e r a t 8 and th e human have any e f f e c t on your a b i l i t y to e x tr a p o la te 9 data from one to the other? 10 A In th e r a t d u rin g th e e a r l i e r s ta g e s o f developm ent- r ; 11 up u n t i l w eaning and even f o r a p e r io d a f te r w a r d s , th e thymus 12 i s ex tre m ely im p o rta n t, and i f you i n t e r f e r e w ith thym ic 13 f u n c tio n s , th e n alm o st c e r t a i n , w e ll, y o u 'r e g o in g t o p ro b a b ly 14 i n t e r f e r e w ith developm ent o f T c e l l s . The human i n f a n t i s 15 p ro b a b ly p a s t th e s ta g e o f maximum dependence, k e e p fin mind 16 t h a t th e thymus s t i l l f u n c tio n s i n a young c h i l d . But th e 17 i n i t i a l f u n c tio n o f d e l iv e r in g coded, to u se y o u r term , i n 18 s t r u c t e d lym phocytes has b een done, and th o s e lym phocytes 19 once o u t i n th e p e r ip h e ry m a in ta in t h e i r own p o p u la tio n . 20 There a r e s t i l l some b e in g p ro c e s s e d by th e thymus, b u t a 21 l o t o f th e e f f e c t , a l o t o f th e T c e l l a c t i v i t y a r i s e s from 22 th e r e p ro d u c tio n o f T c e l l s o u t i n th e lymph nodes* So a 23 good p a r t o f th e work h as b een done,, much o f th e work h as b een 24 done as f a r as th e thymus i s concern ed i n an i n f a n t .
',
-
22
1 Q W ell, does d io x in have an e f f e c t on th e immune system 2 in experim ental animals? 3 A Oh, y e s . 4 Q A ll r ig h t, and what s o r t of e f f e c t does i t have? 5 A W ell, i t h as a g e n e ra l d e p re s s in g e f f e c t on th e immune 6 system . I t has'. a more -- i t has a g r e a te r e f f e c t on c e l l 7 m ediated immunity than i t does on humoral immunity. However, 8 th e r e a re some e f f e c t s on hum oral im m unity. A l o t o f i t 9 depends on what kind of te s ts are done, and th e re are a very 10' la r g e number o f d i f f e r e n t k in d s o f t e s t s a v a ila b le to examine 11 th e f u n c tio n o f th e immune sy stem , and many o f them do show 12 e f f e c t s o f TCDD and th e r e a re some o f co u rse t h a t do n o t. But 13 I th in k t h a t i t i s , as a g e n e ra l s ta te m e n t, y e s , TCDD i s a b le 14' a t s u f f i c i e n t doses to have an e f f e c t on th e Immune sy stem , 15 Q Because o f th e d if f e r e n c e s betw een th e r a t and th e 16 human b e in g in t h i s p a r t i c u l a r p a ra m e te r, th e immune sy stem , 17 how r e l i a b l e i s t h a t d a ta on r a t s i n term s o f p r e d i c t i n g w h a t's 18 g o in g to o cc u r i n a human b ein g ? 19 A W ell, th e r e a r e two k in d s o f in fo rm a tio n . One i s , 20 o f c o u rs e , w hat I j u s t m en tio n ed , t h a t th e c e l l m e d ia te d sy stem 21 i s f a i r l y f u n c tio n a l i n a newborn human. The o th e r i s t h a t th e 22 im p act o f m a te rn a l tr e a tm e n t, t h a t i s tr e a tm e n t o f th e m other 23 p r i o r to b i r t h i n e x p e rim e n ta l an im als does n o t seem to be a s 24 s u b s t a n t i a l as d o sin g a fte rw a rd when th e an im als a r e n u rs in g
23
1 b ec au se i t i s w ell-know n t h a t TCDD i s a b le to f in d i t s way i n t o 2 m ilk , t h a t 's n o t a t a l l a s e c r e t . And th e r e seems to be a 3 g r e a te r e f f e c t on th e r a t s a f t e r b i r t h d u rin g th e p e rio d when 4 th e thymus i s a c t u a l l y s t a r t i n g to do i t s work. So I would 5 su g g e st t h a t th e human i n f a n t i s n o t as v u ln e ra b le to in f e c tio n 6 to f a i l u r e o f immune f u n c tio n , i t i s p r o te c te d by th e m a te rn a l 7 immune system o f co u rse p r i o r to b i r t h , j u s t as th e r a t f e tu s 8 i s , a f t e r b i r t h i t i s in c o n s id e ra b ly -- i t i s much more 9 com petent in term s o f i t s immune f u n c tio n . You haveto keep 10 i n mind t h a t th e e f f e c t on th e immune system in th e -- on 11 th e c e l l m e d iated im m unity a p p a r e n tly i s n o t an e f f e c t on 12 T lym phocytes p e r s e . They d o n 't seem to ^ b e , th e y d o n 't seem 13 to be i n j u r e d , even though th e thymus i s v e ry s e v e r e ly a f f e c t e d , 14 i t i s one o f th e m ost s e n s i t i v e o rg an s to TCDD. 15 Q So th e e f f e c t i s on th e -- i s on th e thymusg la n d 16 and n o t on th e lym phocytes th em selv es? 17 A I t would ap p e ar so . 18 Q What happens to th e -- w hat do th e s tu d i e s i n d i c a t e 19 happens to th e thymus i n anim als? 20 , A W ell, i f you fo llo w alm o st any e x p e rim e n ta l an im al, 21 and young an im als a r e u s u a lly u sed b ecau se th e thymus i s 22 f u n c tio n a l i n young a n im a ls, and th e r e f o r e more v u ln e r a b le , 23 you f in d t h a t a f t e r TCDD tr e a tm e n t, assum ing t h a t th e dose i s 24 s u f f i c i e n t , th e thymus s h r in k s , i t becomes s m a lle r and s m a lle r .
24
One o f th e re a s o n s I t s h rin k s i s b ecau se a l l o f th e lym phocytes
1
t h a t a r e c o n ta in e d i n i t le a v e . Now,, th e r e i s a c o n tin u a l,
2
probably i t i s n 't th a t the lymphocytes le a v e , i t is more than
3
l i k e l y t h a t no new lym phocytes a re coming i n f o r p ro c e s s in g and
4
the ones th a t were going to leave do. I t appears th a t the
5
6 e f f e c t i s on th e , w e ll, th e thymus i s a gland th a t is com prised p rim a rily of e p ith e lia l c e lls . In o th e r words they are c e lls
7
8 in te re s tin g ly enough th a t are r e la te d back in e a rly development to th e c e lls th a t form the sk in . Those are th e c e lls ap p aren tly
9
t h a t a re a f f e c t e d by TCDD. T here i s r e c e n t work t h a t i s
10
d e sc rib e d in Dr. G re e n le e 's Group has been w orking on th is f o r
11
12 q u ite some w h ile . So th e e f f e c t on th e thymus i s r e a l l y to 13 s to p o r i n t e r f e r e w ith th e f u n c tio n o f th e c e l l s t h a t l i n e 14 th e thym us, and th e lym phocytes t h a t i t p ro c e s s e s a r e sim ply 15 n o t b ro u g h t o u t anymore and th e thymus s h rin k s way down.
Q What a r e th e c l i n i c a l m a n if e s ta tio n s o f an im pairm ent
16
o f th e immune system i n an im als?r
17
18 A W ell, th e r e I s a , you know, we a re exposed c o n s ta n tly to every kind of in fe c tio n th a t you can im agine, ju s t about.
19
And we o c c a s io n a lly c a tc h c o ld , we o c c a s io n a lly g e t g a s tr o -
20 !'h
21 e n t e r i t i s , o r we may even g e t an i n f e c t i o n in some t i s s u e . 22 The m a n if e s ta tio n s o f a f a i l e d Immune system w ould be in f e c t i o u s 23 d is e a s e . Now, t h a t s h o u ld n 't be i n t e r p r e t e d to mean t h a t e v e ry 24 I n f e c tio u s i l l n e s s I s a f u n c tio n o f a f a i l e d immune system
25
1 b ecau se we do g e t i n f e c t i o n s . I have b een f i g h t i n g o f f a c o ld 2 f o r th r e e weeks and I h a v e n 1t f u l l y su cceed ed y e t , b u t I'm 3 f i g h t i n g i t o f f . And t h a t c o ld g o t a f o o th o ld , now maybe i f ss 4 my immune system had been p e r f e c t X w ouldn' t have ca u g h t c o ld . 5 But i f i t r e q u ir e s p e r f e c t i o n , th e n we a r e a l l d e f e c tiv e be 6 cause I d o n 't know o f anyone who n e v e r g e ts c o ld s * But once I 7 g o t th e c o ld , th e n a l l o f my immune fu n c tio n s a r e coming to 8 b e a r i n tr y in g to g e t r i d o f i t . So i f we a re lo o k in g a t th e 9 e x p e rim e n ta l c o n te x t, we would do ex p e rim en ts i n w hich we 10 a d m in is te r an i n f e c t i o u s o rg an ism to an anim al and o b se rv e 11 i t s a b i l i t y to resp o n d , and l o t s o f ex p erim en ts l i k e t h a t a r e 12 done. We c o n tin u a lly m o n ito r anim al c o lo n ie s to se e w h eth er 13 th e r e i s an I n c re a s e d in c id e n c e o f t h i s . I f we a r e s e e in g an 14 I n c r e a s e d in c id e n c e , - fa r more an im als w ith th e s n i f f l e s o r 15 O th er k in d s o f d is e a s e , we know t h a t w e'v e e i t h e r g o t a m assive 16 i n f e c t i o n in th e co lony o r t h a t we im a g in a b ly m ight have some v ' 17 k in d o f an immune d y s fu n c tio n . 18 Q W ell, when th e an im als a re t r e a t e d w ith TCDD, does 19 th e immune system j u s t f a i l co m p letely ? 20 A NOj i t ap p e ars t h a t i t d o e s n 't f a i l , b ec au se a t doses 21 -- th e r e a re e f f e c t s , and a g a in th e dose re sp o n se r e l a t i o n s h i p 22 a p p lie s i n immune f u n c tio n j u s t as I t a p p lie s everyw here e l s e . 23 I t ap p e ars t h a t th e r e i s an u p p er l i m i t . I t i s known v e ry c l e a r l y 24 t h a t an im als t h a t d ie o f TCDD i n t o x i c a t i o n do n o t d ie o f i n f e c t i o n .
26
1 Animals t h a t a re m a in ta in e d , i t seems to me t h a t X made some 2 m ention of th is e a r lie r , anim als th a t are m aintained w ithout 3 any m icroorganism s in th e ir environm ent a t a l l are j u s t as 4 s e n s i t i v e to TCDD as an im als m a in ta in e d i n a norm al e n v iro n 5 m ent. Animals t h a t a re m a in ta in e d o v er a l i f e t i m e , we con 6 tin u a lly seem to m ention Dr. K ociba's work, b u t lo ts o f o th e r 7 lo n g term s tu d ie s as w e ll do n o t show any in c r e a s e i n i n f e c t i o u s 8 d ise a se , and even though they d o n 't speak s p e c if ic a lly o f 9 in fe c tio u s disease, in fec tio u s disease causes pathology ju s t 10 l i k e ch em icals do, and e f f e c t s i n th e lungs and so f o r t h would 11 be i d e n t i f i e d in th e p a th o lo g y r e p o r ts on th e s e k in d s o f s t u d i e s . 12 So th e r e i s a c l e a r u p p er l i m i t to th e d eg ree o f im pairm ent 13 t h a t TCDD m ight c a u se . And we can se e t h i s in ex p e rim en ts i n 14 w hich an im als a r e i n f e c t e d and t h e i r m o r t a l i t y i s m easured 15 fo llo w in g TCDD i n t o x i c a t i o n . 16 Q W ell, i s th e r e a no o b se rv e d e f f e c t l e v e l f o r immune 17 resp o n se ? 18 A In ev e ry ex p e rim e n t'w h e re th e r e has b een an ad e q u ate 19 range o f d o se s, no o b serv ed e f f e c t s l e v e l becomes q u ite a p p a re n t, , 20 W hether th e r e a re ex a m in a tio n s o f s p e c i f i c components o f th e 21 immune system o r w h eth er th ey a r e lo o k in g a t th e a b i l i t y o f 22 an im als to w ith s ta n d i n f e c t i o n . 23 Q In c o n n e c tio n w ith w hat you a re d is c u s s in g th e r e , Dr. 24 D ost, have you caused to be p re p a re d a c h a r t d e a lin g w ith t h a t
27
1 subject? 2 A Yes, I have. 3 Q Would th e use o f t h a t c h a r t a s s i s t you in e x p la in in g 4 i t to the jury? 5 A Yes, i t would. 6 (D e fe n d a n t's E x h ib it 1282 was marked f o r i d e n t i f i c a t i o n .) 7 Q A ll r i g h t , s i r , l e t me show you w h a t's been marked as 8 D efen d an t's E x h ib it 1282 and ask you to examine th a f& n d id e n tif y 9 i t f o r me, p le a s e . 10 A This i s a graph t h a t i s p re p a re d from d a ta , t a b u l a r 11 d a ta t h a t was r e p o r te d by a r e s e a r c h e r named T higpen, t h a t 's 12 an u n f o rtu n a te name, in th e jo u r n a l I n f e c tio n and Im m unity, 13 Volume 12, 1975 in w hich he t r e a t e d mice w ith TCDD f o r fo u r 14 weeks and th en in f e c te d them w ith an organism , t h a t i s , w ith 15 a s a lm o n e lla , th e s p e c ie s i s b u rn ed and t h i s i s an organism 16 t h a t when a d m in is te re d to mice a t s u f f i c i e n t doses i s i n f a c t 17 l e t h a l , and t h i s i s one o f s e v e r a l ex p e rim en ts t h a t he d id , 18 b u t i t does r e p r e s e n t th e g e n e ra l fin d in g s in th e s t u d i e s . 19 Q Now, S-bum ? 20 f. A S alm o n e lla , s a lm o n e lla b u rn , s a lm o n e lla i s a v e ry 21 common genus o f m icro o rg an ism s. T his p a r t i c u l a r one i s one 22 t h a t i s u sed in th e la b o r a to r y and i t does ap p ear o u t i n th e 23 g e n e ra l environm ent a l s o . This i s a g a in a dose re sp o n se c u rv e , 24 and ag a in i t looks a l i t t l e funny b ecau se th e dose re sp o n se i s
^`
- .28
1 , so s t e e p . A l.iliu d i,;t h e ..dosage i s e x p re s se d lo g a r ith m ic a lly ,
..
2 t h a t i s . 1, ,1, and 10 m icrogram s o f TCDD g iv e n w eek ly . And
' 3 th e s e a r e - i n young m ice. And w hat we se e h e re i s ;th a t th e r e ~ ^ '
, i.
.o -
4 is an upper lim it to the
. th is is the m o r ta l ity th i s is
i5 th e p e rc e n ta g e o f an im als t h a t d ie d g iv e n t h a t tr e a tm e n t.
6 This is the co n tro l level,the^num ber of anim als, th e-p ercen tag e .
7 o f an im als t h a t w ere ,g iv e n no TCDD w hich succumbs to t h e 'i n f e c
8 t i o n . What we have h e re i s a t t h i s h ig h e s t dose w hich i s
9 c le a rly an upper lim it because you can see a t the next h ig h e st >
10 dose we a r e up h e re on somewhat l e s s th a n 70 p e r c e n t, and th e n
11 we a r e up h e re to 75 p e r c e n t, som ething l i k e t h a t / a r i d t h a t 's
12 as h ig h as th e m o r t a l i t y w ould go. And when w e 'r e up i n t o
. 'i . r
i-
13 doses l i k e t h i s a t , l e t ' s say 50 o r 60 m icrogram s, some o f th e
/"
i-
14 an im als w i l l sim p ly d ie o f 't h e TCDD a t th e lo w est dose t h a t th e y
15 use h e r e , w hich i s a t a b o u t, w h a t's t h a t going to b e , som ething
16 `on th e o rd e r, o f p ro b a b ly .4 m icrogram s p e r k ilo g ra m p e r week -
17 i n f o u r w eekly d o se s, th e r e was no e f f e c t ; In o th e r w ords,
18 t h a t dose o f TCDD d id n o t in c r e a s e th e s u s c e p t i b i l i t y o f th e s e
19 an im als to i n f e c t i o n . And t h a t i s a common f in d in g w h erev er a
20 dose re sp o n se has b een worked th ro u g h .
21 Q What i s a d i r e c t c h a lle n g e t e s t ?
22 A W eil; a d i r e c t c h a lle n g e t e s t , i f you a r e f a m i l i a r
23 w ith a tu b e r c u lin t e s t , w h a t's jd o n e in tu b e 'rc u lin t e s t s i s to '
24 i n j e c t i n t o th e sk in ', I th in k th ey do i t o n .th e fo re arm I f I
i
?
, t
\ . . -
*
--i i .
29
1 remember c o r r e c t l y , in to , Vthe s k i n ,1n o t -under th e s k in , th e s k in 2 is th ic k , you re a liz e , and so. they in je c t r ig h t in to the dermal 3. la y e r o f th e s k in , e i t h e r -- w e ll, u s u a lly i t i s a k i l l e d 4 p r e p a r a tio n o f tu b e r c u lin o rg an ism s, and th e n w hat w e 'r e lo o k in g 5 f o r i s to se e how th e body resp o n d s to i t . Now i f , you have 6 never had tu b e rc u lo sis the. response w ill be minimal because 7 the body has never learned about tu b e rc u lo sis. I f you have had 8' an i n f e c t i o n r e l a t i v e l y r e c e n t l y and i t ' s b een a c t i v e , why th e r e 9 may be a v e ry v ig o ro u s r e s p o n s e . And i f you h ad an i n f e c t i o n 10 tw enty y e a rs ago i t may be somewhat l e s s e r b ec au se th e r e h as 11 been a l o t o f f o r g e t t i n g go in g on. d u rin g t h a t p e r io d o f tim e. 12 So w hat w e 'r e lo o k in g a t h e re a r e in v a s io n o f w h ite c e l l s to 13 a t t a c k t h a t presum ed in v a d e r. 14 Q Now, w hat does, th e l i n e on t h i s c h a r t mean, th e 15 h o r i z o n t a l l i n e i n th e m id d le o f th e c h a rt? 16 A T hat j u s t i d e n t i f i e s th e c o n tr o l l e v e l o f m o r t a l i t y . 17 In o th e r w ords, an im als t h a t w ere n o t g iv e n TCDD b u t w ere g iv en 18 th e in f o rm a tio n , and t h i s i s a c h a lle n g e , to o , f o r t h a t m a tte r , 19 i t i s an in f e c tio u s c h a lle n g e . We a r e g iv e n th e m icro o rg an ism s 20 in 25 p e r c e n t o f th e an im als t h a t w ere n o t t r e a t e d w ith TCDD d ie d 21 o f th e i n f e c t i o n . The no e f f e c t le v e l i s .5 , I d o n 't remember 22 w hat I e s tim a te d i t a t , b u t i t was c a lc u l a te d , 23 Q As .5 m icrogram s p e r k ilo g ram ? 24 ` A Yes, i t i n t e r s e c t s th e .5 .
1.
* jh
y i 1 - \ - ; / .' ,
(
30
1 Q With re s p e c t to the' dose th a t you c a lc u la te d from
I ~ i1
'
2 S tu rg eo n s o i l , would th a t^ d o s e be, s u f f i c i e n t ; to- e f f e c t th e
J
^ ' V " f _ l* i-
.
3 immune system i n t h e 'mice t h a t w ere --
1.
>
4 A As r e f l e c t e d h e r e , 'n o t, t h i s no e f f e c t l e v e l i s a
5 h a lf a microgram, and th is i s a thousandth o f a microgram,
6 t h a t 's a d iffe re n c e o f, w hat, 500 fo ld ..
7 Q So i t w ould be 500 tim es l e s s th a n th e dose n e c e s s a ry
8 to cau se an immune --
9 A . 1 beg your pardon, I'm looking a t th is o th e rn o
10 e f f e c t l e v e l . I m is s ta te d m y s e lf. We h av e , o r hav e I? No,
11 we h av e w ith r e s p e c t to th e dose a t , t h a t we c a l c u l a t e d a t
12 S tu rg e o n , f o r th e sake o f s i m p l i c i t y c a l l t h i s .5, o r change
13 i t up to 5 , and t h a t means t h a t we have a 10,000 f o l d d if f e r e n c e .
14 Q 10,000 f o l d d if f e r e n c e ?
i5 A W ell, th e d if f e r e n c e betw een p o i n t -- we have th e
16 d if f e r e n c e betw een .5 and .00005, e s s e n t i a l l y . - , . , .
*j i
i '*
17. Q So' th e S tu rg eo n dose c a l c u l a t e d by you would be
j
> 1
18 10,000 tim es l e s s th a n t h a t r e q u ir e d to b e g in to show an e f f e c t
19 i n th e m ice?
20 I " A ' In t h i s , d i r e c t c h a lle n g e e x p e rim e n t, y e s i
21 Q Now, when th e a n im a l's immune system i s a f f e c t e d by
22 a s u f f i c i e n t dose o f TCDD, i s t h i s a perm anent e f f e c t ?
23 r A No., i t w i l l re c o v e r, b u t i t ta k e s -- i n ta k e s a
24 s u b s t a n t i a l amount o f tim e i n an e x p e rim e n ta l aniihal:? i t w i l l
31
1 take months,
< .,
2 Q To re c o v e r?
1
3 A To r e c o v e r, ` .j, .
.`
, 'V '. '
~
4 Q But does recovery-bccur? - '
5
A E v en tu ally these, r e tu r n to. n o rm al..
,
,- >
'
- 1
/
; - >* / -v
M" * 1 *
*^ T j
6 THE COURT: '' B efo re you g e t' I n to y o u r n e x t s u b je c t,
7
i s t h i s a good p o in t f o r va s h o r t'b r e a k ?
. -
8 - MR; HEINEMAN.-; Y es, s i r , f i n e .
9 THE COURT: Okay, we w i l l take, a s h o r t b re a k a t t h i s
tim e. I would rem ind you, and th is would go f o r any o th e r breaks
10
11 we ta k e d u rin g th e day,' t h a t you a r e n o t to d is c u s s t h i s m a tte r
12 among y o u r s e lv e s , w ith anyone o u ts id e th e j u r y p a n e l, o r as o f
13 y e t form any o p in io n s cor c o n c lu sio n s ab o u t th e m a tte rs On t r i a l .
14 The C o u rt i s i n r e c e s s .
15 (At t h i s tim e C o u rt was i n r e c e s s '.)
16 MR. HEINEMAN: Your Honor, a t t h i s tim e we w ould move
17 th ad m issio n o f D efen d a n t's E x h ib it 1282.
18 (D e fe n d an t's E x h ib it 1282 was o f f e r e d i n t o e v id e n c e .)
18 THE COURT: A n y ^o b jectio n ?
2C ". '
MR, CARR: None, y o u r H onor/
,21 THE .COURT: A dm itted w ith o u t o b je c ti o n . Thank you.
(D efen d an t's E x h ib it 1282 was ad m itted in to e v id en ce .)
k
*
Q . (By Mr. Heineman) Dr. D ost, i n y o u r rev ie w o f th e
te stim o n y o f LDr. Z ahalsky on b e h a lf o f th e p l a i n t i f f s , do you
32
1 r e c a l l h is t e s t i f y i n g ab o u t a stu d y by a man named Boorman? 2 A Yes. 3 Q L et me show you w h a t's been marked as P l a i n t i f f s ' 4 E x h ib it 260, y e s, i t i s 260. Have you seen th a t b e fo re , s ir ? 5 A I'v e seen a photograph of i t .
6 Q A ll rig h t, is th is the photograph th a t you have seen,
7 sir? 8 A Yes, i t is . 9 (D efen d an t,1s E x h ib it 1283 was marked f o r i d e n t i f i c a t i o n . ) 10 Q What has b een m arked as D e fe n d a n t's E x h ib it 1283, s i r ? 11 I s t h a t a p h o to g rap h o f P l a i n t i f f s ' E x h ib it 260? 12 A Y es, i t is* 13 Q Now, a r e you f a m i l i a r w ith th e Boorman a r t i c l e i t s e l f , 14 s i r ? 15 A Y es, I am. 16 Q And d id you cau se a c h a r t to be made o f some o f th e d a ta 17 from some o f th e d a ta i n th e r e -- ,18 A There i s a t a b l e i n th e p a p e r, and i t was c o p ie d d i r e c t l y 19 from th e , t h a t i s , i t i s n ' t a photocopy o f th e p ag e, b u t th e 20 d a ta h as been co p ied i n e s s e n c e in ^ a t a b l e . 21 Q And b a se d upon y o u r rev iew o f th e te stim o n y o f Dr. 22 Z ah alsk y , i s t h a t th e ta b l e from w hich Dr. Z ahalsky p u rp o rte d 23 to c r e a te t h i s e x h ib it? 24 A Y es, i t i s .
33
1 (D e fe n d a n ts E x h ib it 1284 was marked f o r i d e n t i f i c a t i o n .) 2 Q Now, l e t me show you w h a t's b een marked as D e fe n d a n t's 3 E x h ib it 1284, s i r . Can you examine t h a t and i d e n t i f y i t f o r me, 4 please? 5 A Yes, i t i s a copy o f a ta b le th a t was in an a r t i c l e
6 by Boorman th a t was p u b lis h e d in E nvironm ental H ealth P e r s p e c tiv e s ,
7 and we do n o t have a s p e c i f i c r e f e r e n c e on h e r e , b u t i t i s th e same 8 artic le . 9 Q I t i s ad ap ted , i t ' s tak en d ir e c tly from t h i s Boorman 10 a r t i c l e ? 11 A I t i s ta k e n from th e a r t i c l e and i t i s th e d a ta from 12 w hich Dr. Z a h a ls k y 's c h a r t o r ta b l e was d e riv e d . 13 Q A ll r i g h t . Now, do you a g re e , a i r , w ith th e te stim o n y 14 o f Dr. Z ahalsky w ith r e s p e c t to t h a t c h a rt? 15 A No, I do n o t. 16 Q A ll r i g h t , I wonder i f you w ould s te p up h e r e and 17 e x p la in to th e j u r y why you d o n 't a g re e w ith t h i s c h a r t . 18 A I ' l l p u t t h i s up h e re w here i t can be se e n . The re a s o n 19 t h a t I d o n 't a g re e i s b ecau se some v e ry im p o rta n t d a ta has b een 20 l e f t o u t o f th e c h a r t and th e r e has b een an i n c o r r e c t r e p r e s e n 21 t a t i o n o f th e d o se, and a ls o i n re a d in g th e te stim o n y , I saw no
d e sc rip tio n of the experim ent and the n ature o f the experim ent is o f co n sid erab le im portance because the anim als -- and i t was n e c e ssa ry to go to th e o r ig in a l a r t i c l e , Boorman's a r t i c l e
34
1 i s a review , and they reproduced from an e a r l i e r paper done by 2 th e same group, th e s e d a ta , and in th e e a r l i e r p ap e r they des 3 c rib e more com pletely th e manner o f tre a tm e n t o f th e anim al. 4 What th e y d id was to a d m in is te r to p re g n a n t m ice TCDD sev en days 5 p rio r, I b eliev e , to the b irth of the pups, and then a t seven and 6 fo u rte e n and tw enty-one days fo llo w in g during th e p e rio d when 7 they are n u rsin g , so th a t the anim al, the pups were presumably 8 exposed w hile in the u te ru s, and then exposed through the m ilk, 9 and t h i s a common way o f ex p o sin g v e ry young ro d e n ts i n th e 10 e x p e rim e n ta l c o n te x t, t h a t i s , to t r e a t th e m other and d e l iv e r 11 th e m a te r ia l i n m ilk . There i s a v e ry s e r io u s e r r o r made i n 12 r e p r e s e n tin g th e do sag e. When we sp eak o f p a r t s p e r t r i l l i o n 13 o r m il lio n o r b i l l i o n , we a r e ta lk in g ab o u t c o n c e n tr a tio n . We 14 do n o t u se t h i s r e p r e s e n ta ti o n i n d is c u s s in g d o sag e, b e c a u se th e 15 o n ly th in g t h a t dosage t e l l s us i s how*, much w en t i n t o th e an im al 16 f o r u n i t o f body w e ig h t. I t d o esn f t t e l l us a n y th in g a b o u t th e 17 c o n c e n tr a tio n i n th e l i v e r o r i n bone o r i n th e k id n e y o r th e 18 b r a in o r any o th e r t i s s u e . The c o n c e n tr a tio n s w ith in o rg an s i s 19 g o in g to v a ry w ith th e o rg a n . So t h i s t e l l s n o th in g a b o u t w hat 20 i s , i t d o e s n 't r e p r e s e n t w h a t's h ap p en in g i n th e a n im a l, n e i t h e r 21 does th e dose i n m illig ra m s p e r k ilo g ra m , b u t i t i s th e o n ly 22 in fo rm a tio n we have when we a d m in is te r a ch em ical to an an im al. 23 We d o n 't know w here i t w en t, we know enough ab o u t a ch em ical we 24 can sa y , y e s , so much o f i t i s i n th e l i v e r and th e k id n e y and
35
1 so f o r t h . The o th e r e r r o r w hich i s e x tre m e ly c r i t i c a l , even i f 2 we a c c e p t t h i s id e a a f t e r p a r t s p e r t r i l l i o n as r e p r e s e n tin g th e 3 dose, i f i t were p a r ts p e r t r i l l i o n , i t would mean t h a t th e dose 4 was ,005 micrograms p e r k ilo g ram and .015 microgram s p e r k ilo g ram , 5 a c c e p tin g f o r th e moment t h a t t h i s im proper r e f e r e n c e i s th e way 6 to u se th e la n g u ag e. The f a c t o f th e m a tte r i s t h a t th e doses 7 th a t were adm inistered were a tho u san d fo ld , h ig h er. This is 8 m illigram s p er kilogram , a thousandth of a m illigram per kilogram 9 is one microgram. This is an e ra sa b le -- the dose is one m icro 10 gram p e r k ilo g ra m p e r week. So th e dosage h e r e h as b ee n r e p r e 11 s e n te d as a th o u san d tim es low er th a n i t r e a l l y w as, I d o n 't 12 have a g r e a t d e a l o f ob jectio n * , to changing th e te rm in o lo g y 13 b e c a u se Dr. Z ahalsky was tr y in g to e x p la in th e n a tu r e o f th e s e 14 c e l l s . He d id u se a d i f f e r e n t co n v e n tio n th a n th e a u th o rs d id 15 f o r d e s c r ib in g th e r e s u l t s . He t a l k s ab o u t p e rc e n ta g e o f in 16 h i b i t i o n , t h a t i s , t h a t means how much o f th e a c t i v i t y o f th e 17 th o s e c e l l s was d e c re a se d . W hereas, th e a u th o rs s p e a k .to how 18 much a c t i v i t y was l e f t , . T h a t 's n o t s e r io u s , t h a t 's j u s t a 19 c o n v e n tio n . But I w ant to p o i n t o u t t h a t he i s t a l k i n g a b o u t 20 i n t h i s c a se th e lo s s o f com petency and th e lo s s o f com petency 21 o b v io u sly would be g r e a t e r as th e dosage in c re a s e s * The im por 22 t a n t th in g i s however t h a t h e l e f t o f f th e lo w e s t d o se , he l e f t 23 o f f th e lo w e st d o se. Now, t h i s in fo rm a tio n , t h i s i s th e number 24 o f c e l l s i n th e bone marrow. I t i s n o t j u s t c e l l s making r e d
36
1 blood c e lls , b u t a l l c e lls in th e bone marrow. These a re c e lls 2 th a t w ill ev en tu ally form red blood c e lls as he s ta te s , th ese 3 are c e lls th a t ev en tu ally w ill form macrophages, s im ila r c e lls 4 th a t a c t u a l l y , , I m entioned c e l l s th a t go o u t and su rro u n d , 5 e n g u lf f o re ig n m a te r ia l. The im p o rta n t in fo rm a tio n in th is 6 p a p e r i s th e dose re sp o n se r e l a t i o n s h i p . Now, t h e s e .o t h e r 7 c h e m ic a ls, d o n 't w orry ab o u t -- we sim ply w anted to re p ro d u c e 8 a l l of the m a te ria l so th a t you can see what th e o rig in a l ta b le 9 looks lik e . This is th e d ata th a t w e're concerned w ith , and 10 s u r e ly enough th e r e i s a dose re s p o n s e , one m icrogram p e r 11 k ilo g ra m p e r week, 5, 13. And as you m ight e x p e c t, h e r e i s an 12 e f f e c t , 15 m icrogram s i s a good s iz e dose f o r any a n im a l, any 13 r o d e n t, and h e re o n ly 61 p e r c e n t o f f u n c tio n has b een l e f t , 14 and th e y d e s c rib e th et e s t a t i s t i c a l v a l i d i t y o f th a t* In 15 o th e r w ords, t h a t i s a r e s u l t i n w hich th e y have g r e a t c o n fid e n c e . 16 T his i s a ls o a r e s u l t t h a t th e y have c o n fid e n c e i n , b u t somewhat 17 l e s s b ec au se i t i s n ' t t h a t f a r removed from th e c o n t r o l; These 18 numbers a re p e rc e n ta g e o f c o n t r o l , and t h a t 's th e re a s o n we 19 d o n 't se e a s e p a r a te l i n e f o r th e c o n tr o l in fo rm a tio n ; In t h i s 20 one, th e y d i d n 't d eterm in e th e e f f e c t f o r w h ate v er re a s o n . At 21 th e lo w est dose, b u t a g a in th e r e i s 68 p e r c e n t o f th e f u n c tio n 22 l e f t h e r e , 37 p e r c e n t a t t h i s h ig h e r d o se, w hich i s n o t s u r p r i s i n g . 23 In t h i s c a se th e r e was a more s e v e re e f f e c t even th a n th e o th e r s 24 a t 15 m icrogram s p e r k ilo g ra m . Very l i t t l e e f f e c t a t 5 , and you
37
1 can see h ere th a t they c o n sid er, o r th e s t a t i s t i c s t e l l them 2 th a t 90 p e rc e n t i s so n e a r to a hundred p e rc e n t in th is e x p e ri 3 ment t h a t you c a n 't t e l l th e d if f e r e n c e . And h e re a t one m icro4 gram p e r kilogram is a c tu a lly th e se anim als have more than the 5 norm al amount o f c e l l s , d o n 't ta k e t h a t as meaning t h a t a low 6 dose o f TCDD makes th e bone marrow work b e t t e r . T his i s th e 7 kind of v a ria tio n th a t one is going to expect in an experim ental 8 c o n te x t. What I w ant to show you h e r e , though, i s we have a 9 no e f f e c t h ere -at one microgram p e r kilogram . A p lo t o f a dose 10 re sp o n se cu rv e w hich s u g g e sts t h a t you a r e go in g to have a no 11 e f f e c t l e v e l a t t h i s p o in t, b u t th e d a ta i s n ' t th e r e and a l l we 12 can do i s say t h a t i s su g g e ste d . H ere we c l e a r l y have anno 13 e f f e c t dose w ith g iv e n th e s t a t i s t i c s , th e an im als a r e even 14 b e t t e r th a n c o n tr o ls . A gain, when I se e in fo rm a tio n l i k e t h a t 15 i t d o e s n 't mean th e y a r e b e t t e r , i t means t h a t th e r e i s enough 16 s t a t i s t i c a l v a r i a b i l i t y , b u t g iv e n th e lo c a ti o n o f t h i s d i e 17 p o in t and t h i s p o in t, i t i s v e ry c l e a r t h a t th e r e i s ' a no e f f e c t 18 dose h e r e . So a t th e dose l e v e l t h a t he d id n o t in c lu d e i n 19 h i s te stim o n y th e r e i s c l e a r l y a d o se, a no e f f e c t d o se. 20 ; Q Now, w hat i s th e no e f f e c t le v e l th e r e , Dr. D o st, 21 i s one m icrogram p e r k ilo g ram ? 22 A In t h i s ex p e rim en t, y e s . 23 Q Now, was t h i s an a c u te ex posure? 24 A W ell, i t to o k p la c e o v er a p e r i o d .o f , I th in k , fo u r
38
1 weeks in which th e m a te ria l was ad m in istered p r io r to b ir th of 2 the pups, and adm inistered p e rio d ic a lly a f te r the b ir th of the 3 pups, so th a t th ere were fo u r doses a t th is le v e l and 1 think 4 th e t o t a l p e rio d was over fo u r weeks. 5 Q So would t h a t be c a l l e d a c h ro n ic exposure? 6 A No, i t w ould be -- o f co u rse when you d e f in e th o se 7 te rm s, why everybody h as h i s own d e f i n i t i o n o f w hat i s a c u te , 8 I guess I would c a ll i t , i t is a sh o rt subchronic experim ent, 9 b u t i t could even be c a lle d a long acute experim ent. . I t is a 10 l i t t l e d i f f i c u l t to say when you have t h a t k in d o f a d o sin g 11 sc h e d u le . T h e ir o b je c tiv e was to m axim ize, a t a g iv e n l e v e l 12 o f in p u t th e y w anted to maximize th e ex p o su re to th o s e p u p s. 13 Q Now, w h a t's th e d if f e r e n c e betw een th e no e f f e c t 14 l e v e l o f TCDD d em o n strate d on t h a t d o sin g o f th o s e r a t pup 15 and th e c o n c e n tr a tio n o r th e ex p o su re l e v e l o r th e d o se, 16 p ard o n me, th e dose computed from th e S tu rg eo n in c id e n t? 17 A W ell, w e 'r e b ack a t m icrogram p e r k ilo g ra m a g a in , 18 and I f I remember t h a t would b e , th e d if f e r e n c e would be 19 222, 000. 20 Q 222,000 w hat? 21 A 222,000 f o ld d if f e r e n c e b etw een, t h a t i s t t h e c d o s e tth a t 22 w e 'v e j u s t b een sp e ak in g o f h e re a t one m icrogram p e r k ilo g ra m 23 p e r week, o r w h atev er th e p e r io d would b e , i s b u t one o f th o se 24 d o se s, one o f th o s e fo u r doses i s 222;;000 tim es h ig h e r th a n
39
1 t h i s ex p o su re t h a t we h y p o th e s iz e d . 2 Q From Sturgeon? 3 A From S tu rg eo n . 4 Q Now, and t h a t was a no e f f e c t l e v e l when i t was 5 a d m in iste re d - - when th a t dose, 220,000 tim es more was 6 ad m in istered once a week fo r fo u r weeks? 7 A I t i s my r e c o l l e c t i o n t h a t t h a t was th e s c h e d u le , 8 i t was one time p r io r to the b ir th o f the pups, and then th re e 9 tim es afterw ard. 10 Q Now, Dr. Z ahalsky t e s t i f i e d ab o u t p r o te a s e s . Do 11 you r e c a l l t h a t testim o n y ? 12 A I remember som ething ab o u t I t , y e s . 13 Q He su g g e ste d t h a t p r o te a s e s -- 14 MR. CARR: What p ag e , Counsel? 15 MR. HEINEMAN: 135. 16 MR. CARR: What day? 17 MR. HEINEMAN: May 3 rd . 18 Q He s u g g e s te d t h a t p r o te a s e s a c t on p r o t e i n s and a t t a c k 19 t i s s u e . They a t t a c k p r o t e i n i n t i s s u e , do you r e c a l l t h a t 20 , A Yes. 21 Q Do you a g re e w ith w hat Dr. Z ahalsky s a id ? 22 A Only in p a r t . 23 Q A ll r i g h t . Would you t e l l u s ab o u t y o u r d isa g re e m e n t 24 th e re ?
40
1 A W ell, p r o te a s e s a r e i n f a c t enzymes t h a t b re a k down 2 p r o te in . They do n o t a c t , when you say t i s s u e s , I am assum ing 3 th at is living tissu es. 4 Q Yes, s ir . 5 A They do n o t a c t i n l i v i n g t i s s u e s e x c e p t u n d er e x t r a 6 o rd in a ry circu m st a n c s . They o n ly a c t , l e t me e x p la in . P ro te a s e s 7 a r e p r e s e n t i n a l l c e l l s . Now, I'm u s in g th e term v e ry g e n e r a lly , 8 i t i s my r e c o l l e c t i o n he m en tio n ed a co u p le o f p r o t e i n s i n h e r e . 9 But proteases e x ists in c e lls fo r the purpose of degrading 10 p r o t e i n s , enzymes, o th e r p r o te in s i n th e c e l l when th e y a r e no 11 lo n g e r w orking and have to be d isp o s e d o f . In f a c t , p r o te a s e s 12 a r e r e s p o n s ib le f o r r e g u la tin g th e l i f e t i m e o f c e r t a i n p r o te in s 13 i n th e c e l l , and t h i s i s a v e ry e f f e c t i v e sy stem . I f a p r o t e i n 14 sh o u ld n o t s ta y th e r e to o lo n g , i t w i l l be in th e c o u rse o f tim e 15 depending on i t s s i z e be d egraded by th e p r o te a s e s t h a t a r e 16 alw ays i n th e c e l l . There a re a ls o p r o te a s e s i n c e r t a i n c e l l s 17 in th e c i r c u l a t i o n and t h e i r r e s p o n s i b i l i t y , and th e y a r e p a r t
o f th e immune re s p o n s e , to o , in a se n se b ec au se t h e i r f u n c tio n i s to b re a k down. I f a c e l l e n g u lfs a n o th e r c e l l , i t ' s g o t to dp som ething w ith i t , i t c a n 't j u s t c o n tin u e to h o ld o n to t h a t c e l l . I t ' s g o t to b re a k I t down, i t ' s g o t to d ig e s t i t so th e c e l l e v e n tu a lly , th a t i s p ick ed up e v e n tu a lly i s d ig e s te d . There are a c e rta in amount o f fre e p ro te a se in th e c irc u la tio n as w e ll
they w ill tend to leak out o f c e lls th a t have fa ile d ,
41
1 they w ill leak out of the macrophages th a t are in the tis s u e s
2 and in the blood, ju s t in the normal course of a t tr i tio n of
3 th e c e l l s . So th e r e i s alw ays a l e v e l o f a v a r i e t y o f p r o te a s e s
4 t h a t b re a k down v a r io u s k in d o f p r o t e i n i n th e c i r c u l a t i o n . And
5 i f a c e l l is in , a c e ll i s se v e re ly damaged, fo r example, a
6 lung c e ll, i f a heavy smoker, the c e ll is a liv e and i t is
7 f u n c tio n a l b u t i t i s n o t w e ll . And i n t h a t c irc u m sta n c e a
8 c e ll in th a t m arginal zone between fu n ctio n and nonfunction or
9 l i f e and death could very w ell be a tta c k e d by p ro te a se s th a t
10 c i r c u l a t e . But th e y d o n 't a t ta c k h e a lth y t i s s u e .
11 Q Now, w hat i s th e p u rp o se o f them? As 1 u n d e rs ta n d
i
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12 i t , a p r o te in i s a c h a in o f amino a c id s , d i d n 't you t e l l us
. 13 t h a t th e o th e r day?
_
' ' \\
14 A T h a t's c o r r e c t .
15 Q Okay, now when th e body -- and does th e body t ^ 4l - "
16 -norm ally d is p o s e o f p r o t e i n as an every d ay o c c u rre n c e ?
17 A Oh, c e r t a i n l y , b e c a u se th e y e i t h e r w ear o u t o r th e y
18 a r e no lo n g e r n eed ed , and s in c e we do n o t . r e p a i r a p r o t e i n
19 t h a t i s n ' t w orking p r o p e r ly , th e body d eg rad es i t , makes u se
20 -pf th o se amino a c id s , s t a r t s o v e r a g a in , u se s them f o r w hat
21 e v e r i t n eed s amino a c id s f o r .
22 Q Is i t som ething l i k e u s in g b u ild in g b lo c k s?
23 A W ell, th e s t r u c t u r e o f a p r o t e i n , y e s , i s l i k e b u ild in g
24 b lo c k s . There a r e a r e l a t i v e l y lim ite d number o f amino a c id s ,
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43
X is always leakage and they appear in the c irc u la tio n . 2 Q Now, Dr. Z ahalsky on ab o u t th e same page o r th e n e x t 3 p ag e, I guess 136, t a l k s a b o u t some m a te r ia ls c a l l e d A lpha 1 4 a n t i - t r y p s i n and A lpha 2 m a c ro g lo b u lin , do you remember? 5 A Yes. 6 Q Those term s in th a t testim ony? 7 A Yes. 8 Q Now, do you a g re e w ith w hat he s a i d about th o se 9 m olecules in th a t testim ony? 10 A I t was a l i t t l e d i f f i c u l t to u n d e rs ta n d , b u t b a s i c a l l y 11 no, I don' t a g re e . 12 Q A ll r i g h t ; P o i n t 'o u t to th e j u r y i f you w ould w hat 13 Dr; Z ahalsky s a i d ab o u t th e s e m a te r ia l s and th e a r e a o f your 14 d isa g re e m e n t. 15 A Those two s u b s ta n c e s a re i n h i b i t o r s o f p r o te a s e s and 16 th e y e x i s t in th e c i r c u l a t i o n . They a r e v e ry im p o rta n t com 17 p o n en ts o f th e plasm a p r o t e i n p r o f i l e . In f a c t , th e y 'r e even 18 g r e a t e r q u a n t i t i e s th a n , l e t ' s sa y , th e im m unoglobulins, w hich 19 we depend on f o r immune re sp o n se . T h e ir f u n c tio n i s to i n h i b i t 20 th e a c t i v i t y o f th e s e p r o te a s e s t h a t we have b een t a l k i n g ab o u t 21 t h a t in e v ita b ly escap e from c e l l s . In o th e r w ords, th e y p r o t e c t 22 us from p r o te a s e s . And th e r e a r e a number o f g e n e tic d is e a s e s 23 i n w hich th e s e p r o te a s e i n h i b i t o r s , p a r t i c u l a r l y th e A lpha 1 24 a n t i - t r y p s i n , th e re a s o n i t i s c a lle d a n t i - t r y p s i n i s b ec au se
1 I
\
44
1 i t i s very e f f i c i e n t o f the enzyme try p s in which i s found in
2 c e lls and th a t is a ls o an enzyme th a t i s s e c re te d in to th e
3 d ig e stiv e tr a c t to d ig e st meat. But th ere are g en etic d efects
4 in which th e re is a d eficien cy of the a n ti-try p s in and people
5 w ith th a t d e fic ie n c y seem to have a g r e a te r in c id e n t o f '
6 pulm onary emphysema and p ro b a b ly th e re a so n f o r t h a t i s in th e
7 normal course of a ssa u lts th a t our lungs take, w hether w e're
8 smokers o r n o t, th e re are in ju re d c e lls and those c e lls are
9 v u ln e ra b le o r i f try p s in and o th e r enzymes have access to those
10 c e l l s , t h e r e 's g o in g to be some o th e r k in d o f b re a k down and
11 t h a t r e s u l t s in emphysema. The main th in g a b o u t h is s ta te m e n t
12 i s t h a t he d e s c rib e s a n t i - t r y p s i n more o r l e s s e f f e c t i v e l y , b u t
13 th e n he t a l k s ab o u t th e A lpha 2 m a c ro g lo b u lin w hich i s j u s t
14 a n o th e r k in d o f i n h i b i t o r as b e in g some k in d o f a r e g u l a t o r
15 f o r a n t i - t r y p s i n , and i t i s n o t. He a ls o makes a p e c u l i a r
16 s ta te m e n t t h a t i f you in c r e a s e th e amount o f A lpha 2 m acro
17 g lo b u lin , and I d o n 't know how t h a t i s going to come to p a s s ,
18 b u t i f you in c r e a s e th e amount o f A lpha 2 m a c ro g lo b u lin you
.^
li
19 in c r e a s e th e a c t i v i t y o f th e p r o te a s e s , and i t i s JAist th e
20 o th e r way aro u n d , i t is^ j u s t th e o th e r way aro u n d .
21 Q What do you mean " j u s t th e o th e r way around"?
22 A W ell, i f you in c r e a s e th e amounto f A lpha 2 m acro
23 g lo b u lin , i t s f u n c tio n i s to i n h i b i t th o s e p r o te a s e s , n o t to
24 in c re a s e them. I t p r o t e c t s th e body a g a in s t th e p r o te a s e s .
45
1 Q Well now# i f p ro te a se s don't a tta c k h e a lth y tis s u e ,
2 why does th e body have i n h i b i t o r s , p r o te a s e i n h i b i t o r s ?
3 A W ell, I already mentioned th a t even in th b e st of
4 circum stances c e lls lo se , they break up, th e y ire going to lo se
5 t h e i r -- th e p r o te a s e s , th e enzymes t h a t b re a k down p r o t e i n
i 6
th a t are in sid e the c e ll, th a t are th ere fo r norm ally to dispose
7 of unwanted oarnonfunctional p ro te in s in c e lls , and I th in k I
8 m entioned th e o th e r day about when the l i v e r i s in ju r e d , i t
9 s p i l l s some o f i t s enzymes i n to th e c i r c u l a t i o n . The c e l l s a r e
10 n o t b ro k en , b u t th e y 've become le a k y . The same th in g can happen,
11 a le a k y c e l l presum ably can lo s e i t s p r o te a s e s , i t s t r y p s i n , e t c . ,
12 o r a c e l l t h a t c o m p letely b re a k s up. So t h e r e 's alw ays g o in g to
13 be some l e v e l , even though i n an i d e a l s t a t e th e r e w o u ld n 't be
14 any, t h e r e 's go in g to be some, and we have ev o lv ed t h a t k in d o f
15 p r o t e c t i o n a g a in s t t h a t a c t i v i t y b ec au se we alw ays have c e l l s
16 i n o u r body t h a t a re p erh ap s n o t p e r f e c t l y h e a lth y , and a c e l l
17 t h a t i s m a rg in a lly h e a lth y and h as been in j u r e d i n some way
18 w ould q u ite im ag in ab ly be a f f e c t e d by an enzyme t h a t w i l l b re a k
19 down p r o t e i n . And so we need to p r o t e c t a g a in s t t h a t .
20 Q Now, Dr. D ost, i s th e r e any e v id en ce from an im al
21 s tu d ie s o r from anywhere e l s e t h a t you a re aware o f t h a t TCDD
;
22 a f f e c t s p r o te a s e s o r th e p r o te a s e i n h i b i t o r s A lpha 1 a n t i - t r y p s i n
23 and Alpha 2 m a c ro g lo b u lin ?
24 A I know o f no e v id e n c e , I d o n 't th in k th e q u e s tio n h as
46
1 even been examined. 2 HR. HEINEMAN: Your Honor, i t ' s noon, I n o t i c e , i f 3 i t 's a l l rig h t w ith the Court. 4 THE COURT: Okay, f i n e . We w i l l b re a k f o r lu n c h a t 5 t h i s tim e . We w i l l resume a g a in a t 1 :1 5 , and th e adm onishments 6 th a t I have given you e a r lie r w ill apply during th is break also . 7 The C o u rt i s i n r e c e s s . f o r lu n c h . 8 (At th is tim e Court recessed fo r the noon h o u r.) 9 Q (By Mr. Heineman) Dr. D ost, when we b ro k e f o r lunch 10 we h ad j u s t f i n i s h e d ta lk in g ab o u t th e Boorman s tu d y , and I 11 w anted to ask you a l i t t l e more ab o u t t h a t . Do you r e c a l l 12 t h a t Dr. Z ahalsky t e s t i f i e d t h a t th e e f f e c t o f d io x in on th e 13 immune sy stem c o u ld be i n i t i a t e d by one m o le c u le ,o f d io x in ? 14 A Yes, I r e c a l l t h a t . 15 Q A ll r i g h t . Now, do you a g re e w ith t h a t s ta te m e n t? 16 A No. 17 Q Now, w hat would be th e p o s s i b i l i t y o f an ex p o su re 18 to o r a dose o f one m o lecu le ca u sin g an a d v e rse r e a c t i o n i n 19 an organism ? 20 A No p o s s i b i l i t y a t a l l . 21 Q Why i s t h a t , s i r ? 22 A There a r e a number o f r e a s o n s , even i f th e s u b sta n c e 23 was h ig h ly r e a c t i v e i t is- n e c e s s a ry f o r a r e a c tio n to ta k e p la c e 24, t h a t th e r e be a p o p u la tio n o f m o le c u le s. There would have to be
47
1 a lo t of molecules p rese n t, a re a c tio n th a t involves a sin g le 2 m olecule out of a p o p u latio n , a la rg e population m ight imagin 3 ably take p la ce, b u t th e re are time fa c to rs . M olecules are 4 v e ry , very liv e ly th in g s . They don' t j u s t s i t in one p la c e 5 a l l by th e m se lv e s. And f o r a r e a c tio n to ta k e p la c e , tim e i s 6 n ec essary . Chemical reactio n s are n o t j u s t in sta n ta n e o u s. I t 7 may seem l i k e i t when we lo o k a t them i n a to y c h e m istry s e t 8 and p o u r two c l e a r l iq u i d s to g e th e r and th e y su d d en ly tu r n 9 p u rp le , b u t th e re is a g re a t deal of time involved. A m olecule 10 has to g e t i n to a p o s it io n w here i t can r e a c t , and th e n i t has 11 to be h e ld th e r e f o r some f i n i t e p e r io d i n o rd e r f o r w h ate v er 12 i t i s go in g to i n t e r a c t w ith to r e a c t , f o r th e two o f them to 13 i n t e r a c t to g e th e r . O th erw ise , i t may come i n t o c o n ta c t and be 14 gone a g a in i n an i n s t a n t and more th a n l i k e l y n e v e r to r e t u r n 15 a g a in to t h a t s i t e . 16 Q W ell, a m o lecu le upon e n t e r in g th e body, w hat happens 17 to i t ? Where would i t go? 18 A Anywhere. One m o le c u le , we a r e n o t accustom ed to 19 th in k in g in term s o f one m o le c u le . We a re accustom ed to th in k in g 20 i n term s o f la r g e p o p u la tio n s o f m o le c u le s, and even a dose o f 21 a m icrogram o r a th o u sa n d th o f a m icrogram i s a v e ry , v e ry 22 la r g e number o f m o le c u le s. 23 Q W ell, how does th e f a c t t h a t th e Boorman stu d y e s t a b l i s h e s 24 a no e f f e c t l e v e l a f f e c t Dr. Z a h a lsk y ' s th e o ry t h a t one m o lecu le
48
1 can i n i t i a t e t h i s immune f u n c tio n d i f f i c u l t y ? 2 A W ell, i n terms, o f t h i s p a r t i c u l a r s tu d y , we se e a no 3 e f f e c t le v e l as a microgram p e r kilogram . There a re o th e r stu d ie s 4 lo o k in g a t o th e r p aram ete rs t h a t show somewhat low er no e f f e c t 5 le v e ls - We a re s t i l l ta lk in g ab o u t enormous numbers o f m o le c u le s. 6 Q Now, Dr. Z ahalsky h as s a id , s i r , t h a t when t h i s one 7 m o lecu le would i n i t i a t e t h i s immune f u n c tio n problem , th e n o th e r 8 d io x in m olecules would be re q u ire d to deal w ith th e a d d itio n a l 9 m essenger KNA and r e c e p to r s , do you r e c a l l th a t? 10 A I r e c a l l som ething o f t h a t s o r t , y e s . 11 Q Now, would a sm a ll number o f m o le c u le s, a h a n d fu l o f 12 m o le cu les have any p o s s i b i l i t y o f c a u sin g th e immune f u n c tio n 13 e f f e c t t h a t i s r e f l e c t e d in th e Boorman stu d y ? 14 A Not r e a l l y . Maybe I sh o u ld i l l u s t r a t e w hat I t a l k 15 ab o u t when X say th e r e w ould have to be an enormous number o f 16 m o le c u le s, b ec au se 17 Q A ll r i g h t . 18 A I t happens t h a t we know how many m o lecu les a r e i n a 19 g iv e n amount o f a ch em ical. I t may seem l i k e an a w fu lly e s o t e r i c 20 k in d o f th in g to know, b u t I guess w hat I have to do i s g iv e a 21 b r i e f le s s o n i n c h e m istry . But we a r e ta lk in g ab o u t TCDD, and 22 I w i l l draw th e m o lecu le and show you w hat I mean. E very atom 23 has, a r e l a t i v e mass o r r e l a t i v e w e ig h t. I t i s n ' t v e ry much, b u t 24 i t e x i s t s . And w e'v e g o t a m o lecu le h e re w ith c h lo r in e and
49
1 oxygen, arid carbon and hydrogen. Now,\you remember y e s te rd a y
2 I pointed but th a t a t each of these com ers th ere is a carbon
3 atom and th e r e i s a ls o a hydrogen atom. And we r a r e l y show
4 th e s e i n a s t r u c t u r e u n le s s th e y have some k in d o f m eaning.
5 I t happens th a t the atom ic w eight of hydrogen is l.a n d the
. 6 ato m ic w e ig h t o f carbon i s 12 and o f c h lo r in e i s 35 and a
7 f r a c t i o n and oxygen i s ,16. And i f we w ere to add th o s e a l l
8 up, we would have th e m o le c u la r w e ig h t, so to sp eak , w e ll,
9 i t i s th e m o le c u la r w e ig h t o f 2 ,3 ,7 ,8 TCDD. We have f o u r
10 h y d ro g en s, and th e atom ic w e ig h t i s 1, so t h a t means we have
"rr
t
*
r '\
11
^
a t o t a l o f 4, and we have two oxygens, and th e ato m ic w e ig h t
12 i s 16. So t h a t means 32. Arid we have fo u r c h l o r in e s , and
13 w w i l l c a l l t h a t 35 I t is . a l i t t l e more th a n 35, th e r e i s
; ti
14 a decim al in th e r e , b u t l e t ' s keep th e d ecim als o u t o f i t .
15 So w e'v e g o t fo u r c h lo r in e s , *,,and th e n w e'v e g o t tw e lv e , carb o n s
16 and th e y a r e each 12. S o ,th is comes, o u t 4 tim es 35 i s 140 and
17 12 tim es 12. i s 144, and t h a t adds up .1to 320. Nowy t h a t ' s th e
18 m o le c u la r w e ig h t o f TCDD. : ; And a' man named A v ig o d ril back In
19 th e l a s t c e n tu ry somehow d e te rm in e d t h a t , f o r ev e ry ch em ical 20 compound, I f we ta k e i t s m o le c u la r w e ig h t in gram s, i n o th e r
21 w ords, th e gram m o le c u la r w e ig h t, i f we ta k e 320 grams o f TCDD, 22 i t has i n i t a v e ry , v e ry la r g e number o f m o le cu le^ , as you
23 m ight im ag in e, th e number I s 6.023 tim es 10^3. w hat t h a t
24 means i s th e n s in c e 1 0 ^ i s a t r i l l i o n , t h i s means t h a t th e r e
50
1 i s 600 b i l l i o n t r i l l i o n m o lecu les in t h i s much TCDD. Now, i f 2 we w ere ta lk in g a b o u t, i f we w ere ta lk in g ab o u t p h e n o l, p henol 3 has a m o le c u la r w eig h t o f 94 and we would a r r i v e a t t h i s i n th e 4 same way, 94 grams o f phen o l has th e same number o f m o lecu les 5 i n i t , 180 grams o f g lu c o se has t h i s many m o le c u le s. T his i s 6 a co n stan t fo r a l l m olecules, and i t is a very u se fu l number 7 as you can im agine. I guess to make use o f t h a t -- 8 Q Dr. D o st, l e t me ask you a q u e s tio n . O r ig in a lly when 9 A v ig o d ril came up w ith t h a t number, th e r e c o u l d n 't have been 10 any way f o r him to co u n t t h a t . How in th e w o rld d id he a r r i v e 11 a t i t ? 12 A I'm n o t e x a c tly s u r e . He c e r t a i n l y d i d n 't have 13 in s tru m e n ts to co u n t t h a t . He deduced i t from th e T able o f 14 E lem ents and o th e r in fo rm a tio n known a t t h a t tim e to c h e m is ts . 15 I t was a r a t h e r a s to n is h in g i n t e l l e c t u a l f e a t , r e a l l y . 16 Q Has i t b een e s t a b l i s h e d now as b ein g a c c u ra te ? 17 A I t ' s s t i l l v a l i d , and a p p a re n tly now w i l l rem ain t h a t 18 way. In o th e r w ords, h is c a l c u a l t i o n was c o r r e c t . So i f we 19 w an t, we have 360 grams e q u a l 6.02.3 tim es 102^. 20 ,, Q E xcuse, was t h a t 360 o r 320? 21 A I'm s o r r y , 320. Then 320 m illig ra m s w hich i s a 22 th o u san d l e s s would have tim es 10^0, 300 m icrogram s i s 1 0 ^ , 23 W ell, i f we w anted to f in d o u t how many m o lecu les a re i n one 24 m icrogram , w e ll o b v io u sly we w ould ta k e l/3 2 0 t h , and I d o n 't
51.
1 do a r ith m e tic l i k e t h a t i n my h ea d . What we r e a l l y do i s d iv id e 2 th is by 320. I 'v e .g o t the wrong exponent h e re , I'm so rry , I 3 would be g iv in g you a m illio n more than you r e a lly 'w a n t. . Okay, 4 welcome up w ith a number t h a t i s 1 .8 3 2 2 , we d o n 't c a re to o much 5 ab o u t th e s e l a t e d e c im a ls, tim es 10 m o lecu les i n one rnlcrogram . 6i Now, w e 'r e t a l k i n g about a m icrogram p e r k ilo g ra m , so we a r e 7 ta lk in g h ere about the number of .m olecules -in a kilogram eof 8 mouse, th e r e a r e ab o u t f o r t y m ice to th e k ilo g ra m .. So i f we 9 w anted to know how many m o lecu les w ere i n each o f th e s e m ice, 10 , th e ii we would d iv id e, t h a t number by a p p ro x im a te ly 40, a mouse H w eighs ab o u t 25 gram s. So s in c e we a re ta lk in g i n t h i s e x p e r i 12 ment ab o u t a dosage per. k ilo g ra m , we can le a v e i t a t t h a t . 13 Here i s th e number o f m o le cu les p e r k ilo g ra m i n t h i s e x p e rim en t 14 w hich d id n o t p ro d u ce an e f f e c t . Now, i f we w an ted , l e t ' s s a y , 15 to c o n s id e r how many m o le c u le s, i f o u r no e f f e c t l e v e l h ad been 16. a thousand tim es low er, then th e number would be 1 ,9 , I'm j u s t 17 ro u n d in g t h i s o f f , tim es 10^ , w hich i s 1 .9 t r i l l i o n m o le cu les 18' i n a th o u sa n d th o f a rnlcrogram , w hich i s more th a n I can co u n t 19 i n an a f te rn o o n . 20 . j, Q W ell, how does t h a t r e l a t e to th e Boorman stu d y ? 21 I'm n o t s u re I u n d e rs ta n d .
A I'm s o r ry , th e Boorman stu d y found a no e f f e c t le v e l a t one rnlcrogram p e r k ilo g r a m / So th e "number t h a t we a r e con cern ed ab o u t i s 1 .9 tim es. 10-^- m o le c u le s, . ,, r
52
1' Q So a t no e f f e c t `a t a l l ' i n . t h e an im al th e r e was n e a r ly
2 2 t r i l l i o n m olecules of d io x in /
3 A Per kilogram ?1
4 Q Per kilogram .
5 A Of body w e ig h t.
6 Q A l l . r i g h t . W ell, w hat does t h a t number, how does t h a t
7 e f f e c t what Dr. Zahalsky t e s t i f i e d to as to th e ca u satio n o f th is
8 immune e f f e c t ?
; L"
9 '' 1 A W ell, i t sa y s t h a t one m o lecu le o r even a few o r eveD
10 a 1th o u san d a re n o t l i k e l y to have any e f f e c t o r w i l l n o t have Ah' /
11 e f f e c t . 12 / Q A ll r i g h t , now I b e lie v e y o u 've t e s t i f i e d th e o th e r
r t ' ! ; _r -
13 day t h a t th e Food and Drug A d m in is tra tio n has a r r i v e d a t a no
14 e f f e c t le v e l i n a human b e in g , i n a 70 k ilo g ra m man, i s t h a t 15 r i g h t ?
16 A They have, a r r i v e d a t a c o n c lu s io n . T his i s n o t m easured
17 in human t o x i c i t y s t u d i e s , th is , i s in fo rm a tio n d e riv e d from anim al
18` ex p erim en ts and th ey have d e te rm in e d .o r d ecid ed t h a t . t h e no e f f e c t
19 le v e l in man - - t h a t th ey , can e s t a b l i s h a no e f f e c t l e v e l i n man.
r
20 I
->
Q A ll r ig h t, what i s th a t no e f f e c t le v e l?
21 f A I t i s one. nanogram p e r k ilo g ra m . They u se a f ig u r e o f
22 70 grams j u s t on a g e n e ra l assu m p tio n o f a 70 k ilo g ra m p e rso n .
23 Q So .th a t i n . a 70 k ilo g ra m man, w hich i s w hat you sa y ,
24 150 some pounds?
53
r A 154 pounds.. * *
-,'m - '
2 Q There is* a 70/nanogtam .no e f f e c t le v e l.? '
3
A ^ T h at's c o rre c t.
'
4 Q , A ll r i g h t , how; ma!ny m o lecu les would t h a t be?
5 A W ell, we w i l l m u ltip ly t h i s f ig u r e by 70, and t h a t 's .
-*
-"
^ -< *n ,
6 f o r a 70 k ilo g ra m in d iv i d u a l, i t w ould be 1.31 -- w e ll, l e t ' s
7 c a ll i t 1.32 tim es 1 0 ^ m olecules, which would be tp p u t i t
8 in manageable form, a t r i l l i o n is 1 0 ^ , so w e've g o t 1,32 tim es
9 .or 1,32 thousand t r i l l i o n . 10 Q M olecules?
r. - -
XI ' A M o le c u le s. 12 ' Q Of TCDD?
1
-,
13 A Of TCDD.
r
14 Q A t a no e f f e c t le v e l?
\. 1
15 A . I f we c o n s id e r t h i s to b e a no e f f e c t l e v e l , y e s .
16 Q Now, i n term s o f th e immune system , s ir ,* when you say
17 t h a t an a d v e rse e f f e c t on th e immune system w i l l cauSe a d d i tio n a l
18 i n f e c t i o n s , i s t h a t c o r r e c t?
19 A Y es.
20 V Q What k in d o f in f e c tio n s ? Are you ta lk in g ab o u t a c o ld
21 o r a runny n o se o r w hat k in d o f i n f e c t i o n i s ev id en ced i n an
22 immune su p p re ss e d organism ?
23 A W ell, an o rg an ism t h a t h as a s i g n i f i c a n t - - w ell^
24 to s t a r t w ith th e r e i s a l o t o f ex c ess c a p a c ity . W e're alw ays
54
1. f i g h t i n g o f f in f e c tio n s o f one k in d d r a n o th e r . An in d iv id u a l
2 t h a t has a n o tic e a b ly compromised immune system i s s u f f e r i n g !i'l'"- ?V i m ,
3 prob ab ly continuous a s s a u lts , wounds d o n 't h e a l, pro b ab ly con-
o, '"V ' , ,
\4
' 1 <1
1" . *
;
' 4 tin u o u s r e s p ir a t o r y a ilm e n ts . The o th e r problem i s t h a t th e se
' ^ '
jr
5 tend to p ro g ress, n o t o n ly 'a r they allow ed to take ro o t or
l , J] :r '\ j / / // ^ ^ T'
,
6 ta k e f o o th o ld , b u t " it w ould seem to me t h a t th e y w ould p r o g r e s s .
7 And X th in k t h a t th e , o n c ^ th e - b a r r ie r , so to sp e a k , had been
8 overcome o f o u r ex c ess c a p a c ity f o r immune p r o t e c t i o n , i t w ould
9 seem th a t the in d iv id u a l animal,human o r any anim al presum ably,j
10 o r f o r t h a t m a tte r any o rg an ism t h a t r e l i e s on an immune system
" i_
*
j
',
11 w ould, I th in k , s u f f e r f a i r l y s u b s t a n t i a l i n f e c t i o u s `d is e a s e .
12 Q L et m s t a r t , f i r s t o f a l l , w ith y o u r s ta te m e n t a b o u t ,
13 e x c s s c a p a c ity . What a r e you ta lk in g about?
14
A . W ell, th e immune system has f a r more c a p b il it th a n k iF.
15 i s n e c e s s a ry to h e lp us g e t .o v e r a sim p le c o ld o r to g e t o v er
16 c a se o f pneum onia* Now, to be s u r e , i f th e i n f e c t i o n i s
17 m assiv e enough, i f i t j u s t a b s o lu te l y overw helm ing o r I f th e
18 in d iv id u a l, has a p r o te in - d e f ic ie n c y and i s u n a b le t o 's y n t h e s i z e
19 p r o t e i n , and t h i s i s a s i g n i f i c a n t problem w ith heavy .p r o t e i n
20 mal n u t r i t i o n , p e o p le ca n n o t m an u factu re a n tib o d ie s . I t seems "
21 to me t h a t i f th e f a i l u r e o f ;immune f u n c tio n i s r e a l l y s i g n i
22 f i c a n t i t i s r going to mean p r e t t y n e a r ly a co llap se " o f th e
23 immune system .' There i s a' g r e a t d e a l o f ex c ess c a p a b i l i t y o
24 a v a i l a b l e . If.w e i n t r f e r e w ith p a r t o f th e immune system we
55
1 do n o t n e c e s s a r ily cau se an im als a t l e a s t to succumb to I n f e c tio n .
2 .We can show a l o t o f d i f f e r e n t p a ra m e te rs o f immune f u n c tio n , we
3 can look a t the a b ility o f the r a t, you-can in je c t a challenge
4, I n to th e r a t paw and to se e w h eth er th e paw s w e lls a s - i t o u g h t
5 to b ec au se c e l l s a re b e in g b ro u g h t i n to th e paw, o r we can lo o k
6 a t tit e r s , of a n tib o d ie s .. "
.
? ,r ^ , -r "
4 wr
,.
7 Q Come a g a in now?1 ? 1 v ' '
,
-- " r .
8'
A We can lo o k a t ' th e - amount. o f a n tib o d ie s a v a ila b le in
' . '1
^
;'
. 1\ r . v v ^
-v *
9 the blood.
v: '** v
-
10
Q What i s a t i t e r ? ; ; ;
"
. "i
'
11 A A t i t e r i s a m easurem ent o f th e amount t h a t i s . a v a i l - :
12 a b le f o r r e a c tio n w ith a n ; in f e c ti o u s system . So th e r e i s l o t s
13 o f ways o f -q u a n tify in g d e p re s se d immune f u n c tio n , and we know
14 t h a t TCDD a t s u b s t a n t i a l 'd o s e s d e p re s se s immune f u n c tio n , th e r e - 'j ,
15 i s n ' t any q u e s tio n ab o u t t h a t . Not a l l immune f u n c tio n , th e r e
16.. d o e s n 't seem to be any im pairm ent o f th e re sp o n se to v i r u s e s ,
17 f o r exam ple, to th e e x te n t t h a t la b o r a to r y ex p e rim en ts have
18 d em o n strate d , b u t anim als do n o t d ie o f i n f e c t i o n th a t, a r e
19 h e a v ily a tta c k e d by TCDD, and we know t h a t t h e i r immune f u n c tio n ,
20 m ust be a t l e a s t p a r t i a l l y im p a ire d , you s e e , so
21 Q Now, when you say t h a t th e r e i s ex c ess c a p a c ity , why
22 th e n i f th e Immune f u n c tio n h as more c a p a c ity th a n i s a f f e c t e d .
23 by d io x in e x p o su re , why i s i t t h a t ta k e s so long to f i g h t o f f
24 a cold? Why, i s i t t h a t an I n f e c t i o n can l a s t a w h ile i n an
v
, 1 organism?
2 A - I f I knew why i t took so long to f i g h t o f f a c o ld , L
3 X w ould have p ro b a b ly .become famous long s in c e . T h a t?s a problem
4 t h a t has y e t to be so lv e d . The f a c t rem ain s, though, i s t h a t
5 c o ld s , th e c o ld t h a t I have d id n ' t j u s t sweep i n and w ipe me 1
6 o u t. There were a couple o f days where I was w o rried about th a t,
7 p e rh a p s , an d 1th e n th e r e was a day o r two p erh ap s t h a t I was w o rrie d
8 th a t X wouldn't d ie. But n o n eth eless, th a t in te ra c tio n took
9 p la c e , and i t has p r o te c te d me, and I am h a v in g a l i t t l e tr o u b le
1 i*
* /< i
10 g e t t i n g th e l a s t o f i t r e s o lv e d , b u t i f X d i d n 't have an immune
11 system , I w ould have dropped v e ry q u ic k ly v , Why i t .ta k e s so long
12 i s a good q u e s tio n . C e r ta in ly .th e s e ; th e a n tib o d ie s a r e c o n tin u in g
13 to be p ro d u ced . The c e l l s , t h a t a r e f i g h t i n g o f f s p e c i f i c o rg an ism s,
14 lym phocytes and so f o r t h ; a r e c o n tin u in g to be p ro d u ced . Why
15 th e y c a n 't j u s t s te p in and mop up in s ta n ta n e o u s ly i s som ething
16 t h a t I don' t u n d e rs ta n d .
17 Q I f th e ty p e o f d is e a s e f o r a quenched immune sy stem
18 would be som ething more th a n j u s t a c o ld -- _
19 A Not n e c e s s a r i l y . I g u ess w hat I am sa y in g i s t h a t I f
20 you w ere to red u ce th e f u n c tio n o f some component o f th e immune 21 system by h a l f , l e t 's sa y , by i n t o x i c a t i o n , t h a t d o esn ' t mean
22 t h a t you have e lim in a te d c a p a c ity s u f f i c i e n t to - c a u s e t h a t
23 anim al to succumb to some k in d o f an I n f e c tio n .- There i s
24 r e s e r v e c a p a c ity . We have im p a ire d th e a n im al, b u t we h av en ' t
57
1 im paired i t to the p o in t where h is a b ility to fig h t o ff th is
2 in fe c tio n is re a lly a ffe c te d , and th a t's re a lly very c le a r
3 in a l l o f th e ex p e rim en ta l work, because when anim als a re
4 ch allen g ed w ith a le th a l dose o f, w e ll, I showed a dose response
5 r e l a t i o n s h i p e a r l i e r . At th e h ig h e s t dose o f TCDD, th e r e i s
6 s t i l l su rv iv al. A q u arte r of the anim als survived th is im pact
7 th a t in u n treated animals k ille d a q u arte r of the anim als, and
8 we see i n an im al c o lo n ie s w here TCDD i s b e in g a d m in is te re d to
9 anim als, th e re is no change in th e ir response to th e h o st of
10 in f e c tio u s d is e a s e s w hich e x i s t in m ost an im al c o l o n ie s ;
11
Q W ell, i f th e immune system w ere d e v a s ta te d , w ere
f
12 quenched, as I th in k Dr. Z ahalsky u sed th e te rm , w hat would
13 happen?
14 A We would have no, we would have no p r o t e c t i v e re sp o n se
15 to i n f e c t i o u s d is e a s e , arid t h i s i s th e k in d o f problem t h a t
16 happened. P eo p le who have leu k em ia, who has a bone marrow
17 t r a n s p l a n t , have to go th ro u g h a p ro c e s s by w hich a l l o f th e
18 bone marrow i s d e s tro y e d , u s u a lly by r a d i a t i o n o r by chem otherapy,
19 arid th e n new bone marrow i s tr a n s p la n te d o r , y e s , new bone marrow
20 i s tr a n s p la n te d i n to th o se b o n es, w hich i s a v e ry p a i n f u l p ro c e s s
21 b u t i n t e r e s t i n g l y enough, and t h a t new bone marrow w i l l th e n
22 p r o l i f e r a t e and w i l l form th e s e k in d o f c e l l s t h a t we a r e
23 ta lk in g ab o u t h e re and re p o p u la te w ith norm al w h ite c e l l s . A
24 p erso n who i s in t h a t s ta g e w here th e y have had bone marrow
58
1 d e s tro y e d have had v i r t u a l l y a l l o f t h e i r o th e r immune system
2 destroyed'. They a re v ery v u ln e ra b le to in f e c tio n and i t i s
3 n e c e s s a ry to p r o t e c t them w ith g r e a t c a r e . P eo p le who have
4 organ tra n s p la n ts a re u su a lly m aintained on drugs th a t depress
5 th e immune system v e ry , , v e ry s h a r p ly . There a re many su ch d ru g s,
6 and th e y d e p re s s th e immune sy stem to th e p o in t w here t h a t h e a r t
7 is n o t being re je c te d or th a t kidney is n o t being re je c te d . If
8 a p e rs o n i s n o t f o r tu n a te enough to f in d an org an t h a t i s com
9 p a t i b l e , th e y a r e in a co n tin u o u s s t a t e o f d e p re sse d immune
10 f u n c tio n , c o n tin u o u s ly v u ln e r a b le to m ajor i n f e c t i o n and have
11 to l i v e aev g ry s p e c ia l k in d o f l i f e i n o r d e r to make t h a t
12 p o s s i b le , a g r e a t d e a l o f p r o t e c t i o n , a n t i b i o t i c s a l o t , and
13 so f o r t h . And o f co u rse we o c c a s io n a lly r e a d ab o u t th e c h ild r e n
14 b o m c o m p letely w ith o u t an immune sy stem , and no one h a s s o lv e d
15 t h a t problem y e t , and th e y l i v e i n a b u b b le , and n o t a l l t h a t * A *t,,, , grm
16 lo n g . They have to be c o m p le te ly p r o te c te d from any c o n t a c t
17 w ith o th e r humans o r any..kinds o f o rg an ism s, and t h e i r fo o d i s
1
kM
i
I
if
*
18 s t e r i l i z e d b e f o r e i t is , s e n t i n t o 't h e m (and bo f o r t h . They have
J
i-
*
,,t ,
i ' .* ^ * *
-
* 't
19 no p r o t e c t i o n . '
?9' *
"f r
*
20 b Q So t h a t i f a n 'o rg a n is m ,' an anim al does n o t have a
21 c a t a s t r o p h i c i l l n e s s o r i f a s c r a tc h d o e s n 't become b lo o d 22 p o iso n in g o r i f a m inor i n f e c t i o n d o e s n 't advance to a v e ry
23 s e rio u s i n f e c t i o n , does t h a t i n d i c a t e t h a t th e immune sy stem
24 i s w orking?
59
'1 A Y es. '
2 Q L et me go n e x t, Dr. D o st, to th e k id n e y as an o rg an ,
3 i f I m a y .',D r. S ilb e rg e ld t e s t i f i e d on A p ril th e 12th, 1984,
4 P age197, t h a t th e anim al d e m o n stra te s1t h a t TCDD d e s tro y s k id n ey
5 c e l l s . Do you r e c a l l t h a t te stim o n y ?
6 A Yes.
7 Q Do you a g re e w ith th a t?
8 A No.
9 Q Why n o t?
10 , A ,, W ell, th e an im al d a ta show r e a l l y j u s t th e o p p o s ite ,
11 TCDD a t s u r v iv a b le le v e ls does n o t, have an a p p r e c ia b le e f f e c t
12 on th e kidney, even o v er long p e r io d s o f tim e . I f I may r e f e r
13
again to Kociba's-work and o th e r chronic s tu d ie s , long term
- ;J J
14 s t u d i e s , th e k id n e y i s n o t . s i g n i f i c a n t l y a f f e c t e d . One o f th e
K
15 in t e r e s t i n g ^ a s p e c t s o f the. work t h a t Dr. K ociha d id was t h a t
16 he found t h a t th e n o rm ally o c c u rrin g d e g e n e ra tiv e k id n e y
17, d is e a s e t h a t o cc u rs in a l l o ld an im als was s i g n i f i c a n t l y
18 d im in ish e d a t th e h ig h d o s e so f TCDD. What t h a t m eans, I don' t .
19 know, b u t i t c e r t a i n l y does, mean ,t h a t th e r e was no in c r e a s e d
20 in c id e n c e o f k id n ey t o x i c i t y i n th o s e a n im a ls, n o r was th e r e
21 any a t th e low er d o s e s .,,1' f.,"
' , - -
22 Q Now, how e x a c tly , I w ant to g e t back to th e s e K o c ib a 's
23 s tu d ie s i n j u s t a m in u te,, b u t; how. e x a c tly does th e k id n e y work?
24 You w ere t a l k i n g b e f o r e ab o u t o rg an s in w hich -- t h a t h a n d le
V . . " ':
60
1 la rg e volume o f blood and organs th a t have a l o t o f f a t so th a t 2 TCDD can be s to r e d i n them, Where does th e k id n e y f a l l i n t h a t 3 spectrum o f types o f organs in the body? 4 A W ell, th e kidney is our prim ary ex creto ry organ fo r 5 so lu b le su b sta n c e s, and i t would -- I th in k I ' l l draw a p ic tu re 6 i f I m ight. This by no means i s a p re c is e r e n d itio n o f a kidney 7 tu b u le, but the kidney is com prised o f a la rg e number o f in d iv id u al 8 u n i t s , k id n ey tu b u le s , and th e way th ey work i s th e re i s a f i l t e r , 9 any f a t . The b lo o d b re a k s up in to sm a ll v e s s e ls and i t comes in 10 and th e r e i s j u s t a b ig clump o f v e s s e ls h e re s u rro u n d in g , t h a t 11 i s a k in d o f a cup, and i t i s r e a l l y a f i l t e r , and as th e b lo o d 12 goes th ro u g h th e r e , p ro b a b ly as much as te n p e r c e n t o f a l l o f 13 th e f l u i d i n th e b lo o d a c t u a l l y goes th ro u g h t h a t f i l t e r . The 14 c e l l s d o n 't come th ro u g h , th e la r g e p r o te in s l i k e th e p lasm a, 15 alb u m in , w hich i s a norm al p a r t o f th e p lasm a, and th e immuno 16 p r o te in s t h a t we w ere ta lk in g a b o u t, a l l th o s e la r g e p r o te in s 17 d o n 't go th ro u g h . T his i s a r a t h e r c o n v o lu te d tu b e , b u t I 'm 18 j u s t going to draw i t s t r a i g h t . And i t i s su rro u n d e d by c i r 19 c u l a t i o n . And w hat happens i s t h a t a l l o f th e w a te r, m ost a l l 20 o f th e w a te r, p ro b a b ly 9 9 .9 p e r c e n t o f a l l o f th e f l u i d t h a t 21 w ent th ro u g h th e f i l t e r i s b ro u g h t b ack o u t. But i n th e p r o c e s s , 22 many o f th e m a te r ia l s t h a t we d o n 't w ant to b r in g b ack a r e l e f t 23 in t h a t f l u i d . And m ost o f th e m a te r ia l s t h a t we d o n 't w ant 24 to g e t r i d o f l i k e g lu c o s e , you know, we have l o t s o f g lu c o s e
61
1 in th e b lo o d . Norm ally ab o u t 100 m illio n grams p e r 100 m i l l i l i t e r s 2 and t h a t 's go in g to come th ro u g h here to o , b ec au se i t i s a 3 sm a ll m o le c u le . So i t goes down h e r e and we re a b s o rb i t back 4 o u t a g a in i f we a r e n o rm al. Some p eo p le c a n 't do t h a t v e ry w e ll, 5 b u t we r e c o v e r a l l o f t h a t g lu c o s e , u n le s s w e 'v e j u s t e a te n a 6 candy b a r o r som ething l i k e t h a t , th e r e may be to o much to 7 re c o v e r, and some o f i t w i l l e s c a p e . And th e n th e r e a r e some 8 other c e lls th a t actu ally have the function of secreting certain 9 th in g s i n t o th e u r in e , ammonia and so f o r t h . But anyway, in 10 e s s e n c e , i t i s a f i l t e r fo llo w e d by a scav en g in g u n i t w hich 11 b r in g s m ost o f th e f l u i d and v a lu a b le s t u f f b ack i n , and th o se 12 th o u san d s o f u n i t s a l l converge i n th e c e n te r o f th e k id n e y . 13 And th e n from th e k id n ey th e r e i s a tu b e t h a t goes to th e 14 b la d d e r . So w hat we have h e re i s a f i l t e r , and i t d a ls -- 15 i f we a r e t a l k i n g ab o u t a t o x i c s u b s ta n c e o r any k in d o f a 16 f o r e ig n s u b s ta n c e , i f i t i s a t a l l s o lu b le i n w a te r , I t i s 17 d e l iv e r e d to t h i s p o in t and h o p e f u lly l e f t i n th e u r i n e and 18 d is p o s e d o f . 19 Q Now, th e th in g t h a t you have drawn, th mass o f 20 b lo o d v e s s e ls a t th e to p i s c a l l e d w hat? 21 A I t i s c a l l e d th e g lo m e ru lu s. Z h ad n o th in g to do 22 w ith naming th e s e . 23 Q A ll r i g h t , and th e tu b e i s c a l l e d th e tu b u le ? 24 A T his i s th e k id n ey tu b u le , and i t h as p ro x im al
62
1 segm ent, a c tu a lly i t comes in k in d o f an S sh ap e, and t h i s i s 2 th e d i s t a l segment, and th e re a re d if f e r e n t fu n c tio n s in d i f 3 feren t lengths of th a t tube, but -- 4 Q A ll r i g h t . Now, how many o f th o se o p e r a tin g u n i t s , 5 l e t 's say, the glom erulus and the tubule w ith i t s reab so rp tio n
6 c o l l a r , and t h a t s o r t o f th in g , how many o f th o s e a r e i n th e
7 kidney? 8 A Q uite a few m illio n . 9 Q So th e b lo o d i s c o u rs in g th ro u g h t h i s th in g ? 10 A These a re l i t t l e b i t t y v e s s e ls h e r e , you s e e , th e 11 k id n e y c i r c u l a t i o n h a s d iv id e d i n t o an enormous num ber*of 12 v e s s e l s . T his th in g i s o n ly one c e l l i n th ic k n e s s , you know, 13 th e w a ll o f t h i s tu b u le i s o n ly one c e l l . So th e r e i s r e a l l y 14 a c o l l a r o f c e l l s , you m ight s a y . In o th e r w ords, i f you w ere 15 to -- to o k a tu b e from th e en d , i t would look som ething l i k e 16 t h i s , you s e e . So i t i s a v e r y , v e ry s m a ll s t r u c t u r e . 17 Q And so th e b lo o d c o u rse s th ro u g h th e k id n e y , does 18 th e k id n ey r e c e iv e i t s own b lo o d su p p ly f o r en erg y and l i f e 19 f o r th e c e l l s ? 20 i A N e ll, t h a t 's r i g h t . I t h ap p en s, you s e e , a l l o f 21 th e s e tu b u la r c e l l s r e c e iv e t h e i r c i r c u l a t i o n from th e same 22 b lo o d flo w t h a t i s b r in g in g , i t i s a v e ry e f f i c i e n t sy stem , 23 we d o n 't have to have a w hole s e p a r a te plum bing system i n 24 o rd e r to su p p ly n u t r i e n t to th e k id n e y i t s e l f . The same
63
1 c irc u la tio n th a t brings flu id s to the kidney and c a rrie s wastes 2 i n h e re and comes b ack down h e r e , th e same c i r c u l a t i o n i t 3 b rin g s o u t th e f l u i d t h a t we d o n 't w ant to d isp o s e o f , p ro v id e s 4 the energy resources fo r those c e lls . 5 Q A ll r i g h t . Now, when Dr. K ociba s tu d ie d th e k id n e y ,
6 how many o c c a sio n s d id he do i t ?
7 A I 'm n o t s u re w hat you mean, how many. 8 Q How may s e p a r a te s tu d i e s w ere th e r e i n w hich t h i s was 9 observed? 10 A W ell, th e y d i d n 't f i n d -- he had done two s t u d i e s , 11 one t h i r t e e n week stu d y t h a t I th in k I m en tio n ed y e s te r d a y , 12 and th e n th e long term , l i f e t i m e stu d y t h a t th e y d id . 13 Q Was one o f them i n 1976? 14 A The s h o r t one was r e p o r te d i n 1976. The lo n g e r one 15 was r e p o r te d i n 1978, and th e y had a s u b s t a n t i a l number o f 16 an im als a ta e a c h e x p e rim e n ta l l e v e l , o f c o u rs e . 17 Q Now, w hat d id th e 976 stu d y show w ith r e s p e c t to 18 k id n e y e f f e c t s i n an im als? 19 A I t d i d n 't show any k id n e y p a th o lo g y t h a t I r e c a l l . 20 h Q Were th e r e any b lo o d c h e m istry r e s u l t s ? 21 A Oh, y e s . They d id a v a r i e t y o f b lo o d c h e m is try , 22 and th e y d i d n 't f i n d , e x c e p t -- I b e l ie v e a t th e h ig h e r 23 dose th e y found some e l e v a t io n s i n le v e l s t h a t w ould s u g g e s t 24 t h a t m ight be i n t e r p r e t e d upon th e k id n e y , b u t th e r e c o u ld
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1 also have been e ffe c ts upon the liv e r , too.
2 Q Now, you say th e h ig h e r doses?
3 A They u se d a maximum dose i n t h a t s tu d y , i n th e 1976
4 study, o f one microgram p er kilogram p er day.
5 Q A ll r i g h t , was t h a t h ig h enough to cause d e a th in
6 some o f th e an im als?
7 A I t c e rta in ly did.
8 Q And a t t h a t h ig h d o se, th e r e w ere some b lo o d c h e m istry
*
9 re su lts, is th at right?
v
10 A Yes.
11 Q Did th e y do any k id n e y p a th o lo g y on t h a t o c c a sio n ?
12 A Y es.
13 Q Did th e y f in d any p a th o lo g ic a l?
14 A To my r e c o l l e c t i o n th e y found no p a th o lo g y a t a l l .
15 They found no change in th e a r c h i t e c t u r e o r th e shape o f th e
16 k id n e y c e l l s ,
17 Q A ll r i g h t , so t h a t i n 1976 K ociba d id n o t show t h a t
18 TCDD d e s tro y s k id n e y c e l l s ?
19 A T h a t's r i g h t , and th e same outcome em erged i n th e
20 stu d y r e p o r te d i n 1978 w here th e y had m a in ta in e d i t f o r two
21 y e a r s .
22 Q A ll r i g h t , now th e 1978 was a c h ro n ic stu d y f o r
23 th e l i f e t i m e o f th e anim al?
24 A Yes.
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1 Q And how d id th e k id n e y e f f e c t i n th e 1978 stu d y 2 r e la te between the dosed and the undosed animal? 3 A W ell, X th in k I m entioned a s h o rt tim e ago th a t the 4 normal incidence of degenerative kidney ailm ents, a l l , p a r ti 5 cu larly r a ts , as they age, have a su b sta n tia l incidence of 6 k id n e y damage. The nephron h e r e , t h i s u n i t t h a t we have been 7 ta lk in g about w ill ju s t sim ply f a l l , and th ere w ill be areas 8 in th e kidney where is sc a r tis s u e and a l o t of the number
\ 9 of functional u n its decreases w ith age. 10 Q T hat n a t u r a l l y o c c u rs? 11 A T h at o c c u rs i n any, i t o cc u rs i n a l l o f u s , o f 12 c o u rs e , b u t i t i s q u ite v i s i b l e and e a s i l y q u a n t i f i a b l e i n 13 e x p e rim e n ta l an im als b ec au se we a r e exam ining t h e i r p a th o lo g y 14 a l l o f th e tim e . And th e i n t e r e s t i n g th in g i n t h a t p a r t i c u l a r 15 stu d y was t h a t th e in c id e n c e o f t h a t k in d o f d e g e n e ra tiv e 16 image d e c re a se d a t th e h ig h e s t d o se, and d e c re a s e d to an 17 e x t e n t t h a t was s t a t i s t i c a l l y s i g n i f i c a n t . 18 Q D ecreased i n th e an im al t h a t had r e c e iv e d th e h ig h e s t 19 doses o f TCDD? 20 A Yeah, I'm a f r a i d s o . 21 Q W ell, i s th e r e any e v id e n c e i n th e anim al d a ta t h a t
th e k id n e y i s a d v e rs e ly a f f e c t e d by th e 2 ,3 ,7 ,8 TCDD exposure? A W ell, re a lly only a t doses th a t have caused the
anim al to c o lla p se , and sometimes n o t th en . T h a t's where i t is
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1 seen, is a t very, very high doses where the anim al is probably 2 m ortally affected. 3 Q At below th a t high dose? 4 A I t appears th a t th e re i s n ' t an a p p re c ia b le im pact on 5 the kidney. 6 Q A ll r i g h t , s i r . Now, Dr. D o st, I ' d l i k e to t a l k n e x t 7 about cancer. I wonder i f you would d efin e cancer fo r u s. 8 A Cancer is a co n d itio n in which c e lls have lo s t th e ir 9 reg u latio n , u su ally have regressed in th e ir d iffe re n tia tio n . 10 By t h a t , I mean t h a t i n a l l o f o u r c e l l s b e g in th e same, and 11 as we d e v e lo p , c e l l s become, we u se th e term d i f f e r e n t i a t i o n , 12 and t h a t I th in k i s s e lf - e x p l a n a t o r y . E v e n tu a lly we have to 13 have a l i v e r and we have to have bones and we have to have 14 k id n e y s , and as th e anim al d e v e lo p s, c e l l s b e g in to ta k e 15 d i r e c t i o n tow ard the: fo rm a tio n o f th e v a r io u s o rg a n s , and so 16 as th e embryo d e v e lo p s, th e c e l l s become more and more and 17 more d i f f e r e n t u n t i l f i n a l l y we have a co m plete o rganism 18 w ith arms and f i n g e r n a i l s and eyes and a l l o f th e s t r u c t u r e s , 19 b u t th e y a l l s t a r t e d w ith s i m i l a r c e l l s . I n th e c a se o f tumor 20 che l l s , th e r e i s a ten d en cy to go b ack p a r t way down t h a t 21 la d d e r and lo s e th e d i f f e r e n t i a t i o n . Now, th e y d o n 't go a l l 22 th e way b ack to th e p o i n t w here th e y a r e j u s t l i k e i n i t i a l 23 em bryonic c e l l s , b u t c a n c e r i s a c o l l e c t i o n th e n o f c e l l s 24 t h a t i s re p ro d u c in g by i t s e l f and form ing u s u a lly a tumor
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1 t h a t h a s some form , and i t h as b een removed p r e t t y much from 2 the form al reg u la tio n th a t th e body e x e rts on i t s e l f . These 3 c e l l s have become autonomous. They a re se lf-g o v e rn in g to a 4 larg e ex ten t, so th a t they are no longer c o n tro lla b le and they 5 grow and grow and grow a t a slow r a t e . But they grow a l l by 6 them selves, w ith o u t r e g u la tio n by th e r e s t o f th e body* 7 Q Now, when you say r e g u la tio n , how i s i t t h a t th e body 8 re g u la te s a c e l l 's growth? 9 A W ell, we a r e r e a l l y a v e ry w e ll- o r g a n iz e d system 10 and n o t o n ly does th e body r e g u l a t e each c e l l ' s g ro w th , b u t 11 th e com panions, a l l o f th e c e l l s i n th e l i v e r have a common 12 m is s io n , a l l o f th e c e l l s o f a g iv e n ty p e , and th e y e x e r t 13 r e g u l a t i o n on one a n o th e r. When th e y lo s e t h a t r e g u l a t i o n , 14 w e ll , c e l l s t h a t have become a c a n c e r c e l l may s t a r t . t o i t i 15 m u lt ip ly , and i t ' s going to form a l a r g e r and l a r g e r u o lo n y 16 o f c e l l s t h a t e v e n tu a lly i t i s in d e p e n d e n t o f th e c e l l s t h a t 17 a r e aro u n d i t . But we r e g u l a t e o u r c e l l s , we have hormones 18 t h a t tu r n c e l l s on and o ff* I f we w ant to ca u se su g a r to be 19 ab so rb ed by c e l l s , we r e l e a s e i n s u l i n and t h a t t e l l s c e l l s 20 to ab so rb th e s u g a r. And t h a t i s a form o f r e g u l a t i o n t h a t 21 a r i s e s from a c e n t r a l p o i n t . T hat p a r t i c u l a r phenom enaodoesn*t 22 have a n y th in g to do w ith c a n c e r e x c e p t when you have a c a n c e r 23 o f th e o rg an t h a t p ro d u ces i n s u l i n , and th e r e m ight v e ry w e ll 24 be f o r a w h ile a g r e a t d e a l o f i n s u l i n p ro d u ced so t h a t we have
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1 caused a problem elsew here in the body. G enerally, th e problem 2 w ith cancer is i t reaches a p o in t where i t invades and in te r 3 f e r e s w ith th e f u n c tio n o r some o f th o s e c e l l s may b re a k o f f 4 and go to o th e r p a r ts o f th e body and co n tin u e to reproduce 5 by them selves w ithout re g u la tio n . 6 Q W h at's th e name o f th e l a t t e r c o n d itio n you j u s t 7 described? 8 : A T h a t's a phenomena known as m e ta s ta s is , m - e - t - a - s - t - a - 8 - i - s , 9 and i t sim ply means th e movement o f th e c e lls from t h e i r o r ig in 10 to some o th e r l o c a t i o n . 11 Q A ll r i g h t . Now, when you a r e ta l k i n g ab o u t c e l l s 12 d e v e lo p in g , each c e l l n u c le u s c o n ta in s , you m en tio n ed a l i b r a r y 13 o f m a te r ia l? 14 A Yes. 15 Q C a lle d w hat? 16 A DNA. 17 Q Now, b u t you say i t s t a r t s , th e human embryo s t a r t s 18 from th e jo in in g o f two c e l l s , r i g h t ? 19 A T h a t's r i g h t , i t s t a r t s from th e f e r t i l i z a t i o n o f an 20 egg, and we have one c e l l , i t d i v i d e s ,i n t o two i d e n t i c a l c e l l s , 21 and th e y d iv id e . E a rly bn, you have a s t r u c t u r e 't h a t lo o k s 22 r a t h e r l i k e a r a s p b e r r y t h a t ' s com prised o f a s u b s t a n t i a l 23 number o f c e l l s t h a t a r e k i l n i d e n t i c a l 1. 24 Q Now, i s th e DNA i n th e n u c le u s i n each o f th e s e c e l l s
69
1 th e same? 2 A Yes. 3 Q Now, w hat a b o u t when th e d i f f e r e n t i a t i o n b e g in s to 4 o ccu r, when c e l l s s t a r t to become p a r t o f th e bone o r p a r t of 5 the fo o t or skin or h a ir or w hatever i t is? 6 A A ll o f th e in fo rm a tio n i s t h e r e , b u t by some p ro c e s s 7 th a t i s n 't re a lly w ell understood, c e lls only u tiliz e p a rt of 8 the inform ation, the p a rt th a t's Im portant to th a t function. 9 So i f we have a l i v e r c e l l , t h a t i s a l l th e same in fo rm a tio n 10 t h a t k id n e y c e l l s h av e , b u t th e y d o n 't u se th e p a r t t h a t 's 11 r e l a t e d s t r i c t l y to th e k id n e y c e l l s , you s e e . 12 Q And th e k id n ey c e l l d o e s n 't u se th e p a r t o f th e DNA 13 c e l l t h a t r e l a t e s to w hat th e l i v e r p a r t s use? 14 A E x a c tly . 15 Q So th e DNA i s th e same i n ev e ry c e l l ? 16 A Y es, I t i s a q u e s tio n o f w hich book we ta k e o f f th e 17 s h e l f * 18 Q Now, w hat does DNA have to do w ith can cer? 19 A Cancer i s g e n e r a lly b e lie v e d to s t a r t w ith some k in d 20 o f an in ju r y o r a le s io n on DNA, so t h a t DNA i s changed and 21 n o t r e p a i r e d , and t h a t t h a t change rem ain s and somehow i t
i
remains p a rt o f the g en etic equipment in the c e ll, and u ltim a te ly is trig g e re d to develop in to a tumor c e ll. I t i s a very long, complex p ro cess. I t d d e s n 't j u s t happen.
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1 Q Have you develop a c h a r t w hich would a s s i s t you in 2 explaining this? 3 A I have a ch art X use in le ctu res fo r th a t very 4 purpose. 5 (D efen d an t's E x h ib it 1285 was marked f o r i d e n t i f i 6 c a tio n *) 7 Q Dr. D o st, l e t me show you w h a t's b een marked as 8 D efen d a n t's E x h ib it 1285 and ask you to examine th a t and 9 id e n tif y i t f o r me, p le a s e . 10 A I t i s a -- i t i s th e c h a r t t h a t X b ro u g h t w ith 11 me t h a t X u se in d e s c r ib in g th e p r o c e s s . Xt i s a g e n e ra l 12 d e s c r ip tio n o f th e p ro c e s s by w hich a c e l l e v e n tu a lly 'c a n 13 become a tum or. And as i t p o in ts o u t, i t i s h y p o th e tic a l 14 s t e p s , t h i s i s g e n e ra te d n o t from a s p e c i f i c body o f r e s e a r c h 15 d a ta , b u t i t r e p r e s e n ts th e p r e s e n t u n d e rs ta n d in g o f th e 16 p r o c e s s . A ll r i g h t -- 17 Q Would you s te p up and use i t , p le a s e ? I am i n t e r e s t e d 18 i n th e term " h y p o th e tic a l." 19 A W ell, no one has d eterm in ed f u l l y th e p ro c e s s by 20 ^ h lc h c a n c e r i s ca u se d . T h at p ro b a b ly i s th e m o st, p ro b a b ly 21 i t i s th e m ost h e a v ily i n v e s t i g a t e d problem i n b io lo g y a t th e 22 p r e s e n t tim e i s to t r y to e s t a b l i s h j u s t how c a n c e r i s ca u sed . 23 There i s a g r e a t d e a l known a b o u t how i t i s ca u sed , th e r e i s 24 a g r e a t d e a l known ab o u t in te r m e d ia te s t e p s . B ut u n f o r tu n a te ly
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1 th e pro cess seems to vary, w ith c e rta in k in d s o f can cer, and i t 2 a l s o , and a ls o th e r e are- some v e ry s p e c i f i c and im p o rta n t p ie c e s 3 o f in fo rm atio n th a t sim ply a re n o t y e t known, so i t would be 4 presum ptuous to say t h a t t h i s i s th e way can cer happens. I t 5 does, I think, rep resen t q u ite properly the general scope and 6 sequence o f th e p r o c e s s . So th e s e a re a l l , th e s e a r e g e n e r a l i 7 z a tio n s . They do n o t r e l a t e to some s p e c i f i c tumor ty p e , n o r 8 do th e y r e l a t e to any s p e c i f i c p a t t e r n t h a t someone has l a i d 9 o u t. But th e f i r s t th in g t h a t h ap p en s, you spoke o f DNA, and 10 th e f i r s t th in g t h a t happens i s a p p a re n tly a m u ta tio n o f some 11 o th e r damage to DNA i n th e c e l l , and t h a t a f f e c t s some p a r t 12 o f th e g e n e tic a p p a ra tu s . Now, th e r e i s a c o n c ep t o f th e 13 oncogene t h a t h as c r e p t i n t o th e p o p u la r p r e s s , and i t i s a 14 v e ry im p o rta n t is s u e o f d is c u s s io n among c a n c e r r e s e a r c h e r s , 15 and t h a t i s th e id e a t h a t th e r e a r e segm ents o f DNA, g e n e s, 16 t h a t by some m ischance w ith v e ry m inor changes can be c o n v e rte d 17 i n t o a gene t h a t a c t u a l l y le a d s to c a n c e r. In o th e r w o rd s, th e 18 id e a i s t h a t we have i n a l l o f o u r c e l l s genes t h a t r e q u ir e 19 v e ry l i t t l e m o d if ic a tio n to to u ch them o f f and cau se them to 20 ta k e c o n tr o l o f th e c e l l , so to sp e a k , and cause c a n c e r. In 21 any c a s e , th e r e s t i l l has to be some k in d o f a change i n th e 22 DNA to s t a r t t h i s w hole p r o c e s s . And t h a t s te p p ro b a b ly i s 23 i r r e v e r s i b l e . But o th e r th in g s have to happen f o r i t to be 24 e x p re s s e d , and when I sa y , by e x p r e s s , I mean o th e r th in g s have
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1 to happen f o r i t to ta k e e f f e c t . Now, one o f th e th in g s t h a t 2 apparently have to happen is th a t the c e ll has to d iv id e, and 3 i n d iv id in g i t , c o p ie s o f i t s DNA f o r th e second g e n e ra tio n 4 c e l l . So t h a t th e r e i s two i d e n t i c a l s e t s o f DNA. And i f 5 t h a t h ap p en s, th e s p e c u la tio n i s t h a t p ro b a b ly i t ta k e s two 6 c e l l d iv is io n s i n o rd e r to do t h a t f o r t h a t to be c o p ie d . So 7 a t th e same tim e X w ant to s te p b ack f o r j u s t a moment# We 8 have ta lk e d ab o u t a DNA l e s i o n , t h a t i s i r r e v e r s i b l e . ` T here 9 i s damage o c c u rrin g to DNA a l l o f th e tim e i n th e c e l l , n a t u r a l l y , 10 and as- a r e s u l t o f e v e ry th in g t h a t we do i n o u r. en v iro n m en t. 11 And t h i s le s io n h as to s u r v iv e t h i s r e p a i r p ro c e s s ,: and I 12 p ro b a b ly w i l l have an o p p o r tu n ity to d is c u s s th e id a o f DNA 13 r e p a i r a l i t t l e l a t e r . 14 Q Dr. D o st, excu se me, when you t a l k a b o u t o u r e n v iro n 15 m ent, w hat a r e you r e f e r r i n g to s p e c i f i c a l l y ? 16 A D ie t, e v e ry th in g we do, o u r u l t r a v i o l e t r a d i a t i o n 17 from th e su n , o u r environm ent i s k in d o f h o s t i l e p la c e , and 18 we a r e ta k in g i n a l l o f th e tim e a l o t o f g e n e t i c a l l y a c t i v e 19 m a te r ia l i n o u r d i e t . I f you w ere to go th r o u g h .e v e ry th in g 20 t h a t you e a t and i d e n t i f y a l l o f th e su b s ta n c e s t h a t a r e 21 known to have some k in d o f g e n e tic a c t i v i t y as d eterm in ed 22 i n m ic r o b ia l m u ta tio n t e s t s and so f o r t h , you w ould soon be 23 s e a rc h in g tr y in g to f in d o u t w hat i n th e w o rld you c o u ld e a t 24 t h a t w i l l s u s t a i n you w ith o u t c r e a tin g ;arproblem . A p p a re n tly ,
r
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1 we a r e n o t c r e a tin g problem s by e a tin g o u r d a i ly m eals b ec au se 2 our l i f e span is increasing* but the fa c t remains th a t th ere 3 are lo ts and lo ts and lo ts of m aterials in the n a tu ra l environ 4 ment, both p h y sical and chem ical, th a t has cap acity to a t le a s t 5 th e o re tic a lly cause in ju ry . Not only th a t, w ith in the c e ll, 6 th e in te r c e llu la r environm ent is a lso very v io le n t and. th e re 7 are lo ts of very re a c tiv e m olecular species created in the 8 normal metabolism in the c e ll. Perhaps I can ta lk about th a t 9 a l i t t l e l a t e r , too^ 10 Q When you t a l k ab o u t g e n e t i c a l l y a c t i v e , w hat do you 11 mean? 12 A X mean i t has th e a b i l i t y to i n t e r a c t w ith DNA. 13 Q Now, w e'v e h e a rd t a l k h e re a b o u t DNA and RNA, and 14 m essengers and a l l t h a t s o r t o f th in g . What does a l l t h a t 15 mean? I s DNA a rib b o n o r i s i t a p ie c e o f -- 16 A DNA i s , okay, DNA i s th e p rim ary s o u rc e o f In fo rm a tio n . 17 The RNA t h a t we have ta lk e d a b o u t, th e r e a r e a number o f k in d s 18 o f RNA, b u t th e o n ly number o f RNA t h a t we have p a r t i c u l a r 19 co n cern ab o u t i s t h a t i s made as a copy o f a segm ent o f DNA 20 and goes o u t in to th e c e l l to th e rib o so n e o r to en d o p lasm ic 21 r e tic u lu m to form a p a t t e r n f o r th e s y n th e s is o f some p r o t e i n , 22 DM i s a v e ry , v e ry , v e ry lo n g m o le cu le. There a r e p ro b a b ly 23 i t i s com prised o f p a i r s . I may have to s to p and go b ack 24 now and th e n b ecau se t h i s i s a l i t t l e comple x , b u t DNA, you
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1 may have h ea rd o f th e double h e lix ,w h ic h was a book t h a t 2 Watson and C rick w rote about t h i r t y y ears ago when they 3 f i n a l l y d isc o v e re d w hat th e c o n f ig u r a tio n o f DNA was. 4 Q And won a Nobel p r iz e ? 5 A And th ey won a Nobel p r i z e . The DNA, I guess th e 6 b e s t way to d e s c rib e i t i s to th in k o f a z ip p e r, an o rd in a ry 7 z ip p e r . And you know t h a t a z ip p e r has te e th and th ey come 8 to g e th e r l i k e so . W ell, DNA i s s e t up as p a i r s o f b a s e s , 9 and I d o n 't need to describ e what these bases a re , but th ere 10 a re fo u r k in d s in DNA. And I g uess we co u ld say t h a t th o se 11 fo u r b a s e s a re th e l e t t e r s o f th e g e n e tic a lp h a b e t, and you 12 can im agine t h a t th e re i s an u n im ag in ab le -- t h a t 's n o t v e ry 13 good language - - th e r e i s an enormous number o f p o s s ib le 14 co m b in atio n s b ecau se th e s e m o lecu les a re v e ry , v e ry lo n g . 15 They c a rr y an enormous amount o f in fo rm a tio n . And i f we th in k 16 o f t h i s as b e in g , as th r e e , fo u r l e t t e r s in t h i s a lp h a b e t, and 17 th e re a re words in th e DNA and th e y a re a l l th r e e l e t t e r s long 18 and each o f th o se words codes f o r an amino a c id . Now, when we 19 go th ro u g h t h i s p ro c e s s o f in d u c in g , w hich w e'v e ta lk e d a b o u t, 20 th e DNA opens up and we form RNA as I 'v e j u s t m en tio n ed . And 21 t h a t RNA c a r r i e s th e same code, t h i s s e r i e s o f th r e e l e t t e r 22 w ords. I t i s v e ry long and i t codes f o r th e s e th o u san d o r 23 two thousand amino a c id s t h a t come to g e th e r to make t h i s p r o te in . 24 Q So i t u n z ip s i t ?
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1 A U nzips, h a l f o f i t comes a p a r t and th e ENA i s co p ie d 2 o f f th e o th e r h a l f , o r i t comes a p a r t and RNA i s c o p ie d o f f o f 3 one h a l f . But i f you th in k a b o u t t h i s z ip p e r , f o r e v e ry 10 4 te e th i t has taken one tw ist, th is thing is tw isted , and I 5 mention the number o f b ases. There are in th e c e ll about, 6 betw een 10$ and 10* b a se p a i r s . In o th e r w ords, i n each c e l l 7 th e r e a r e , l e t ' s sa y , 10 words o r l e t t e r s , how w ould I p u t 8 th a t? No, we w i l l d iv id e i t in to th r e e and make w ords. We 9 w ill say th e re are 3 tim es 10^ words o f in fo rm atio n in each 10 c e l l . T h a t's 30 m il lio n , p ro b a b ly more th a n t h a t , b u t th e 11 p o in t i s t h a t i n th e s e r a t h e r i n t e r e s t i n g th r e e l e t t e r words 12 a re th e codes f o r a l l o f th amino a c id s . And so i t i s th e 13 sequence on th e DNA t h a t governs th sequence o f s y n th e s is 14 o u t i n th e c e l l . A ll r i g h t , , i f th e r e i s an e r r o r , i f th e r e 15 i s an e r r o r in t h a t se q u en ce , th e n i n c o r r e c t in fo rm a tio n i s 16 going to be tr a n s m itte d . Now, t h a t may n o t make any d i f f e r e n c e , 17 i t may be a s e r io u s e r r o r o r a b ig chunk o f DNA i s l o s t and 18 th e c e l l d ie s . T h at, f o r t u n a t e l y , i s n o t an unhappy circu m 19 s ta n c e e x c e p t in th e n erv o u s sy stem , b u t i t i s h ap p en in g a l l 20 th e tim e . In t h i s s i t u a t i o n we have in v e r te d some k in d o f an 21 i n c o r r e c t m essage. A ll r i g h t , i t may s ta y t h a t way f o r e v e r 22 and n o th in g e v e r happen, b u t i f th e c e l l d iv id e s b e f o r e i t i s 23 r e p a i r e d , t h a t le s io n may be th ic k i n th e gene so t h a t i t s ta y s 24 th e re and th e c e l l th en i s h e a rin g t h a t m essage. W ell th e n ,
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1 th e r e a r e a number o f p ro c e s s e s t h a t go fo rw ard t h a t come in t o 2 th e c a te g o ry o f p rom otion and enhancem ent. Some o f th e s e words 3 are used in terch an g eab ly , sometimes you fin d p ro g re ssio n used 4 h ere and sometimes tr a n s la tio n . This q u estio n o f c e ll tra n s 5 fo rm a tio n f a l l s up h e re aways, b u t l e t me r e a d t h i s to you. 6 " I t is assumed th a t s p e c ific p h y sio lo g ic a l or biochem ical 7 phenomena are re q u ire d to f a c i l i t a t e th e e x p re ssio n .o f the 8 i n i t i a t i n g e v e n t." Now, t h a t may be c e l l i n j u r y , th e r e a r e 9 c e r ta in chem icals th a t a re known to promote th is e f f e c t . I 'v e 10 m entioned th e p h o rb o l e s t e r s th e o th e r day when we w ere showing 11 t h a t c h a r t on th e d if f e r e n c e betw een p rom otion and i n i t i a t i o n . 12 TCDD, p h e n o lb a r b ito l, smoke co n d e n sa te s h av in g been s u g g e ste d 13 as p ro m o te rs. And th e p ro m o tio n p ro c e s s a p p e a rs to be r e v e r s i b l e 14 a t l e a s t i n i t s e a r l y s ta g e s , in o th e r w o r d s ,if i t does ta k e 15 p la c e b e f o r e some o f th e o th e r e v e n ts going on, why th e n t h a t 16 c e l l may j u s t r e v e r t b ack to n o rm al. I t a ls o i s c l e a r l y dose 17 dependent and c l e a r l y th e r e i s argum ent ab o u t w h e th e r i t i s 18 a th r e s h o ld o r n o t, b u t i t I s v e ry c l e a r l y dose d ep en d en t. 19 Then, th e n e x t s te p i s w hat is . c a l l e d c e l l tr a n s f o r m a tio n , and 20 th e c e l l changes i t s w hole g e n e tic makeup i n some f a s h io n and 21 I t does d i f f e r e n t th in g s th a n i t u sed to do. I t becomes 22 j u v e n i l e , i f you w i l l . I t r e v e r t s b ack to a l e s s d ev elo p ed 23 form . And one o f th e problem s t h a t i t may o r may n o t be 24 r e c o g n iz a b le as a s tr a n g e c e l l , as a d i f f e r e n t c e l l , and i f i t
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1 I s r e c o g n iz a b le , th e n th e immune system w i l l c le a n i t up. And 2 t h i s i s p ro b ab ly going on i n o u r b o d ie s a l l th e tim e . From 3 t h i s p o in t forw ard, th o se c e l l s when they reproduce they c a rry 4 a l l o f th ese bad c h a r a c te r is tic s w ith them, and they b eg in to 5 re p ro d u c e . I f th e y do evade th e immune s u r v e il la n c e and they 6 could very w ell rep ea t c h a ra c te ris tic s o f the normal c e ll th a t 7 a r e s u f f i c i e n t t h a t th e immune, th e c e l l u l a r immune c e l l s , th e 8 k i l l e r c e lls d o n 't reco g n ize them as being tumor c e lls o r n e a rly 9 tumor c e l l s . W ell, so th e r e i s a p r o g r e s s io n h e r e w here we go 10 from a g e n e t i c a l l y i n j u r e d c e l l to a c e l l t h a t h as to be f u r t h e r 11 s tim u la te d in some f a s h io n . And th e n from t h a t wer go to c e l l s 12 t h a t a re p ro d u c in g , changing t h e i r c h a r a c te r and e v e n tu a lly 13 becom ing a s u f f i c i e n t p o p u la tio n o f c e l l s t h a t e i t h e r th e y 14 form a tum or t h a t can be se en o r th e y may form a tum or i n th e 15 bone marrow t h a t i s re c o g n iz e d a s , sa y , leu k em ia o r som ething 16 l i k e ' t h a t , w hich i s a c i r c u l a t i n g tumor i n e f f e c t . T here i s no 17 m ass, no h a rd tum or. I d o n 't know w h eth er I have made t h a t 18 s u f f i c i e n t l y c l e a r , i t i s a v e ry muddy p r o c e s s , and th e r e a 19 l o t o f d i f f e r e n t -- th e r e i s a l o t o f Ja l t e m a t i v e i i d e s c r i p 20 tio n s t h a t can be I n s e r te d . 21 Q I 'm i n t e r e s t e d in th e i n i t i a t i n g e v e n t th a t, y o u 'v e 22 g o t th e r e a t th e to p . I'm n o t s u re I u n d e rs ta n d how t h i s 23 o c c u rs . I s t h i s som ething t h a t h as to come in from th e 24 o u ts i d e , o r i s i t som ething t h a t n a t u r a l l y o c c u rs in th e c e l l
78
1 b e c a u se you ta lk e d ab o u t DNA r e p a i r and e r r o r s i n th e c e l l b e f o r e , 2 A W ell, DNA i s , to s t a r t w ith , we have an enormous 3 amount o f m a te r ia l t h a t has to be m a n u fa ctu red , and we c o n s id e re d 4 t h a t th e m an u factu re o f DNA i n th e c e l l i s a v e ry a c c u r a te p r o c e s s . 5 But the e rro r ra te is about one in every m illio n base p a irs . W ell, 6 I 'v e alread y ta lk e d about a s itu a tio n in which w e've got w hat, 7 10^, t h a t 's 100 b i l l i o n , I beg your p ard o n , a 100 m illio n b ase 8 p a i r s i n th e c e l l . And i f we a r e , a c t u a l l y th e number i s h ig h e r 9 th a n t h a t , i t i s p ro b a b ly a b o u t 300 m il lio n . And i f we h av e, 10 i f we have an e r r o r r a t e o f one i n a m i l l i o n , t h a t means t h a t 11 f o r th e grow th o f a s in g l e c e l l th e r e w i l l have b een 300 e r r o r s 12 j u s t i n t h a t c e l l a lo n e . J u s t i n m a n u fa c tu rin g DNA. And t h a t 13 has to be c o r r e c te d . There i s a r e p a i r p ro c e s s t h a t in s p e c ts 14 DNA. I t i s f a s c i n a t i n g . I t l i t e r a l l y in s p e c ts i t , l i k e go in g 15 o v er a s t r i n g o f bead s and f in d d e f e c tiv e s p o ts , s l i c e s them 16 o u t and f ix e s them. Now, t h a t i s n o t a p e r f e c t p r o c e s s . Some 17 r e p a i r may have b een m issed o r th e r e m ight be a m is ta k e made in 18 th e r e p a i r p r o c e s s , b u t i f you c o n s id e r t h a t th e r e a r e 300 19 e r r o r s i n a g iv e n c e l l and i f , oh, i n th e l i v e r th e r e i s some 20 th in g i n ex c ess o f one b i l l i o n c e l l s p e r gram, so l e t ' s say 21 10^ c e l l s p e r gram f o r a gram o f l i v e r , w hich i s maybe w ould 22 be a p ie c e t h a t s i z e , p e rh a p s , th e r e would be 300 b i l l i o n e r r o r s 23 com m itted i n th e p ro c e s s o f m a k in g ,th e DNA f o r th o s e c e l l s . 24 So t h i s p ro c e s s i s g o in g on a l l th e tim e o f e r r o r i n s y n th e s is .
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1 The o th e r th in g th a t i s happening i s th a t, I ' l l j u s t w rite th a t 2 down, th e o th e r th in g t h a t 's happening i s , as I s a id , th e c e l l 3 i s a v e ry v i o l e n t p la c e and th e r e a r e some v e ry v i o l e n t r e a c tio n s 4 going on in the c e ll in the process of g en eratin g energy. There 5 a re some v e ry r e a c t i v e oxygen atoms t h a t a r e g e n e ra te d i n th e 6 c e l l , and most o f them are trap p ed and a re , d o n 't cause any 7 harm, b u t enough o f them g e t away t h a t th e re i s a v ery s i g n i 8 f i c a n t and m easu rab le amount o f DNA damage by some o f th e s e v e ry 9 r e a c t i v e oxygen s p e c ie s in s i d e th e c e l l . And p e ro x id e , hydrogen 10 p e ro x id e i s made in s i d e th e c e l l , and th e r e a r e enzymes t h a t g e t 11 r i d o f t h a t s t u f f . There a re t r a p s t h a t c o l l e c t i t , b u t some o f 12 i t in e v ita b ly e scap e s and th e r a t e o f in ju r y o f c e l l u l a r DNA 13 can be m easured by th e c o l l e c t i o n o f some o f th e p r o d u c ts , b e 14 cau se as I s a i d , th e y s l i c e o u t p ie c e s o f DNA and b r in g them, 15 th e y a r e e x c r e te d i n t o th e b lo o d and i n t o th e u r in e and th e y 16 a r e d is p o s e d o f . P eo p le i n B ruce Ames' la b a t B e rk ley have 17 done some i n t e r e s t i n g work on t h i s , and w hat th e y f in d i s th o se 18 p ro d u c ts emerge a t a r a t e t h a t i n d i c a t e s c e l l damage to th e 19 e x t e n t o f 320 DNA damaging e v e n ts p e r c e l l p e r day* And when 20 you m u ltip ly t h a t by th e number o f c e l l s In th e body, w hich h as 21 been e s tim a te d , th e r e a re v a r io u s e s tim a te s , b u t th e one I 22 g e n e r a lly u se i s 6 tim es 10*^ c e l l s , w hich i s 60 t r i l l i o n c e l l s 23 i n th e body. Then we have an enormous number o f e v e n ts ta k in g 24 p la c e i n th e c e l l ev e ry day in th e body, ev ery day t h a t can
;-
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1 be p o t e n t a i l l y le a d in g to g e n e tic damage o f some k in d . V ir tu a ll y
2 a l l a r e r e p a ir e d . Now, i f th e sy stem was a b s o lu te ly p e r f e c t ,
3 we m ig h t n o t have c a n c e r. But u n f o r tu n a te ly th e sy stem i s n o t
4 a b s o lu te ly p e r f e c t and th e r e a re in e v ita b ly some e r r o r s i i n DNA,
5 some damage to DNA t h a t p e r s i s t s . Now, a l s o , e x t e r n a l f u n c tio n s .
6 I 'v e ta lk e d ab o u t some o f th e mutogens t h a t a r e i n n a t u r a l sub
7 s ta n c e s , b la c k p ep p er, b r o c c o li, and when I say b r o c c o li, I d o n 't
8 mean to i s o l a t e i t , X th in k we can say t h a t ab o u t m ost v e g e ta b le s ,
9 d o n 't worry about i t . Carcinogens, b en a'a'p y ren e in autom obile"
1.0 e x h a u s t, o r g a s o lin e , b en zen e, l o t s o f chem ical c a rc in o g e n s
11 t h a t can a ls o r e s u l t in t h i s DNA damage t h a t s e rv e s as th e
12 i n i t i a t i n g s te p . I f th e c o n c e n tr a tio n i s h ig h enough I t can
13 cause th e s e k in d s o f e v e n ts , to o . So we have a l l k in d s o f
14 th in g s h ap p en in g , v e ry h ig h r e p a i r r a t e t h a t presum ably i s
15' n o t a b s o lu te ly p e r f e c t , so you can s e e , th e r e i s an i n c r e d i b l e
16 amount o f a c t i v i t y going on i n th e c e l l a l l o f th e tim e t h a t
IV r e l a t e s to th e p o t e n t i a l f o r DNA damage i n t h i s i n i t i a t i n g
18 e v e n t.
19 THE COURT: Mr, Heineman, I s t h i s a good p o in t f o r
1
20 $ s h o r t b reak ?
4
21 MR. HEINEMAN: Yes, s i r , i t w i l l be f i n e .
22 THE COURT: We w i l l ta k e a s h o r t b re a k a t t h i s tim e.
23 The adm onishments t h a t I gave you e a r l i e r w i l l ap p ly d u rin g
24 t h i s b re a k a ls o .
(At th is tim e C ourt was in r e c e s s .)
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1 (Defendant's Exhibit 1286 through 2 1289 were marked for Identification.)
3 BY MR. HEINEMAN: 4 Q Dr. Dost, for the record, I have had these sheets 5 marked. Defendant's Exhibit 1286 Is the calculation of the gram
6 molecular weight of dioxin.
7 A That's correct.
8 Q Showing of Avigodril's number at the bottom?
9 A Yes.
10 THE COURT: What was that, 1286? 11 MR. HEINEMAN: 1286, your Honor. 12 Q 1287 Is the calculation of the total number of cells.
13 A Total number of molecules. 14 Q I'm sorry, total number of molecules, and what? 15 A Well, I made an error when I calculated the number. 16 First, we are talking about the number of molecules In one 17 microgram, and then the number If we are talking about a 18 thousandth of a microgram, and then I carried that calculation to 19 the number of molecules that there would be in a 70 kilogram person
20 and I had made an error In the exponent and I had a thousand times 21 more than It should have been, so It should have been 132 trillion 22 rather than 132 thousand trillion. This is not like Senator
23 Dirksen's famous saying, "A billion here and a billion there and 24 you are talking real money" We need to be a little more precise.
-4
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1 So the number Is 132 trillion, not 132 thousand trillion. 2 Q Okay, and that's exhibit 1287?
3 A If you want, I will put per 70 kilogram person. 4 Q All right. Then I have marked asDefendant's Exhibit 5 1288 the drawing you did of the--
6 A Kidney tubule.
7 Q Kidney tubule. You referred to it as a nephron?
8 A A nephron, yes, that's the word for the whole unit. 9 Q And 1289 Is the calculation you did for what, sir? 10 A For the number of natural errors in the formation of 11 the DNA In cells as cells grow, as tissue grows, and then a 12 statement of Dr. Bruce Ames' finding of about 320 DNA or finding
13 the residue that has arlsed from the repair of about 320 damaging 14 events per cell per day, and the number below is the number of 15 cells in the estimated in the human body, 60 trillion. Perhaps I 16 should identify that number so in the future-- 17 Q All right, thank you. 18 MR. HEINEMAN: Your Honor, at this time we would 19 offer into evidence Defendant's Exhibits 1283 through 1289.
20 (Defendant's Exhibits No. 1283 to 1289 were 21 offered into evidence.) 22 MR. CARR: The Plaintiffs have no objection to these
23 exhibits, your Honor. 24 THE COURT; Fine, they are all admitted without
83
1 objection. Thank you. 2 (Defendant's Exhibits 1283 to 1289
3 were admitted into evidence.) 4 Q Dr. Dost, if you would resume your place there by 5 your chart, dlscussing^the mechanisms of cancer, you did some
6 detail on the initiation, and to make sure I understand i t , the
7 initiation can be an internal event?
8 A It is an internal event in.the cell.
9 Q All right. But it can be caused perfectly naturally
10 without any outside Intervention at all? 11 A It can be caused, well, we assume that all of the 12 events that take place in the cell that damage DNA also have the
13 capacity to cause a DNA mutogen. Now, how does that relate to the 14 event of initiation that you are describing? 15 A Well, first I should point out that a cell has 16 initiated does not behave any differently in the organism than a 17 cell that has not been initiated. So its function has not 18 changed, its function hasn't changed, and it appears that even 19 when it divides, while that Initiating lesion has stayed in the
20 cell and then carried to the next cell, the daughter cells also 21 will not function any differently than the normal cell. So 22 we have got a cell here whose nature hasntj rCallycbeeh'ichaiiged in
23 terms of what it does in the organism. Other events are necessary 24 to make that happen
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1 You asked about mutation. Mutations are changes 2 In the genetic material of a cell such that the cell In either
Its next generation or possibly some subsequent generation will
3
be altered. Generally when we are speaking at the cellular level,
4
we are talking about a change that appears in the next generation.
5
6 If we are speaking about the whole animal, then we would be 7 discussing a change that is Induced in germinal tissue in a 8 reproductive organ, and then again there Is some complex genetics 9 involved here. It could very well be that offspring of the parent 10 with damage might be similarly affected or If it Is a recessive 11 change of some kind, It may appear several generations down the 12 family tree, in an adult organism. Now! we use, it is thought
13 and again pretty universally thought, that mutation, the event 14 that leads to mutation are rather similar to the events that 15 lead to the initiation of cancer. And it is possible for us 16 to study those kinds of events In the laboratory. Microorganisms 17 are very handy for this sort of thing, cultured cells. So 18 there are a variety of laboratory, of fruit flies, there are a 19 variety of systems available In the laboratory for learning
20 whether a chemical has the capacity to Interact with 'DNA, It 21 happens that there Is a good correlation between the ability of 22 a chemical to interact with bacteria and other bench-top kinds
23 of tests and the ability of chemical to cause cancer. Generally 24 chemicals that do not cause mutation does not cause the initiating
85
1 kinds of events . 2 Q Well then, as I understand mutation or mutogenicity
3 ie not the same thing that you are talking about here? 4 A The initial event is probably very similar, just 5 the initial event. The only difference is that i t 's likely
6 that in mutogenosis there is sufficient change that the next
7 generation of the cell is changed, you see, and imaginably we
8 could have cells in, let's say, the liver that had a minor
9 mutation, and they functioned a little differently, and we
10 may accumulate a few cells of that sort in the liver, but it 11 wouldn't matter because it isn't transmitted to the next 12 generation, and those cells d o n 't necessarily make a tumor,
13 they are just a little bit different from the cells, the normal 14 cells of the liver, and that presumably happens frequently.
15 But the carcinogenic event may be sufficiently
16 specific that it only can be transmitted in this fashion to the 17 next generation of cells. The point about mutogenicity, though, 18 is that apparently the damage in the cell is about the same, 19 and there is an empirical correlation, by that I mean the
20 experimental evidence shows that chemicals that aie ruutogens 21 are frequently carcinogens. Okay, now, precisely what is 22 happening at the level of DNA in the two cases to link them
23 is not clearly known, but we know that DNA damage takes place 24 in both instances, and that there is a correlation. Chemicals
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1 A That has been demonstrated by again, Dr. Poland 2 and D r G u n t h e r . Poland has used, has done experiments in
3 which he has used labeled TCDD and has found no evidence of 4 binding to DNA, much evidence of binding to protein and other 5 structures, but none to DNA.
6 Q Now, Dr. Sllbergeld in her testimony here
7 stated that TCDD--
8 MR. CARR: What page, counsel, and day?
9 MR. HEINEMAN: April the 12th, Pages 120 and 149.
10 Q States that TCDD causes cancer itself. It is the 11 most powerful chemical carcinogen identified, and is a complete 12 carcinogen. Do you recall her testimony in that regard?
13 A Yes, I do. 14 Q Do you agree with that, sir? 15 A N o , I don't agree that it is a complete carcinogen. 16 Q And on what basis do you disagree? 17 A I've already discussed really the evidence that 18 shows that TCDD is in fact a promoter and it does not 19 interact with DNA either by direct chemistry or by any ability
20 to cause a significant mutation incidence, and also by studies 21 that have been done that show that in organisms that are treated 22 with TCDD there is no increase in the DNA repair activity,
23 which is a measurable, a measurable factor in cells. 24 Q Now, at the same time in her testimony she suggested
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1 that the Koclba studies show quite clearly that exposure to 2 TCDD alone causes cancer, and that's why it Is a complete
carcinogen. Do you agree with that statement?
3
4 A No, I don't. Vhat the Koclba studies show In
5 that light is that at high doses, at very high doses up at a
6 point where theranimal Is suffering really substantial injury,
7 there is a lot of effects on the liver, particularly In those
8 animals, that the Incidence of a variety of tumors changes, a
9 variety of tumors, and when I say change, I mean just that.
10 There were Increases In the number of hepatocellular carcinomas 11 and also nodules In the liver that probably are reversible by 12 the definitions of most pathologists. There were many kinds
13 of tumors. Now, you must understand that in any population of 14 animals there Is going to be a high population of tumors, a 15 high Incidence of tumors of any kind. We all know unfortunately 16 in our population that most cancers appear In.older people. 17 The same is true In laboratory animals. It Is Interesting that 18 in those experiments the Incidence of many kinds of tumors, 19 particularly those of the endocrine system of the glands,
20 actually It decreased statistically. In other words, it w a s n 't 21 so much an Increase In total numbers of tumors, but an 22 alteration In the whole tumor pattern. In another study that
23 was done In the National Toxicology Program, there was an 24 increase In some tumors In the thyroid at the highest dose.
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1 When the National Toxicology Program study was done, Instead of 2 feeding TCDD In the diet., ,t:hey introduced it by stomach tube
3 once a week. Long term ,,experiment,, they gave the rats theit
4 treatment once a week on,.ar* .exact basis with a stomach tube*
5 Those rats did not get the .somewhat-- rthe slight Increase in
6 tumors of the palate and,, th& ,,nasal passages and the upper
7 respiratory tract that Kpciba's.rats did. The reason for that
8 is because the material ,,was..not in contact with those areas.
9 Rats when they eat tend .to.Inhale.quite a bit of food, particularly
10 when it is in that powdejr ,,form .as is used in these kinds of
11 experiments. The fact that these tutors occurred at the sie of
12 contact is suggestive of,,. promotion, particularly when we compare
13 with the studies that we.re -done ,on .the .skin, TCDD has .also been
14 tested as a skin carcinogen again.in the National Toxicology
15 Program, and the only tumors, that they found were at high doses
16 again, and they were a tumor^called ,a fibrosarcoma of the skin.
17 It is a connective tlssu,e stumor, and there were no other tumors
18 anywhere in the animals., .That, too, is suggestive of a promoting
19 response rather than i n i t i a t i o n a n d promoting at the site of
20 application.
,, ... ........
21 MR. HEINEMAN: Ajt this pqint, we would move the admission
22 of Defendant's Exhibit 129.0.,.
23 (Defendant *s.^Exhlbit ,1290 was offered into
24
e v i d e n c e .)
,,_
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1 MR. CARR: No object^on, ,,your Honor.
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2 THE COURT: Okay,, 12$0 1%,admitted without objection.
3 Thank you.
4 (Defendant *s.tExhibit ,,129Q. .was admitted
5 into evidence.) ^ ,, _ _
6 Q Now, Dr. Dost, I/d lj.ke o talk to you again about some
7 of Dr. Zahalsky's testimony,, .and in that connection, I'd like to
8 show you what's been marked ,,as Plaintiff's Exhibit 253. Have
9 you seen that before, sir?. . .
10 A I 've seen pictures o it.,
11 Q All right. Or a. picture ,,of it?
12 A Yes.
,. . . . . . .
13 (Plaintif f 1s,,..Exhibit ,129 \ was marked
14 for Identification.),
15 Q I 'll now show yo,,a w h a t 's ,,been marked as Defendant's
16 Exhibit No, 1291 and ask. .if .that *s .a blow-up of a photograph of
17 Plaintiff's Exhibit 253.,,. _
18 A Yes, it appears ,,to rapreaent.it,
19 Q Now, Dr. Dost, you have read, have you not, sir, the
20 testimony of Dr. Zahalskyvln .which .this chart was discussed? 21 A Yes, I have. ,, ,,. ,, . 22 Q All right. Do you agree,,with that testimony and this
23 chart?
.
24 A No, I do n 't, reaJLly.,
it
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1 Q All right. Would yot| .please come up and explain to the 2 Jury why you disagree with ,,the .chart and .the testimony relating
3 to it.
4 A This as I recall, appeared in Dr. Zahalsky's testimony at
5 a time when he was dlscu.ssing ,,effects on the Immune system, and
6 if I remember correctly began talking ,,about this representation,
7 and I guess that I would, ignore ,,that and take for granted his
8 representation that this, scheme .represents the way that dioxin
9 causes cancer, and it is..pearly incorrect. There is, the
10 schematic that w e 1re dealing .with here-11 MR. CARR: Mr. Meineman, ,,what,,would be the date the 12 testimony was taken and .thepage number that he is referring to?
13 MR. HEINEMAN: I^t starts.at 101, ,,1 think, and following
14 that, X think is where hje ,,starts talking about it.
15 MR. CARR: What ,date7,
16 MR. HEINEMAN: May th,e 3t,d._ 17 A Well, the scheme... that^ s represented here is clearly a
18 scheme that is apparently ..related .through the .induction of
19 microsomal enzymes, the J?45.Q enzymes. . And what we have here,
20 this looks very much Ilka a ,,diagram that D r , Elsen published at
21 one time or a part of thatdiagram, but.ITm not prepared to say
22 that it was adapted from it ,directly. ,, When TCDD or any of a
23
' ** t >' "
*" ,T'
*1 '
number of other polyaromatic hydrocarbons, and we've discussed
24 this earlier, I believe,,; when it ,,moves into the cell in order to
N
\J
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1 cause the induction process ,,,it :*has to have a way of starting 2 the process, and just by. tts;elf ^in the cell it can't do that.
3 It attaches to a r e c e p t o r ,I d o n !t li^e the term recognizer, 4 because that implies tha.t ,,this is so .specific that it Identifies 5 each molecule that comes..intQ .the cell. .The fact is that this
6 receptor will bind a very .large variety of substances. It happens
7 that it binds TCDD more ,.intensely than it does any other, and I
8 think that's just an accjldent of^the .very .flat structure of TCDD.
9 It turns out to have provided .us with an excellent tool, however, 10 for studying this process -Qf,,induetion, .and it has led to an 11 understanding of the way^ that all of these complex chemicals cause 12 induction. The two join, and.they form a complex between TCDD and 13 the receptor, and that receptor moves into the nucleus of the cell. 14 Now, this receptor is no.t .put .on .the .surf ace of the cell, it is 15 free within the cytosol jar .the body of the cell. That's why they 16 call it a cytosolic receptor. It is in the cytosol of the cell. 17 And it is correct that it ,,does mq.ye rom the cell body into ,,the 18 nucleus of the cell. Bet-it .starts CtP .in here. The T C D D vcomes 19 into the cell and Joins ,,with .the receptor to form this complex,
20 and the complex then moves ,,into.the nucleus. . There is a whole 21 series of these, and I suppose ,,that we should bring them all 22 up one by one.
23 Q Let me hand you,, sir K what's previously been marked as 24 Plaintiff's Exhibit 254,,. 255, ,256, and 257, and ask you if you have
i
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107
1 seen those documents before, 2 A These represent ,,the RhotQgraph's that I have seen and
3 apparently they are In the ,,s^queijce that they originally appeared. 4 (Defendant's,,,,,Exhibits, 129>2 to 1295 were marked 5 for identification,), ,, ,,
6 Q Now, sir, would ..you ^ompqre these photographs by number
7 with the Plaintiff's exhibit ,to be sure I've got them right?
8 Defendant's Exhibit 1292, is .a photograph of what, sir?
9 A Of Plaintiff's Exhibit 254.
10 Q All right, , ,,.
11 A And this Is the ,,next ,,step. ,,
12 Q And Defendant's .Exhibit 1293 is a copy of what, sir?
13 A Let's see, is a ,,copy wof Plaintiff's 255.
14 Q All right, sir. ,, And .Plaintiff 'sExhibit 12-- I'm sorry,
15 Defendant's Exhibit 1294. is a .copy ,,of--
16
A Plaintiff's 256.^
_
17 Q And Defendant's rExh ih.it 1295?
18 A Plaintiff's 257.^
19 Q Now, would you like t;o gQ .through these one at a time?
20 A Maybe in order to maintain sequence I will put this over
21 on this easel. This complex.does in fact go into the nucleus
22 and it interacts with the,,genetic material in the nucleus I
23 don't know what really is meant by .a critical target here, because 24 what is Interacted with J.s a .segment of DNA, and this complex, I
108
presume when he says nuclear^complex ,,does not act as a message.
1
The message, what It does.It.cause In some fashion, causes the 2
DNA in a particular part,.of the .cell,. In a particular part of the
3
genome to go through the.; process of opening up and making a
4
copy of messenger RNA. .Nq w , lt.ls a fact, yes, that it does
5
result In the making of .messenger.. .We .are talking here about a 6
form of nucleic acid called.RNA, and It la very similar to the
7
DNA. Then the messenger.8. the mess.enger RNA goes to the structures 8
in the cell that manufacture .protein, .the endoplasmic reticulum,
9
10 and they provide instructions to m a k e .certain protein, and In a 11 sense, I suppose we could.say this is correct, except that as he 12 has it depicted here and^as.he,,states,It here, they are making 13 new receptors. If I understand that,, he means recognizer as,o 14 representing the receptor .or TCDD. , That ,,does not happen. What 15 is made are the group of^ enzymes that.we have discussed at various 16 times, the P450 e n z y m e s t h e ,other .non-P450 enzymes that we saw 17 illustrated on that charjt .of ,,Dr. Eisen's. I think yesterday, but 18 we do not, the cell does, not .make additional receptors, they do 19 not go to the outside lining ,,of .the cell .or the cell wall. I am 20 not sure what is m e a n t , a n d ,,they don*.t react with dioxin except 21 that those are the very .enzymes.that,,are responsible for changing 22 dioxin, metabolizing it J9Q ,,that it c$n be disposed of. They are 23 not a new set of receptors that ate ,,here in the cell body which 24 interact with TCDD. Thcre,,,is a clear limit to the number of those
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109
1 receptors. There are a few .compounds that are known to be able 2 to cause the formation oj: ..additional receptors to a limited
3 extent, TCDD is apparently ..not one of them. And these proteins 4 that we are talking about v.that .are ..formad do not go to the 5 outside of the cell. They .got t they are attached to this 6 endoplasmic reticulum, where .they are synthesized, They stay
7 there and are held in place. . These .are .what are known as
8 membrane bound enzymes as.differentiated from some of the
9 enzymes that I mentioned .an another .occasion that are soluble
t
10 that can actually leak o.ut ,q the .cell. These P450 enzymes
11 can't leak out of the cell ,,because they are stuck on larger
12 structures that can't leave.. Now, apparently Dr. Zahalsky
13 believes that these TCDD^, ,these, .new TCDD receptor complexes go
14 back through the same cycle again, ,,according to this diagram,
15 but then they do something .dlf fo r i e n t A c c o r d i n g to his
16 description, they react vand ,form ,,reactive intermediates. There .
17 are some real problems with .that .because to begin with the metabolism
18 of chemicals that might ,,or .might .not take place and might proceed
19 to "reactive intermediatnsV takes place out in the cell body on
20 that endoplasmic reticulum. ,,..There .are no such reactions taking
21 place., in the nucleus. Tjie .nucleus .has other kinds of work to do.
22 So somehow or other we have .two cycles described here that start
23 out exactly alike and t h e n .ere presumed to end up differently with
24 a different function in .the ^ecor^d cycle. So this part of the
1 - t-
110
1 description Is really completely ..incorrect. Ue don't have
2 formation of new receptors * .There is a great deal of binding 3 of TCDD to the proteins Jin .the .liver .cell, particularly the 4 endoplasmic reticulum, b,ut ,,it .is nonspecific. It Is where most 5 of the TCDD that is storjsd.in the ,,liver ends up and it is 6 just stuck there, it is jxot ..really ,,doing .anything in particular, 7 and it can be taken back, out fit a .biochemistry lab so we can 8 tell where it is. There, is ,,just a .very small amount of receptor. 9 Measurements that have b.een made indicate that no more than one 10 percent, and probably much .less ,,than that, of the TCDD in the 11 liver cell actually has Jbqund to a receptor. The rest of it is 12 all bound nonspecifically .to ,,other proteins. So we don't have 13 reactive intermediates f o r m e d ,in the nucleus, and this complex 14 between the receptor and,, dioxin _has ,nqthing to do with the 15 metabolism of dioxin except,as that receptor causes the formation 16 of the enzymes that eventually .can .break d.ioxin or it can alter 17 dioxin so that it can bea,,disposed o f , 18 The last chart sjays t,hat vnqw ,the .reactive intermediates 19 bind to critical targets.. .. He know that TCDD and its metabolites 20 do not bind to DNA. The,, implication here is that it has bound to 21 DNA inside the nucleus of ,,the cell. This has been demonstrated 22 not to occur. It has also .been demonstrated that we do not have 23 a problem of reactive intermediates that are thrashing around in 24 the cell, which is 3-methylcholanthrene or dimethyl bez anthracene
111
1 the reactive Intermediates of those substances do a great deal
2 of harm and they do bind,.anywhere they can find a point in their
3 path. The Implication la ,,this.is what results in toxicity, and
4 we've discussed this I think sufficiently to show that there isn't 5 a relationship between reactive, ,,intermediates and their binding
6 to toxicity because we d,,on*t h a v e .reactive intermediates as
7 part of the system. The,,implication is .also that this initiates
8 the process by which cancer cells, cancer is caused, starting
9 with the initiation in the .nucleus of ,,the cell, and it also
10 results in transformation* ,, .And I am .not sure what the relationship
11 of these two events is. ,,.We .would have to concede that TCDD is
12 capable of the promotion.process which, depending on definition,
13 may or may not include transformation. ,^But it has nothing to
14 do with the initiation process .So, the ,,representation that this
15 scheme is a description jaf ,,the .way that TCDD causes cancer is,I
16 think, erroneous, entirety .erroneous , I don't know where it arose.
17 THE COURT: Gentlemen cquld ,,1 see you at the bench for
18 a minute, please?
19 (At this time an ,,off-the-record conference was
20 held at the ,,bench.) , . .
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21 THE COURT: Ladl,es and gentlemen, we are going to break
22 for the day at this time..,. .We .will resume again tomorrow morning
23 at 9:30. I would remind, you as ,,1 do for any overnight break that
24 you are not to read, listen to,.or.watch anything about this case
112
1 in particular or subject matter in .general In any of the media, 2 print or electronic. Tl\ank,,yqu for your attention and cooperation. 3 Court is adjourned. We *will.start .at .9 :30 tomorrow morning. 4 (At this time Court tjas adjourned for the day.)
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1 STATE OF ILLINOIS 2 COUNTY OF ST. CLAIR
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8 I, Patricia A. Gandy,, CSR>, RPR, Official Court Reporter
9 in and for the Twentieth, .Judicial ^Cltcuit, and the Official Court
10 Reporter who transcribed^the above-styled cause had on November 5,
11 1985, do hereby certify ,,that .the foregoing transcript of proceedings
12 is a true, correct and complete transcript of the proceedings had
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14 DATED this 10th jday qf November, 1985.
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8 I, RICHARD P. GOLDEN^ERSH, .Circuit Judge in and for the
9 Twentieth Judicial Circuit, .hereby certify that the above is a
10 true and correct transcript .Qf ,tbte proceedings had in the case 11 captioned: FRANCES E. RJ3MNER, ,et ,al, , ,v. .MONSANTO COMPANY, 12 Cause.No. 80-L-970, heard .on .November.5 , 19 85. 13 DATED this _____ _ day, of _November, 1985.
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18 RICHARD JP. QOLDENHERSH, Circuit Judge
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