INFLUENCE OF THE ADRENAL CORTEX ON BODY WATER DISTRIBUTION AND RENAL FUNCTION.

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G audino, N ic o la s Mario H o ra cio , 1918I n f lu e n c e o f th e ad ren al c o r te x on body w a ter d i s t r i b u t i o n and r e n a l - fu n c tio n ,, New Y ork, l9l|-9« 39 t y p e w r it t e n le a v e s * d ia g r s . 29cm. T h e s is (PhoD .) - New York U n iver­ s i t y , G raduate S c h o o l, 19!?0* B ib lio g r a p h y : p . 3 1 -3 9 • C50718 )

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THIS DISSERTATION HAS BEEN MICROFILMED EXACTLY AS RECEIVED.

LIBRAR2 O f m TORK UfllYERSlTT m n ® i n h eioh is

INFLUENCE ON

BODY

OF

THE

ADRENAL

WATER

D I S T R I B U T I O N

AND RENAL

N.

FUNCTION

MARIO

H.

«»*

GAUDINO

A d is s e r ta tio n i n th e departm ent o f Physiology subm itted to th e f a c u lty o f th e G raduate School o f A rts and S cience o f New York Uni­ v e r s i t y i n p a r t i a l f u lf illm e n t o f th e requirem ents f o r th e deg ree o f D octor o f P hilosophy.

f

"

CORTEX

**r " ^

ftflKWnWT.EDGMKHT

I w ish to ex p ress my in d eb ted n ess to Dr. Homer W. Smith f o r providing f a c i l i t i e s and guidance f o r th e p ro se c u tio n o f t h i s v o rk , to Dr. Domingo Gamez f o r h is v a lu a b le advice and c r itic is m , to D r. Marvin F. L e v itt f o r h i s h e lp and companionship in accom plishing th e a c tu a l v o rk , and to a l l th o se t h a t one vay o r an o th er had an in flu e n c e i n i t s f i n a l com pletion.

2

o

\ o

^

INFLUENCE OF THE ADRENAL CORTEX ON BODY EATER AND RENAL FUNCTION TABLE OF CONTENTS

Page INTRODUCTION

....................................................................... . . . . .

REVIEW OF THE LITERATURE...................................................

1

3

In flu en ce o f th e A drenal Cortex on th e Body W ater D is trib u tio n • • • • • « » • • • . • .

3

In flu en ce o f th e Adrenal Cortex on. Renal Function

5

METHODS

.......................................................................................................

7

RESULTS

.........................................................................................................

9

A otion o f PesQgycortiooBteroneA cetate

. . . . . . . .

9

A ction o f Adrenal C o rtic a l E x t r a c t ......................... . E f fe c t o f T o tal Adrenalectomy DISCUSSION SUMMARY

15

............................

• . 15

....................................................................... . . . . . . . ..............................................................................

BIBLIOGRAPHY............................................................................

25 29

31

UfTRQDUCTIOH

A r e l a t i o n between th e a d re n a l o o rte x and th e d is tr i b u ti o n o f body w a ter has been recognised sin ce th e f i r s t dem onstration t h a t a d re n al in s u ffic ie n c y was a s s o c ia te d w ith a marked degree o f h aao co n o e n tra tio n .

The su ggestio n t h a t t h i s hem occncantraticn

n ig h t be a tt r i b u t e d to a d eerease in plasm a volune was l a t e r confirm ed by d i r e c t measurement and by th e circum stance t h a t n o m a l plasma volume was re s to re d w ith adequate d e so x y co rtico stero n e therapy* The a d m in is tra tio n o f c o r ti c a l e x tr a c t to anim als in a d re n al in s u ffic ie n c y was follow ed by hem odilutian and d iu r e s is even though no w a ter was in g e s te d .

Swingle and h is c o lla b o ra to rs suggested

t h a t a d re n a le o to a ise d anim als were unable t o m obilize f l u i d s from th e i n t e r s t i t i a l and o e ll sp ac e s, and t h a t th e hormone e f fo o te d th e r e d is tr ib u tio n of. f lu id s in th e organism .

According to Harrop

f l u i d en tered e e l l s during in s u ffic ie n c y and was re le a s e d by th e ra p y . The e x tr a c e llu la r spaoe, a s judged by th e volume of d is t r ib u ti o n o f o h lo rid e and th io c y a n a te , was decreased a f t e r adrenalectom y and r e s to re d by hormone th e ra p y .

M oreover, th e p ic tu r e o f a d re n al

in s u ffic ie n c y was sim ulated by p e rito n e a l d ia ly B is , a prooedure whioh reduoes e x tr a c e llu la r f l u i d volume. As a consequence o f th e changes n o te d in serum e le e tr o l y te p a tte r n d u rin g a d re n al in s u ffic ie n c y , a tte n tio n was focused on th e sim ultaneous e le c tr o l y te and w a te r changes in th e t i s s u e s .

C alcu la ­

tio n s of th e tis s u e w a ter compartments, based upon th e assum ption t h a t e h lo rid e i s lim ite d to e x tr a o e ll u la r d i s t r i b u t i o n , re v e ale d a

s h i f t of w ater from th e i n t e r s t i t i a l in to th e i n t r a o e l l u l a r space* M arshall and D avis, by measuring th e e x c re tio n o f phenol s u lfa a e p h th a le in , u rea and c r e a tin in e , dem onstrated a d ecrease in re n a l fu n c tio n follow ing oomplete adrenalectomy*

The subsequent

use of more p re c is e measures o f re n a l a c t i v i t y confirm ed th e l a t t e r observation*

I n u lin , c r e a tin in e , u rea and d io d ra s t c le a ra n c e s

as w e ll as d io d ra s t Tm were found to be reduced a f t e r adrenalectom y and re s to re d a f t e r adequate hormone th e ra p y . In view o f th e above evidence, i t seemed d e s ir a b le to measure ohanges in a l l th e body w a ter compartments and to c o rre la te them w ith m o d ificatio n s in re n a l fu n c tio n observed sim u ltan eo u sly in anim als s u b je c t to a l t e r a t i o n s of ad ren al fu n c tio n .

Previous methods fo r

th e e stim atio n o f th e e x tr a c e llu la r space used e i t h e r th io c y a n ate o r c h lo rid e , b o th o f which substances a re now known to e n te r c e l l s in a v a ria b le p ro p o rtio n .

To overcome th i s o b je o tio n a more a c c u ra te

method ( in u lin spaoe ) f o r th e d e term in atio n o f th e e x tr a o e llu la r volume was d e v ise d .

Coupled w ith th e use of heavy w qter (DgO) as a

measure of th e t o t a l body w a te r, a c c u ra te e stim a te s of i n t r a c e l l u l a r volume were f e a s i b l e .

The use o f ra d io a c tiv e sodium (9a 24 ) o r

potassium (X42 ) , a ffo rd e d means f o r d e te rmining th e i n t r a o e l l u l a r d is tr ib u tio n of th e se io n s .

"3" REVIEW OP THE LITERATURE

In flu en ce of th e a d re n al o o rtex on th e body w ater d is t r i b u t i o n .

Since th e f i r s t o b serv atio n s o f a d re n al

in s u ffic ie n c y in anim als and in man. th e e x iste n c e o f a marked degree of hem oooncentration and a h ig h h em ato crit count were recognized a s an o u tstan d in g m a n ife sta tio n of th e d is e a s e (3*24*33,35*57,62).

These changes were c o n si­

dered to be an in d ic a tio n o f a dim inished plasma volume* a concept t h a t was l a t e r confirm ed by d ir e c t measurement w ith v a rio u s methods ( t o t a l exsanguination* v i t a l red* T -1824).

The plasma volume was decreased 50 p e r c en t or

more from th e c o n tro l v alues (6*53*57*60,62).

A fte r desoxy-

c o rtic o s te ro n e therapy* th e blood volume of p a tie n ts w ith A ddison's d is e a s e rev ealed a marked in c re a se (6 2 ).

The same

e f f e o t was observed in adrenalectom ized dogs up to values above p re o p e ra tiv e c o n tro ls (6*53).

m th e dog* th e maximum

a c tio n of d e so x y c o rtic o stero n e was reached about th e seventh day and tended to d isap p ear under prolonged tre a tm e n t (5 3 ). C o rtin was l e s s e f f e c tiv e in th e r e s to r a tio n of th e plasma volume and re q u ire d th e a d d itio n o f sodium o h lo rid e f o r com­ p le te n o rm a lis a tio n . (6 ) . The w ithdraw al o f th e o o r tio a l hormone in ad renalectom ised dogs was follow ed by a n eg ativ e w ater balance* b u t th e lo s s o f f l u i d was n o t enough t o account f o r th e d e e lin e in blood volume (5 9 ). e sta b lish m e n t o f th e tre a tm e n t determ ined an immediate blood d i­ lu tio n and a s ig n if ic a n t d iu re s is* although no w a ter was in g e ste d (20*58).

Re­

The su ggestion was made by Swingle and c o lla b o ra to rs t h a t anim als in a d re n al in s u ffic ie n c y a re unable to m o b ilise f lu i d s from th e i n t e r s t i t i a l and o e l l spaces (6 8 ).

A ccording to Barrop (20)* th e

i n t e r s t i t i a l f l u i d e n te re d th e o e lls d u rin g in s u ffic ie n c y and was re le a s e d by th e hormone therapy*

The improvement of th e hemocon-

c e n tr a tio n as a consequence o f th e hormone tre a tm e n t was indepen­ d ent o f th e plasma le v e ls o f sodium and c h lo rid e and le d Swingle and c o lla b o ra to rs (55) to p o s tu la te t h a t th e hormone e ffe c te d the r e d is tr ib u tio n o f f lu i d s in th e organism . Harrop (20) and W inkler* Ekinton and Eisenman (65) studying one dog re s p e c tiv e ly * found t h a t th e e x tr a o e llu la r space* as measured by th e volumes of d is tr ib u tio n o f th io c y a n a te and ra d io a o tiv e chloride* was deoreased a f t e r adrenalectom y and re s to re d w ith hormone th e ra p y . The e f f e c t o f d eso x y c o rtic o stero n e was more in te n s e when used alone th an when combined w ith c o r tin (6 5 ).

C o rtin n o rm alises th e th io -

oyanate space b u t th e ooupling w ith d e so x y o o rtico ste ra n e r e s u lts in an in c re a se o f th a t space above normal v a lu es ( 6 ) .

In p a tie n ts w ith

A d dison's disease* th e volume o f d is tr ib u tio n o f th io o y an ate i s a ls o en larg ed a f t e r d e so x y c o rtic o stero n e th erap y (3 5 ). The in tr a p e r ito a e a l in je c tio n o f gluoose* a prooedure t h a t i s considered to reduce e x tr a c e l lu la r f l u i d volume (9)* reproduces th e p ic tu r e o f a d re n al in s u ffic ie n c y * in c lu d in g th e hem oconoentration and th e decrease in th e th io o y a n a te space (14*20*42*54). The c h a r a c te r is tic changes observed in th e plasm a e le o tr o l y te p a tte r n d u rin g a d re n al in s u ffic ie n c y (5*4*55) d ire o te d th e a tte n tio n t o th e sim ultaneous e le o tr o ly te and w ater m o d ifica tio n s in th e t i s s u e s .

The t o t a l w a ter oont e n t o f th e liv e r* musole and

o th e r ti s s u e s in o reased a f t e r adrenalectom y (50*51)*

By assuming

t h a t a l l th e e h lo rid e i s e x tra o e llu la r* th e tis s u e w a ter ocmpartments were c a lc u la te d durin g a d re n a l in su ffic ie n c y * re v e a lin g a s h i f t o f w a ter from th e i n t e r s t i t i a l in to th e i n t r a o e l l u l a r space (8«17*26»43).

According to H arriso n and Darrow (8*17)* th e con­

c e n tr a tio n of th e e x tr a c e llu la r e le c tr o ly te i s th e p r in e ip a l f a c to r in th e c o n tro l of w a ter exchange in a d re n al in su ffic ie n c y * th e inw c re ase i n th e i n t r a o e l l u l a r w a ter being th e consequence o f th e de­ crease in th e plasma and i n t e r s t i t i a l f l u i d c o n c e n tra tio n s o f sodium and c h lo rid e*

When Upjohn* s o o r tio a l e x tr a c t was given th e

t o t a l w ater decreased*the e x tr a o e ll u la r f l u i d b ein g f u r th e r reduced w ith a sim ultaneous f u r th e r expansion o f th e i n t r a c e l l u l a r volume (42)*

Conversely* d e so x y c o rtic o stero n e in normal anim als produoed

a s h i f t o f w a te r from th e i n t r a o e l l u l a r in to th e i n t e r s t i t i a l space (4 1 ). The above evidence su p p o rtin g th e eonoept t h a t d u ring ad ren al in s u ffic ie n c y th e re i s a decreased volume o f e x tra ­ c e l l u l a r f l u i d w ith a probable in c re a s e in th e i n t r a c e l l u l a r w a ter i s based on th e e stim a tio n of th e e x tr a o e llu la r spaoe w ith th io ­ cyanate o r o h lo rid e .

Both o f th e se su b stan ces a re known to e n te r

th e c e ll s in a v a ria b le p ro p o rtio n (1*2*7*36*37)* e s p e c ia lly in c e r ta in p a th o lo g ic a l c o n d itio n s (44*45*46)*

Thus* th e re i s no

c e r ta in ty th a t c a lc u la tio n s b ased an th e assum ption t h a t th e se substanoes measure e x tr a o e llu la r space a r e a c tu a lly p re s e n tin g th e tru e p ic tu r e o f th e events concerned* In flu en c e o f th e a d re n a l o o rte x on re n a l fu n c tio n s

M arshall

and Davis (39)* in 1916* m entioned an in o re a se in th e u re a co n ten t

-6 -

of th e blood, a f t e r complete a d ren aleo tcn y and dem onstrated t h a t th e cap a o ity o f th e kidney f o r e x c re tin g u re a , c re a tin in e and p h e n o lsu lfa a e p h th a le in was dim inished*

The in o re a se in non

p r o te in n itro g e n has been c o n s is te n tly observed in anim als as v e i l a s m man {21*32*23* 3 4 ). The more p re o is e measures o f kidney fu n o ticn developed sub­ seq u en tly confirmed th e g en eral deorease in r e n a l a e t i v i t y a f t e r a d re n ale o tc o y .

la dogs and c a t s , th e c r e a tin i n e , in u lin and u re a

c lea ra n c es a re reduced, e s p e c ia lly d u ring s e rio u s in s u ffic ie n c y (19*63,67).

D io d ra st olearance and d io d ra s t Tm were re p o rte d

dim inished in two dogs (63)*

Complete r e s o tr a tia n o f the

c re a tin in e and u rea c le a ra n c e s was o b tain ed w ith c o r ti c a l hormone th e ra p y (19)*

The re tu rn to normal o f th e in u lin and d io d ra s t

clea ra n c es and o f d io d ra s t Tm has been stu d ie d w ith a combined tre atm en t of o o rtio a l e x tr a c t, d eso x y o o rtio o stero n e and Preloban (6 3 ). The in je c tio n o f U pjohn's c o r ti c a l e x tr a c t in one normal dog in c re ase d th e in u lin o le a ra n c e , decreased th e d io d ra s t Tm and d id n o t change the d io d ra s t clearan o e (27)*

In normal cats*

d eso x yoorticosterone th e ra p y does n o t modify s ig n if ic a n tly th e c re a tin in e clearanoe (6 6 ). The dim inution in th e re n a l fu n c tio n was a ls o p re s e n t in p a tie n ts w ith A ddison's d ise a se d u rin g in s u ffic ie n c y (38*61)* The most prom inent re d u c tio n was found in th e i n u lin o le aran c e and in the glucose Tm. Ae th e d io d ra s t clearan o e was le s s m odified th e f i l t r a t i o n f r a o tio n d ecreased . m a in tain ed .

D io d ra st

was g e n e ra lly

D esoxyoortioosterone tre a tm e n t te m p o ra rily re s to re d

th e s e fu n ctio n s* b u t had no n o tio n on normal in d iv id u a ls (6 1 ).

-7 -

METHODS E x t r a c e l lu la r space wcs m easured a s thevolum e o f d is tr ib u tio n of in u lin .

T h e to ts l i n u l i n re c o v e re d , a f t e r th e

c e s s a tio n o f a c o n s ta n t in tra v e n o u s e q u i lib r a t in g in f u s io n , c o rre c te d f o r u r in a r y d e la y tim e , and d iv id e d by th e plasma, c o n c e n tra t.'o n a t th e moment o f th e in t e r r u p t i o n o f th e in f u s io n , e q u a ls th e volume o f d i s t r i b u t i o n o " in u l i n (1 2 ). T o ta l body w ater was determ in ed by th e space o f d i s t r i b u t i o n o f heavy w ater (D2O ).

A known amount o f 99*8 se r

c e n t pure DgO was in j e c te d in tr a v e n o u s ly .

T hat amount was

c - le u la t e d to g iv e a plasma, c o n c e n tr a tio n o f a p p ro x im ate ly 0 .1 0 p e r c e n t D2O.

Blood sam ples were drawn 2 and 3 h o u rs l a t e r , when

com plete e q u i l i b r a t i o n was a s s u r e d .

The bloods were an aly zed f o r

D2O c o n c e n tr a tio n by th e f a l l i n g drop method o f K esto n , E itte n b e r g and Schoenheimer (2 8 ), a cc o rd in g to th e fo llo w in g p ro ced u re:

th e

blood sample kps c h i lle d w ith carbon d io x id e and th e n d i s t i l l e d i n v acu o .

The d i s t i l l a t e was p a s s e d th io u .h th e com bustion tu b e

d e s c rib e d (28) and a g a in condensed.

T his sample o f w ater was

k e p t in th e r e f r i g e r a t o r u n t i l th e moment o f th e d e te rm in a tio n , when i t was r e d i s t i l l e d f o r a t h i r d tim e .

The s ta n d a rd s o lu tio n s

were s im i la r l y p assed th ro u g h th e com bustion tu b e and f i n a l l y d is tille d .

The d e n s ity o f th e unknown end s ta n d a rd s o lu tio n s was

determ ined by th e v e l o c ity o f f a l l o f a drop o f f ix e d volume in an in m is c ib le medium ( o - f lo r o to l u e n e ) , k e p t a t a c o n s ta n t te m p e ra tu re w ith in 0.001°C .

The c o n sta n c y o f th e s iz e o f th e d ro p was a s s u re d

by th e u se o f a s in g le dro p Levy p i p e t t e .

To im prove th e acc u racy

-8 o f th e d e te rm in e .tio n , th e unknown sam ples were dropped r.etvreen s ta n d a rd s o f a p p ro x im ate ly th e same d e n s ity . cu rv e was c o n s tr u c te d , p l o t t i n g th e tim e a g a in s t p e r c e n t o f D2O.

A c a lib ra tio n

r e c i p r o c a l o f th e f a l l i n g

An example o f th e v a lu e s u sed in

p l o t t i n g such a cu rv e i s th e fo llo v d n g : Per cen t o f DoO

Time in seconds ( l )

l/T im e

0 .0 0

125.0

.0080

0 .0 5

9 7 .0

.0103

0 .1 0

7 9 .8

.0125

0 .1 5

6 6 .8

.0150

0 .2 0

5 7 .9 .

.0173

0.2 5

5 0 .7

.0197

The c o n c e n tra tio n o f th e unknown sam ples was determ in ed from such a c u rv e .

I f th e an im al had been in j e c te d w ith D2O in

p re v io u s e x p erim en ts, a c o n tr o l blood sample (B0 ) was ta k e n b e fo re th e d e te rm in a tio n and an aly zed in th e same way.

The amount o f D2O

c o n ta in e d i n t h i s c o n tr o l sample - a s s u b tr a c te d from th e 2 (Bi) and 3 (B2) h o u r sam ples.

The t o t a l amount i n j e c t e d , d iv id e d by th e

c o r r e c te d plasm a c o n c e n tr a tio n in c c . e q u a ls th e volume o f d i s t r i b u t i o n in c c .

A sample o f th e c a l c u la tio n i s th e fo llo w in g :

Sample

Time (seco n d s)

Average tim e

1/T

D20 p e r c e n t

B0

9 2 .4

9 2 .6

9 2 .6

9 2 .5

.0108

.061

Bl

6 0 .1

6 0 .2

60 .3

6 0 .2

.0166

.184

B2

6 0 .2

6 0 .2

6 0 .3

6 0 .2

.0166

.184

T o ta l amount in j e c t e d : Bi

_

B0

:

9 .9 6 c c .

.184. - .061

( ! ) A verage o f 3 o r A d ro p s.

:

.123

p e r c e n t D2O

-8 a Volume o f d i s t r i b u t i o n : 9 .9 6 .123

:

8,100 c c .

I n t r a c e l l u l a r volume v/as c a lc u la te d as th e d if f e r e n c e between th e DgO and th e i n u l i n s p a c e s .

The plasma volume

was determ ined v d th T-1824 (15) end th e th io c y a n a te sp ace was m easured by th e p ro ced u re o f C ra n d a ll and Anderson (.7). I n u lin c le a ra n c e (Cjn ) , p-am ino’n ip p u ric a c id c le a ra n c e (Cp^jj) and maximal tu b u la r e x c re to ry c a p - c ity foi* p -am in o h ip p u rate (Tmp^g) were d eterm in ed d u rin g th e e q u i l i b r a t i n g i n u l i n in f u s io n fo llo w in g th e sta n d a rd p ro ced u re o f Smith and h i s co-w orkers (5 2 ).

The chem ical

a n a ly s e s were done w ith th e method o f H a rriso n (16) f o r i n u l i n and o f B ra tto n and M a rsh a ll (5) m o d ified (52) f o r PAH. Plasma sodium end p o tassiu m were m easured w ith sn i n t e r n a l l y com pensated Perkin-ELm er flam e p h o to m eter. The t o t a l c a tio n in th e body was d eterm in ed by "in v iv o " d i l u t i o n o f Na2^ o r K^2 .

These is o to p e s were g iv e n in

a s in g le i n j e c t i o n i n such a manner a s to c o in c id e in tim e w ith th e o th e r body volume d e te r m in a tio n s .

The r a d i o a c t i v i t y

o f th e plasm a was m easured w ith a G e ig er-M u ller co u n te r on a 3 h o u r sam ple f o r Na24 and on a 9 hour sample f o r

■8b-

X

(3 1 ,3 2 ).

The number of counts re ta in e d , o o rre c te d f o r *

u rin a ry l o s s , d iv id ed by th e oounts p e r ®Eq. in th e plasm a, equals th e t o t a l o a tia n in th e body*

The i n t r a c e l l u l a r c a tio n

was c a lc u la te d from th e d iffe re n c e between t o t a l o a tic n and e x tr a c e llu la r c a tio n ( e x tr a o e llu la r volume tim es plasma c o n c e n tra tio n ) and th e average i n t r a c e l l u l a r c a tio n concen­ t r a t i o n determ ined by d iv id in g t h i s l a s t fig u r e by th e i n t r a c e l l u l a r w ater (3 1 ,3 2 ).

The plasma HHf was determ ined by

th e F o lin and WU method (1 1 ), and plasma s p e c if io g ra v ity w ith th e copper su lp h ate method (48)*

The blood p re s su re was

measured d ir e o tly in th e fem oral a r t e r y (Tyoos gauge) and th e h em ato crit determ ined by th e u su al procedure* O bservations were made an norm al, tr a in e d u n a n e sth e tise d female dogs in a 24 hour f a s tin g s t a t e , and an id e n tic a l p ro ce­ dure f o r c lea ra n c es and spaoe d e term in atio n s was follow ed in each experim ent.

A t l e a s t th re e com plete c o n tro l experim ents o f t h i s

type were perform ed, and th en th e anim als were d iv id ed in to th re e I groupst one was tr e a te d w ith d eso x y o o rtio o stero n e a c e ta te (DCA) , £ th e seoond w ith a d re n a l c o r t i c a l e x tr a c t and th e t h i r d was adrenalectom ised and m aintained e x c lu s iv e ly on DCA, s tu d ie s b ein g made b e fo re and a f t e r w ithdraw al o f DCA*

In th e f i r s t two groups,

th e dogs were follow ed u n t i l th e p h y sio lo g io a l v a ria b le s had r e ­ tu rn e d to c o n tro l values* 1*

Ik e desox y o o rtio o stero n e a c e ta te used was su p p lie d by c ib a Pharm aceutical P ro d u c ts, Summit, lew J e rs e y , and by RocheOrgancn i n c . , H u tle y , Hew J e rse y

C* P a r t o f th e Upjohn1s a d re n a l c o r ti c a l e x tr a c t u t i l i s e d was su p p lie d by th e Upjohn Company, Kalamazoo, M ichigan.

-9 -

RESULTS A otion o f degoxyoorticoB terone a o e ta te i Three normal dogs were in je c te d in tra m u sc u la rly w ith 20 o r 30 mg. o f d e so x y o o rticoaterone a o e ta te (DCA) in o i l d a ily f o r 3 weeks* 4 to 6 s e r ie s of o b serv atio n s b ein g made during t h a t p e rio d (T able I* F ig u re 1 ) . The most pronounced and e a r l i e s t response n o ted was a ohange in th e d is tr ib u tio n of body w ater*

Whereas th e t o t a l body w ater r e ­

mained n e a r ly oanstant* f a l l i n g ab o u t 10 p e r c e n t from th e c o n tro l value* th e i n t r a c e l l u l a r volume p ro g re s s iv e ly deoreased w hile th e e x tr a c e llu la r volume expanded*

T his f l u i d s h i f t was ap p aren t on

th e seoond day o f tre a tm e n t (dog 3* F ig u re 1) and a tta in e d i t s mxTiwnnn qq th e 10th o r 11th day when th e i n t r a c e l l u l a r volume in dogs 1* 2 and 3 dim inished 31* 28 and 39 p e r oent* re s p e c tiv e ly * and th e e x tr a o e ll u la r space in o re a se d 60* 51 and 62 p e r oent* The volume o f d is tr ib u tio n of th io c y a n a te q u a lit a t iv e ly tended to fo llo w th e e x tr a c e llu l a r space* as measured by in u lin * b u t no c o n sta n t r e la tio n was m aintained between them*

Plasma

volume in g en eral d id n o t change s ig n ific a n tly * d e s p ite th e la rg e tr a n s f e r s o f w ater in th e o th e r compartments} dog 2* however* sheared an ir r e g u la r increm ent o f about 30 p e r o en t from c o n tro l v alu es w ith no d e f in ite r e la tio n to th e ohanges in e x t r a c e l l u l a r flu id * At th e same tim e t h a t th e r e d is tr ib u tio n of body w a ter ooourred* th e r e was a p ro g re ssiv e in o re a se in f i l t r a t i o n r a te o f 37* 40 and 55 p e r o ent i n dogs 1* 2* and 3 re sp e c tiv e ly * an in o re a se whioh was f a i r l y w e ll c o rre la te d w ith th e expansion of th e e x tr a o e llu la r space* th e peak ohanges c o in cid in g in time*

I f I YJ-.tLcj, 1 EFFECT 0 ? ;£SQ ?rC O ?.?IC O STER O ?S

! VOLT 13

OF

D Ioff IBUTION

(lite rs )

E00

1

INULIN

a

2 .9 9

10 .2 6

7 .2 7

4 .6 0

1 .0 6

b

3 .0 4

10.45

7 .4 1

4 .8 6

1 .0 0

c

3 .0 0

10.19

7 .1 9

4 .4 9

.9 9

T reatm ent w ith 20 n g . DCA d a i l y

4

3 .6 9

-

-

4 .7 6

1.05

10

A. 82

9 .9 9

5.17

5.51

.9 8

40 mi’, d a i l y th e fo llo w ­ in g 6 d ay s.

16

3 .2 7

8.27

5.00

4.7 0

1 .0 7

23

2 .9 4

9.75

6 .8 1

5.00

1 .05

C o n tro ls .

16 ds7S,

d2°

INTRASON C -ALULAE V.AT2R

DAY

i x ? z ?jl z n t

T-1S24

i A f te r d i s c , orea

53

2 .8 5

10.37

7 .5 1

4.5 3

1 .0 5

C o n tro ls

a

3 .0 1

11.17

8 .1 6

5.22

1 .1 6

b

3 .0 6

11.25

8 .1 9

4 .9 7

1 .1 7

2

2 .9 4

11 .0 4

8 .1 0

5.5 6

1 . 15!

oiu^nt.

2 •

r

1

T reatm ent w ith 30 mr. DCA d a i l y '

3

3 .3 8

6

3 .8 6

10.90

7 .0 3

9

4 .1 9

11.50

7 .5 1

6.1 7

1.-15

13

4 .3 6

11.10

6.7 4

7.61

16

4 .5 4

10.55

5-81

-

1 .2 5 f 1 .5 3

6.09

1 .0 0 1 .4 2

5

i

i ! ! 1j

\

>

*

j

21

4.C0

10.63

6 .6 3

34

to•

1

11.30

7 .4 7

5.69

j

i j

. V k'

' A f te r d is c .

....6.95 ‘

1 .4 0

1 .3 4

• -i

'5 1I

210

5.51

.93

.0

234

5.00

4 .7 0

1.07

• j-

200

9 .7 5

6 .8 1

5.00

1.05

.7

217

2.85

10.37

7 .5 1

4.53

1.05

,2

220

362,

3 .0 1

11.17

3 .1 6

cy • a► .•?“»

.0

a

1 .1 6

b

3 .0 6

11.25

8 .1 9

4 .9 7

1 .1 7

c

2 .9 4

11.04

8.10

5 .5 6

1 .1 5

3

3.33

-

6.09

1 .0 0

6

3 .3 6

10.90

7 .0 3

9

4.19

11.50

7 .3 1

6.17

1 .1 5

13

4 .3 6

11.10

6 .7 4

7.61

1 .2 5

16

4 .5 4

10.35

5 .8 1

21

4 .0 0

10.68

6 .6 8

c • ->P ny*

~'r 9

303.

34

11.30

7 .4 7 9 .0 0 . _

-

5-69

6.95 3.10

1 .4 2

1 .5 3 1 .4 0

1 .3 4 ■>

TA i. • J.

-*

.2-70,

12.70

b

3.68

12 .4 2

3 .7 4

5 .0 4

1.25

c

3 .5 7

12.00

8.43

6.00

1.19

a ' ,1'

2

4 .1 7

10.90

6.73

6.9 2

1 j I . 02!

7 1 /1

217.

5

4 .9 2

1 1 .8 4

6.92

6 .8 7

1 .1 8

7 3 .0

2^

11

5 .5 4

11.00

5 .4 6

7 .3 0

1 .2 6 ;

17

4 .7 3

12 .9 0

8 .1 7

7 .0 1

.99

24

3 .9 2

12.40

3 .4 8

7 .5 6

1 .3 4 :

.A

T reatm ent 1 t h 50 mg. ECA d a i l y

i •

...

5 .

..

H

187.

o r

•

2

. . -

191.

^

• w

f*? ;

Z;"-

10. z t

56.5

1 3 .7

4 .0

112

32.0

U .6

4 .2

103

.1 3 . 226.

3 3 .6

1 5 .6

3 .8

110

*. *

210.

56.5

9-5

-

410

•.« *

254.

3 5 .2

1 0 .6

3 .0

104

200.

4 0 .0

1 2 .0

3 .0

105

217.

30.3

1 1 .1

2 .9

102

220.

30 .0

1 5 .8

4 .2

102

r0J

3.62.

2 6 .2

21 .0

3 .7

113

.0

321.

25.1

2 1 .7

3 .9

116

•'V

278.

3 1 .2

2 6 .3

3 .8

109

'

240.

3 2 .2

-

3*9

114

.0

365.

2 6 .0

2 2 .6

2 .7

116

314

3 1 .0

5-3

2 .5

116

:.o

394.

3 1 .2

1 7 .5

2 .2

115

•8

3 -4 .

*8

1 3 .4

2 .1

10a

-5

4^3 *

2 7 .0

2 8 .0

2 .5

11c

32.5

2 4 .4

4 .2

ir

«-■’ *

^ ^ • 'i

9« •

10.45

-

8.27

'

5.17

j

5.00

N

6.81

5.03

10.37

7 .5 1

4.53

8 .1 6

5.22

11.25

8.19

11.04

8.10

—

-

6.09

10.90

7.03

—

11.50

7 .J 1

6 .1 7

11.10

6.74

7.61

10.35

5.81

10.63

6.68

t

j-

9.75

11.17

•

• -v*

10.19

9.99

.*

4 .9 7 5 .5 6

,

-

1. l.l; l.lf 1. 1.42 1.15 1.25 1.53

5 .6 9

1 .4 0

-

■ ).0

,

303.

-i * o 3

7 .4 7

6.95

1 .3 4

: .70

9.00

3.10

l.I v

12.42

3 .7 4

5 .0 4

1•

( _.D

187.

5 2 .0

1 9 .4

3*3

12.00

8.43

6.00

1.19

5 7 .0

191.

2 9 .8

2 0 .0

5 .9

10.90

6.73

6 .9 2

1.02

7 1 .0

217.

3 2 .7

1 3 .0

3 .6

1 1 .8 4

6.92

6 .8 7

1.18

7:,o

242.

^ 0 .2

1 8 .0

2 .5

11.00

5.46

7 .3 0

9 : .2 1

256.

3 6 .0

1 5 .0

2 .0

12.90

8 .1 7

7 .0 1

.99

257.

3 1 .1

1 6 .8

2 .8

12.40

8.48

7 .5 6

1.34

.1.0' f " -.4

251.

5 4 .4

1 8 .8

2 .8

1.26

• *

>

-1 1 -

Doe 3

Percentile deviation

from, control Values

I

Total

Body Water

Intra­ cellular Water

11 -*0«-

1

+4o r

.1

I

1

I

I

Extra­ cellular Water

¥•60 +40 ■

I' 2

F igure 1*

4

A ction o f dog. The expressed values in

I

Cue. Tm>AH

'

DC.A. 3 0 n f./d a y

6

8

/O 10. Dcuya

/4

lb

IB HO 22 a*

d e so x y co rtico stero n e a o e ta te in a normal r e s u lts o f dog 3 re p o rte d in Table I a re as p e r c e n tile d e v ia tio n s from th e o o n tro l r e la tio n to tim ea

•12-

The re n a l plasma flow a ls o in c re a se d , b u t to a le s s e r ex ten t* w ith a consequent in o re a se in th e f i l t r a t i o n fra c tio n *

Tmp^H,

on th e c o n tra ry , was reduoed to 26* 74 and 32 p e r cent* w ith o u t any obvious c o r r e la tio n w ith th e o th e r ohanges. The serum potassium f e l l to a l i t t l e above h a l f o f i t s c o n tro l value* b u t no ohanges were d eteo ted in plasma sodium* Plasma s p e c ific g ra v ity and th e h e m ato crit b o th d eo lin ed s l i g h t l y during th e p e rio d of treatm ent* w h ile no s ig n if ic a n t v a ria tio n s were observed in the mean blood p re s su re o r in th e plasma non­ p r o te in n itro g e n concentration* In each o f th e do^s th e m o d ific a tio n s n o tic e d as a con­ sequence of th e tre a tm e n t augmented p ro g ressiv ely * reach in g a maximum approxim ately an th e e lev e n th day* and th e n tended to r e tu r n to normal d e sp ite th e c o n tin u a tio n o f th e in je c tio n of DCA and d e s p ite an in o re a se in dosage in dog 1*

The plasma

potassium c o n ce n tra tio n was th e s o le e x cep tio n in t h a t i t r e ­ v e rte d only p a r t i a l l y a f t e r having reached i t s low est lev el* Two to s ix weeks a f t e r th e tre a tm e n t had been discontinued* a l l th e anim als had re tu rn e d to normal* The changes in t o t a l body sodium were stu d ie d in dog 1 (T able II ) *

D uring tre a tm e n t w ith DCA th e t o t a l body sodium

in c re a se d by 30 p e r o ent over th e o o n tro l value* w ith a sim ultaneous in c re a se in th e average i n t r a o e l l u l a r c o n ce n tra tio n o f t h i s ion* T o tal potassium was measured in dog 3 (Table I I I ) and a t th e h e ig h t o f th e response to th e treatm ent* when th e serum potassium was 2 mEq* p e r l i t e r * i t had d ecreased 12 p e r o e n t; sin c e th e i n t r a c e l l u l a r w ater had decreased by 59 p e r oent* i n t r a o e l l u l a r c o n c e n tra tio n had in c re a se d s ig n if ic a n tly *

-1 3 TABIE I I DOG I

, E xperim ent

Day

C o n tro l T reatm en t w ith 20 mg. DCA d a i l y 16 days 4.0 mg. d a i l y th e fo llo w ­ in g 6 days A f te r tr e a tm e n t

T o ta l Sodium mEq

In tra o e llu la r Sodium mEq

Average In tra c e llu la r I n t r a c e l l u l a r Sodium Con­ W ater c e n tra tio n m Eq/l cc.

606

170

7,2 9 0

22

785

284-

5,0 0 0

5 6 .8

750

304

6 ,8 1 0

4 4 .7

642

210

7 ,5 1 5

2 8 .0

16 23

30

-1 4TABIE I I I

DOG 3

E xperim ent

T o ta l P o tassiu m ' Day- mEq

C o n tro l T rea tm e n t w ith 30 mg. DCA d a i l y

11

In tra c e llu la r P o tassiu m mEq

A verage In tra c e llu la r I n t r a c e l l u l a r P o tassiu m W ater C o n c e n tra tio n cc. m Eq/l

962

948

8 ,7 5 0

108

845

834

5 ,4 6 0

152

-15-

A otion of a d re n al o o r tio a l e x tra c t* Three n o ra a l dogs (dogs 4 , 5 and 6) were tr e a te d d a ily w ith 15 oo. of Upjohn*s b e e f ad re n al c o r tic a l e x tr a c t f o r 15 days* and 2 o r 3 observa­ tio n s were made d u ring th a t p e rio d (Table 17, F igure 2 ) . The ohanges obtained d if f e r e d markedly from th o se produced by DCA and involved body w a ter d is tr ib u ti o n w ith o u t a l te r a t i o n s in re n a l fu n c tio n or plasma e le c tr o ly te le v e ls*

C o rtio a l e x tr a c t

oaused an in c re a se in th e i n t r a c e l l u l a r f lu i d volume and in t o t a l body w a te r, w ith l i t t l e or no change in th e e x tr a o e ll u la r spaoe* The enlargem ent of th e i n t r a c e l l u l a r space amounted to 48 , 46 and 38 p e r c e n t o f c o n tro l values and was ap p aren t on the th i r d day of th e tre a tm e n t ( F ig u re 2 )•

The e x tr a c e llu la r space d id n o t

ohange i n dogs 4 and 5 and in o re a se d m oderately (23 p e r oen t) in dog 6*

T hiocyanate volume follow ed th e se v a r ia tio n s i n a somewhat

i r r e g u la r manner.

Plasma volume d id n o t show any s ig n if ic a n t change.

There was no marked change in

o r CpAH, alth o u g h th e

form er again showed a f a i r c o r r e la tio n w ith th e e x tr a o e llu la r volume,

TipATT in c re ase d alm ost 74 p e r oent in dog 4 and in th e

o th e r two anim als f i r s t decreased and subsequently ro se above c o n tro l v a lu e s . Plasma sodium, potassium and n o n -p ro te in n itro g e n oanoent r a t i o n d id n o t vary t o any s ig n if ic a n t e x te n t, n o r d id th e hemat o o r i t o r th e mean blood p re s s u re .

Ho d eterm in atio n s o f th e t o t a l

o a tio n in th e body w ere made. The m o d ific a tio n s in th e i n t r a o e l l u l a r spaoe induced by o o r tio a l e x tr a c t alBo tended to d isap p e a r under prolonged tre a tm e n t, B ffe o t of t o t a l adrenaleotom yt

Two normal dogs (F igures

f TABLE EFFECT OF TOTAL ADRENAL CORTICJ VOLUMES OF '

DISTRIBUTION ''

LOT a? ■ti

(lite rs ) DOG

u

INTRA-_ _ SCN INTRACELLULAR. WATER______

INULIN

a

2 .3 7

6 .6 9

4 .3 2

3.9 0

b

2 .4 5

6 .8 0

- 4 .3 5

4.65

c

2 .5 3

7.0 0

4 .4 7

4 .9 8

T reatm ent w ith 15 cc. o f e x tra c t d a ily .

5

2.62

9 .1 1

6 .4 9

4 .7 9

12

2 .8 7

8 .6 2

5 .7 5

A f te r d is c , tr e a tm e n t.

47

2 .5 1

7 .1 0

4 .5 9

4 .7 0

C o n tro ls .

a

1 .9 0

6.13

4 .2 3

3.05

b

2 .0 5

6 .4 0

4 .3 5

3 .5 7

c

2 .2 0

6 .6 1

4 .4 1

3 .8 8

3

1 .9 8

8.30

6 .3 2

3.9 0

6

2 .0 3

7 .3 6

5.3 3

3 .7 8

11

1 .9 6

6 .8 6

4 .9 0

3.10

A f te r d is c , tre a tm e n t

19

2 .1 6

6.2 0

4 .0 4

3.39

C o n tr o ls .

a

,270

7 .8 5

5.15

5 .1 8

b

2 .9 0

8.1 5

5.25

6.00

C o n tr o ls .

T reatm en t w ith 15 c c . o f e x tra c t d a ily .

6

DgO

DAY

EXPERIMENT

T reatm ent w ith 15 c c . o f e x tra c t d a ily

4

3 .5 6

8 .7 0

5.1 4

5.1 6

10

3 .8 9

11 .1 0

7 .2 1

7 .2 1

T - I 824

58.0

i

105.0

TABLE EFFECT OF TOTAL ADRENAL CORTICi VOLUMES OF

fcCT IK NORMAL DOGS

DISTRIBUTION

v

PLASM

RENAL FUNCTION

BLOOD ■ PRESSURE*

(lite rs ) Ds °

INTRASCN GJSLLUEIHP WATER

T-1824

.904

>.69

4 .3 2

3 .9 0

,.80

4.3 5

4.65

'.0 0

4 .4 7

>.11

. c in jSg/min.

CPAH cc/m in .

F .F . p .c e n t

^PAH mg/min.

K m E q ./l.

mm. o f H i

----6 7 .6

192.

3 5 .1

1 1 .7

3 .8

117

I

1 .0 2

6 6 .9

193.

3 4 .6

1 0 .9

3 .7

115

1

4 .9 8

.9 6

6 8 .6

191.

3 5 .9

1 2 .8

3 .9

122

1

6 .4 9

4 .7 9

1 .0 5

6 6 .4

248.

2 6 .8

2 0 .5

3 .8

114

I

(.62

5 .7 5

-

1 .0 4

7 5 .2

225.

3 3 .4

1 4 .0

4 .0

125

I

MO

4 .59

4 .7 0

.9 2

5 8 .0

160.

3 6 .2

11.3

4 .0

120

I

pi

>.13

4 .2 3

3.05

.534

#M

175.

2 2 .7

8 .4

4 .0

120

I

>.40

4 .3 5

3 .5 7

.570

4 2 .2

172.

2 4 .5

7 .8

3 .9

117

I

>.6l

4 .4 1

3 .8 8

.636

54 .5

163.

33 .3

4 .0

117

I

k.30

6 .3 2

3 .9 0

.710

124.

3 7 .3

6 .6

4 .4

-

'.3 6

5.33

3 .7 8

.580

161.

2 9 .7

7 .0

4 .0

-

.8 6

4 .9 0

3.10

.598

4 6 .3 i» I- 4 7 .7 1r. ,1 1 4 3 .3

116.

3 7 .3

1 0 .9

4 .1

-

.2 0

4 .0 4

3.39

.538

- t 5 2 .4

146.

35-9

5 .6

3 .7

-

:

6 .0

jts, 7.85

5.15

5 .1 8

.81

m ,

320.

3 5 .3

1 2 .0

4 .2

$.15

5.25

6 .0 0

.* ■

> -& )5.0

280.

3 7 .5

1 0 .0

4 .0

.70

5 .1 4

5 .1 6

dA'•“

1 0 5 .0 9

316.

3 3 .2

5 .0

3 .1

l.ie

7 .2 1

7 .2 1

233.

3 8 .1

1 4 .0

3 .0

1 ,^

‘J

Ip

P 39*0

-1 7-

D og s

p

fQ O -

l■

.

ll

Intracellular Water

*20

Percentile deviation

from

control -values

+40r *20

Total t ± BocUf Water

Extracellular W ater -20

*20

Cin -2 0 L

J

**0

*20

-

I

20l

15cc o f Adrenal extract/ day 0

F igure 2*

4.

A ction o f dog* The expressed values in

6

8

10

Days

J

10

I

14

I

16

I

18

7~mPAH

L.

OO

th e a d re n a l o o r ti e a l e x tra o t in a normal r e s u lts of dog 6 tak en from Table IV a re a s p e r c e n tile d e v ia tio n s from th e o o n tro l r e la tio n to time*

-18 -

3 , 4 and Table 7 ) were com pletely adrenaleotom ized in two s ta g e s and m aintained w ith 10 and 15 mg. o f DCA d a ily (dogs 7 and 8)** While s t i l l under tre a tm e n t and showing no sig n s o f a d re n al in s u ffic ie n c y , an experim ent was perform ed 6 days a f t e r complete adrenalectom y in dog 8 and a f t e r 21 days in dog 7 .

In b o th

anim als th e changes from p re o p e ra tiv e v alu es were s im ila r to th o se observed in normal dogs tr e a te d w ith DCA, i . e . , i n t r a ­ c e l l u l a r w a ter was reduoed and th e e x tr a o e ll u la r space was e n la rg e d .

C ^ and Cp^H b o th in c re a se d , w ith an increm ent in

th e f i l t r a t i o n f r a c tio n , and TmpAH f e l l s l i g h t l y in dog 8*

Plasma

potassium d e o lin e d as in th e non-qperated dogs tr e a t e d w ith DCA. Subsequently su p p o rtiv e th e ra p y was in te rru p te d and th e anim als were stu d ie d in d i f f e r e n t p e rio d s o f in s u ffic ie n c y *

In

dog 7 an experim ent was perform ed an th e 4 th day a f t e r c e s s a tio n of th e ra p y ; i t was tr e a te d ag ain f o r 2 days and re-exam ined a f t e r a second p e rio d o f 7 days w ith o u t su p p o rtiv e th e ra p y . experim ent was done 8 days a f t e r resum ption of therapy*

A fin a l In dog

8 experim ents were perform ed 6 and 8 days a f t e r DCA in je c tio n s were discontinued* The ohanges observed in th e s e two dogs became more con­ spicuous as th e degree of a d re n a l in s u ffic ie n c y increased*

D is­

placem ent o f th e body f l u i d was ag ain one o f th e most prem inent f e a tu r e s .

The i n t r a c e l l u l a r w ater in c re ase d p ro g re s s iv e ly up to

15 and 30 p e r c e n t above th e p re o p e ra tiv e o o n tro l, w h ile th e e x tra

E*

The dogs were m aintained an t h e i r u su al d ie t* c h lo rid e was added.

No sodium

TABLE EFFECT OF ADESMLEC1 VOLUMES OF

DISTRIBUTION

(lite rs ) DOG

EXPERIMENT

DAY

INULIN

C o n tr o ls .

a

2.68

8 .5 0

5 .8 2

A. 20

b

2 .5 2

8 .3 0

5 .7 8

4 .4 4

A d ren ale cto raized , tre a te d w ith 15 mg. DCA d a i l y .

21

3 .7 4

8 .3 5

4 .6 1

5.70

T reatm ent in t e r r u p te d

4

2 .4 5

1 0 .9 0

8.4 5

6 .8 4

7

.97

8 .1 0

7 .1 3

4 .0 7

T reatm ent resumed 10 mg.DCA d a i ly C o n tr o ls .

INTRASCN CELLULAR WATER

T-1S

1.61

8.10

6 .4 9

5 .4 0

a

4 .1 0

10 .7 0

6.60

5.60

1.

b

4 .5 0

1 1 .3 0

6 .8 0

6 .8 3

1.

5 .5 8

1 2 .0 0

6.42

8.00

1.

6

2 .9 8

1 1 .7 0

8 .7 2

7 .2 3

1

8

1.88

9.84

7 .9 6

A d ren ale cto m iz e d ,tr e a ­ te d w ith 10 mg.DCA d a i l y T reatm en t in t e r r u p te d

D20

Formal

dogs

RENAL

m {min.

PLASMA

FUNCTION

CPAH c c /m in .

F *F * p .c e n t

TmpAR mg/min.

K m E q ./l.

BLOOD PRESSURE

Na m Eq/l.

NPN mg/100

mm. o f Hg

i,o

214.

3 2 .7

1 5 .5

A.5

152 .

1 8 .8

125

fh °

240 .

3 0 .4

1 7 .0

4 .5

153-

1 7 .2

126

l.9

255-

3 2 .1

16.6

3 .0

153 .

20.1

130

).0

238 .

33.8

11.6

6.0

141.

4 5 .0

102

i.8

110.

2 4 .4

8 .3

5 .0

153.

7 0 .6

64

).9

196 .

26.0

9 .1

3 .2

141.

1 6 .0

97

2.1

308.

2 9 .9

2 2 .4

4 .0

161.

1 4 .0

122

9.1

296 .

3 3 .5

2 6 .4

4 .0

163.

1 6 .0

118

s .o

350.

3 3 .1

1 6 .5

2 .7

148.

2 3 .0

120

^•5

298.

2 3 .4

10 .6

6 .7

137.

4 7 .6

112

-2 0-

Doo 7

-20 -40 tS O |-

I

a ---- ™

I

Hit deviation

from

control veUtue

rott

-40

-60

I

ixtranUular Water

'in. -20

-40 -60

Tm P.A.H.

O C .A iS m f/d m f

a

F igure 3*

i4

1

!

I

Ckiy

day D.CA.IOmf/datf.

— ,— M*i— i— >— , o^ vns>vv^ i_,

/»

/a

flc

aa M 36 a t to sa D o yj o tfttr adrtnalnsbom ij.

S4

St

51

E f fe c t o f t o t a l adrenalectom y on dog V. The r e s u lts of Table V a re expressed as p e r c e n tile d e v ia tio n s from th e c o n tro l valu es in r e la tio n to time*

-2 1 -

Dog 8 Total Body Water

-20 ♦20

♦20

Percentile deviation from control Values

Intracellular Water

T

-20

~Extracellular Water -2 0 - 40 -60 +20

-20

-4 0 -60

O

I

-20

-40 -6 0

D.CA.iomf/datf.

60

iW W N W vM i 2

Figure 4o

4 - 6

8

i ie

t o tffii 12

(4

16

DaufS a fte r adrenalectomy.

i

i

i»

10

E ffe o t o f t o t a l adrenalectom y on dog 8« The r e s u l ts o f Table V a re expressed as p e re e n tile d e v ia tio n s from th e c o n tro l v alu es in r e la tio n to tin e *

-22 -

c e l l u l a r volume declined. 59 and 61 p e r c e n t.

T o tal body w ater

decreased in one dog and remained unchanged in th e o th e r, and th e re fo re th e i n t r a c e l l u l a r oompartment a t th e h e ig h t o f ad ren al in s u ffio ie n o y re p re se n te d from 81 to 88 p e r c en t of th e t o t a l w ater of th e organism ( F igure 5 ) ,

Plasma volume remained r e ­

markably c o n sta n t a t m ild degrees o f in s u ffio ie n o y , a t a tim e when th e changes in th e o th e r w a ter compartments were a lre a d y prom inent, and only decreased in th e l a s t s ta g e s , f a l l i n g about 35 p e r oent*

As th e e x tr a o e llu la r volume c o n tra c te d m arkedly,

e s p e c ia lly in th e l a t e r stag e s o f in s u ffio ie n o y , th e plasma v o l­ ume, d e s p ite an a b so lu te d e c re a se , corresponded to o n e -h a lf (dog 8) and th re e -q u a rte rs (dog 7) o f th e e x tr a o e llu la r volume* Renal fu n c tio n a ls o showed marked changes*

C ^ , Cp^g

and IttpATT s e r e a l l markedly and p ro g re s s iv e ly reduced*

A lthough

a l l th re e fu n c tio n s seemed to d ecrease in approxim ately th e same p ro p o rtio n , th e d ecrease in Cp^g was s l i g h t l y le s s than th e r e ­ d u ctio n in C jn , and th e re was th e re fo re a f a l l in th e f i l t r a t i o n fra c tio n . The u su a l chemical and p h y sic a l sig n s o f ad re n al i n s u f f i ­ ciency were p re s e n t in th e bloods

th e plasma potassium in c re a s e d ,

th e sodium d ecreased , although n o t very m arkedly, th e n o n -p ro te in n itro g e n ro s e , and th e s p e c if ic g ra v ity and h e m ato crit increased* The blood p re s su re was reduced, p a r t i c u l a r l y in th e te rm in a l stages* These a l t e r a t i o n s were more conspiouous, theiomore sev ere th e in s u ffio ie n o y .

INTRACELLULAR WATER fVo of Total Body W aH r)

•23-

90i

90

TO

60

Cbntrol

SO »

40

D.C.A. to m g /d a y

WITHOUT TREATMENT

o

Z

\

\ i t

DAYS

F igure 6*

I n t r a c e l l u l a r w a te r, ex p ressed as percen tag e o f t o t a l body w a te r, in an adrenaleotam ised dog (dog 8)* In ­ flu e n c e of DCA th e ra p y to th e l e f t o f th e v e r t i c a l l i n e j in flu e n c e o f p ro g re ssiv e a d re n al in s u ffio ie n o y to th e r ig h t o f th e v e r t i o a l lin e *

In dog 7 , when tre a tm e n t w ith DCA was re e s ta b lis h e d f o r a week, th e plasma potassium and HFN re v e rte d to norm al, b u t th e sodium remained low, and th e a lte r a t io n s in body w ater and re n a l fu n c tio n were only p a r t i a l l y re s to re d to normal* T hioeyanate space during a d re n a l in s u ffio ie n o y tended to deorease s l i g h t l y , b u t i t d id n o t fo llo w th e i n n l ia space and a t tim es exceeded th e l a t t e r by s e v e ra l fold* The g en eral appearanoe of body dogs was good throughout and only in th e te rm in a l hours o f in s u ffic ie n c y d id th ey show in c re a s in g d e b i l i t y , vom iting, and d iarrh ea*

T h e ir w eights

decreased 2 kg* during th e development o f sev ere in s u ffic ie n c y (7 th day in dog 7 and 8 th day in dog 8)*

Both anim als went in to

c r i s i s and d ied s e v e ra l hours a f t e r th e com pletion o f th e l a s t experim ent, d e s p ite in te n s iv e hormone and s a lin e therapy*

-2 5 DISCUSSKW

From th e above r e s u l t s i t ie e v id e n t t h a t th e a d re n a l c o rte x has a prim ary in flu e n c e on th e e q u ilib riu m d is tr ib u tio n o f f l u i d be­ tween th e e x tr a o e llu la r and i n t r a c e l l u l a r oompartments*

DCA produced

a d ecrease in th e i n t r a o e l l u l a r spaoe and an in c re a se o f th e e x tra ­ o e llu la r space, whereas adrenalectom y was follow ed by an in c re a s e in i n t r a c e l l u l a r space and a decrease in e x tr a o e llu la r space*

C o rtic a l

e x tr a o t th e ra p y induced a marked in c re a s e in th e i n t r a o e l l u l a r space w ith no s ig n if ic a n t m o d ifica tio n o f th e e x tr a c e llu la r volume*

The

d is s im ila r a c tio n s of DCA and c o r t i c a l e x tr a c t, as re p o rte d h e re , r e f l e c t a p o s s ib le d u al a c tio n on th e p a r t o f th e p h y s io lo g ic a l 1 hormones* These marked changes in body f l u i d d is tr ib u tio n m ight be considered secondary t o ohanges in c a tio n c o n c e n tra tio n in th e e x tr a o e llu la r compartment, b u t s e v e ra l f a c ts seem to r e f u te th i s in te r p r e ta tio n *

During DCA th erap y i n normal dogs, th e re was a

s h i f t o f w a te r from th e o e l l s a t a tim e when th e plasma sodium was unchanged, and t h i s s h i f t occurred d e s p ite an in c re a s e in i n t r a c e l l u l a r c a tio n c o n c e n tra tio n (T ables I I and I I I ) *

At th e

h e ig h t of ad re n al in s u ffic ie n c y in dog 7 , th e re was a pronounced s h i f t of w a te r in to th e c e l l s d e s p ite th e unchanged plasma sodium c o n c e n tra tio n (Table V, F igure 5)*

F u rth e r i t has been p re v io u s ly

shown t h a t hem odilution and hem oeoncentration r e la ti v e to plasm a p r o te in a re independent o f th e plasma le v e ls of sodium and e h lo rid e (65,56)*

The increm ents in i n t r a c e l l u l a r space noted d u rin g

p ro g re ssiv e a d re n a l in s u ffic ie n c y were f a r g r e a te r than what m ight 1* The a d re n a l o o r tio a l e x tr a c t used does n o t c o n tain s ig n i f ic a n t q u a n titie s o f DCA (8St©9) •

/

— 26—

be expected, from a sim ple p ro p o rtio n a l response to th e de­ c re ase in plasm a sodium c o n c e n tra tio n .

F in a lly in th e normal

dogs in je c te d w ith c o r tic a l e x tr a c t, th e i n t r a o e l l u l a r space expanded 40 p e r c en t w ith no change in plasma c a tio n co n cen tra­ ti o n . Assuming t h a t osm otic e q u ilib riu m between plasma and tis s u e s o b ta in s a t a l l tim e s , th e s e f a c ts in d ic a te t h a t v ario u s le v e ls of a d re n a l a c t i v i t y in flu e n c e th e o sm o tio ally a c tiv e c o n s titu e n ts w ith in th e c e l l , o th e r th an sodium and potassium , so as to a l t e r th e e ffe o tiv e i n t r a o e l l u l a r osmotic p r e s s u r e . A lte r n a tiv e ly , i f osmotio e q u ilib riu m between o e lls and e x tra ­ o e llu la r f l u i d does n o t o b ta in , i t would seem n eo essary to assume t h a t tis s u e o e lls g e n e ra lly a re capable o f c o n d itio n in g th e d is tr ib u tio n of w a ter independently o f osmotio p re s s u re , a r a th e r im p la u sib le s u p p o sitio n .

In e i t h e r view , th e e x tra ­

o e llu la r spaoe would a c t as a re s e rv o ir in to o r ou t of whioh w ater m ight move in a s s o c ia tio n w ith re c ip ro c a l changes in i n t r a ­ c e l l u l a r w a te r c o n te n t.

In n e ith e r case i s i t d e a r why th e

volume of th e e x tr a c e llu la r spaoe i s n o t p re serv e d as i t i s in th e normal anim al. The plasma volume remained f a i r l y c o n sta n t d u rin g DCA tre a tm e n t, d u ring th e in je c tio n s o f o o r tie a l e x tr a c t, and d u rin g m ild degrees o f ad re n al in s u ffic ie n c y , as oampared to mnoh g re a te r changes in th e o th e r compartments.

The ohanges in

plasma volume do n o t appear to be a s e n s itiv e index o f e x tra ­ o e llu la r spaoe.

In sev ere degrees o f in s u f f ic ie n c y , th e plasma

volume deoreased only 56 p e r c e n t whereas th e e x tr a c e llu la r spaoe deoreased to such an e x te n t t h a t i t was only s l i g h t l y g re a te r than

-2 7 -

th e plasma volume*

This l a t t e r ciroum stanoe seems c h a r a c te r is tic

of th e shoek-lilce s ta t e of te rm in a l ad ren al in su ffio ien o y *

I t is

p o s s ib le t h a t th i s defio ien o y in i n t e r s t i t i a l f lu i d has g re a te r p a th o lo g ie im p lic a tio n than th e re d u c tio n of plasma volume* Judging by our experim ents, th io o y an ate space measures a volume v a ria b ly in te rm e d ia te between e x tr a c e llu la r spaoe and t o t a l body w a te r, and i s , a t b e s t , only a q u a lita tiv e assessm ent of th e former*

In a d ren al in s u ffic ie n c y alm ost no o o rr e la tio n

e x is te d between th e th io o y an ate and in u lin spaces* Throughout th e observ atio n s here re p o rte d , th e r a te of glom erular f i l t r a t i o n q u a lita tiv e ly follow ed th e changes in e x tr a c e llu la r volume*

Eenal plasma flow m anifested v a r ia tio n s

in th e same d ir e e tia n as glom erular f i l t r a t i o n , b u t o f a le s s e r magnitude*

The n a tu re o f th e r e la tio n between e x tra ­

o e llu la r volume and glom erular f i l t r a t i o n in th e dog i s n o t known, b u t our d a ta would suggest t h a t i t i s n o t m ediated through th e ad ren al gland sin ce i t p e r s i s t s a f t e r t o t a l adrenalectom y.

Nor

is th e f i l t r a t i o n r a t e c o rre la te d w ith plasm a volume, plasma p ro ­ t e i n c o n ce n tra tio n or blood p re s s u re exoept in l a t e in s u ffio ie n o y where th e blood p re s su re was s u b s ta n tia lly reduced* Tm showed a s ig n if ic a n t f a l l b o th during DCA tre a tm e n t PAH and d u rin g p ro g re ssiv e ad ren al in su ffio ien o y * The e f f e o ts o f e o r tio a l e x tr a c t on

w ere irre g u la r*

Xn th e dogs in je o te d w ith DCA and w ith ad ren al e o r tio a l e x tr a o t, th e peak responses oocurred a f t e r some 10 days of tre a tm e n t and then g ra d u a lly su b sid ed , d e s p ite th e c o n tin u a tio n o f in je c tio n s and even do u b lin g o f dosage in one of th e DCA dogs*

-2a -

The so le exoeption to t h i s r e f r a c to r y phenomenon was n o ted in th e plasma potassium c o n c e n tra tio n during DCA tre a tm e n t which remained low when a l l th e o th e r measurements had re v e rte d alm ost com pletely,

This re f r a c to r y s ta t e has been

observed by s e v e ra l a u th o rs (2 5 ,4 7 ,5 3 ) f o r o th e r p h y s io lo g ic a l ohanges produced by DCA o r e o r tio a l e x tr a c t,

Whether i t

r e s u lts from in h ib itio n of form ation of p i t u i t a r y a d re n o o o rtic o tro p h ic hormone, th e form ation of antihorm ane, o r th e d e s tru c ­ tio n o r n e u tr a liz a tio n of th e in je c te d hormone by th e a d re n al gland is n o t know.

The l a s t named hy p o th esis seems l i k e l y in

view of th e f a o t t h a t th e DCA e f f e c t s p e r s is te d in th e a d re n a le c tcm ized dog a f t e r 3 weeks o f tre a tm e n t. D uring DCA tre a tm e n t, u rin e flow measured only in th e p re -in fu s io n p e rio d (TJ0) in c re a se d from normal v alu es o f le s s than 1 oe. p e r minute to 2 - 5 c c , p e r m in u te.

This o iro u n -

stan c e may be a dem onstration o f th e " d ia b e te s in s ip id u s - lik e syndrome* re p o rte d by o th e rs (1 0 ,4 0 ,4 9 ,6 6 ,6 8 ), U n fo rtu n a te ly , body w eig h ts were m easured u n d er such i r r e g u l a r c o n d itio n s t i r t th e changes found were n o t s ig n ific a n t. -

.

-29-

SUMMARY 1*

Sim ultaneous measurements o f e x tr a o e llu la r f l u i d

Tolume ( in u lin sp ao e ), t o t a l body w a ter (D2O sp a c e ), i n t r a 24 42 o e llu la r f l u i d volume (by o a le u la tio n ) , Ha and K volumes, plasma volume, th io o y an ate spaoe, and re n a l fu n c tio n (Cjq , CpAK, TmpAH) , were made in dogs during tre a tm e n t w ith desoxyo o rtio o ste ro n e a c e ta te (DCA.) (3 d o g s), ad ren al c o r t i c a l e x tra o t (3 dogs) and during p ro g re ssiv e a d re n al in s u ffic ie n c y (2 dogs)* 2*

DCA oaused a d ecrease in i n t r a o e l l u l a r f l u i d volume

and an expansion of th e e x tr a o e ll u la r space*

Cjq and CpAH

in c re a se d d u ring th erap y w h ile TntptTT deoreased*

The i n t r a ­

c e l l u l a r c o n ce n tra tio n s o f sodium and potassium were both e le v a te d a t th e peak of response to therapy* 3*

C o rtic a l e x tr a c t oaused an in c re a se in i n t r a o e l l u l a r

f l u i d volume w ith no a l t e r a t i o n in th e e x tr a c e l lu la r space*

No

c o n s is te n t in flu en c e on re n a l fu n c tio n was observed during ex­ t r a c t treatm ent* 4*

During ad ren al in s u ffic ie n c y , th e i n t r a o e l l u l a r volume

was en larg ed w h ile th e e x tr a c e l lu l a r space co n tracted *

C ^ , Cp^g,

TBp£g decreased* 5*

The r e la tiv e constancy o f th e plasma volume as compared

to much g r e a te r v a ria tio n s in th e e x t r a c e llu la r space in d ic a te s t h a t plasma volume i s n o t a s e n s itiv e index o f a lte ra tio n s , in th e le tte r. Thiooyanate space does n o t fo llo w th e e x tr a c e ll u la r space a s measured by in u lin *

— 30"

6*

The e f f e o ts o f DCA. end o o rtio a l e x tr a c t a re

tr a n s i e n t , a l l p h y s io lo g ic a l v a ria tio n s observed te n d in g t o re tu r n towards normal d e s p ite continued treatm en t*

-31-

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-37-

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4

-3 9-

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LIBRAE! OP HEW YORK OHIVERSITT UNIVERSITY HEIGHTS

Am. J . l e d . ,