# OAR Box 1131 Prepped by Ollie Stewart

Document

Number: 

# 133)11-1-41 0

Docket Number: 

# A-83-2 2

Printed 4/21/2008 2:34:12 PM Sheet 133 of 162 p. 1P. 2 .-iiWr^;>«nr-i<fefc*'Uj l qiii»i*iiJ^ ^ NAJU R l*V O L . 23 6 MARC H 1 0 197 2 

## iV C 

# ^34 1 T/o Wher e th e sandstone ha s remaine d burie d an d natura l weatherin g ha s notye treache d into it , th e sulphu

r content decrease s fro m severa l tenths o f

a pe r cen tnea r the coal t o les s tha n one tent

h o f a pe r cen tnea r th e to p o f

the sandstone . Ther e is n o region simila

r t o th e highly leached weathere d zon e observe d i n th e same sandstone whe n itcontacts th e surface . Th e patter n i s shown in Fig . 1 h. A fe w o f th e roc k columns sampled t o 1 6 m sho w n o high sulphu r zon e and hav

e chroma s highe r tha n 2 . Mild geological foldin g a tthes e location s ha s resulted i n roc k fractur e wit h subsequen tweatherin

g reachin g greate r depths . I

n suc h cases , a stud y o f the fres h hig h wall s resulting fro

m mining operation s verifie s suspecte d latera

l penetration o f

weatherin

g fro m fractures, indicated b

y iron oxid e chroma

s enclosin g a

n interior o f massive , low-chroma sandston e a tdepths below th e 6 m weathere

d zon e indicated in Fig . 1 . Thi s latera l penetra tion o f weathering , extending inwar d severa l centimetres fro

m th e vertica l fissures , vividly illustrates th e "weatherin g front " (ref . 6) . Evaluatio n o f

chroma s i n suc h bicoloure d specimens consistentl y distinguishes betwee

n hig h sulphur an

d lo w sulphur portion s o f the fragment

s (Tabl e 1) . Tabl e 1 SulDhu

r i n Relatio n t o Chrom

a i n Si x Natura l Bicoloure

d Fragment s o f Lowe r Mahonin

g Sandstone * Fragmen tChrom a 1 1 2 2 3 3 4 4 5 5 6 6 4

0 3 0 6 0 6 0 4 0 6 2 Positio n in fragmen t Nea r surface Interio r Nea r surface Interio r Nea r surface Interio r Nea r surfac

e Interio r Nea r surface Interio r Nea r surface Interio r Sulphur (% )

< 0.00 1 0.17 5 0.04 8 0.28 8 0.06 0 0.19 8 < 0.00 1 0.03 5 0.03 2 0.14 0 0.00 4 0.01 0

* Groun d t o <25 0 \im; colou

r evaluate d o n dr y sample . Highe r chroma s penetrat e severa l centimetre

s fro m beddin g o r fractur e planes, lo

w chroma s typif y interiors. "Althoug h th e presenc

e o f sulphides , othe r reduce

d chemica

l species , and lo w chroma (grey) coloratio n migh tindicat e th e effect s o f th e water table , t!i

e 6 m dept

h fro m th e presen

tland surfac e is well abov e th e presen tpermanen

twate r table . The evidenc e fo r oxidatio n t

o greate r depth s o n fractur

e planes confirm s tha ta permanen

twate r tabl e i s notinvolved . Pro longe d saturation o f

th e sandston e i s a likely conditio n within th e roc k tha tha s retaine

d its lo w chroma colou r an d pyrite . Thi s conditio n i s relate

d t o roc k porosity , positio n in th e Mcction , recharg e b y meteori c water , and roc k chemistry . Details o f thes e influence

s have no tbee

n studied , bu

ttheir resultan tinteraction s evidently have maintained a n oxidatio n potentia l lo w enough t

o preserv e pyrit e in th e lo w chrom

a rock . Th e inheren tlo w fertilit

y o f soi l develope d fro

m Lowe r Mahonin g sandstone 1 i s readil

y understoo d in vie

w o f th e .iropertie s o f th e pre-weathere

d paren tmaterial . Th e solubil izatio n an d subsequen

tleaching o f

all bu tth e mostacid insoluble compound s b

y th e sulphuric acid forme

d b y oxidatio n if th e pyrit e in th e weathering sandstone have lef troc k whic h i s almostbarre

n o f plan tnutrients . Measurement s o f availabl e plan tnutrien tcation s throughou

tth e depths studie

d indicat e mens e remova l o f essentia l plan tnutrient

s fro m th e 6 m oxidize d zone . Thi s stron g aci d leaching durin g th e early >tage s o f soi l developmen

taccount s fo r lo w bas e saturation level s an d moder n classification o f

thes e soil s a

s Dystrochrept

s o r Udults 1

. The usua l metho d o f

spoil disposa

l in thi s are

a i s backfilling iin d covering wit

h materia l favourable fo

r planne d land use , usuall y afforestation , forag

e growth , or specialt

y uses . The ota l sulphur and chroma profile s i n near-surfac

e Lowe r Masonin g sandston e indicat

e tha tther e ar e adequat e volumes o f

pyrite-fre e roc k materia l t o plac e i n the oxidatio n zon e o f spoi l heap s t o avoi d acid pollution . Bu tnutrient s mus tb e adde d befor e vigorous ' vegetatio n ca

n b e established . On th e othe r hand , th e bul

k o f potentially toxi c lo w chroma materia l belo w a dept

h o f 6 m constitute s a formidable barrie r t o revegetatio n and a likely prolonged source o f aci d wate r pollutio n unles s protected fro m oxidatio n b

y buria l o r othe r means . Studies ar

e unde r wa y t o determin

e weatherin

g o f spoi l materials , rat

e o f plan tnutrien trelease , an d th e progres s o f soi l propert

y genesis , whic

h ar e expecte d t o proceed differentl y in fracture

d spoil materials tha

n in stratifie

d sedi ment s in situ. Thi s wor k was supported b

y the Environmenta

l Protectio n Agency , Wate

r Qualit y Office . WALTE

RE . GRUBE , jun . RICHAR D M . SMIT

H EVERET T _M . JENCK S RABIND , College of

Agriculture and

Forestry, West Virginia University, Morgantoyvn, West

Virginia 2650 6 Received Jul

y 23 ; revise d Novembe r 16,1971 . 

# TUSli 8 

# -. !

m I 3 

# 189 2 l U

1 White . I . C , Bull. US Geol. Surv., No . 65 (189] 3 Grimsley , G . P. , Iron Ores, Salt and Sandsh Geol . Surv. , 4 (1909) . * Jackson , M . L. , Soil

Chemical

Analysis (Prentice-Hall , 1958) . * Munsell

Soil ColorCharts (Munsel l Colo r Co. , Inc. , Baltimore , Maryland) . 5 Temple , K. L , an

d Koehler , W. A. , Drainage from Bituminous Coal Mines, WestVirginia Eng . Exp . Sta . Res . Bull . No . 2 5 (1954) . 4 Oilier , C , in Weathering (American Elsevier , Ne w York , 1969) . 1 Soil Surve

y Staff , Placement

of Series

— Northeast

Region, USDA SC S (Northeas tRegiona l Technica

l Service Center , Uppe r Darby , Pennsylvania , 1969) . _ 

## DOCKE T



# VJ* r BIOLOGICA L SCIENCE S 

# Defective Synthesi s o

f c c an d( 3 Globi n Chain s in LeadPoisonin g TH E cell s o f

man y tissues , including bone , re d cells , liver , brai

n an d nerves , absor

b lead 1 whic

h accumulate s chiefl

y o n th e mitochondria

l membrane . Her

e i tdisturb s mitochondria l metabolism an

d produces th e swelling an d disruption reveale

d b y electron microscopy 2

. Th e mostobviou s metaboli c defec t i s the inhibitio n o f

th e synthesi s o f

hae m (an d porphyrin) . Lea d inactivates a

tleastthre

e mitochondria

l enzyme s in th e hae m pathwa

y ( 8 aminolevulini c aci d synthetase3 , 5-amino laevulini c dehydrase 3 , and hae

m synthetase*) . Thi s aspec to f lea d toxicit y (reviewe

d i n ref . 5) reduce

s hae m productio n an

d lead s t o th e accumulation and excessive excretio n o f severa l porphyri n intermediates . Lea

d seems t o affec

tre d cell s chiefl y b y causin g increase

d destructio n and interfering wit h haemoglobi n synthesis , a s judged b

y th e presenc

e o f ring sideroblast

s i n th e bone marro

w an d hypochromia o f

th e re d cells . I n norma l re d cell precursor

s ih e synthesi s o f

hae m and globi n is finely balanced , bu ti n diseases suc

h as sideroblasti c anaemia

7 an d iron deficienc

y anaemia (R . Barr

, A . V. Hoff

bran d an d J. M. W. , unpublished) a defec ti n hae m synthesi s lead s t o a disturbance o f

globi n synthesis . A previou s repor t indicate d tha tglobin synthesi s wa

s defective i

n lea d poisoning 8 , bu tn o report

s hav e deal twit h the synthesi s an

d assembl y o f a an d B chains , whic

h w e hav e no w studied . Reticulocyte-ric

h bloo d samples fro

m 3 y r ol d twins , wh o ha d develope

d lea d poisonin g afte

r eating ol d paint, wer e prepare d an d incubate

d fo r 6 0 mi n wit h 3 H-leucin e an

d th e cell s wei c divide

d int o tw o equa

l aliquots . Fro

m on e aliquo t w pre p Me d "imour** " "lobi; - ! b x ' 2'J'I!" " the c)! s <1!rcc'.! y t o 

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. 4fifil )

60 S O 10 0 12 0 I J O Fractio n No . Fig . 1 Patter n o f

radioactivit y o f

protei n fraction s o f

lysat e o f twi n 1 separate d b y 'G-10 0 Sephadex ' chromatography . , Absorbanc e -280;nm ; •—• , radioactivit y (d.p.m. ml.-') . Not e tha tth e radioactivit

y associate

d wit h th e dextra n blu e pea k (arrow ) i s nearl y a s grea ta s tha tassociate d wit h haemoglobi n peak . aci d acetone . Fro

m th e other aliquotw

e mad e "pure " globi n b y separatin

g the haemoglobin fro m othe r proteins b

y chromatograph

y o f a lysat e o f washe d cell s o n 'G-10 0 Sepha dex' . Th e radioactivit

y an d absorbanc

e o f th e protei n fractions o f the lysate eluted fro

m th e column wer

e measured . Finally , th e pur e haemoglobin fractio

n wa s concentrated b

y ultra filtration and th e globi n precipitate d b y addition o f

acid acetone . The a an

d f J chains fro

m impur e an d pur e globin s wer e separate

d b y chromatograph

y o n CMC-2 3 using 8 M urea-phosphat

e buffers' . The radioactivit y associate d wit

h th e a an d f 3 chain protei

n peak s wa s measured , an

d the n th e total radioactivit

y o f eac h pea k wa s determine

d (Table 1) . Th e synthesi s o f

a an d P chain s o f

their reticulocytes was measure d in vitro, a

tthe tim e th e patient

s firstpresente

d an d 6 an d 1 2 week s later while the

y wer e recoverin

g afte r treatmen

t wit h penicillamine . Whe n firststudied th e patients ' haemoglobi n level s wer e 7. 0 an d 7. 6 g 10

0 ml.- 1 an

d bloo d lea

d levels 9 8 u g 10 0 ml.- 1

. The radioactivit

y o f the protei

n fraction s o f the cell s separate

d by chromatograph

y o n a 'G-10 0 Sephadex* i s shown in Fig . I . (Only th e result s o f on e twi n ar e shown : the othe r twin' s result

s ar e identical. ) Tw

o importan

tfeature s ar e shown . First, the radioactivit

y o f th e voi d column pea

k is nearl y a s grea ta s th e radioactivit

y o f the pur e haemoglobin fraction . (Thi s pea k contain s ribosomes an d apoferriti

n an d it s radio activit y is normall

y onl y one-twentiet

h t o one-fifteent

h o f the radioactivit

y associate d wit

h th e haemoglobi n fraction. ) 80 0

r

I 20 0 .1 0 Fractio n No . Fig . 2 Firs texperiment

. Patter n o f radioactivit

y a s associate d wit h th e a an d Pchain s o f twi n 1, separate

d o n CMC-23 . , Absorbanc e 28 0 nm ; •—• , "impure " globi n (c.p.m . ml.-') ; O O . "pure " globi

n (c.p.m . ml.-') . Note : (I ) i n "impure " globi n larg e amoun to f "y" pea k radioactivity ; Pchai

n radioactivit y > a chai

n radioactivity ; (2

) i n "pure " globi

n "y" pea k ha s disappeared ; fal

l i n P chai n radioactivit

y bu tal l a chai n radioactivit

y recovered

. P. 3

NATUR E VOL

. 23 6 MARC H 1 0 1 9 I Tabl e 1 Labora t

Hb( g 10 0 ml."' ) MCH C (% ) Reticulocyte s (% ) Bloo d lea d (n g 10 0 ml. " UbF(% ) HbA,(% ) M b electrophoresi s ory Inv c

' ) istigation s o n

Twi n I 7. 3 28. 0 10. 0 98. 0 3. 2 1. 3 A Patient s

Twi n 2 k 7. 6 27. 5 11. 5 98. 0 3. 0 1. 8 A Second , the amoun

to f radioactivit

y associate d wit

h haem o globin fractio

n (3,00 0 d.p.m . ml. -1 ) is ver y low fo r areticulocyi

i counto f 10% . Togethe r thes e findings indicat e indirectl y th a th e synthesi s o f

the haemoglobi n tetrame r i s abnormal . Dire

c evidence fo

r thi s wa s obtaine d fro

m studie s o f

the synthesi : o f th e isolate d a an

d fi . chains .

o

• 6 o a:

h raclion No

. Fig . 3 Secon d experiment

. Patter n o f radioactivit

y o f aan d P chainso f (wi n I . Note : (1 ) i

n "impure " globi

n fal l o f "y" pea k activit y an d relativ e increas e i n P chai n an d a chai n radioactivity ; (2

) mos to f th e p an d a chai n radioactivit

y recovere d fro m "pure " globin . The radioactivit

y associate d wit

h th e a an

d ( 3 chains isolate

d fro m the impur

e globi n preparations i s shown in Fig . 2 . Mos t o f th e radioactivit

y wa s found i n a pea k i n fron to f the P chai

n pea k i n the region corresponding t o th e y chain . Thi s pea

k i s called "y" . The radioactivit y o f th e P chain was les s tha n thi s initia l pea k bu tsignificantly greate

r tha n tha tassociated wit h th e a chain peak. Th e ratio s o f th e radioactivit

y of th e a : P+ "Y " chains wer

e 0.23 an d 0.2 5 in twi n 1 an d twi n 2, respectively , and th e ratio s o f th e radioactivit

y o f th e a: p chains alone , 0.6 0 an

d 0.61 . The patter

n o f radioactivit

y associated wit

h the a an

d P chains isolate d fro

m pur e globi n preparation

s showe d severa

l differences . Th e "y " pea k o f radioactivit

y ha d disappeared and ther

e was a fal

l in th e radio activit y o f th e P chains . Virtuall

y all th e radioactivit

y asso ciated wit h the a chai n i n th e impur e globi n sample , however , was recovere

d in th e o chai n isolate d fro m th e pur

e globin . Si x week s late r th e patter

n o f radioactivit

y obtained afte

r elutio n o f the impur

e globi n preparatio

n show s a fal

l i n radioactivit y associate d wit

h th e "y " pea

k and a relative increas

e in th e radioactivit

y associate d wit

h the P an.

d a chains (Fig . 3) . Th e patter

n o f radioactivit

y o f th e chains isolate d fro

m th e pur e globin preparatio

n was ver

y simila r excep ttha

tth e "y" pea k wa s missing and th e specific activit y o f th e P chain wa

s reduced . Th e ratio s o f

radioactivit

y o f a : P + "y " peak

s mad e fro m th e impur e globi n wer

e 0.65 an d 0.6 8 an d the o : P ratio

s 0.8 3 an d 0.87 . Th e findings o f

th e final experiments , 12 week s later , ar e show n i n Fig . 4 . The radioactivit y o f the "y " pea

k ha d disappeare

d an d the radioactivit

y o f the a an

d P chai n peak s fro m bot h th e pur e an d impur e globi n preparation

s wa s ver y similar . Th e rati o o f th e radioactivit

y o f a: P + "y " chains wa

s 0.8 9 an d 0.9 0 an

d th e a : p rati o 0.93 an

d 0.96 . Fig . 5 shows the ratio s o f radioactivit

y o f th e a: P + "Y

" chain s mad

e fro m impur e globi n preparation

s fro m eac h twi n

UP M «iMman.B,n.wiJM-tfmmJHUg w "mmrtmSTJyWii CT^.^».».fiSiBP. 4

NATUR E }/0 L 23 6 MARC H 1 0 197 2

.0. 8 0. 7

f 0. 6 g 0. 5

" * 0. 4 0. 3 0. 2 0. 1 .Gradien t



### - 4 r

1 0 

# j - 

# I \ 

# / v i \ i \ 

# J 

# L ^ V

2 0 3 0 4 0 5 0 Fractio n No . 80 0 70 0 1 E 60 0 g 500 - S "iviiyoacl 20 0 « 10 0

Fig . 4 Third experiment . Pattern o f radioactivit

y o f a an d P chains , twi n I . Note : absence o f "y" radioactivit

y an d nearly equal amounts o f

p an d a chai n activity , mos to f whic h i s recovere d fro

m th e "pure " globin . plotte d agains tth e haemoglobi n values . A s th e haemoglobi n level s rise, an d th e bloo d lea d level s fall , th e a : P + "y " rati o become s nea r t o 1.0 . Thes e findings indicat e tha ti n th e reticulocyte s o f th e - patient s globi n synthesi s i s disordered . I n th e initia l experi ments , th e P chai n pea k o f th e impur e globi n preparatio n wa s mor e radioactiv e tha n th e a chai n peak . Becaus e th e chain s wer e isolate d fro m whol e cell s thi s finding indicate s tha tth e tota l synthesi s o f

p chain s wa s greater tha n tha to f a chains . Bu tradioactivit y o f

th e a chai n pea k wa s virtuall y th e sam e i n th e impur e an d pur e globi n preparations , whic h indicate s tha tnearl y al l th e a chain s synthesize d combin e wit h P chain s an d hae m t o for m haemoglobin . Th e finding tha tth e radio activit y o f

th e P chai n pea k wa s les s i n th e pur e globi n prepara tion s agai n suggest s tha tther e wa s a relativ e exces s o f P chain s whic h existe d a s a fre e (uncombined ) pool . Th e natur e o f

th e "y " pea k i s uncertain . A simila r pea k o f activit y ha s bee n foun d i n preparation s mad e fro m marro w cell s culture d withou thaem' 0,11

. Th e pea k wa s originall y though tt o b e du e t o th e synthesi s o f tru e y chains 10 , bu t recen tanalysi s ha s show n i tt o b e a protei n o f a large r mole cula r weigh twit h a differen tpeptid e sequence 11

. A n alterna tiv e suggestio n i s tha ti ti s apoferritin 11

. Ou r dat a suppor t thi s a s w e wer e unabl e t o demonstrat e thi s pea k i n th e pur e preparation s an d als o foun d a larg e amoun to f radioactivit y associate d wit h th e apoferriti n pea k afte r 'G-10 0 Sephadex ' Chromatography . Moreover , th e leve l o f H b F (a 2 Pj ) i n th e tw o patient s wa s neve r greate r tha n 3 % an d highe r level s woul d hav e bee n expecte d i f thi s pea k wa s tru e o f

y chain . A simila r althoug h lesse r degre e o f defectiv e synthesi s i s see n i n tw o othe r condition s du e t o hae m deficiency , namel y sideroblasti c anaemia 7 an d iro n deficienc y anaemi a (R . Barr , A . V . Hoffbran d an d J . M . W. , unpublished) . I ti s temptin g

?•:• !• . V j 2

•:> • 

### f. V i^

. *

a 

### x » • 0

> N • >cti 73dio 5(al o C o 0.9 0. 8 0. 7 0. 6 0. 5 0. 4 0. 3 0. 2 0. 1 Bloo d leve l (/' g 100ml.-' ) 9

8 4 5 2 0 ;, . 7.

0 8-

0 9.

0 10.

0 11.

0 V H b value s < g 100ml.' 1 )

- Fig . 5 Correlatio n between ratio s o f tota l radioactivit

y o f th e «***» : P + "y " chain s wii h haemoglobi n value s o f bot h twins . Bloo d --•• V lea

d level s ar

e als o indicated . S.Tv.ir . 3 ; C.twir . T 7 3 t o postulat e tha tlead , i n additio n t o it s undoubte d effec t o n hae m synthesis , directl y affect s th e synthesi s and/o r assembl y o f th e globi n chains . I n suppor to f thi s conclusion , whe n reticulocyte s ar e incubate d wit h lea d in vitro polysoma l dis aggregatio n occurs 12

. Thi s ha s als o bee n observe d i n reti culocyte s wit h hae m (iron ) deficiency , however , an d th e disordere d synthesi s i n lea d poisonin g ma y b e wholl y du e t o hae m deficiency . I ti s uncertai n whethe r protei n synthesi s i s disturbe d i n othe r cell s bu ti n reticulocyte s th e abnorma l synthesi s o f globi n chain s seem s t o b e a facto r i n th e pathogenesi s o f th e anaemi a o f lea d poisoning . J . M . WHIT E MR.C Group on Haemolytic Mechanisms, Royal Postgraduate Medical

School D . R . HARVE Y Department

of Child Health, Hammersmith Hospital, Ducane Road, London W\2 Receive d Jul

y 13 ; revise d Septembe r 20 , 1971 . * Barry , P. S . I. , an d Mossman , D . B. , Brit. J. lndust. Med.,11, 33 9 (1970) . 2 Bessis , M. , an

d Breton-Gorius , J. , Blood, 19 , 63 5 (1962) . 1 Dresel , E . I . B. , and Falk , J . E. , Biochem. J., 63 , 8 0 (1965) . 4 Goldberg , A. , Ashenbracker , H. , Cartwright, G . E. , an d Win trobe , M . M. , Blood, 11 , 82 1 (1956) . 5 Goldberg , A. , Seminars Haematol., 5 , 42 4 (1968) . 6 Waldron , H . A. , Brit. H. lndust. Med., 23 , 8 3 (1964).

_ 7 White , J . M. , Brain , M . C.an d Ali , M . A . M. , Brit. J. Haematol., 20,263(1971) . • Kasenaar , A. , Morel! , H. , and London , I . M.,J. Biol. Chem. ,229, 423(1957) . 9 Clegg , J . B. , Naughton , M . A , an d Weatherall , D . J. , J. Mol. JB/O/.,19,91(1966) . 1 0 Gabuzda , T . G. , Melvin , M. , Silver , R. K. , and Sachs , C , Proc. Thirteenth Congr. Intern. Soc. Haematol., 8 (1970) . 1 1 Wood , N. , Whittacker , J . A. , Clegg , J . B. , an d Weatherall , D . J. , Absl. Brit. Soc. Haematol., 19 (1971) . 1 1 Trakatellis , A . C , Heinle , E. , Montjar , M. , Axelrod , A . E. , an d Jensen , W . K , Arch. Biochem. Biophys., 112 , 8 9 (1965) . 

# Transmissio n o f

Scrapi e t o th e Cynomolgu s Monke y (Macaca fascicularis) SCRAPI E ha s bee n transmitte d t o th e cynomolgus , o r crab eating , monke y (Macaca fascicularis) wit h a n incubatio n perio d o f

mor e tha n 5 y r fro m th e tim e o f intracerebra l inoculatio n o f

scrapie-infecte d mous e brain . Th e anima l develope d a chroni c centra l nervou s syste m degeneration , wit h ataxia , tremo r an d myoclonu s wit h associate d sever e scrapie lik e patholog y o f

intensiv e astroglia l hypertroph y an d proli feration , neurona l vacuolatio n an d statu s spongiosu s o f gre y matter . Th e strai n o f scrapi e viru s use d wa s th e eight h passag e i n Swis s mic e (NIH ) o f a Compto n strai n o f

scrapi e obtaine d a s nint h intracerebra l passag e o f th e agen ti n goa t brain , fro m D r R . L . Chandle r (ARC , Compton , Berkshire). Th e monkey , a juvenil e cynomolgus , wa s inoculate d o n Apri l 18 , 1966 , wit h a 10 % suspensio n o f th e scrapie-infecte d mous e brai n i n phosphat e buffere d physiologica l saline , p H 7.4 , whic h ha d bee n clarifie d b y centrifugatio n a t3,50 0 r.p.m . fo r 3 0 min . Th e anima l receive d 0. 2 ml . o f suc h suspensio n b y eac h o f five routes : intracerebral , intravenous , intraperitoneal , intramuscula r an d subcutaneous . Thi s suspensio n ha d a n LD5 0 infectivit y titr e o f 10~ 7 S ml." 1 b y intracerebra l inoculatio n i n mice . Fiv e year s an d five month s afte r inoculation , th e anima l develope d sligh tataxi a an d tremor s o f th e hea d an d extremities . Thes e sign s becam e progressivel y mor e sever e wit h generalize d bod y tremor s mor e prominen tdurin g feedin g an d o n agitation . U y C suck s afic i onse tliiei c v>a: > pionouncc u locomoto r ataxi a an d cl^msinc ^ ir. handlin g foo d an d myocloni c jerk s wc i