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A Study on Red Blood Cells

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U n p u b l i s h e d t h e s e s s u b m i t t e d f o r t h e M a s t e r ’ s and
D o c t o r ’ s d e c r e e s and d e p o s it e d in the N o rth w e s te rn U n iv e rs ity '
L i b r a r y a r e o p e n f o r i n s p e c t i o n , b u t a r e t o be u s e d o n l y w i t h
due r e g a r d to th e r i g h t s o f th e a u t h o r s .
Bibliographical
r e f e r e n c e s may b e n o t e d , b u t p a s s a g e s ma y b e c o p i e d o n l y w i t h
t h e p e r m i s s i o n o f t h e a u t h o r s , and p r o p e r c r e d i t must be
g iv en in s ubseque nt w r i t t e n or p u b lis h e d work.
Extensive
copying o r p u b l i c a t i o n of the t h e s i s in whole o r in p a r t
r e q u i r e s a l s o t h e c o n s e n t o f t h e Dean o f t h e G r a d u a t e S c h o o l
of Northwestern U niversity.
T h is t h e s i s by.
h a s b e e n u s e d by t h e f o i l
a t t e s t t h e i r acceptance of
its
the
p e r s o n s , whose s i g n a t u r e s
above r e s t r i c t i o n s .
A L i b r a r y which borrow s
p a tro n s i s expect©d to secure
NAME AND ADDRESS
t h i s t h e s i s f o r u s e by
the s i g n a t u r e of each u s e r .
DATE
n orthw est : ™
u n iv e r s i t y
A STUDY ON RED BLOOD CELLS
A DISSERTATION
SUBMITTED TO THE GRADUATE SCHOOL
IN PARTIAL EULEILLMENT OE THE REQUIREMENTS
f o r tL e d e g r e e
DOCTOR OE PHILOSOPHY
DEPARTMENT OE PHYSIOLOGY
BY
HERMAN SAUL 1IGODSKY
CHICAGO, ILLINOIS
JULY, 1940
ProQuest Number: 10308443
All rights reserved
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a n o te will indicate th e deletion.
uest.
ProQuest 10308443
Published by ProQuest LLC (2017). Copyright o f th e Dissertation is held by th e Author.
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Table of Contents
In tro d u c tio n
1
P a rt I
T he E f f e c t o f W H is D ie t on t h e Red B lo o d
C e lls
o f M o n k ey s, S w in e, an d Dogs
R e v ie w o f t h e L i t e r a t u r e
3
E x p e r im e n t a l M e th o d s
19
R e s u lts
20
D is c u s s io n
42
C o n c l u s io n s
47
B ib lio g ra p h y
48
P a r t IX
T he E f f e c t o f G a s t r i c J u i c e a n d L i v e r E x t r a c t
A d m i n is t e r e d t o t h e P r e g n a n t M o th e r,
o n t h e E r y t h r o c y t e s o f New born R a t s
R e v ie w o f t h e L i t e r a t u r e
51
E x p e r im e n ta l M eth o d s
55
R e s u lts
56
D is c u s s io n
59
C o n c lu s io n s
61
B ib lio g ra p h y
63
P a rt I I I
The P r e s e n c e o f t h e A n t i - P e r n i c i o u s Anem ia E a c t o r
i n a n E x t r a c t o f P e t a l B o v in e L i v e r s
R e v ie w o f t h e L i t e r a t u r e
65
M e th o d s
66
R e s u lts
67
D is c u s s io n
69
Summary
71
B ib lio g ra p h y
73
P a r t IV
T he E f f e c t o f I n d o l e o n t h e Red B lo o d C e l l s
o f N o rm al a n d G a s t r e c t o m i z e d Dogs
R e v ie w o f t h e L i t e r a t u r e
76
M e th o d s
87
R e s u lts
89
D is c u s s io n
91
C o n c lu s io n s
92
B ib lio g ra p h y
96
P a rt V
T he E f f e c t o f a G o ld b e r g e r D i e t on t h e G ro w th
a n d Red B lo o d C e l l s o f R a ts
R e v ie w o f t h e L i t e r a t u r e
100
E x p e r im e n ta l M eth o d s
104
R e s u lts
105
D is c u s s io n
109
C o n c lu s io n s
110
B ib lio g ra p h y
111
E p i lo g u e
112
A c k n o w le d g e m e n ts
114
V ita
115
P u b lic a tio n s
116
S
(
!
1
!
In tro d u c tio n
M any a t t e m p t s h a v e b e e n m ade t o d e te r m i n e t h e e t i o l o g y o f p e r n i c i o u s
|
j a n e m ia .
A lth o u g h a r e m i s s i o n o f t h e d i s e a s e m ay b e r e a d i l y o b t a i n e d , t h e
1 c a u s e r e m a in s a m y s t e r y .
C a s t l e d h y p o th e s is a s to th e e tio lo g y o f th e
d is e a s e i s to d t h e o r e t i c a l and in c o m p le te .
i( t h e p h e n o m e n a .
I t f a i l s t o a c c o u n t f o r m any o f
No o t h e r s a t i s f a c t o r y t h e o r y a s t o t h e b a s i s o f t h e d i s e a s e
J!
| h a s b e e n p ro p o u n d e d .
i
|
I
I f i t w e re p o s s i b l e t o r e p r o d u c e t h e d i s e a s e i n a n im a ls o r t o
e s t a b l i s h th e n a tu r e o f t h e s u b s ta n c e r e s p o n s i b le f o r r e m is s io n s o f th e
d i s e a s e , i t i s n o t u n l i k e l y t h a t t h e e t i o l o g y w o u ld f o l l o w s h o r t l y . Many
II
j| i n v e s t i g a t o r s h a v e a tt e m p t e d t o r e p r o d u c e t h e d i s e a s e w i t h o u t s u c c e s s . T h e r e
I
ji
ii
i s a t t h e p r e s e n t tim e no know n m eth o d f o r a s s a y i n g a n t i - p e r n i c i o u s a n e m ia
jj m a t e r i a l i n a n im a ls *
P e r n i c i o u s a n e m ia p a t i e n t s m u st s t i l l b e r e l i e d up o n
j f o r th e a s s a y o f th e e f f e c t i v e m a te r ia l.
ji
l|
|!
The f o l l o w i n g s e r i e s o f e x p e r im e n ts w as p e r f o r m e d i n a n a t t e m p t t o
|p r o v i d e a n im a ls s u i t a b l e f o r t h e a s s a y o f a n t i - p e r n i c i o u s a n e m ia m a t e r i a l .
PART I
The E f f e c t o f W i l l s D i e t o n t h e Red B lo o d C e l l s
o f M onkeys, S w in e a n d Dogs
I
E x p e r im e n ts d e s ig n e d t o r e p e a t t h e w ork o f W i l l s o n m onkeys a n d
i
j: e x te n d e d t o sw in e a n d g a s t r e c t o m i z e d d o g s w e re u n d e r t a k e n t o r e p r o d u c e
t r o p i c a l m a c r o c y t i c a n e m ia fo u n d i n t h e P a r E a s t .
T h i s d i s e a s e i s know n t o
re s p o n d to a u to ly z e d y e a s t , l i v e r an d l i v e r e x t r a c t s .
T he b lo o d p i c t u r e
I
is
s i m i l a r t o t h a t o f p e r n i c i o u s a n e m ia i n many r e s p e c t s .
|
I
R e v ie w o f t h e L i t e r a t u r e
W h ile i n v e s t i g a t i n g t h e p r o d u c t i o n o f s t o n e s i n t h e u r i n a r y b l a d d e r
j i n r a t s , M e C a rris o n ( l )
if
|! o f e p id e m ic d r o p s y .
i n 1986 n o t e d t h a t h i s a n im a ls d e v e lo p e d som e f o r m
T he d i s e a s e w as m a rk e d b y a s p l a y e d , u n c e r t a i n g a i t ,
|i
| e d e m a to u s s w e l l i n g o f t h e f e e t a n d t a i l , p u n c t a t e h e m o r r h a g e s an d a s e v e r e
i!
!| a n e m ia i n t h e s e r a t s .
A t p o s t m o rtem t h e c o n d i t i o n h a d some r e s e m b la n c e s
'
|| t o p e r n i c i o u s a n e m ia .
The d i e t w h ic h h e w as u s i n g c o n s i s t e d o f t i n n e d
| S c o t c h o a tm e a l , l i n s e e d m e a l, t i n n e d c o r n f l o u r ,
ji
| p h o s p h a te a n d d i s t i l l e d w a t e r .
so d iu m c h l o r i d e , c a lc iu m
W i l l s (2 ) i n a s t u d y on p e r n i c i o u s an e m ia o f p r e g n a n c y com m ented
j
| o n M e C a rr is o n * s w o rk a n d s u g g e s t e d a m ic r o b ic o r i g i n o f t h e d i s e a s e s i n c e i t
o c c u r r e d m o re f r e q u e n t l y when t h e r a t s w e re h o u s e d i n l a r g e c o l o n i e s .
S in c e
j 10 c c . o f m il k w o u ld p r o t e c t t h e r a t s a g a i n s t t h e a n e m ia , s h e s u g g e s t e d
j t h e p o s s i b i l i t y o f a V ita m in A an d C d e f i c i e n c y .
S he s t a t e d t h a t a f u n d a ­
m e n t a l c a u s e o f a n e m ia i n p r e g n a n c y w as a V ita m in A a n d C d e f i c i e n c y s i n c e
j t h e s e tw o w e re l a c k i n g i n t h e d i e t o f I n d i a n women. M any o f t h e s e women
j
j d e v e lo p a m a c r o c y t i c a n e m ia o f p r e g n a n c y .
( T h a t t h e c o n d i t i o n M e C a rr is o n
j r e p o r te d i s n o t a C d e fic ie n c y i s
s u p p o rte d by th e w e ll e s ta b lis h e d f a c t
|| t h a t r a t s do n o t show a n y m a ld e v e lo p m e n t o r d i s e a s e w hen p l a c e d on a C
4
i!!
I d e fic ie n t d ie t.
I t i s p re s u m e d t h a t V ita m in C i s m a n u f a c t u r e d , a n d m ade
i
!! a v a i l a b l e t o t h e r a t , b y t h e i n t e s t i n a l f l o r a .
T h a t t h e d i s e a s e i s a V ita m in
j!
! A d e f i c i e n c y i s q u e s t i o n a b l e s i n c e h e d i d n o t r e p o r t t h e u s e o f V ita m in A
i
I
i n t r e a t i n g th e a n im a ls .)
The f i r s t c o m p re h e n s iv e r e p o r t b y W i l l s o n t h e " P e r n i c i o u s A nem ia
I o f P r e g n a n c y " a s s e e n i n I n d i a a p p e a r e d i n 193 0 ( 2 ) ,
I
| c r i b e d i s w e l l know n i n I n d i a .
The c o n d i t i o n d e s -
T h e a u t h o r e x c lu d e d a l l t h o s e c a s e s w h ic h
j!
j| w e re c o m p lic a te d b y tap e w o rm s o r m a l a r i a .
T he sym ptom s o f t h e d i s e a s e a r e
i
I a s e v e r e a n e m ia , p u f f y f e e t , a n k l e s , a n d f a c e , w e a k n e s s , lo w b l o o d p r e s s u r e ,
I
I and u s u a lly a h i s t o r y o f f e v e r d u rin g th e d is e a s e .
I t is d iffe re n tia te d
ji fro m p e r n i c i o u s a n e m ia b y t h e a g e a n d s e x i n c i d e n c e , a b s e n c e o f j a u n d i c e ,
j!
|| a n d t h e a b s e n c e o f b i l i r u b i n fro m t h e u r i n e .
T he b lo o d p i c t u r e c o n s i s t s o f :
ij!
||
l . M a c r o c y t o s i s w i t h lo w r e t i c u l o c y t e c o u n t s a n d a v a r y i n g d e g r e e o f
II
:|
a n is o c y to s is .
jj
b a s o p h ilia .
ji
I
|
i
j
D u rin g r e c o v e r y t h e r e i s p o l y c h r o m a s i a , p u n c t a t e
S o m etim es t h e r e a r e l a r g e
a n d a t o t h e r t im e s n o r m o b l a s t s a r e p r e s e n t .
2 . V a r i a b l e w h i t e b lo o d c e l l p i c t u r e .
When t r e a t m e n t i s b e g u n t h e
jj
b lo o d p i c t u r e c h a n g es w ith an in c r e a s e
'i
ij
n u m b er o f g r a n u l o c y t e s .
!<
n u m b er o f n u c l e a t e d f o m s
i n t o t a l and r e l a t i v e
ij
3 . D e c r e a s e d p l a t e l e t n u m b ers d u r i n g a c t i v e s t a g e s o f d i s e a s e .
!|
4 . N orm al q u a n t i t a t i v e Van d e n B e rg h r e s u l t s .
ij
!i
||
5 . A l t e r e d b lo o d c h e m i s t r y .
||
M
T h e r e i s n o r e a l a c h l o r h y d r i a a s s e e n i n p e r n i c i o u s a n e m ia .
The
j re s p o n s e t o l i v e r i n tre a tm e n t o f th e d is e a s e i s s im i la r to t h a t o f p e r ­
il
i jn ic io u s a n e m ia .
T he a u t h o r s t a t e s t h a t t h e r e i s no e v id e n c e o f a to x e m ia a s
I
jjth e c a u s e o f t h e c o n d i t i o n .
A s im ila r c o n d itio n i s
s e e n i n men i n t h e Bombay
|
jja r e a .
Ji
j!
C o n tin u in g h e r s t u d i e s o n t h e p e r n i c i o u s a n e m ia o f p r e g n a n c y ,
W i l l s (3 ) r e p o r t e d t h a t t h e d i e t o f t h e h o s p i t a l c l a s s o f women i n Bombay
j w as d e f i c i e n t i n c a l o r i e s , t o t a l a n im a l f a t a n d p r o t e i n , f r e s h f r u i t a n d
J
v e g e t a b l e s , V i t a m in s A a n d C, a n d s a l t s .
c o n s id e r a b ly .
T he V ita m in B c o n t e n t v a r i e d
She s u g g e s t e d t h a t a r e l a t i v e d e f i c i e n c y o f V i t a m in s
A a n d C o r some f a c t o r a s s o c i a t e d w i t h t h i s d e f i c i e n c y a r e r e s p o n s i b l e
o r c o n c e r n e d i n t h e e t i o l o g y o f t h e p e r n i c i o u s a n e m ia o f p r e g n a n c y a n d
t h a t V ita m in B a n d f a t p r o b a b l y p l a y some p a r t .
I n th e tre a tm e n t o f th e
c o n d i t i o n , W i l l s ( 4 , 5 , 6 ) r e p o r t e d t h a t i r o n a n d a r s e n i c w e re o f no v a lu e
w h i l e l i v e r , l i v e r e x t r a c t , o r m a r m ite ( a n a u t o l y z e d y e a s t p r o d u c t )
w e re o f d e f i n i t e v a l u e a n d a b o u t e q u a l l y e f f e c t i v e i n c u r i n g t h e m a c ro ­
c y t i c a n e m ia a n d r e l i e v i n g t h e o t h e r sym ptom s.
u s u a l l y d i d n o t r i s e a b o v e 3 ,2 5 0 ,0 0 0
H ow ever t h e r e d c e l l
w i t h a lo w
C.I.
in
count
th e t r e a t e d
p a t i e n t s ( 2 ) b u t m any so c a l l e d h e a l t h y women o f t h e same c l a s s h a d c o u n ts
o f o n l y 3 ,0 0 0 ,0 0 0 .
A d i e t s i m i l a r t o t h e d i e t o f t h e women s u f f e r i n g fro m t r o p i c a l
m a c r o c y t i c a n e m ia w as p r e p a r e d a n d f e d t o m onkeys ( 6 , 7 ) .
The d i e t c o n ­
s is te d o f:
P o lis h e d r i c e
30 %
G hi
10
W h ite b r e a d
3 2 ,5
W h ite p u m p k in
10
W heat c h a p a t t i
1 2 .5
M eat
5
To t h i s w as a d d e d 10 gm s. o f o r a n g e j u i c e o r 20 gm s. o f c a r r o t s
o ? -to m a to e s p e r m o n k ey .
S u p p le m e n ts w h ic h w e re a d d e d w e re 2 c c . c o d
liv e r
o i l d a i l y , two m onkeys r e c e i v e d 60 c c . o f m il k , a n d some r e c e i v e d 1 gm.
o f y e a s t p o w d er d a i l y .
T w elve m onkeys s u r v i v e d 66 t o 4 3 6 d a y s .
m onkey w h ic h l i v e d 436 d a y s d i d n o t d e v e lo p a n a n e m ia a l t h o u g h i t s
w as t h e sam e a s o n e w h ic h d e v e lo p e d a s e v e r e a n e m ia .
One
d ie t
T he r e s u l t s by
1 g ro u p s w e re :
B ro u p I :
176 d a y s .
D i e t a n d 10 gm s. o r a n g e j u i c e .
A l l f o u r w e re d e a d by
T h e r e w as n o c a u s e f o r d e a th s e e n a t a u t o p s y .
A l l d e v e lo p e d
some d e g r e e o f a n e m ia , t h e r e d b lo o d c e l l c o u n t b e in g a b o u t 4 . 5 — 5 .0
m i l l i o n s p e r c u . mm. a t d e a t h .
■Group I I :
B a s a l d i e t a n d 20 g m .s o f to m a to e s o r c a r r o t s p l u s
2 c c . cod l i v e r o i l .
The r e s u l t s w e re t h e sam e a s i n G roup I b u t t h e
a n im a ls s u r v i v e d 200 d a y s .
T h is g ro u p d e m o n s t r a t e s t h a t t h e d i e t was
n o t Y ita m in A o r C d e f i c i e n t .
G roup I I I :
B a s a l d i e t p l u s 10 c c . o r a n g e j u i c e a n d 1 gm. c o n ­
c e n t r a t e d y e a s t p o w d e r.
T h r e e a n im a ls w e re u s e d i n t h i s g r o u p .
T hey g a v e
t h e sam e r e s u l t s a s t h e p r e v i o u s tw o g r o u p s and i t w as c o n c lu d e d t h a t
t h e Y ita m in B i n t a k e w as
G roup IY :
B asal
t o o s m a ll o r o f no b e n e f i t .
d i e t p lu s 2 c c . cod l i v e r o i l and o th e r r a t i o n s .
T h re e a n im a ls w e re u s e d i n t h i s g ro u p .
A ll th r e e liv e d 465 d ay s.
One a n im a l r e c e i v e d y e a s t ; t h e r e d b lo o d c o u n t f e l l
A la rg e
t o 3 .5 m i l l i o n s .
d o s e o f y e a s t a s " M a rm ite " w as g i v e n a n d t h e a n e m ia w as c u r e d a n d
g ro w th r e s t o r e d .
One a n im a l r e c i e v e d m i l k , 60 c c . p e r d a y , d o u b le f r u i t r a t i o n s ,
b u t no y e a s t .
A f t e r t h i r t y w eek s t h e a n im a l d i d n o t grow a n d becam e
a n e m ic .
The t h i r d a n im a l r e c e i v e d 60 c c . o f m il k d a i l y , d o u b le f r u i t
r a tio n s and y e a s t.
No a n e m ia d e v e lo p e d i n t h i s a n im a l a n d g ro w th was
n o r m a l.
A f o u r t h a n im a l on a l a r g e m il k r a t i o n , w i t h o u t y e a s t ,
s p o n ta n e o u s c u r e o f t h e a n e m ia .
show ed a
The anemia which the monkeys developed was of a macrocytic type
w ith a s h i f t i n th e P r ic e - J o n e s c u rv e to th e r i g h t .
T h e r e w as a n i s o c y t o s i
M y e l o b l a s t s w e re so m e tim e s p r e s e n t a t t h e h e i g h t o f t h e a n e m ia , b u t norm o­
b l a s t s w e re m ore commonly s e e n .
The c o l o r i n d e x a p p r o a c h e d 1 . 0 a t t h e
h e i g h t o f t h e a n e m ia , b u t w as a ro u n d 0 . 7 5 when t h e a n im a ls w e re n o r m a l.
A t a u to p s y (7 ) t h e r e w e re h e m o s i d e r i n d e p o s i t s i n t h e l i v e r a n d s p l e e n
b u t t h e r e w as no f a t t y
d e g e n e r a t i o n o f t h e l i v e r o r h e a r t m u s c le a s s e e n
i n p e r n i c i o u s a n e m ia .
jj
I n o r d e r t o d e te r m i n e w h ic h s u b s t a n c e c o n t a i n e d i n M a rm ite was
1 r e s p o n s i b l e f o r t h e c u r e o r p r e v e n t i o n o f t h e m a c r o c y t i c a n e m ia i n t h e
m o n k ey s, t h e f o l l o w i n g s u b s t a n c e s w e re u s e d ( 8 ) :
si 1 .
Y e a s t p o w d e r, 30 gm s. d a i l y
I n a c tiv e
In a c tiv e
I 2 . W a te ry y e a s t e x t r a c t s
|j
5 . W a te ry y e a s t e x t r a c t s a f t e r f i v e h o u r s o f a u t o c l a v i n g
In a c tiv e
jj
4 . U n f la v o r e d M a rm ite , 30 gm s. d a i l y
A c tiv e
|| 5 . B o i l e d M a rm ite tw o m in u te s w i t h e q u a l w e ig h t o f w a t e r
(a )F iltra te
A c tiv e
(b )R e s id u e
In a c tiv e
I! 6 .A u to c la v e d M a rm ite (B d e s t r o y e d )
A c tiv e
j 7 . A l k a l i n e s o l u t i o n o f a u t o c l a v e d M a rm ite
L ess a c tiv e
jl
8.80% a l c o h o l e x t r a c t o f M a rm ite
ji
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i
(b )R e s id u e
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| 9 . A c id c l a y V ita m in
an d B^ 4 gm s. d a i l y
i! 1 0 . V ita m in Bg fro m e g g w h ite
In a c tiv e
In a c tiv e
From t h e s e r e s u l t s W ills c o n c lu d e d t h a t t h e a c t i v e f a c t o r i n M a rm ite
w as n o t V ita m in Bp o r Bg .
F u r t h e r e x p e r i m e n t s ( 9 , 1 0 , 11) i n t o t h e n a t u r e o f t h e s u b s t a n c e
r e s p o n s i b l e f o r t h e c u r e o f t r o p i c a l m a c r o c y t i c a n e m ia g a v e t h e f o l l o w i n g
re s u lts :
( 9 , 1 0 , 11)
1 .2 2 7 gm s. o f f r e s h l i v e r b y m o u th f o r s i x d a y s
(■
1
J 2 .C a m p o lo n ( l i v e r e x t r a c t p a r e n t e r a l l y ) a c t i v e i n
jl
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i!
A c t iv e
A c t iv e
Ij
p S .A n a h e m in ( r e f i n e d l i v e r e x t r a c t ) a c t i v e i n
I!
(I p e r n i c i o u s a n e m ia
f;
;i4 .L a lo n d s l i v e r e x t r a c t ( r e f i n e d l i v e r e x t r a c t )
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5 . C am polon an d y e a s t f r a c t i o n e d w i t h ammonium s u l f a t e
( a ) (NH4.) 53SO4 s o l u b l e p o r t i o n fro m cam polon
a c t i v e i n p e r n i c i o u s a n e m ia
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( b ) ( NH^ ) 2S0 4 s o l u b l e p o r t i o n fro m y e a s t
'!
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(d )
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I n a c tiv e
I n t h e r e l a t i o n o f t h e f a c t o r t o t h e V ita m in B*? co m p lex t h e
f o l l o w i n g w as d e te r m in e d :
11 . 5 g a s . o f M a rm ite
2.80% a l c o h o l s o l u b l e f r a c t i o n
A c t iv e
A c t iv e
9
,j 3 . S o l u b l e a n d i n s o l u b l e p o r t i o n s o f 90$ a n d 1 0 0 $
a l c o h o l f r a c t i o n s show ed w h o le o r p a r t i a l d e s t r u c t i o n o f a c t i v i t y
4 . D r ie d b r e w e r s y e a s t
In a c tiv e
5 .0 .1 $ a c e t i c a c id e x t r a c t o f f r e s h b re w e rs y e a s t
( a ) 90$ a lc o h o l s o lu b le f r a c ti o n
A c t iv e
(b )9 0 $ a lc o h o l in s o lu b le f r a c t i o n
A c t iv e
| 6 . ! F r a c t i o n a t i o n o f (NH^)gSO^ s o l u b l e p o r t i o n o f 8 0 $
a lc o h o l e x t r a c t o f f r a c t i o n 5 w ith 90$ and 95$ a lc o h o l
(a )S o lu b le p o r tio n
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(b )In s o lu b le p o r tio n
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7 . 1 'u l l e r s e a r t h f i l t r a t e
V ita m in
o f fra c tio n 5 fre e of
and f l a v i n
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8 .H e p a t o f l a v i n 0 . 5 mgm. d a i l y
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9 . W heat germ e x t r a c t s , v e r y l a r g e d o s e s
A c t iv e
A n o th e r v i ta m i n d e f i c i e n c y i n m onkeys w ho u ld b e d e s c r i b e d h e r e .
D ay, L a n g s to n , a n d S h u k e rs (1 2 ) r e p o r t e d t h a t m onkeys on a d i e t o f c a s e i n ,
w h o le w h e a t, p o l i s h e d r i c e ,
26 t o 81 d a y s .
s a l t s and o ra n g e j u i c e d ie d i n
N orm al c o n t r o l s on t h e b a s a l d i e t p l u s b r e w e r s y e a s t s u r v i v e d
f o r m ore t h a n 200 d a y s .
le u k o p e n ia .
cod l i v e r o i l ,
The a f f e c t e d a n im a ls d e v e lo p e d a n a n e m ia and
T h e r e w e re u l c e r a t i o n s a ro u n d t h e gum s, t h e t e e t h f e l l o u t i n
som e, a n d t h e r e w as a s e v e r e d i a r r h e a .
I n some a n im a ls t h e r e d b lo o d
c e l l c o u n t f e l l b e lo w t h r e e m i l l i o n an d t h e h e m o g lo b in b e lo w 3 gm s.
T hat
t h e d i s e a s e i s n o t m onkey p e l l a g r a was shown by t h e r e s u l t s w hen 10 t o 50
m ga. o f n i c o t i n i c a c i d w as a d d e d t o t h e d a i l y d i e t ( 1 4 ) .
T he m onkeys
d e v e lo p e d a c y t o p e n i a , e s p e c i a l l y a l e u k o p e n i a , i n s p i t e o f t h e a d d i t i o n
o f n i c o t in i c a c id .
R i b o f l a v i n w as w i t h o u t r e s u l t s i n p r e v e n t i n g t h e d i s e a s e .
The a u t h o r s c o n c lu d e d t h a t t h e r e i s
tra c t,
some f a c t o r i n y e a s t a n d l i v e r e x ­
o t h e r t h a n t h e know n v i t a m i n s , w h ic h i s n e c e s s a r y f o r n o rm a l
h e m a t o p o i e s i s i n m o n k e y s,
^ o r t h i s v ita m in th e a u th o r s p ro p o se d th e
name V ita m in M.
H a r r i s (1 5 ) u s e d a s i m i l a r d i e t i n p r o d u c in g m onkey p e l l a g r a :
c o r n , p e a m e a l, c a s e i n , c o t t o n s e e d o i l ,
V ita m in s A a n d D.
s a l t s , a n d o ra n g e j u i c e and
H i s m onkeys d e v e lo p e d a n o r e x i a , d i a r r h e a , p r o g r e s s i v e
w e a k n e s s , e m a c ia t io n , d e n u d a ti o n o f f u r a n d a r o u g h , d r y ,
s c a ly s k in .
The d i s e a s e c o u ld b e c u r e d o r p r e v e n t e d b y t h e a d d i t i o n t o t h e b a s a l d i e t
o f 1% d r i e d y e a s t , L i l l y ' s l i v e r e x t r a c t # 3 4 3 , w h o le w h e a t r e p l a c i n g
c o r n i n t h e d i e t o r 1 0 - 3 0 mgms. o f n i c o t i n i c a c i d p e r d a y ( 1 6 ) .
d i e t s u p p l i e s a d e q u a te a m o u n ts o f V ita m in Bg.
The
R ib o f la v in i s w ith o u t
e f f e c t i n c u r i n g o r p r e v e n t i n g t h e sy n d ro m e .
S w ine h a v e a l s o
s e r v e d a s e x p e r i m e n t a l a n im a ls i n d e te r m i n in g t h e
e f f e c t o f V ita m in B a n d t h e a n t i - p e r n i c i o u s a n e m ia f a c t o r on r e d b lo o d
c e lls .
McGowan a n d S i n c l a i r (1 7 ) fo u n d t h a t sw in e k e p t on a r a t i o n
c o n s i s t i n g o f c o r n , f i s h m e a l, a n d d r a f f d e v e lo p e d a n a n e m ia , j a u n d i c e , a n d
l i v e r d am ag e.
T he a n e m ia w as m a c r o c y t i c i n t y p e .
red and c e llu la r .
The b o n e m arro w s w e re
F o l lo w in g rav* l i v e r f e e d i n g s , t h e r e w as a n i n c r e a s e
i n r e t i c u l o c y t e s a n d t h e a n e m ia d i s a p p e a r e d .
M i l l e r a n d R hoads r e p o r t e d
i n 193 5 (1 8 ) t h a t siwine p l a c e d on a m o d i f i e d G o ld b e r g e r d i e t d e v e lo p e d
a c h l o r h y d r i a , a n e m ia , a n d s t o m a t i t i s .
Some o f t h e a n im a ls d i e d s u d d e n ly
b e f o r e o r s h o r t l y a f t e r b lo o d c h a n g e s o c c u r r e d .
A ll o f th e r e s t o f th e
a n im a ls d e v e lo p e d sym ptom s n o t u n l i k e c a n i n e b l a c k - t o n g u e .
A c h lo rh y d ria
a p p e a r e d i n f o u r t o s i x t e e n w eeks i n t h e g ro u p i n w h ic h a m a c r o c y t i c
j a n e m ia d e v e lo p e d i n f o u r t o e l e v e n w e e k s .
I n t i i e g ro u p w h ic h d e v e lo p e d a
I m i c r o c y t i c a n e m ia , a c h l o r h y d r i a a p p e a r e d i n f o u r t o t e n w e e k s .
T h re e
o f t h e a n im a ls w h ic h d e v e lo p e d a m a c r o c y t i c a n e m ia a n d o n e o f t h e m ic I
| r o c y t i e g ro u p show ed s i g n s o f m o to r w e a k n e s s o f t h e l i m b s .
T he l i v e r s o f t h e a n e m ic h o g s w e re e x t r a c t e d a n d t h e e x t r a c t
w as g i v e n i n t r a v e n o u s l y t o a t y p i c a l p e r n i c i o u s a n e m ia p a t i e n t f o r t e n
days.
T h e re w as no h e m a t o l o g i c a l r e s p o n s e .
An e x t r a c t o f n o im a l sw in e
l i v e r s p r o d u c e d a r e t i c u l o c y t e p e a k o f 2 1 . 2$ o n t h e n i n t h d a y w i t h a r i s e
i n t h e r e d b lo o d c e l l c o u n t a n d h e m o g lo b in .
A s i m i l a r e x p e r im e n t w i t h
a n e x t r a c t o f t h e a n e m ic l i v e r s e q u i v a l e n t t o
t
650 gm s. o f t h e f r e s h l i v e r s
| w as g i v e n t o a n o t h e r p e r n i c i o u s a n e m ia p a t i e n t f o r t h i r t e e n d a y s w i t h
!
| no r e s p o n s e .
T h e re w as a r e t i c u l o c y t e p e a k o f 9 .2 $ o n t h e f i f t h d a y w hen
!
jj n o rm a l s w in e l i v e r e x t r a c t w as u s e d , b u t t h e r e w e re o n ly s l i g h t im p ro v e j B e n ts i n t h e r e d b lo o d c e l l a n d h e m o g lo b in v a l u e s .
ij
ij
G a s t r i c j u i c e fro m t h e a c h l o r h y d r i c a n e m ic sw in e was o b t a i n e d
i|
jlf o llo w ln g t h e a d m i n i s t r a t i o n o f h i s t a m i n e .
T h is j u i c e (1 5 0 c c . ) was
j
in c u b a t e d w i t h 12 gm s. o f "V egex" a n d a d m i n i s t r e d t o a p e r n i c i o u s
a n e m ia p a t i e n t d a i l y f o r t e n d a y s .
v a lu e s d e c lin e d d u rin g t h i s p e r io d .
The r e d b lo o d c e l l a n d h e m o g lo b in
H ow ever when n o rm a l sw in e g a s t r i c
J j u i c e a n d 12 gm s. o f trV egex” w e re i n c u b a t e d a n d f e d , t h e r e w as a r e t i j c u l o c y t e p e a k o f 2 8 .2 $ on t h e e i g h t h d a y w ith a r i s e i n r e d b lo o d c e l l
jan d h e m o g lo b in v a l u e s .
i
J
B i r c h , C h ic k , a n d M a r t i n (1 9 ) u s i n g a m o d if ie d G o ld b e r g e r d i e t
jp ro d u c e d a s i m i l a r syndrom e b u t d id n o t r e p o r t t h e h e m a t o l o g i c a l d a t a .
f
C h ic k , M a c ra e , M a r t in a n d M a r t in ( 20)
a ls o f a i l e d to r e p o r t
jh e m a to lo g ic a l d a t a on sw in e f e d a m o d if ie d G o ld b e r g e r d i e t .
H o w e v e r., t h e y
12
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| r e p o r t e d t h a t a f u l l e r s e a r t h a d s o r b a t e o f y e a s t a t pH 1 . 3 , a 95%
l|
ji a l c o h o l e x t r a c t o f y e a s t , o r 60 mgms. o f n i c o t i n i c a c i d d a i l y w e re
:j
j| s u f f i c i e n t t o c u r e o r p r e v e n t t h e
sy n d ro m e .
A m a c r o c y t i c a n e m ia w as r e p o r t e d by W in tr o b e (2 1 ) t o o c c u r i n
j!
j
s w in e r e c e i v i n g a d i e t c o n s i s t i n g o f c a s e i n , l a r d ,
m ix tu re , and a s c o r b ic a c id .
cod l i v e r o i l , m i n e r a l
I f t h r e e o r m o re gm s. o f y e a s t p e r k i l o -
|g ra m o f b o d y w e ig h t w e re f e d w i t h t h e b a s a l d i e t ,
a n e m ia d i d n o t o c c u r .
!
j I n a l l b u t o n e o f t h e s w in e r e c e i v i n g s m a l l e r
j | o c c u r r e d o n t h e f o l l o w i n g s u p p le m e n ts :
a m o u n ts o f y e a s t , a n e m ia
ji
(a)
F o u r sw in e r e c e i v i n g o n l y t h i a m i n — s l i g h t a n e m ia 3 , s e v e r e 1 ,
J
(b )
F i v e sw in e r e c e i v i n g t h ia m i n a n d r i b o f l a v i n — s l i g h t a n e m ia
P
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T h r e e sw in e r e c e i v i n g t h ia m i n a n d n i c o t i n i c a c i d — s e v e r e
a n e m ia i n a l l 3 ,
(d ) H h r.ee
ii
j|
sw in e r e c e i v i n g t h i a m i n , n i c o t i n i c a c i d ,
r i b o f l a v i n — a l l becam e s e v e r e l y
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and
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t h e a n e m ic a n i m a l s , f o u r r e c e i v e d l a r g e d o s e s o f y e a s t a n d
I
■ o b ta in e d c o m p le te r e l i e f o f sym ptom s.
One a n em ic a n im a l r e c e i v e d a
1
P r e fin e d l i v e r e x t r a c t a n d a n o th e r a c ru d e r l i v e r e x t r a c t b u t th e s e had
I,
i no e f f e c t on t h e a n e m ia .
I n n o n e o f t h e anem ic a n im a ls w as a p e r s i s t e n t
j
| a c h lo r h y d ria o r h y p e rb ilir u b in e m ia o b se rv e d .
T he a u t h o r c o n c lu d e d fro m
I
j
t h e s e d a ta t h a t , u n d e r th e d i e t a r y c o n d itio n s u s e d ,
ii
jj
s u b s t a n c e w h ic h o c c u r s i n b r e w e r s y e a s t i s n e c e s s a r y f o r n o rm a l b lo o d
j f o r m a ti o n
some u n i d e n t i f i e d
i n s w in e .
As r e g a r d s t h e t r e a t m e n t o f p e r n i c i o u s a n e m ia w i t h y e a s t , y e a s t
j
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e x tra c ts ,
a n d v a r i o u s 1161111)e r s o f t h e V ita m in B c o m p le x , t h e r e i s a
g r e a t d if f e r e n c e o f o p in io n .
C o m p eten t o b s e r v e r s h a v e r e c o r d e d r e s u l t s
w h ic h a r e b o t h f a v o r a b l e a n d u n f a v o r a b l e .
The f o l l o w i n g t a b l e i s a
c o m p ila tio n o f t h e r e s u l t s o b ta in e d w ith t h e s e s u b s ta n c e s :
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d ie t.
S i n c e some o f t h e s u b s t a n c e s u s e d b y W i l l s w e re n o t a v a i l a b l e ,
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T he r i c e w as c o o k e d w i t h a n e q u a l v o lu m e o f w a t e r u n t i l s o f t .
The s h o r t e n i n g a n d m a tz o s m e a l w e re t h e n a d d e d w i t h c o n s t a n t s t i r r i n g .
The s a l t m i x t u r e , c o d l i v e r o i l ,
e n t i r e d i e t t h o r o u g h l y m ix e d .
ad lib itu m .
a n d to m a to j u i c e w e re a d d e d a n d t h e
A1 1 o f t h e a n im a ls o n t h e d i e t w e re f e d
The a n im a ls w e re f e d o n c e a d a y a n d w i t h t h e e x c e p t i o n o f
20
Ithe rats, coprophagy was not prevented.
The d i e t w as f e d t o f i v e y o u n g s w in e , s i x m o n k ey s (M acacus r h e s u s ) ,
f o u r g a s tr e c to m iz e d d o g s, and f i f t e e n r a t s .
One o f t h e m o n k ey s, tw o o f
t h e d o g s , a n d f i v e o f t h e r a t s r e c e i v e d m a rm ite (V eg ex ) i n a d d i t i o n t o
th e b a s a l d ie t.
The m onkeys a n d d o g s r e c e i v e d f i v e g ram s o f t h e s u p p l e ­
m en t d a i l y f o r e a c h a n im a l.
m o d ifie d t o c o n ta in
The d i e t o f t h e r a t s r e c e i v i n g m a r m ite w as
o f t h e s u p p le m e n t.
P o u r r a t s w e re u s e d a s n o rm a l
c o n t r o l s on a d i e t o f P u r i n a dog c h e c k e r s .
B lo o d f o r d e t e r m i n a t i o n s o f r e d b lo o d c e l l c o u n t s , h e m o g lo b in ,
q n d h e m a t o c r i t v a l u e s w as o b t a i n e d fro m t h e a n t e c u b i t a l v e i n i n t h e d o g s
a n d m onkeys an d im m e d ia te ly p l a c e d i n s m a ll v i a l s c o n t a i n i n g d r y h e p a r i n .
B lo o d w as a llo w e d t o r u n f r e e l y i n t o
e a r o f th e p ig s .
s i m i l a r v i a l s fro m a s l i t i n t h e
R ed b lo o d c e l l c o u n t s w e re m ade i n t h e u s u a l m a n n e r.
H e m o g lo b in d e t e r m i n a t i o n s w e re p e r f o r m e d a c c o r d i n g t o t h e Newcomer m e th o d
u s in g a s ta n d a rd iz e d g la s s d is c i n th e c o lo r im e te r .
H e m a to c r it d e t e r -
!j m in a tio n s w e re m ade a c c o r d i n g t o t h e m eth o d o f Rosajbn ( 3 9 ) .
A t t e m p ts w e re m ade t o o b t a i n g a s t r i c j u i c e s a m p le s fro m t h e
s w in e f o l l o w i n g t h e i n j e c t i o n o f h i s t a m i n e , b u t a l l o f t h e s a m p le s
o b t a i n e d w e re c o n ta m in a t e d w i t h l a r g e a m o u n ts o f b i l e , t h e g a s t r i c
a n a l y s e s w e re o m i t t e d .
The g ro w th r a t e o f t h e r a t s p l a c e d on t h e d i e t w as u s e d a s a n
i n d i c a t i o n o f t h e V ita m in B c o n t e n t .
R e s u lts
l lh e r e s u l t s o f t h e v a r i o u s g ro u p s o f a n im a ls on t h e d i e t a t
t a b u l a t e d i n T a b l e s I t o X II i n c l u s i v e .
I
1!
i)
Rats (Tables 1 and II)
Si
T he s u r v i v a l tim e o f t h e f i f t e e n r a t s p l a c e d on t h e d i e t w as t e n
Sj
^days t o a d u l t h o o d .
One o f t h e r a t s , Num ber 1 2 , on t h e u n s u p p le m e n te d d i e t
ii
I's u r v iv e d o v e r 1 5 0 d a y s b e f o r e t h e d i e t w as d i s c o n t i n u e d .
In a l l in s ta n c e s
jr
.
j'of t h e r a t s on t h e u n s u p p le m e n te d d i e t , t h e r e w as no a p p r e c i a b l e i n c r e a s e
jjin w e ig h t e v e n a f t e r t h e y h a d b e e n on t h e d i e t f o r e i g h t y d a y s . Number 12
i1
i
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<1
jo f t h e r a t s t h e r e w as no e v id e n c e o f a c r o d y n i a .
One r a t , N um ber 5 , showed
j,
iisome p a r a l y s i s o f t h e h in d l i m b s , b u t i n n o o t h e r i n s t a n c e w as a n y p a r a l y s i s
jn o te d .
i-
j
Of t h e f i v e r a t s p l a c e d on t h e d i e t s u p p le m e n te d w i t h m a r m ite , two
1s u r v i v e d o v e r 230 a n d 150 d a y s r e s p e c t i v e l y .
t h e f i f t e e n t h an d t w e n t i e t h d a y s an d t h e f i f t h
Two o f t h e s e r a t s d ie d b e tw e e n
r a t su rv e d f i f t y
d ay s.
In
| a l l i n s t a n c e s t h e a n im a ls g a in e d w e ig h t v e r y s lo w ly an d d i f f e r e d m a r k e d ly
'fro m t h e c o n t r o l s .
Even a t t h e end o f 150 d a y s , t h e i r w e ig h ts w e re 170
iand 202 gram s w h ic h w as f a r b e lo w t h a t o f n o rm a l a n i m a l s .
T a b le I I i l l u s t r a t e s t h e l a c k o f g ro w th i n t h e r a t s on t h e s u p p le ­
m e n te d a n d u n s u p p le m e n te d d i e t .
The a v e r a g e g a i n p e r r a t f o r e a c h f i v e
•day p e r i o d f o r s i x t y d a y s i s 1 4 .6 gram s f o r t h e n o rm a l r a t an d 2 . 4 gram s
f o r t h e r a t s on t h e s u p p le m e n te d d i e t w h ile t h e r a t s on t h e u n s u p p le m e n te d
d i e t show no c h a n g e i n w e i g h t.
The d i e t a p p e a r s t o b e d e f i c i e n t i n some g ro w th p r o m o tin g s u b ­
s t a n c e f o r t h e r a t w h ic h i s n o t r e s t o r e d e v e n on t h e a d d i t i o n o f a u t o l y z e d
iy e a s t t o a l e v e l o f 3$ i n t h e d i e t .
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T a b le I I
W i l l s D i e t V ita m in A s s a y on R a ts
A v e r a g e s o f T a b le I
W eig h t i n Grams
on D i e t
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
D ie t
31
30
29
29
30
28
29
27
27
27
27
29
30
29
29
30
29
D i e t & M a rm ite
29
32
34
34
35
37
39
41
43
45
43
48
57
56
60
64
63
C o n tro ls
28
40
53
78
93
111
128
144
146
177
185
204
212
25
I
>j
;|
!!
M onkeys ( T a b le s I I I ,
IV , V, a n d V I)
I|t
|
T he m onkeys s u r v i v e d 9 4 ,9 4 , 9 6 , 1 0 0 , 1 2 8 , 1 3 0 , 152 d ay s*
m onkey, N um ber 3 4 , w as p l a c e d on a n o rm a l d i e t a f t e r 130 d a y s .
t h e m onkeys l o s t w e ig h t s lo w ly o n t h e d i e t .
One
A ll o f
T hey d i d n o t e a t t h e d i e t
Iw e l l , o n s e v e r a l o c c a s i o n s r e f u s i n g t o e a t f o r o n e o r two d a y s . T h e r e
(i
II
jlwas no s u d d e n f a l l i n w e ig h t b e f o r e d e a t h ( T a b le I I I ) .
|i
A l l o f t h e m onkeys d e v e lo p e d a m o d e r a te l y s e v e r e h y p o c h ro m ic a n e m ia
|j
Jlw hile o n t h e d i e t .
The a n e m ia w as s lo w ly p r o g r e s s i v e i n a l l c a s e s e x c e p t
(iNumber 12 w h ic h sh o w ed a su d d e n a n e m ia a f t e r s e v e n t y - t h r e e d a y s .
The
ji
[one m onkey, Num ber 4 1 , w h ic h r e c e i v e d v e g e x i n a d d i t i o n t o t h e b a s a l d i e t
s d ie d a t 1 5 2 d a y s a n d show ed t h e same l o s s o f w e ig h t a n d a n e m ia a s t h o s e
S|
^ r e c e i v i n g t h e u n s u p p le m e n te d d i e t .
A t t h e tim e o f d e a t h t h e r e d b lo o d
c e l l c o u n ts v a r i e d fro m 3 .2 8 t o 4 . 4 6 , t h e h e m o g lo b in v a l u e s fro m 6 .8 3
[to 1 1 .1 8 a n d h e m a t o c r i t v a l u e s fro m 2 4 .5 t o 4 3 .5 .
ii
j
A t t h e tim e o f d e a th t h e m onkeys a p p e a r e d e m a c ia te d .
They d i d n o t
ii
jh p p e a r t o s u f f e r fro m a n o r e x i a b e f o r e d e a t h , e a t i n g t h e d i e t a p p a r e n t l y
f
las w e l l a s a t a n y o t h e r tim e d u r i n g t h e s t u d y . T hey a l l d e v e lo p e d a s e v e r e
l!
ijd ia r r h e a w h ile o n t h e d i e t , b u t n o t a r r y s t o o l s w e re o b s e r v e d . The
(^.utopsy p r o t o c o l s o f N um bers 4 0 , 4 1 , 1 2 , 28 a n d 35 f o l l o w :
t
!'!
il
A u to p s y P r o t o c o l s
Ib
1■
j w i l l s D i e t M onkey N um ber 40
W e ig h t, 4 p o u n d s .
I;
jj
© a a c i a te d m onkey, t h e r e a r e no e x t e r n a l s i g n s e x c e p t f o r e v id e n c e
|o f d i a r r h e a .
26
A l l o f t h e o r g a n s a r e g r o s s l y n o rm a l e x c e p t f o r t h e t r a n s v e r s e
c o lo n w h ic h i s i n j e c t e d ,
c o lo n a n d i s i n j e c t e d .
T he om entum i s w ra p p e d p a r t i a l l y a ro u n d t h e
T he c u t s u r f a c e o f t h e c o lo n r e v e a l s a t h i c k e n e d
- w a ll w i t h a g r e e n i s h m u c o id m em brane o v e r t h e e p i t h e l i u m .
T h i s m em brane
c a n n o t b e rem o v e d w i t h o u t t e a r i n g t h e m u c o s a .
T he f e m o r a l b o n e m a rro w i s
f a t a n y w h e re i n t h e m a rro w .
o f th e bone.
e x c e e d i n g ly r e d w i t h no e v id e n c e o f
T he m arro w e x t e n d s w e ll i n t o t h e d i a p h y s e s
The c o r t e x o f t h e b o n e i s a p p a r e n t l y t h i n n e d .
W ills D i e t M onkey Num ber 4 1 .
R e c e iv e d V e g e x .
W e ig h t, 3 p o u n d s .
V e ry e m a c ia te d m onkey w i t h s i g n s o f d i a r r h e a b e i n g t h e o n l y e x t e r n a l
s ig n .
I
j
A ll o f th e organs appear normal excep t fo r th e co lo n which i s
|in je c te d throughout most o f i t s le n g th .
The cecum i s markedly blu e in
Jcolor and surrounded by th e omentum which i s in j e c te d .
Upon opening the
■ in te stin e , th e mucous membrane i s r e d -y e llo w in c o lo r .
The il e o - c e c a l
!v a lv e protrudes f i v e mm. in to th e colon and i s very b lu e in c o lo r .
The
f i r s t and m iddle p a r ts o f th e colon are markedly in je c te d and th ick en ed .
jThe mucous membrane appears n e c r o t ic .
The lymph nodes ly in g along the
Jcolonic v e s s e l s are en larged .
|
T he lo w e r end o f t h e fe m u r on b o t h s i d e s i s
Is h a f t a t t h e e p i p h y s e a l j u n c t u r e .
s e p a r a t e d fro m t h e
The b o n e i s d r y a n d a p p e a r s n e c r o t i c ,
j!
!ino r e d n e s s , n o m o i s t u r e , a n d no e x u d a te .
jl
I'to th e same d e s c r ip tio n .
i|
I
i
T he h e a d o f t h e h u m e ri a n s w e r
27
W i l l s D i e t M onkey Num ber 12
J
W eight, 6 pounds.
No e x t e r i o r s i g n s e x c e p t f o r e v id e n c e o f d i a r r h e a .
A l l o f t h e o r g a n s a p p e a r n o rm a l e x c e p t f o r t h e c o l o n .
o u te r s u r f a c e o f t h i s o rg an th e r e i s an a re a 5 x 7
in c o lo r.
On t h e
cm s. w h ic h i s b l u e r e d
On e x a m in a tio n fro m t h e lu m e n , t h i s i s s e e n t o b e t h e s i t e
of
tw o e r o s i o n s 1 . 5 cm s. a n d 3 cm s. s q u a r e a b o u t 7 cm. fro m t h e i l e a l o p e n in g .
T h e re i s a n o t h e r a r e a i n t h e m id d le s e c t i o n o f t h e c o lo n w h ic h show s some
e p it h e li a l e ro s io n .
T he lym ph n o d e s a lo n g t h e c o l o n i c v e s s e l s a r e
e n la rg e d and d is c o lo r e d .
The b o n e m a rro w o f t h e f e m o r a l b o n e s a p p e a r s d a r k r e d i n c o l o r
t h r o u g h o u t t h e b o n e a n d e x te n d s w e l l i n t o t h e d i a p h y s i s .
W ills D i e t Monkey N um ber 2 8 .
W e ig h t, 3§ p o u n d s .
The o n l y e x t e r i o r s i g n i s
e v id e n c e o f a d i a r r h e a .
A l l o f t h e o r g a n s a r e n o rm a l e x c e p t f o r t h e c o l o n .
A s m a ll
se g m e n t o f t h e c o lo n 2 x 2 cm. i n a r e a i s a d h e r e n t t o t h e l a t e r a l a b d o m in a l
w a l l i n t h e lo w e r l e f t a b d o m in a l q u a d r a n t .
om entum w h ic h i s h e a v i l y i n j e c t e d .
a re e n la r g e d and d is c o lo r e d .
S u r r o u n d in g t h i s i s t h e
The lym ph n o d e s a l o n g t h e c o l o n i c v e s s e l s
On o p e n in g t h e c o lo n t h e m ucous m em brane
a p p e a r s r e d - y e l l o w i n c o l o r an d i s
s l i g h t l y t h i c k e n e d i n some a r e a s .
T h e r e a r e m any e r o s i o n s o f t h e m u c o s a .
T he f e m o r a l b o n e m arro w i s d a r k r e d i n c o l o r th r o u g h o u t t h e b o n e
and e x te n d s w e ll i n t o th e d ia p h y s is .
28
;I
jW
1i
|
ills D i e t M onkey N um ber 35
W e ig h t, 3 . 5 p o u n d s .
E x t e r i o r l y th e o n ly s ig n i s
th a t o f a d ia rrh e a .
A l l o f t h e o r g a n s a p p e a r n o rm a l e x c e p t t h e c o l o n .
The c o lo n i s
m a r k e d ly u l c e r a t e d .
M ic r o s c o p i c E x a m in a tio n
. W i l l s D i e t M onkey Num ber 28
C o lo n :
^ d e m a to u s m ucous m em brane i n m any a r e a s .
The m u c o s a l
c e l l s a r e g o n e a n d o n l y c o n n e c t i v e t i s s u e a n d s tro m a
I
a re l e f t .
T h e r e a r e s e v e r a l l a r g e a r e a s w h e re t h e r e
i s n o t h i n g b u t in f la m m a to r y c e l l s an d e x u d a te
j
re p la c in g
m ucous m em brane a n d e x te n d i n g down t o a n d i n t o
■
t h e su b m u c o sa .
T h is i s a n a c u t e i n f la m m a to r y p r o c e s s
w i t h n e c r o s i s o f t h e m ucous m em brane.
I
j
Lymph N ode:
E v id e n c e o f a c u t e l y m p h a d e n i t i s .
j
J
C o n t a in s many l a r g e s i n g l e n u c l e a t e d p h a g c y t i c c e l l s ,
L iv e r:
K u p fe r c e l l s l a r g e .
L i v e r c e l l s a p p e a r t o h a v e u n d e rg o n e
some c lo u d y s w e l l i n g , b u t n u c l e i a p p e a r n o r m a l.
Spl e en:
j
i
I
I
N o rm a l.
B one m arrow :
M any n o r m o b l a s t s .
o r no f a t .
E v id e n c e o f m uch a c t i v i t y .
The g r a n u l a r s e r i e s a r e r e l a t i v e l y
|
I
I
||
in c r e a s e d o v e r th e e r y th r o c y tic s e r i e s .
I
|
a re o f th e u n d iffe re n tia te d ty p e .
i!
tj
|i
ii
ii
L ittle
n o t a s im p l e h y p e r p l a s i a .
T h is i s
The p r e d o m in a n t c e l l s
“'Wills Diet Monkey Number 12
C o lo n :
^ d e m a to u s ; m u c o sa a b s e n t ; no e v id e n c e o f in f l a m m a t o r y c e l l s
b u t m uch n e c r o s i s .
L iv e r:
S p le e n :
Same a s N um ber 28 b u t m ore d e g e n e r a t i o n .
N o rm a l.
Lymph N o d e s :
E d e m a to u s ; l a r g e a r e a s d e s t r o y e d .
n u c le a te d c e l l s .
Some l a r g e s i n g l e
D oes n o t a p p e a r so a c t i v e a s
N um ber 2 8 .
Bone m arro w ;
Some f a t b u t l e s s i n am ount t h a n n o r m a l,
fia tio o f
r e d b lo o d c e l l t o w h ite b lo o d c e l l fo rm s i s a b o u t
n o r m a l.
N o t a s c e l l u l a r a s N um ber 2 8 .
W il ls D i e t Monkey N um ber 35
C o lo n :
L a r g e a r e a w h e re m u co sa i s d e s t r o y e d a n d in f la m m a to r y
a n d e x u d a t i v e c e l l s a r e p r e s e n t e x t e n d i n g down t o a n d
i n t o t h e su b m u c o sa .
is
e d e m a to u s .
I n a d j a c e n t a r e a s t h e m ucous m em brane
T h e re i s
some edem a a n d r o u n d c e l l
i n f i l t r a t i o n in to th e se ro sa n e a r th e b ase o f th e u lc e r .
L iv e r:
K u p fer c e l l s la r g e .
E v id e n c e o f p o s t m ortem a u t o l y s i s
h i d e s a n y p a t h o l o g i c a l c h a n g e s w h ic h may b e p r e s e n t .
Spl een:
N o rm a l.
ii
j!
fe lls
ii
D i e t M onkey N um ber 40
Bone M arrow :
M id -fe m u r:
W id e ly d i l a t e d b l o o d - f i l l e d s i n u s o i d s .
E a t v a c u o l e s o c c u p y a p p r o x i m a t e ly 10 t o 20$ o f t h e
a r e a r e p r e s e n t e d by t h e s e c t i o n s .
T h e re i s d i f f u s e
h y p e rp la s ia o f u n d if f e r e n tia te d la r g e b a s o p h ilic
50
c e l l s r e s e m b l i n g h e m o c y t o b l a s t s , n e u t r o p h i l i c m y e l­
o c y t e s a n d m e ta m y e lo c y te s , w i t h s u p p r e s s i o n o f t h e
e ry th ro b la s tic
e le m e n t s .
F o c i o f n u c le a te d e r y th ­
r o c y t e s a r e e x tr e m e l y s m a ll a n d fe w .
nu m b er o f m a t u r e n e u t r o p h i l s ,
A m o d e r a te
e o s i n o p h i l i c m y e l o c y te s
a n d m e ta m y e lo c y te s a r e p r e s e n t i n t h e t i s s u e .
N orm al
m e g a k a r y o c y te s a r e r e l a t i v e l y n u m e ro u s .
N o te :
T h i s i s a s t r o n g l y h y p e r p l a s t i c m arro w i n v o l ­
v i n g c h i e f l y t h e ste m c e l l s ,
w i t h a te n d e n c y t o
d i f f e r e n t i a t e i n th e d i r e c t i o n o f g ra n u lo c y te s
{ i . e . a le u k e m o id p i c t u r e ) .
I n sum m ary, t h e o n l y p a t h o l o g i c a l e v id e n c e o f d i s e a s e i n t h e s e
m onkeys i s a n a c u t e u l c e r a t i v e in f la m m a to r y c o n d i t i o n o f t h e c o lo n *
The
bone m a rro w p i c t u r e s a r e c o m p a tib le w i t h a n in f la m m a to r y c o n d i t i o n .
N one o f t h e m onkeys d e v e lo p e d a s i g n i f i c a n t m a c r o c y t i c a n e m ia
b e fo r e d e a th .
T h e r e w as no e v id e n c e o f t h e t r o p i c a l m a c r o c y t i c a n e m ia .
11 o f th e monkeys dyin g w h ile on th e d ie t d ie d o f an u lc e r a t iv e c o l i t i s .
S w ine ( T a b le s V I I , V I I I , IX , X, a n d X I)
The s u r v i v a l t im e s o f t h e s w in e on t h e d i e t w e re 1 3 2 , 1 3 7 , 1 5 0 ,
58 and one p laced on normal d ie t 1 5 4 days.
The swine a te th e d ie t w e ll
j^t f i r s t , but by 125 days showed sig n s o f a n o rex ia .
In Table VIJ i t m i l
e seen th a t Number 1 and Number 3 in crea sed in w eight fo r the f i r s t 97
a ys and th en began to lo s e w eight slo w ly .
Numbers 4 , 5, and 6 showed
jjuaximum w eight a t 46 days, and d e c lin e d s t e a d ily u n t i l death.
The swine
L id not develop s c a ly s k in s a t any tim e du rin g th e experim ental period
31
T a b le I I I
W i l l s D i e t M onkeys
W eig h t i n P o u n d s
D ays o n
D ie t
0
8
16
30
45
61
73
82
96
112
124
138
#40
#41
#12
#28
#34
#35
5 .5
5 .5
5 .5
5
5
5 .3
8
4 .5
5 .0
5 .3
5 .3
5 .3
5 .0
5 .3
4 .8
4 .5
4 .8
4 .0
5 .3
5 .5
5 .3
5 .3
4 .5
4 .5
4 .0
Dead
1 00 d a y s
7 .8
8 .0
7 .3
7 .0
6 .5
D ead
194d ay s
4 .3
4 .5
4 .0
4 .0
3 .8
Dead
96 d a y s
5 .3
4 .5
4 .0
4 .8
4 .0
4 .0
N orm al
d ie t
130 d a y s
3 .8
D ead
128
days
D ead
152 days
32
T a b le IV
W i l l s D i e t M onkeys
R ed B lo o d C e l l s X 1 0 ^ p e r Cu. Mm.
D ays on
D ie t
0
8
16
30
45
61
73
82
96
112
124
138
.
#40
#41
6 .8 6
7 .1 6
6 .7 3
7 .8 5
6 .3 9
6 .5 4
6 .3 5
6 .8 8
4 .6 6
4 .4 6
5 ,3 6
6 .6 6
6 .2 8
5 .1 0
4 .1 0
#12
#28
#34
#35
5 .8 4
5 .5 6
5 .5 3
5 .9 4
5 .4 6
5 . 86
4 .7 9
4 .9 7
3 .7 5
3 .2 8
5 .7 0
5 .1 0
4 .8 0
4 .9 8
3 .3 0
4 .7 9
5 .8 5
5 .1 2
5 .0 7
3 .8 6
4 .2 4
4 .3 3
4 .6 2
4 .5 5
3 .9 0
33
T a b le V
W i l l s D i e t M onkeys
H e m o g lo b in Gram s p e r 100 c c .
D ays o n
D ie t
0
8
16
30
45
61
73
82
96
112
124
138
#40
#41
#12
#28
#34
#35
1 3 .4 9
1 3 .2 3
1 3 .0 6
1 1 ,8 8
1 1 .5 2
1 2 .9 0
1 1 .8 8
1 1 .9 5
1 1 .4 8
1 1 .0 5
1 1 .0 0
1 2 .1 3
1 2 .3 6
1 2 .7 5
1 1 .1 8
1 2 .3 0
1 1 .3 0
1 1 .1 2
1 1 .1 8
1 1 .8 8
1 0 .0 5
9 .1 0
7 .8 6
6 .8 3
1 0 .1 5
9 .9 5
8 .8 9
9 .0 9
1 0 .0 5
8 .7 6
1 2 .0 1
1 1 .6 1
1 0 .0 0
1 0 .8 9
9 .4 9
1 0 .5 6
9 .8 6
9 .9 5
7 .7 4
9 .4 5
8 .9 8
34
T a b le 71
W i l l s D i e t M onkeys
H e m a to c r i t v a l u e s p e r c e n t c e l l s
D ays o n
© ie t
0
8
16
30
45
61
73
82
96
112
124
138
#40
#41
#12
#28
#34
#35
4 8 .0
46. 5
4 7 .0
4 3 .5
4 2 .5
4 7 .0
4 3 .7 5
4 6 .5
4 3 .7 5
4 2 .0
4 4 .0
4 5 .5
4 5 .5
4 7 .0
4 1 .0
4 6 .0
4 0 .0
4 1 .0
3 7 .0
4 7 .5
3 7 .0
4 3 .0
3 0 .0
2 4 .5
3 6 .5
3 3 .5
3 1 .5
3 2 .0
3 7 .5
3 7 .5
4 3 .5
4 3 .0
3 6 .0
3 8 .0
3 2 .0
3 6 .5
4 3 .5
3 4 .0
3 1 .0
3 3 .0
3 4 .5
j n o r w e re t h e i r g a i t s u n u s u a l . H o w ev er, N um bers 4 , 5 , a n d 6 d i d b e g i n t o
i
| l o s e t h e i r t e e t h a b o u t th e 1 1 1 th d a y .
T h is w as n o t i c e d w hen t h e w ooden
j g a g u s e d t o k e e p t h e i r m o u th s o p e n d u r i n g t h e p r o c e s s o f p a s s i n g t h e s to m a c h
i
jj t u b e w as p l a c e d i n t h e i r m o u th s .
j lin e .
The t e e t h w o u ld b r e a k o f f a t t h e gum
T h e r e w as no s u b s e q u e n t b l e e d i n g fro m t h e m o u th .
Num ber 6 b ecam e
m a r k e d ly s w a y -b a c k e d a b o u t t h e 1 2 5 t h d a y a n d r e m a in e d so - u n t i l d e a t h .
[
As s e e n i n T a b l e s T i l l ,
IX , X, a n d X I, t h e r e d b lo o d c e l l s o f t h e
v a r i o u s a n e m a ls d e s c r i b e d a n e r r a t i c
A s e v e r e a n e m ia d i d n o t d e v e lo p i n a n y o f t h e a n i m a l s .
The l o w e s t
e r y t h r o c y t e c o u n t a t d e a t h w as N um ber 6 , 5 .0 7 m i l l i o n .
The te n d e n c y
i n a l l c a s e s w as t o
jj
c o u r s e d u r in g t h e e x p e r i m e n t a l p e r i o d .
d e v e lo p a m i l d h y p o c h ro m ic a n e m ia ,
T h e r e w e re no n o t i c e a b l e g r o s s a b n o r m a l i t i e s a t a u t o p s y i n a n y
j o f t h e a n im a ls e x c e p t f o r t h e a b s e n c e o f m o s t o f t h e t e e t h i n N um bers 4 ,
5 , a n d 6 a n d t h e sway b a c k o f Num ber 6 .
M ic r o s c o p i c F i n d in g s
i W i l l s D i e t S w ine N um ber 4
I
H
j
K id n e y :
M ark ed a u t o l y s i s o f t u b u l e s b u t t h e r e i s e v id e n c e o f
[
3
I
!
!
fa tty
S p le e n :
!
L iv e r:
changes.
M uch c o n n e c t i v e t i s s u e ; o t h e r w i s e seem s n o r m a l.
S e v e re f a t t y
d e g e n e ra tio n .
j W i l l s D i e t S w in N um ber 5
!
S p l e e n : N o rm a l.
I
jI
j;
L iv e r:
M o d e r a t e l y s e v e r e am ount o f f a t t y d e g e n e r a t i o n m o s t p r e d o m in a n t
a t c e n te r o f lo b u le s .
c o n n e c tiv e t i s s u e .
I n c r e a s e d am ount o f
Ki dney:
S to m a c h :
N o rm a l.
I n f l a m m a t o r y c o n d i t i o n i n su b m u co sa e x t e n d i n g a lo n g t h e
b lo o d v e s s e l s i n t o t h e m u s c u l a r i s .
T h is i s a s u b a c u te
i n f l a m m a t i o n s i m i l a r t o t h e c o lo n s o f t h e m o n k ey s.
W i l l s D i e t S w in e N um ber 6
K id n e y :
N o rm a l.
S p le e n :
N o rm a l.
T he m jfltro s c o p ic f i n d i n g s a r e c o m p a t i b l e w ith t h e d i e t a r y d e f i c i e n c y ,
T he s w in e on t h e W i l l s d i e t , t h e n , f a i l e d t o d e v e lo p a m a c r o c y t i c
a n e m ia , a n d , i n fa c & , f a i l e d t o d e v e lo p a s e v e r e a n e m ia o f a n y k i n d b e f o r e
d e a th .
T he W i l l s d i e t a s m o d i f i e d w as a p p a r e n t l y d e f i c i e n t i n som e su b ­
s t a n c e w h ic h i s n e c e s s a r y f o r t h e g ro w th o f s w in e .
G a s t r e c to m iz e d D ogs ( T a b le X I I , X I I I )
T h e f o u r g a s t r e c t o m i z e d d o g s w e re p l a c e d on t h e m o d i f i e d W i l l s
d i e t i n th e hope t h a t t h e added l o s s o f th e
sto m a c h a n d i t s
s e c re tio n s
m ig h t l e a d t o a m a c r o c y t i c a n e m ia .
T he s u r v i v a l tim e o f t h e tw o d o g s on t h e m o d if ie d W i l l s d i e t
jj w i t h o u t t h e s u p p le m e n t w as 1 4 7 d a y s , a n d t h e o t h e r dog w as s a c r i f i c e d a t
j100 d a y s f o r a n o t h e r g a s t r e c t o m y p r o b le m .
I
O f t h e tw o d o g s o n t h e d i e t
j p l u s v e g e x , o n e d i e d i n 192 d a y s , a n d t h e o t h e r w as p l a c e d o n a n o im a l
d i e t a f t e r 268 d a y s.
One o f t h e tw o d o g s t h a t r e c e i v e d n o s u p p le m e n t t o t h e b a s a l d i e t
(N um ber 7) g a in e d w e ig h t d u r i n g t h e f i r s t m o n th on t h e d i e t ,
and th e n l o s t
jl
jj w e ig h t s l o w l y , f a l l i n g t o 19 p o u n d s a t d e a t h o r 3 p o u n d s b e lo w h i s i n i t i a l
T a b le V I I
W i l l s D i e t S w ine
W e ig h t i n P o u n d s
D ays o n
D ie t
#1 P .
0
11
46
58
62
69
90
97
104
111
125
152
7 8 .5
8 1 .5
8 9 .0
7 2 .0
8 0 .5
8 6 .0
8 7 .5
8 9 .5
8 7 .5
8 5 .5
8 7 .5
8 6 .5
146
155
223
# 4 M.
#5 P .
#6 P .
7 5 .0
8 1 .5
1 0 0 .0
9 9 .0
1 0 0 .0
1 0 1 .0
1 0 6 .5
1 1 0 .0
1 0 5 .5
1 0 3 .5
1 0 1 .0
9 9 .0
8 4 .0
8 1 .5
9 2 .5
8 4 .0
8 5 .0
8 5 .0
8 5 .5
8 6 .5
8 4 .0
8 3 .5
8 1 .5
7 8 .5
5 1 .5
5 3 .0
5 7 .0
5 5 .0
5 6 .0
5 5 .0
5 5 .0
5 3 .5
5 3 .5
5 0 .5
4 9 .0
D ie d
132 days
8 6 .5
8 6 .0
9 2 .0
7 5 .5
8 3 .0
8 5 .0
8 1 .5
8 1 .5
7 9 .5
7 7 .0
7 4 .0
7 2 .0
8 8 .0
9 8 .0
7 1 .0
8 8 .5
9 6 .0
D ie d
150 d a y s
P l a c e d on
n o m al d ie t
154 days
200
#3 P .
P l a c e d on
n o rm a l d i e t
154 d a y s
D ie d
158 d a y s
D ie d
137 d a y s
38
T a b le V I I I
W i l l s D i e t S w in e
R ed B lo o d C e l l s X 10® p e r C n. Mm.
D ays on
D ie t
0
46
58
62
69
90
97
104
111
125
132
146
153
273
#1
#3
#4
#5
#6
7 .0 4
6 .9 9
6 .8 1
6 .5 2
7 .1 0
7 .1 0
5 .5 5
7 .2 2
7 .6 5
8 .0 5
8 .4 2
6 .1 5
5 .8 5
7 .9 5
7 .0 2
7 .4 7
5 .5 3
6 .5 2
8 .2 0
7 .5 5
7 .6 2
7 .4 0
8 .8 9
8 .0 5
7 .6 0
9 .0 7
7 .2 5
5 .5 4
6 .3 7
5 .4 9
5 .7 5
6 .7 5
6 .0 5
6 .4 0
5 .5 7
5 .7 5
5 .1 8
5 .3 0
5 .1 2
4 .8 4
6 .8 0
5 .4 5
6 .4 7
5 .6 5
5 .2 5
7 .6 5
5 .5 5
5 .2 0
5 .2 8
5 .0 7
8 .2 7
6 .5 4
6 .1 7
6 .7 7
6 .2 5
5 .8 0
7 .6 0
6 .8 5
8 .1 8
5 .1 5
39
T a b le IX
W i l l s D i e t S w ine
H e m o g lo b in G ram s p e r 100 c c .
D ays o n
D ie t
0
46
58
62
69
90
97
104
111
125
132
146
153
273
#1
#3
#4
#5
#6
1 1 .0 0
1 0 .5 6
1 2 .9 0
1 1 .3 6
1 0 .3 0
9 .1 6
9 .8 8
9 .7 1
1 2 .9 8
1 1 .1 8
1 0 .4 5
1 1 .3 6
1 1 .2 4
1 1 .3 6
1 2 .5 9
1 2 .2 2
1 0 .0 0
1 1 .4 8
1 1 .7 5
1 0 .2 0
1 4 .3 1
1 2 .0 1
1 2 .0 1
1 2 .4 4
1 3 .0 6
1 2 .1 3
1 4 .4 1
1 3 .4 0
1 2 .9 0
1 2 .4 4
1 1 .1 2
9 .1 7
9 .0 9
1 0 .2 5
1 1 .8 8
9 .1 0
9 .7 6
9 .2 4
8 .5 2
9 .0 9
9 .8 7
8 .9 3
7 .8 6
1 0 .3 5
8 .5 7
8 .5 7
7 .8 6
1 1 .0 0
9 .3 3
1 0 .0 0
1 2 ,4 4
1 2 .0 1
1 3 .8 4
9 .6 3
8 .8 9
T a b le X
W i l l s D i e t ' S w in e
H e m a to c r i t p e r c e n t c e l l s
D ays o n
D ie t
0
46
62
69
90
97
104
U1
125
132
146
153
273
#1
#3
H
#5
#6
3 3 .0
3 4 .5
3 5 .0
3 7 .0
3 5 .5
3 2 .0
3 8 .0
3 5 .0
3 6 .0
3 4 .5
3 6 .0
3 4 .0
4 6 .5
4 0 .0
3 9 .0
4 1 .0
4 2 .0
3 9 .0
4 1 .5
4 1 .5
4 6 .5
4 6 .0
4 1 .0
4 0 .0
3 5 .5
2 7 .0
3 2 .5
3 1 .5
3 2 .2
3 3 .5
3 7 .0
3 3 .5
3 0 .0
2 8 .0
2 7 .0
2 8 .0
3 3 .0
3 3 .0
3 4 .5
3 1 .0
2 6 .5
3 4 .0
2 6 .0
2 8 .5
2 8 .0
4 1 .0
3 9 .0
4 1 .0
3 1 .0
3 2 .5
3 7 .5
3 8 .0
4 4 .0
3 2 .0
2 9 .5
41
T a b le XX
'W ills D i e t S w ine
A v e ra g es
D ays o n
D ie t
0
11
46
58
62
69
90
97
104
111
125
132
146
153
N um ber
s w in e
5
5
5
5
5
5
5
5
5
5
5
4
3
2
W e ig h t
7 5 .1
7 6 .7
8 6 .1
7 7 .1
8 0 .9
8 2 .4
8 5 ,2
8 4 .3
8 2 .0
8 0 .0
7 8 .6
8 4 .0
8 5 .5
9 2 .2
R ed b l o o d
c e lls
Hemog lo b in
Hemato c rit
6 .5 4
1 1 .2 1
3 4 .8
6 .8 3
5 .9 0
6 .4 1
6 .8 0
6 .3 5
6..9S
6 .5 2
7 .1 3
6 .3 4
6 .6 2
6 .7 8
6 .5 5
10-. 40
3 5 .6
1 0 .5 0
1 0 .6 8
1 0 .3 6
1 0 .8 3
1 0 .8 8
1 1 .7 6
1 0 .2 3
1 0 .4 5
1 1 .0 9
1 0 .6 6
3 6 .6
3 4 .6
3 3 .4
3 6 .4
3 5 .4
3 6 .8
3 4 .0
3 3 .0
3 4 .6
3 4 .8
jw e ig h t.
N um ber 8 l o s t w e i g h t s lo w ly fro m t h e b e g in n i n g o f t h e d i e t u n t i l
n d e a th , l o s i n g 6 p o u n d s i n 100 d a y s .
Of t h e tw o d o g s r e c e i v i n g v e g e x , o n e
-|(Number 4 ) g a in e d w e ig h t s t e a d i l y o v e r a p e r i o d ©f 26® d a y s , g a i n i n g
!j
;|8 p o u n d s i n a l l .
The o t h e r d o g , N um ber 6 , d i e d a f t e r 192 d a y s on t h e
j!
ijd ie t, h a v in g l o s t 9 .5 pounds d u rin g t h i s p e r io d .
ij
A l l o f t h e d o g s e x c e p t N um ber 4 d e v e lo p e d a m o d e r a te m i c r o c y t i c
ij
'(hypochromic anemia w h ile on th e d i e t .
i|
Number 4 v a r ied co n sid e ra b ly during
]|th e e x p e r i m e n t a l p e r i o d , b u t d i d n o t d i f f e r a p p r e c i a b l y a t t h e e n d o f t h e
l[
~
I*
•jp e rio d fro m t h e i n i t i a l f i g u r e s .
j!
jj
;l
A l l o f t h e d o g s d e v e lo p e d d i a r r h e a w h i l e on t h e d i e t .
j|o f Num ber 4 w e re v a r i a b l e , u s u a l l y b e i n g se m i s o l i d .
!|
The s t o o l s
The p e r i o d b e f o r e
j!d e a th i n N um ber 6 a n d 8 w as c h a r a c t e r i z e d by a n o r e x i a .
jj
j
A t a u t o p s y , a s i d e fro m e v id e n c e o f a d i a r r h e a an d e m a c i a t i o n ,
itt h e r e w e re n o n o t i c e a b l e e v id e n c e s o f g r o s s p a t h o l o g y .
;j
j
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When t h e w h o le o f M is s wi l l s
w ork i s c o n s i d e r e d , t a k i n g i n t o
'a c c o u n t t h e d e s c r i p t i o n o f t r o p i c a l m a c r o c y t i c a n e m ia , t h e m onkey
a n e m ia , t h e d i e t s o f t h e i n d i v i d u a l s i n v o l v e d , a n d t h e d i e t s g i v e n t o
: t h e m o n k e y s, o n e o f t h e m o st o u t s t a n d i n g f e a t u r e s i s t h e s m a ll am o u n t
3!I
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The d i e t g i v e n t o t h e m onkeys
:j
jb y W i l l s i s n o t e n t i r e l y d e v o i d o f n i c o t i n i c a c i d , b u t c o n t a i n s , c e r t a i n l y ,
•m in im a l a m o u n ts o f t h i s s u b s t a n c e .
T h at th e d i e t i s o b v io u s ly d e f i c i e n t
i n o t h e r e s s e n t i a l s u b s t a n c e s s u c h a s V ita m in B-j_ c a n n o t b e d e n i e d .
H ow ever
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43
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|j
Ijam o u n ts o f n i c o t i n i c a c i d c o n t a i n e d i n t h e d i e t w e re s u b m in im a l i n comii
I p a r i s o n t o t h e d a i l y n e e d s o f t h e m onkey.
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i|
ij
p a t s tu d ie s .
The f a i l u r e o f r a t s t o grow on t h e d i e t a s r e p o r t e d h y
j|
jiW ills (4 0 ) o r t o becom e a n e m ic i s
c o n s i s t e n t v rith o u t own r e s u l t s i n t h i s
jl
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j
An i n d i r e c t p i e c e o f e v id e n c e f o r t h e l a c k o f n i c o t i n i c a c i d i n
jth e d i e t c a u s a t i v e o f t r o p i c a l m a c r o c y t i c a n e m ia i s
se en in th e tre a tm e n t
!
|o f t h e d i s e a s e . Y e a s t i s known t o c o n t a i n l a r g e a m o u n ts o f n i c o t i n i c a c i d ,
}
jps i s l i v e r .
The i n a b i l i t y t o c u r e t h e d i s e a s e w ith h i g h l y r e f i n e d l i v e r
Jj
p r e p a r a ti o n s - - w h i c h a r e v ita m in f r e e — and t h e r a p id c u re w ith c ru d e r
|j
i j e x t r a c t s — w h ic h c o n t a i n l a r g e a m o u n ts o f V ita m in B c o m p le x — i s s i m i l a r
i!
jito t h e e x p e r i e n c e s o c c a s i o n e d i n t h e t r e a t m e n t o f p e l l a g r a .
The d i s -
fl
i t r i b u t i o n o f t h e c u r a t i v e s u b s t a n c e i n t h e v a r i o u s f r a c t i o n s o f y e a s t an d
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l i v e r i s t h e sam e a s t h a t t o b e e x p e c t e d w e re t h e a g e n t n i c o t i n i c a c i d .
ij
jphe f a c t t h a t V i t a m in s A,
C, D, E a n d h e p a t o f l a v i n w e re n o t a c t i v e
■in d u r i n g t h e d i s e a s e p o i n t s a l s o
to w a rd some m em ber o f t h e
ijPhe d i e t i s a d e q u a t e i n Bg a s sh o w n b y t h e r a t
c o m p le x .
s t u d i e s i n w h ic h no a c r o d y n i a
i
ijie v e lo p e d e v e n w h en t h e r a t s w e re f e d t h e d i e t f o r o n e y e a r .
j|
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t
T h a t t h e d i s e a s e i s n o t d u e t o V ita m in "M" d e f i c i e n c y i s a p p a r e n t
i
Sjfrom t h e d e s c r i p t i o n o f t h e tw o d i s e a s e s .
The l a c k o f m o u th l e s i o n s ,
th e
jj.onger p e r i o d o f s u r v i v a l , a n d t h e l a c k o f as s e v e r e a n a n e m ia i n t h e M i l s
jjlie t m onkeys c o m p a red t o t h e V ita m in "M" d e f i c i e n t a n im a ls b e s p e a k s a g a i n s t
tj
t h e tw o b e i n g i d e n t i c a l .
H o w ev er, i t
i s n o t i m p o s s i b l e t h a t t h e d i e t may
46
be p a r t i a l l y
d e fic ie n t in t h is
s u b s ta n c e ,
if it
e x is ts .
W h e th e r o r n o t t h e g a s t r e c t o m i z e d d o g s d e v e lo p e d b l a c k to n g u e on
t h e d i e t i s a m oot p o i n t .
E x c e p t f o r t h e m o u th l e s i o n s ,
r e s e m b le d i n e v e r y r e s p e c t t h e b l a c k to n g u e s y n d ro m e .
t h e a n im a ls
S in c e th e g a s tr e c ­
to m iz e d d o g s h a v e a v e r y s h o r t i n t e s t i n a l p a s s a g e tim e a n d r e a d i l y d e v e lo p
a d i a r r h e a , i t i s n o t u n l i k e l y t h a t t h e m o u th l e s i o n s m ig h t n o t a p p e a r
s i n c e t h e d e f i c i e n c y m ig h t becom e m o re p r o n o u n c e d i n l e s s tim e t h a n i n t h e
n o rm a l d o g s d u e t o p o o r a b s o r p t i o n .
T h is m ig h t p o s s i b l y l e a d t o d e a t h
b e f o r e t h e m o u th l e s i o n s c o u ld d e v e lo p .
in w e ig h t,
T he s u d d e n t e r m i n a l a b r u p t l o s s
e m a c i a t i o n , f o u l d i a r r h e a , a n d s u d d e n d e a th a r e v e r y s i m i l a r ,
i f n o t i d e n t i c a l , w ith b la c k to n g u e .
T h a t t h e c o n d i t i o n i s c u r a b l e w ith
a u to l y z e d y e a s t w as d e m o n s t r a t e d .
!
B e c a u s e i n o u r d i e t t h e s u b s t i t u t i o n o f m a tz o s m e a l f o r w h ite b r e a d ,
J e w e l s h o r t e n i n g f o r g h i , a n d t h e o m is s io n o f w h e a t c h a p a t t i a n d m e a t
l o w e r s t h e V ita m in B c o n t e n t , we m ig h t w e l l e x p e c t t h e sym ptom s o f t h e
d i s e a s e i n o u r m onkeys t o b e m o re s e v e r e .
H o w ev er,
we a r e u n a b le t o a c c o u n t
f o r t h e i n a b i l i t y t o p r o d u c e a m a c r o c y t i c a n e m ia a s d e s c r i b e d bjr W i l l s .
A s i d e fro m t h e b l o o d p i c t u r e , t h e s i g n s an d sym ptom s o f t h e m onkey d i s e a s e
as d e s c r ib e d by W ills a r e v e ry s i m i l a r to th e o n es o b se rv e d i n t h i s
e x p e rim e n t.
The f a i l u r e o f t h e m onkey Humber 41 t o r e s p o n d t o t r e a t m e n t
w ith v e g e x i s p r o b a b l y d u e t o a n i n s u f f i c i e n t f e e d i n g o f t h e m a t e r i a l .
The s u r v i v a l t i m e s o f o u r m onkeys on t h e d i e t c o r r e s p o n d s w e l l w i t h t h e
s u r v i v a l t i m e s o f W i l l s m o n k e y s.
I n r e g a r d t o t h e s w in e , t h e f a i l u r e t o p r o d u c e a m a c r o c y t i c
a n em ia d o e s n o t n e c e s s a r i l y r u l e o u t t h e l a c k o f n i c o t i n i c a c i d s i n c e
R h o ad s a n d M i l l e r r e p o r t e d t h a t t h e i r s w in e d e v e lo p e d m i c r o c y t i c and
l o r m o c y t i c a n e m ia s a s w e l l a s m a c r o c y t i c o n e s .
H o w e v e r, we f a i l e d t o
o t h e r e v id e n c e o f b l a c k t o n g u e , a s i d e fro m d i a r r h e a ,
It is
see
th a t th e y r e p o r t.
t o b e re m e m b e red , h o w e v e r, t h a t o u r d i e t w as o n l y r e l a t i v e l y
d e f ic ie n t in n ic o tin ic a c id .
T h is m ig h t a c c o u n t f o r t h e d i f f e r e n c e s
b e tw e e n m o n k ey s, s w in e a n d d o g s f e d a n a lm o s t a b s o l u t e l y d e f i c i e n t d i e t
an d t h o s e f e d o u r d i e t .
T h e r e a r e o t h e r p o s s i b i l i t i e s a s t o t h e n a t u r e o f t h e s u b s ta n c e
a b s e n t fro m t h i s d i e t .
I t i s r e g r e t t a b l e t h a t W ills h a s n o t t r i e d ,
to
o u r k n o w le d g e , n i c o t i n i c a c i d p e r s e i n t h e t r e a t m e n t o f t h e d i s e a s e a s
s e e n i n m an a n d m o n k e y s.
C o n c lu s io n s
M onkeys, s w i n e ,
d ie t a s used in t h is
I
The d i e t i s
an d g a s t r e c t o m i z e d d o g s f e d t h e m o d if ie d W i l l s
e x p e r im e n t d i d n o t d e v e lo p m a c r o c y t i c a n e m ia s .
d e f i c i e n t i n some s u b s t a n c e n e c e s s a r y f o r g ro w th a n d
m a in te n a n c e o f t h e s e a n i m a l s .
I
W h e th e r o r n o t t h i s s u b s t a n c e i s p r e s e n t i n a u t o l y z e d y e a s t
Ic a n n o t b e s t a t e d .
T he m o d if ie d W i l l s d i e t i s n o t s u f f i c i e n t t o m a i n t a i n n o rm a l
g ro w th i n im m a tu re r a t s .
B ib lio g ra p h y
I ( l ) M c C a r r i s o n , R . , I n d . J . M ed. R e s . 1 4 : 8 9 5 , 1 9 2 6 .
| ( 2 ) W i l l s , L . , a n d M e h ta , M .M ., I n d . J . M ed. R e s . 1 7 : 7 7 7 , 1 9 3 0 .
( 3 ) W i l l s , L . , a n d T a l p a d e , S . N . , I n d . J . M ed. R e s . 1 8 : 2 8 5 , 1 9 3 0 .
| ( 4 ) W i l i s , L . , L a n c e t 1 : 1 0 5 9 , 1931
(5) li ll s f
B r i t . M ed. J r . 1} 1 0 5 9 , 1 9 3 1 .
( 6 ) W i l l s , L . , a n d B i l m o r i a , H . S . , I n d . J . M ed. R e s . 2 0 : 3 9 1 , 1 9 3 2 .
( 7 ,) W i ll s , L . , a n d S t e w a r t , A ., B r i t . J .
Exp. P a th . 16: 44 4 , 1935.
( 8 )W ills , L ., L a n c e t 224: 1 2 8 3 , 1933.
( 9 ) W i l l s L . , C l u t t e r b u c k , P .W ., a n d E v a n s , B .D .F ., B iochem J .
31 : 2 1 3 6 , 1 9 3 7 .
( 1 0 ) W i l l s , L . , C l u t t e r b u c k , B .W ., a n d E v a n s , B . D . F . , L a n c e t 2 3 2 , 1 9 3 7 .
( 1 1 ) W ills , L . , and E v an s, B .D .F ., L a n c e t 2: 416, 1938.
( 1 2 ) D ay, P . L . , L a n g s to n , W. C . , a n d S h u k e r s , C . F . , J . R u t r . £ :
637, 1935.
(1 3 )D a y , P . L . , L a n g s to n , W. C. a n d D a rb a y , W. J . , P r o c . S o c . E xp. B i o l ,
a n d M ed. 3 8 : 8 6 0 , 1 9 3 8 .
( 1 4 ) L a n g s t o n , W. C . , D a rb y , W. J . , S h u k e r s , C . F . , D ay, P . L . , J . E xp.
M ed. 6 8 : 9 2 3 , 1 9 3 8 .
( 1 5 ) H a r r i s , L . J . , B io ch em . J . 3 1 : 1 4 1 4 , 1 9 3 7 .
( 1 6 ) H a r r i s , L . J . , B io c h e m . J .
32: 1 0 , 1938.
(17)M cG ow an, J . P . , a n d S i n c l a i r , R. D ., E d in b u r g h , M ed. J .
38 : 4 0 5 , 1 9 3 1 .
| ( l 8 ) M i l l e r , D .K ., a n d R h o a d s , C . P . J . , C l i n I n v e s t , 1 4 : 1 5 3 , 1 9 3 5 .
( l 9 ) B i r c h , T . W ., C h ic k , H . , a n d M a r t i n , C . J . , B iochem . J .
31: 2065, 1937,
|{ 20) C h ic k , H . , M a c ra e , T . F . , M a r t i n , A .J - P * , a n d M a r t i n , C . J . , B iochem
J . 32: 8 4 4 , 1938.
2 1 ) W in tr o b e , M .M ., S a m te r , M ., a n d L i s c o , H . , J o h n s H o p k in s H o sp . B u l l .
64: 399, 1939.
2 2 ) C ohn, E . J . , M in o t, G .R ., A l l e s , G .A ., a n d S a l t e r , W .T ., J . B i o l .
Chem. 7 7 : 3 2 5 , 1 9 2 8 .
49
2 3 ) D a v id s o n , S . , L a n c e t 2 2 1 : 6 3 , 1 9 3 1 .
2 4 ) D a v id s o n , S . , L a n c e t 22 3 : 9 1 9 , 1 9 5 2 .
2 5 )T Jn g le y , C .C ., L a n c e t 2 2 1 :
63, 1931.
2 6 ) U n g le y , C .C ., L a n c e t 2 2 3 ; 8 6 7 , 1 9 3 2 .
2 7 )U n g le y , C .O ., Q u a r t. J". M ed. 2: 3 8 1 , 1 9 3 2 .
2 8 ) U n g le y , C .C ., a n d J a m e s , G .V ., Q u a r t. J . M ed. 3 : 5 2 3 , 1 9 3 4 .
2 9 ) S t r a u s s , M .B ., a n d C a s t l e ,
W .B ., New l a g . J . M ed. 207: 5 5 , 1 9 3 2 .
3 0 ) S t r a u s s , M .B ., a n d C a s t l e ,
W .B ., L a n c e t 2 2 3 : 1 1 1 , 1 9 3 2 .
3 1 ) G o o d a l l , A ., L a n c e t 223: 7 8 1 , 1 9 3 2 .
3 2 ) W H is , L . a n d N a i s h , A ., L a n c e t 224: 1 2 8 6 , 1 9 3 3 .
3 3 ) L a s s e n , H. C .A ., a n d L a s s e n ,
H .E .,
J . M ed. S c i . 1 8 8 :4 6 1 , 1 9 3 4 .
3 4 ) R u s s e l l , H .K ., A nn. I n t . M ed. 7 : 1 3 9 8 , 1 9 3 4 .
3 5 ) S p l e s , T . D . , P a y n e , W ., a n d C h in n , A .B ., P r o c . S o c . E xp. B i o l , an d Med.
32: 3 28, 1934.
3 6 ) W in tr o b e , M .M ., Am. J . M ed. S c i * , 1 9 7 : 2 8 6 , 1 9 3 9 .
3 7 ) S y d e n s t r i e k e r , V . P . , S c h m id t, H .L . J r . ,
Am. J . M ed. S c i . 1 9 7 : 7 5 5 , 1 9 3 9 .
G - e e s lin , L . E , , a n d W eav er, J . W . ,
3 8 ) H a n s e n - P r u s s , O .C ., N . E n g . J . M ed. 2 1 8 : 1 0 5 0 , 1 9 3 8 .
3 9 )R o s a h n , P . D . , P r o c . S o c . E x p . B i o l , a n d M ed. 2 8 : 4 9 1 , 1 9 3 1 .
50
P a rt I I
T he E f f e c t o f
G a s tr ic J u ic e and L iv e r E x tr a c t
A d m i n is t e r e d t o t h e P r e g n a n t M o th e r ,
o n t h e E r y t h r o c y t e s o f N ew born R a t s
51
!
I n c o m p a r is o n t o t l i e a d u l t b lo o d v a l u e s ,
t h e f e t u s e s a n d n e w b o rn
iiof some a n im a ls h a v e a lo w e r y t h r o c y t e c o u n t , m a c r o c y t o s i s , a n d h i g h c o l o r
!
in d e x .
T h is i s
e s p e c ia lly tr u e o f th e r a t .
T he b lo o d p i c t u r e o f t h e new­
b o r n r a t re s e m b e d t h e h e m a t o lo g i c c o n d i t i o n s f o u n d i n p e r n i c i o u s a n e m ia i n
m an.
If
t h i s c o n d i t i o n i n t h e n e w b o rn r a t c o u ld be c o r r e c t e d by su b ­
s t a n c e s u s e d i n t h e t r e a t m e n t o f p e r n i c i o u s a n e m ia , i t m ig h t s e r v e a s a
c o n v e n ie n t m e th o d f o r t h e a s s a y o f a n t i - p e r n i c i o u s a n e m ia m a t e r i a l .
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R e v ie w o f t h e L i t e r a t u r e
T he p a r e n t e r a l a d m i n i s t r a t i o n o f c o n c e n t r a t e d n o rm a l human a n d
hog g a s tr ic
Mann ( l ,
j u i c e t o t h e p r e g n a n t r a t w as r e p o r t e d b y S t a s n e y , H i g g i n s , a n d
2) t o c a u s e a r e d u c t i o n i n t h e m ax im a l s i z e o f t h e e r y t h r o c y t e s
(of t h e n e w b o rn pu p s..
T he r a t s r e c e i v e d i n t r a p e r i t o n e a l l y d a i l y o n e C c.
Jof n o r m a l, p o o l e d , g a s t r i c j u i c e c o n c e n t r a t e d t o l / l O t h o f t h e o r i g i n a l
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T h e i r r e s u l t s u s i n g n o rm a l h o g g a s t r i c j u i c e w e re s i m i l a r t o t h e
.
above.
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T h i s sam e m e th o d w as u s e d b y S c h l i c k e (3 ) t o i n v e s t i g a t e t h e
je ffe d t o f g a s tr ic
j u i c e o b t a i n e d fro m p a t i e n t s s u f f e r i n g fro m p e r n i c i o u s
i
ja n e m ia , a c h l o r h y d r i a , a n d g a s t r i c c a r c in o m a . One t o t h r e e c c . o f t h e
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[ |v a r i o u s t y p e s o f g a s t r i c j u i c e w e re g i v e n b y s to m a c h t u b e d u r i n g t h e l a s t
tw o w e e k s o f p r e g n a n c y .
The d i a m e t e r s o f f i f t y o f t h e r e d c e l l s w e re
m e a s u r e d fro m p h o t o g r a p h s .
T he g a s t r i c
j u i c e fro m p e r n i c i o u s a n e m ia p a t i e n t s
d i d n o t a f f e c t t h e m ean d i a m e t e r o f t h e b lo o d c e l l s o f t h e nev /b o rn p u p s
w h i l e n o r m a l, n e u t r a l i z e d n o m a l , a c h l o r h y d r i c , a n d g a s t r i c c a rc in o m a
g a s t r i c j u i c e r e d u c e d t h e m ean d i e m t e r a n d c o r p u s c u l a r vo lu m e o f t h e r e d
b lo o d e e l l s o f t h e n e w b o rn p u p s .
When v e n t r i c u l i n w as f e d t o p r e g n a n t r a t s a t a l e v e l o f 2 0 $ o f t h e
d i e t , B r i e s e a n d H i g g i n s (4 ) r e p o r t e d t h a t t h e r e w as a r e d u c t i o n i n t h e
[ L a rg e s t d i a m e t e r o f t h e e r y t h r o c y t e s o f t h e n e w b o rn p u p s i n p r o p o r t i o n
t o t h e l e n g t h o f t i m e t h e m o th e r h a d b e e n o n t h e d i e t b e f o r e d e l i v e r y .
jT here w as a s h i f t i n t h e m e d ia n o f t h e e r y t h r o c y t e d i a m e t e r s t o a s m a l l e r
s iz e .
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D ays o f g e s t a t io n on d ie t
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The a u t h o r s c o n c lu d e d t h a t v e n t r i c u l i n c o n t a i n s some s u b s t a n c e
L h i c h when p r e s e n t i n t h e f e t a l o r g a n is m u s u a l l y a c c e l e r a t e s t h e m a t u r a t i o n
Ii
jjof t h e b lo o d c e l l s i n t h e d e v e l o p i n g f e t u s .
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jito ry t h a n o t h e r s t o t h e a c t i o n o f t h e a n t i a n e m i a s u b s t a n c e .
S
T h e s e c o n c l u s i o n s w e re p a r t i a l l y
s u b s t a n t i a t e d by J o n e s (8 ) .
When
[ p re g n a n t r a t s w e re g iv e n a d i e t c o n t a i n i n g 20$ v e n t r i c u l i n , n e i t h e r t h e
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l e s s v e n t r i c u l i n , b o t h t h e m ean c e l l a n d m ean n u c l e a r d i a m e t e r s o f t h e
p r i m i t i v e e r y t h r o b l a s t s i n e l e v e n d a y o l d r a t em broys w e re d e c r e a s e d .
J o n e s b e l i e v e s t h e l a c k o f e f f e c t a t t h e 2 0 $ l e v e l w as d u e t o i n t e s t i n a l
irrita tio n
o r t o x i c i t y r e s u l t i n g i n i n t e s t i n a l h u r ry and d i a r r h e a .
T h is
l e d t o i m p e r m e a b i l i t y o r t o m a l a b s o r p t i o n o f t h e a n ti a n e m i a s u b s t a n c e .
I f l i v e r e x t r a c t w e re u t i l i z e d i n t h i s m e th o d , i n s t e a d o f g a s t r i c
j u i c e , B r u n e r ( 6 ) r e p o r t e d t h a t t h e b lo o d c e l l v a l u e s o f y o u n g r a t s
w h o se m o th e r s h a d b e e n i n j e c t e d w i t h l i v e r e x t r a c t b e f o r e a n d d u r i n g
p r e g n a n c y w e re fo u n d t o b e n o t s i g n i f i c a n t l y d i f f e r e n t fro m c o n t r o l v a l u e s
a t c o m p a r a b le a g e s a f t e r b i r t h .
B lo o d c e l l v a l u e s o f yo u n g r a t s d i r e c t l y
i n j e c t e d w i t h l i v e r e x t r a c t up t o t h e f o u r t e e n t h day a f t e r b i r t h w e re
fo u n d n o t t o b e s i g n i f i c a n t l y d i f f e r e n t fro m c o n t r o l v a l u e s a t c o m p a ra b le
ages a f t e r b ir th .
U s in g t h e o p p o su m , on t h e o t h e r h a n d , S t a s n e y a n d B u rn s (7 )
j j i n j e c t e d n o rm a l g a s t r i c j u i c e a n d l i v e r e x t r a c t i n t o im m a tu re opposum s f o r
'L h r e e t o f o u r t e e n d a y s w h i l e t h e y w e re i n t h e m a t e r n a l p o u c h .
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J
| c u la r ly w ith a p o te n t l i v e r e x tr a c t .
I n one c a se th e y succeeded in
i n j e c t i n g t h e e x t r a c t i n t o o n e u t e r i n e h o r n ; t h e r e d b lo o d c e l l s o f t h e
f e t u s e s o f t h e i n j e c t e d h o r n d i d n o t d i f f e r s i g n i f i c a n t l y fro m t h o s e o f
th e f e t u s e s o f t h e u n in je c te d h o rn a t te rra .
j
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!
ju s e d i n t h i s e x p e r i m e n t . The p r e g n a n t r a t s w e re p l a c e d i n i n d i v i d u a l
jc a g e s a n d f e d a d i e t o f P u r i n a dog b i s c u i t s an d t a p w a t e r a d l i b .
I
T he b lo o d o f t h e p u p s was c o l l e c t e d a n d s t u d i e d u s u a l l y w i t h i n
[two h o u r s a f t e r b i r t h .
The b lo o d w as o b t a i n e d b y c u t t i n g o f f t h e t a i l .
■ E r y th r o c y te c o u n ts w e re m ade i n t h e u s u a l m a n n e r.
i
H e m o g lo b in
j a i n a t i o n s w e re p e r f o r m e d a c c o r d i n g t o t h e Newcomer m e th o d .
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d e t e r m i n a t i o n s w e re m ade on t h e p o o l e d s a m p le s o f t h r e e p u p s .
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ii
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ii
l i n a n a r e a o f t h e sm e a r w h e re t h e c e l l s w e r e d i s c r e t e a n d n o t t o u c h i n g o n e
Ii
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T he g a s t r i c j u i c e w as o b t a i n e d fro m tw o n o rm a l men f o l l o w i n g
t h e i n j e c t i o n o f o n e mgm. o f h i s t a m i n e .
T he j u i c e was o b t a i n e d f r e s h
e a c h d a y a n d c o n c e n t r a t e d t e n t i m e s i n v a c u o a t 3 7 ° C ., n e u t r a l i z e d
1w i t h 0 . 1 N so d iu m h y d r o x id e a n d I n j e c t e d .
f
! l C c. i n t r a p e r i t o n e a l l y d a i l y .
s to m a c h t u b e r e c e i v e d 3 C c.
and n o t
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T he r a t s r e c e i v i n g t h e g a s t r i c j u i c e b y
o f f r e s h n o rm a l
n e u t r a l i z e d . The g a s t r i c
g a s t r i c j u i c e u n c o n c e n tra te d
ju ic e o b ta in e d
w as t e s t e d
f r e e a c i d a n d a t n o t im e w as f r e e a c i d a b s e n t .
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T he l i v e r e x t r a c t u s e d was R e t i c u l o g e n ( L i l l y )
^ C o n c e n tr a te d ( C h a p p e l 's ) ;
ja n e m ia .
re c e iv e d
each d ay f o r
and L iv e r E x tra c t
b o t h w e re a c t i v e i n t h e t r e a t m e n t o f p e r n i c i o u s
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f
j]
'i
j|
ji n u m b e rs a n d h e m o g lo b in v a l u e s i n t h e n ew b o rn r a t s w hose m o th e r s h a d
ii
i1
j| r e c e i v e d g a s t r i c j u i c e p a r e n t e r a l l y an d by s to m a c h t u b e , t h e i n c r e a s e s
it
j a re n o t p r o p o rtio n a l to th e d u r a tio n o f tre a tm e n t.
I n no i n s t a n c e w as
I t h e r e a s i g n i f i c a n t i n c r e a s e o r d e c r e a s e i n t h e m ean r e d b lo o d c e l l d i a m e t e r s .
|i
fj
T he n e w b o rn r a t s w hose m o th e r s r e c e i v e d l i v e r e x t r a c t d i d n o t h a v e
|| r e d b l o o d c e l l a n d h e m o g lo b in v a l u e s w h ic h v a r i e d w i d e l y fro m t h e c o n t r o l s .
JjThe m ean r e d c e l l d i a m e t e r s w e re n o t s i g n i f i c a n t l y d i f f e r e n t fro m t h e
ji c o n t r o l s i n a n y o f t h e t w e lv e t r e a t e d g r o u p s .
The r a n g e i n r e d b lo o d
|j
i| c e l l d i a m e t e r s i n t h e s e g r o u p s w as r e d u c e d s l i g h t l y fro m t h a t i n t h e
j
Jj c o n t r o l s .
i!
i;
!;
ji
i
;j
D is c u s s io n
I n t h e f o l l o w i n g t a b l e t h e e r y t h r o c y t e v a l u e s o f n e w b o rn r a t s
l a s d e te r m i n e d b y v a r i o u s o b s e r v e r s a r e l i s t e d .
Ii
F rom t h i s t a b l e i t i s e v i d e n t t h a t t h e c o n t r o l v a l u e s e s t a b l i s h e d
!i
[ i n t h i s e x p e r im e n t f a l l w e l l w i t h i n t h e r a n g e o f v a l u e s f o u n d b y o t h e r
ji
ij o b s e r v e r s .
l|il
jj
S t a s n e y a n d H i g g i n s r e p o r t e d o n ly t h e l a r g e s t d i a m e t e r s a n d m ean
ij c o r p u s c u l a r v o lu m e o f t h e r e d b lo o d c e l l s i n t h e n ew born r a t s w hose m o th e r s
I
j jr e c e iv e d n o rm a l human a n d h o g g a s t r i c j u i c e b u t d i d n o t r e p o r t t h e m ean
fd ia m e te rs .
T h e s e d i a m e t e r s w e re m e a s u re d fro m p h o to g r a p h s o f t h e s m e a r s .
[The c e l l s w e re s e l e c t e d a t ran d o m a n d t h e l a r g e s t d i a m e te r o f t h e c e l l
j:
jjwas m e a s u r e d , i . e . ,
ijth an t h e s h o r t .
i f t h e c e l l w as o v a l , t h e l o n g a x i s w as u s e d r a t h e r
T h i s m e th o d a p p e a r s t o b e t o o a r b i t r a r y t o b e u s e d i n
ii
[ 'd e te r m in in g t h e e f f e c t o f a n t i - p e r n i c i o u s a n e m ia m a t e r i a l . H ow ever, m
j;
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61
d e c lin e s s i g n i f i c a n t l y w ith th e d u r a tio n o f tre a tm e n t.
T he sam e m e th o d o f c a l c u l a t i n g t h e l a r g e s t d i a m e t e r w as u s e d by
B r ie s e and H ig g in s .
c o r p u s c u l a r v o lu m e s .
T h ey f a i l e d t o i n c l u d e i n t h e i r t a b l e o f d a t a t h e m ean
I f t h e s e a r e c a l c u l a t e d fro m t h e d a t a t h e y g i v e , t h e
ire s u lts a re :
D ays o f
tre a tm e n t:
M .C .V .
C o n tro ls
7 days
10 d a y s
14 d a y s
21 d a y s
1 4 4 .3
1 3 9 .7
1 4 4 ,3
1 1 5 .7
1 4 1 .4
W ith t h e e x c e p t i o n o f t h e g ro u p t r e a t e d f o r t h e l a s t f o u r t e e n d a y s
|of p r e g n a n c y , t h e m ean c o r p u s c u l a r v o lu m e s a r e e s s e n t i a l l y t h e same and
j!
jjresem ble t h e d a t a w h ic h w e re o b t a i n e d i n o u r e x p e rim e n t*
S i n c e a ch a n g e
jjln m ean c o r p u s c u l a r d i a m e t e r o r m ean c o r p u s c u l a r v o lu m e d o e s n o t i n v o l v e
!l
j(a.s m uch p e r s o n a l e le m e n t , we b e l i e v e them t o b e m ore r e l i a b l e t h a n t h e
!
la r g e s t d ia m e te rs .
U s in g t h i s
a s a b a s i s , we c a n n o t a g r e e w i t h B r i e s e an d
|j
R i g g i n s t h a t v e n t r i c u l i n c o n t a i n s some a n t i - a n e m i c s u b s ta n c e w h i c h p ro m o te s
I
j p a t u r a t i o n o f r e d b lo o d c e l l s i n t h e f e t u s .
||
B e c a u s e o f t h e v a r i o u s r e s u l t s o b t a i n e d by d i f f e r e n t o b s e r v e r s
ipn t h e e f f e c t o f t h e a d m i n i s t r a t i o n o f g a s t r i c
j u i c e and l i v e r e x t r a c t t o
lihe p r e g n a n t m o th e r on t h e r e d b lo o d c e l l s o f n e w b o rn r a t s ,
j i e n t s u p o n w h ic h t h e d a t a a r e b a s e d ,
and th e m ea su re-
we b e l i e v e t h i s m eth o d i s n o t r e l i a b l e
(jfor t h e d e t e r m i n a t i o n o f a n t i - p e r n i c i o u s a n e m ia s u b s t a n c e s .
I!
l
K
C o n c l u s io n s
. A d m i n i s t r a t i o n o f n o rm a l hum an g a s t r i c j u i c e t o p r e g n a n t r a t s b y i n t r a jj p e r i t o n e a l i n j e c t i o n o f c o n c e n t r a t e d j u i c e a n d by s to m a c h t u b e o f
j li n c o n c e n t r a t e d j u i c e i s w i t h o u t e f f e c t on t h e s i z e o f t h e r e d b lo o d c e l l s o f
jl
iip d w b o rn r a t s .
j!2 .
A d m in is tra tio n o f l i v e r e x tr a c t , p o te n t i n th e tre a tm e n t o f p e rn ic io u s
j
j
a n e m ia , t o p r e g n a n t r a t s i s w i t h o u t e f f e c t u p o n t h e s i z e o f t h e r e d
!
b l o o d c e l l s o f t h e n e w b o rn r a t s .
'i
|0 .T h e m e th o d o f a d m i n i s t e r i n g s u b s t a n c e s known t o b e p o t e n t i n t h e t r e a t j
I m e n t o f p e r n i c i o u s a n e m ia o r c o n t a i n i n g a n t i - p e r n i c i o u s a n e m ia f a c t o r s
!
t o p r e g n a n t r a t s a n d d e t e r m i n i n g t h e d i m i n u ti o n i n s i z e o f t h e r e d b lo o d
j
c e l l s o f t h e n e w b o rn r a t s i s u n r e l i a b l e .
j!
B ib lio g ra p h y
ii
j!
j |( l ) S t a s n e y , J . , H i g g i n s , G .M ., P r o c . S t a f f M e e tin g s o f Mayo C l i n i c 1 2 :
, 490, 1937.
I
J(2 ) ^ t a s a e y , J”. , H i g g i n s , G .M ., a n d M ann, E . C . , P r o c S t a f f M e e tin g s Mayo
|
C lin ic 12: 399, 1937.
( 3 ) S c h l i c k e , C . P . , P r o c . S t a f f M e e tin g s Mayo C l i n i c 1 4 : 1 4 5 , 1 9 3 9 .
^ 4 ) B r i e s e , E . , a n d H i g g i n s , G .M ., A n a t. R e c . 73: 1 0 5 , 1 9 3 9 .
;(5} J o n e s , O . P . , A n a t . R e c . 7 6 : P 3 4 , 1 9 4 0 ,
j ( 6 ) B r u n e r , H .D ., P r o c . S o c . E x p e r . B i o l , a n d Med* 4 1 : 2 6 0 , 1 9 3 9 .
j
(7 )S ta s n e y , J . ,
544, 1939.
j |( 8 ) J o l l y , J . ,
|
I
J o lly ,J .,
a n d B u r n s , E . L . , P r o c . Soc . E x p e r . B i o l , a n d M e d ., 4 2 :
Comp. R e n d . S o c . de B i o l . 6 0 : 5 6 4 , 1 9 0 6 .
I b id .,
62: 1 3 6 , 1909.
( 9 ) J o l l y , J . , Comp. R e n d . S o c . de B i o l . 8 2 : 8 0 0 , 1 9 1 9 .
!(1 0 )C h is h o lm , R .A ., Q u a r t . J . E xp. P h y s i o l . 4 : 2 0 7 , 1 9 1 1 .
ii
jl( 1 1 ) ^ u r e , B ., K ik , M .C ., a n d W a lk e r, D . J . , J . N u t r . 1 : 2 9 9 , 1 9 2 9 .
ji
l( 1 2 ) K in d r e d , J . E . , a n d C o r e y , E . L . , A n a t. R e c . 4 7 : 2 1 3 , 1 9 3 0 .
jl{13)Q nith C . , J . P a t h , a n d B a c t . 3 5 : 7 1 7 , 1 9 3 2 .
i,
! ! ( l4 ) W ills , L . , a n d M e h ta , M .M ., I n d . J . M ed. R e s . ±Q: 3 0 7 , 1 9 3 0 .
!!
! j( l5 ) B r u n e r, H .D ., V an de Ewe, J . , an d C a r l s o n , A . J . , Am. J . P h y s i o l . 1 2 4 :
jj
620, 1938.
|i(1 6 )B ru n e r, H .D ., P r o c . S o c . E x p e r . B i o l , a n d Med. 4 1 : 2 6 0 , 1 9 3 9 .
!j( 1 7 ) M in t r o b e , M .M ., K i n s e y , R . E . , B l o u n t, R .E . a n d T r a g e r , W ., Am. J .
jj
M ed. S c i . , 1 9 3 : 4 4 9 , 1 9 3 7 .
64
P a rt I I I
T he P r e s e n c e o f t h e A n t i p e r n i e i o u s Anemia.
F a c to r i n an E x tra c t o f F e ta l
B o v in e L i v e r s
T he f a i l u r e o f l i v e r e x t r a c t i n j e c t e d i n t o t h e p r e g n a n t r a t t o
a f f e c t t h e m ean c o r p u s c u l a r d i a m e t e r o f t h e r e d b lo o d c e l l s o f t h e n e w b o rn
r a t g a v e r i s e t o t h e h y p o t h e s i s t h a t t h e f e t a l l i v e r m ig h t a l r e a d y c o n ­
t a i n t h e a n t i - p e r n i c i o u s a n e m ia s u b s t a n c e .
I n o r d e r t o d e te r m in e t h e
a n s w e r t o t h i s q u e s t i o n , an e x t r a c t o f f e t a l c a l v e s l i v e r s was p r e p a r e d .
W h e th e r t h i s a n t i - p e r n i c i o u s a n e m ia s u b s t a n c e , i f p r e s e n t i n t h e f e t a l
liv e r,
i s m a n u f a c tu r e d i n t h e f e t u s i t s e l f ,
o r p a s s e s fro m t h e m a t e r n a l
c i r c u l a t i o n to th e f e t a l c i r c u l a t i o n th ro u g h th e p la c e n ta i s a m a tte r o f
c o n je c tu re .
I n c o r r e la tin g th e re sp o n se to l i v e r th e ra p y w ith a c h y lia g a s tr ic a
i n p e r n i c i o u s a n e m ia p a t i e n t s , C a s t l e a n d h i s c o w o rk e rs ( 5 - 6 ) d e m o n s tr a te d
t h a t t h e g a s t r i c j u i c e o f p e r n i c i o u s a n e m ia p a t i e n t s i s r e l a t i v e l y d e f i ­
c i e n t i n some f a c t o r w h ic h i s p r e s e n t i n n o rm a l g a s t r i c
ju ic e .
On t h e
b a s i s o f t h e s e f i n d i n g s , C a s t l e p r o p o s e d h i s t h e o r y c o n c e r n in g t h e m echan­
ism o f t h e p r o d u c t i o n o f p e r n i c i o u s a n e m ia .
T h is t h e o r y h a s b e e n a m p ly
c o n f ir m e d ( 9 - 1 1 , 1 5 - 1 5 , 1 7 , 2.0, 2 2 ) . C a s t l e s t a t e d t h a t t h e i n t e r a c t i o n o f
some s u b s t a n c e i n t h e g a s t r i c j u i c e
p rin c ip le
(e x trin s ic
( in tr in s ic fa c to r)
f a c t o r ) p r o d u c e s a s u b s ta n c e w h ic h i s a b s o r b e d a n d
s to re d in th e l i v e r (1 6 ).
A c c o r d in g t o t h i s t h e o r y , a d e f i c i e n c y o f t h e
l i v e r f a c t o r may r e s u l t i n s e v e r a l w ay s:
in th e d i e t ;
an d some d i e t a r y
(b )la c k o f th e i n t r i n s i c
(a )la c k o f th e e x tr in s ic f a c to r
f a c to r in th e g a s tr ic
ju ic e ;
(c )fa ilu re
o f a b s o r p t i o n o f t h e e n d p r o d u c t o f t h e i n t e r a c t i o n o f t h e a b o v e tw o
fa c to rs .
To t h e s e m ig h t b e a d d e d tw o m o re m eans by i^ h ic h a m a c r o c y t i c
a n em ia m ig h t b e p r o d u c e d , s.s s u g g e s t e d b y G oldham er( 7 ) ;
L iv e r to
( d) i n a b i l i t y o f t h e
s t o r e t h e a n t i p e m i c i o u s a n e m ia s u b s t a n c e ( 2 3 a ) ; a n d ( e ) i n a b i l i t y
Df t h e b o n e m arrow t o u t i l i z e t h e a n t i p e r n i c i o u s a n e m ia f a c t o r ( 1 2 ) .
66
A c c o r d in g t o C a s t l e 's t h e o r y , t h e l i v e r f a c t o r c a n b e p r o d u c e d
o n ly by th e i n t e r a c t i o n o f th e i n t r i n s i c and e x t r i n s i c f a c t o r s .
fe tu s is
The
e n t i r e l y l a c k i n g i n t h e e x t r i n s i c f a c t o r e x c e p t f o r t h e a m n io tic
f l u i d a n d t h e s m a ll am ount o f d e t r i t u s w h ic h c o l l e c t s i n t h e g a s t r o - i n t e s tin a l tr a c t.
I t i s u n l ik e ly t h a t e i t h e r o f th e s e s e rv e a s s o u r c e s o f th e
e x trin s ic f a c to r .
C o n s e q u e n tly , t h e f e t u s e i t h e r m u st n o t r e q u i r e t h e
l i v e r f a c t o r o r r e c e i v e s i t fro m t h e m o th e r t h r o u g h t h e p l a c e n t a .
If
C a s t l e 's t h e o r y d o e s n o t a p p l y t o t h e f e t u s , t h e a n t i p e r n i c i o u s a n e m ia
s u b s t a n c e m ay b e fo rm e d he n o v o b y t h e l i v e r o r some o t h e r o r g a n .
T he p u r p o s e o f t h e p r e s e n t i n v e s t i g a t i o n i s t o d e te r m in e w h e t h e r
t h e f e t a l l i v e r c o n t a i n s t h e a n t i p e r n i c i o u s a n e m ia f a c t o r .
I t s p resen ce
i n s i g n i f i c a n t a m o u n ts w ould s u g g e s t t h a t t h e f e t u s r e q u i r e s t h e f a c t o r
an d f u r t h e r m o r e w o u ld i n d i c a t e
(a s s u m in g C a s t l e ' s t h e o ^ r t o b e t r u e ) t h a t
t h e m o th e r m u s t p r o d u c e e x t r a q u a n t i t i e s o f t h e f a c t o r t o p r o v i d e b o t h
h e r s e l f and t h e f e t u s .
T h is p o s s i b i l i t y w o u ld h a v e c o n s i d e r a b l e s i g n i f i c a n e
i n v ie w o f t h e f a c t t h a t a m a c r o c y t i c a n em ia so m e tim e s a c c o m p a n ie s p r e g ­
nancy.
M e th o d s
An e x t r a c t o f f e t a l b o v i n e l i v e r s w as p r e p a r e d by a m eth o d known
jto y i e l d a p o t e n t e x t r a c t when a p p l i e d t o l i v e r s o f a d u l t a n i m a l s .
E ach
ijcc. o f t h e p r e p a r e d e x t r a c t w as e q u i v a l e n t t o 100 gm. o f t h e f r e s h l i v e r s .
T h e e x t r a c t c o n t a i n e d 7 * 74$> t o t a l s o l i d s .
j
f
T h re e p e r n i c i o u s a n e m ia p a t i e n t s , o n e i n s e v e r e r e l a p s e , w e re
j t r e a t e d w i t h t h e f e t a l l i v e r e x t r a c t b y o n e o f u s ( O .R .) a t t h e Cook C o u n ty
lio s p ita l.
R ed a n d w h i t e b lo o d c e l l c o u n t s , r e t i c u l o c y t e c o u n ts , a n d hemo­
g l o b i n d e t e r m i n a t i o n s "were m ade i n t h e u s u a l m a n n e r.
R e s u lts
C a s e 1 . — M .B r .( N o . 2 1 9 9 ) , a w h i t e f e m a le , a g e d . 5 3 , w as a d m i t t e d
to th e h o s p ita l, J a n u a ry 14, 1938.
T h is woman h a d b e e n d i s c h a r g e d s i x
m o n th s b e f o r e , a f t e r b e i n g t r e a t e d f o r p e r n i c i o u s a n e m ia .
re c e iv e d l i v e r s in c e h e r d is c h a rg e .
She h a d n o t
On a d m i s s i o n , h e r b lo o d p r e s s u r e was
9 2 /6 4 , te m p e ra tu re 1 0 0 ° F . , p u ls e 1 0 0 .
B lo o d e x a m in a tio n r e v e a l e d
1 ,9 6 0 ,0 0 0 r e d b l o o d c e l l s p e r c .m m ., p o i k i l o c y t o s i s , a n i s o c y t o s i s , 4 6 $
h e m o g lo b in ( S a h l i ) ,
a n d 2400 w h i t e b lo o d c e l l s p e r c . mm.
was p e r n i c i o u s a n e m ia i n s e v e r e r e l a p s e .
T he d i a g n o s i s
T r e a tm e n t c o n s i s t e d o f i n t r a ­
m u s c u la r i n j e c t i o n s o f 0 . 5 c c . o f t h e e x p e r i m e n t a l e x t r a c t f o r s e v e n t e e n
d a y s, 0 .5 c c . a g a in on th e n in e te e n th d ay .
On t h e t w e n t i e t h d a y 2 e e .
w e re a d m i n i s t e r e d a n d 2 c c . w e e k ly t h e r e a f t e r f o r two w e e k s .
d o s e a g e w as 15 c c .
The t o t a l
A t t h e e n d o f t h i r t y f o u r d a y s o f t r e a t m e n t , t h e b lo o d
p i c t u r e w as 4 ,2 7 0 ,0 0 0 r e d b lo o d c e l l s p e r c . mm., 1 1 ,0 0 0 w h i t e b lo o d c e l l s
jp e r c . m m ., w i t h 7 9 $ h e m o g lo b in .
T h e re w as m a rk e d s u b j e c t i v e im p ro v e m e n t,
p h e w as d i s c h a r g e d , F e b r u a r y 1 , 1 9 3 8 , 17 d a y s a f t e r a d m i t t a n c e .
I
C a se 2— M .B. (N o. 2 5 7 0 ) , a c o l o r e d f e m a l e , a g e d 6 1 , w as a d m i t t e d
o th e h o s p ita l, Ja n u ary 16, 1938.
On a d m i s s io n , h e r b lo o d p r e s s u r e was
|l 3 0 / 5 4 , t e m p e r a t u r e 1 0 0 .6 ° F . , p u l s e 1 0 4 .
S he c o m p la in e d o f d i a r r h e a ,
p r o j e c t i l e v o m i t i n g , b lo o d i n s t o o l s , an d p a r e s t h e s i a s o f t h e h a n d s .
P h y s ic a l e x a m in a tio n r e v e a le d a p a l e ,
sm o o th , s o r e t o n g u e ; t h e l e f t
b a t e l l a r , a b d o m in a l a n d e p i g a s t r i c r e f l e x e s w e re a b s e n t .
An e x a m i n a t i o n o f
th e g a s t r i c c o n t e n t s f o l l o w i n g a n E w ald t e s t m e a l r e v e a l e d no f r e e a c i d .
T he b l o o d e x a m i n a t i o n r e v e a l e d 1 ,1 1 0 ,0 0 0 r e d b lo o d c e l l s a n d 3800 w h ite
b lo o d c e l l s p e r c . m m., h e m o g lo b in 2 5 $ ( S a h l i ) .
T h e re was m a rk e d a n i s o c y t o s i
68
jand p o i k i l o c y t o s i s .
T he d i a g n o s i s w as p e r n i c i o u s a n e m ia .
T r e a tm e n t
( C o n s is t e d o f i n t r a m u s c u l a r i n j e c t i o n s o f 1 .5 c c . o f t h e e x p e r i m e n t a l
e x t r a c t d a i l y f o r n in e d a y s, th e n e v e ry t h i r d day f o r f iv e i n j e c t i o n s .
At
t h e e n d o f t h i s t im e t h e p a t i e n t r e c e i v e d 5 .6 c c . o f a p o t e n t h o r s e l i v e r
e x tra c t.
The t o t a l d o s a g e w as 21 c c . o f t h e e x p e r i m e n t a l e x t r a c t .
The
p a t i e n t im p ro v e d m a r k e d ly u n d e r t r e a t m e n t ; t h e p a r e s t h e s i a s d i s a p p e a r e d
e n tire ly .
A t t h e t i m e o f t h e c h a n g e fro m f e t a l l i v e r t o t h e h o r s e l i v e r
c o n c e n t r a t e , a b l o o d e x a m in a tio n r e v e a l e d 4 , 0 9 0 ,0 0 0 r e d b lo o d c e l l s a n d
^400 w h i t e b lo o d c e l l s p e r c . m m ., a n d 65$ h e m o g lo b in .
C a s e 3— B . J .
(N o. 6591 ), a w h i t e f e m a le , a g e d 6 8 , w as a d m i t t e d
;o t h e h o s p i t a l , F e b r u a r y 6 , 19 3 8 .
l
On a d m is s io n , h e r b lo o d p r e s s u r e w as
30 / 7 0 , t e m p e r a t u r e 9 9 . 5 ° P . , a n d p u l s e r a t e 9 6 .
l i a r r h e a an d s w e llin g o f b o th f e e t .
S he c o m p la in e d o f
P h y s i c a l e x a m i n a t i o n r e v e a l e d an em ac-
. a t e d o l d woman, le m o n y e l l o w i n c o l o r w i t h a p a l e a t r o p h i c to n g u e , a n d a
.a r g e u m b i l i c a l h e r n i a .
T he b lo o d e x a m in a tio n r e v e a l e d 1 , 3 6 0 ,0 0 0 r e d b lo o d
j e l l s a n d 5600 w h i t e b l o o d c e l l s p e r c . mm., a n d 34 $ h e m o g lo b in ( S a h l i ) .
'•h e d i a g n o s i s w as p e r n i c i o u s a n e m ia ,
ea'tmeii'b c o n s i s t e d o f i n t r a m u s c u l a r
i n j e c t i o n s o f th e e x p e rim e n ta l e x t r a c t , 1 .5 c c . d a i l y f o r t h i r t e e n d a y s ,
a n d 1 . 5 c c . on t h e s i x t e e n t h d a y .
The e x p e r i m e n t a l e x t r a c t h a v in g b e e n
e x h a u s te d , a p o t e n t h o r s e l i v e r c o n c e n t r a t e w as g i v e n on t h e e i g h t e e n t h
Lay o f t r e a t m e n t .
A t t h e tim e o f t h e i n j e c t i o n o f t h e l a s t d o se o f t h e
e x p e rim e n ta l e x t r a c t ,
b l o o d e x a m i n a t i o n r e v e a l e d 2 ,7 7 0 ,0 0 0 r e d b lo o d c e l l s
lipid 450 0 w h i t e b l o o d c e l l s p e r c . m m ., a n d 4 9 $ h e m o g lo b in .
T he r e s p o n s e o f a l l t h r e e p a t i e n t s i s t y p i c a l o f t h e r e s p o n s e
f o llo w in g th e i n j e c t i o n o f a l i v e r e x t r a c t c o n ta in in g th e a n ti p e r n i c io u s
a n e m ia f a c t o r .
D is c u s s io n
T h e p ro m p t r e t i c u l o c y t e r e s p o n s e s t o g e t h e r w i t h t h e r e t u r n t o
n o rm a l b l o o d p i c t u r e s g i v e u n d i s p u t a b l e p r o o f o f t h e p r e s e n c e o f t h e a n t i p e r n i c i o u s a n e m ia f a c t o r i n t h e f e t a l b o v i n e l i v e r s .
I f C a s t l e Ts t h e o r y
a s to th e n e c e s s i t y o f b o th t h e e x t r i n s i c and i n t r i n s i c f a c t o r s f o r th e
f o r m a t i o n o f t h e a n t i p e r n i c i o u s a n e m ia s u b s t a n c e i s a s s u m e d t o b e t r u e ,
t h e n we c a n m o s t r e a s o n a b l y a c c o u n t f o r t h e p r e s e n c e o f t h i s
s u b s ta n c e i n
th e f e t a l l i v e r o n ly a s th e r e s u l t o f i t s p a s s a g e th ro u g h th e p la c e n ta ,
s i n c e t h e f e t u s i s n o t s u p p l i e d w ith t h e e x t r i n s i c f a c t o r .
A c a r e f u l s e a r c h o f th e l i t e r a t u r e r e v e a le d o n ly f o u r p r e v io u s
R e p o rts c o n c e rn in g th e u s e o f f e t a l l i v e r i n th e tre a tm e n t o f p e r n ic io u s
janem ia.
B e r g lu n d , W a tk in s a n d J o h n s o n {£) r e p o r t e d t h a t f e t a l b o v in e
( l i v e r , f e d a s t h e d e s i c c a t e d m a t e r i a l e q u i v a l e n t t o 300 gm. o f f r e s h
i v e r d a i l y , w as a c t i v e i n t h e t r e a t m e n t o f o n e c a s e o f p e r n i c i o u s a n e m ia ,
b w e v e r , t h e r e t i c u l o c y t e c o u n t , w h ic h i s i n p a r t a m e a s u re o f t h e b o n e
jnarrow r e s p o n s e t o t h e a n t i p e r n i c i o u s a n e m ia f a c t o r , d i d n o t r i s e a b o v e
G o ld h a m e r, I s a a c s a n d S t u r g i s ( 8 ) p r e p a r e d a n e x t r a c t fro m t h e
l i v e r o f a s e v e n m o n th hum an f e t u s .
T w en ty c c . o f t h i s e x t r a c t , r e p r e -
e n t i n g 65 gm. o f t h e f r e s h t i s s u e , w e re a d m i n i s t e r e d i n a s i n g l e d o s e .
'he r e t i c u l o c y t e r e s p o n s e r e a c h e d a maximum, o f 1 7 .6 $ o n t h e s e v e n t h d a y ,
1 th o u g h t h e h e m o g lo b in d e c l i n e d a n d t h e r e d b lo o d c e l l c o u n t re m a in e d
a lte re d .
On t h e b a s i s o f t h i s r e s u l t t h e a u t h o r s c o n c lu d e d :
"T he a c t i v e
p r in c i p le n e c e s s a ry f o r th e m a tu r a tio n o f re d c e l l s i s p r e s e n t in th e
i v e r a t l e a s t tw o m o n th s b e f o r e b i r t h . "
is
B ecause th e r e t i c u l o c y te count
i n i t s e l f no m e a s u r e o f t h e p o t e n c y o f a l i v e r e x t r a c t ,
th e s e r e s u l t s
ja re n o t c o n c l u s i v e .
I t i s w e l l know n t h a t t h e i n j e c t i o n o f i n a c t i v e p r o t e i n
s u b s ta n c e s and c e r t a i n d r u g s w i l l c a u se a r e t i c u l o c y t e , r e s p o n s e .
Spon­
t a n e o u s r i s e s i n t h e n um ber o f r e t i c u l o c y t e s a s h i g h a s t h a t r e p o r t e d by
G o ld h a m e r e t a l . h a v e b e e n s e e n i n o u r c l i n i c .
T he same c r i t i c i s m
a p p lie s
to th e one p a t i e n t o b se rv e d by B e n c e (l).
W in tr o b e , K i n s e y , B lo u n t a n d T r a g e r (£ 4 ) p r e p a r e d e x t r a c t s o f
f e t a l p ig l i v e r s i n th e age g ro u p s o f second t h i r d ,
s i x t h , and l a s t s i x t h o f p re g n a n c y .
th ird th ird , f i f t h
A l l o f t h e s e e x t r a c t s w e re w i t h o u t
e f f e c t i n d o s e s o f 1 t o 10 c c . w hen a d m i n i s t e r e d t o p e r n i c i o u s a n e m ia
p a tie n ts .
je x tr & c t.
T h e s e p a t i e n t s r e s p o n d e d s u b s e q u e n t l y t o a c o m m e rc ia l l i v e r
We c a n n o t a d e q u a t e l y a c c o u n t f o r t h i s a p p a r e n t d i f f e r e n c e i n
t h e l i v e r f a c t o r c o n t e n t o f t h e l i v e r s o f f e t a l c a l v e s a s co m p ared w i t h f e t a l
p ig s .
The m a rk e d d i f f e r e n c e s i n p l a c e n t a l s t r u c t u r e i n t h e cow a n d p i g
may a c c o u n t f o r a d i f f e r e n c e i n p e r m e a b i l i t y o f t h e p l a c e n t a s t o t h e l i v e r
fa c to r.
The a b s e n c e o f t h e l i v e r f a c t o r fro m t h e l i v e r s o f f e t a l p i g s
Ip u g g e s ts t h a t t h e f e t u s i s u n a b l e t o fo rm t h i s f a c t o r de n o v o .
I t a ls o
s u g g e s t s t h a t t h e l i v e r f a c t o r w h ic h i s e f f e c t i v e i n t h e t r e a t m e n t o f
p e r n i c i o u s a n e m ia i s n o t r e q u i r e d b y t h e f e t u s f o r t h e m a t u r a t i o n o f r e d
D lood c e l l s .
T h is l a t t e r s u g g e s t i o n i s
f u r t h e r s u b s ta n t ia te d by th e r e s u l t s o f
W igodsky an d I v y ( 2 1 ) w i t h t h e i n j e c t i o n o f a p o t e n t l i v e r e x t r a c t i n t o
[p re g n a n t a l b i n o r a t s i n t h e a t t e m p t t o r e d u c e t h e r e d b lo o d c e l l s i z e o f
ibhe n e w b o rn .
The i n j e c t i o n o f a l i v e r e x t r a c t o f known p o te n c y i n t o
■ pregnant r a t s a s m any a s f o u r t e e n d a y s b e f o r e d e l i v e r y e x e r t e d no s i g -
f
j i i f i c a n t e f f e c t u p o n t h e r e d b l o o d c e l l s o f t h e n e w b o rn .
lin t h e r a b b i t h a v e b e e n r e p o r t e d b y W in tro b e e t a l .
S im ila r r e s u l t s
( 2 3 b ) . ^ t a s n e y et_ a l .
I ( 1 8 ,1 9 ) h a v e d e m o n s t r a t e d t h a t a c o n c e n t r a t e o f n o rm a l hum an o r s w in e
jg a s tric
j u i c e , w hen i n j e c t e d i n t r p e r i t o n e a l l y i n t o p r e g n a n t r a t s ,
red u c e d
t h e r e d b lo o d c e l l d i a m e t e r s o f t h e n e w b o rn p u p s*
A ssu m in g t h a t t h e a n t i p e r n i c i o u s a n e m ia f a c t o r w i l l p a s s fro m t h e
c i r c u l a t i o n o f t h e m o th e r t o t h a t o f t h e f e t u s a n d w i l l b e s t o r e d i n t h e
f e ta l liv e r, i t
i s p l a u s i b l e , a s s u g g e s t e d b y W in tr o b e a n d S h u m a ck e r (2 5 b )
t h a t t h e m o th e r * s s to r f e o f t h e l i v e r f a c t o r may b e so d e p l e t e d b y f e t a l
s t o r a g e o f t h e f a c t o r a s t o c a u s e a m a c r o c y t i c a n e m ia d u r i n g p r e g n a n c y .
Summary
l . T h e p r e s e n c e o f t h e a n t i - p e r n i c i o u s a n e m ia p r i c n i p l e i n f e t a l b o v in e
l i v e r s i s d e m o n s t r a t e d b y t h e f a c t t h a t e x t r a c t s o b t a i n e d fro m t h i s
s o u r c e p r o d u c e d t y p i c a l , p r o m p t, an d d e f i n i t e r e m i s s i o n s i n t h r e e
p e r n i c i o u s a n e m ia p a t i e n t s ,
j s . S in c e
r
|
t h e f e t u s i s c o m p l e te l y d e f i c i e n t i n t h e e x t r i n s i c f a c t o r o f
C a s tle ,
t h e a n t i - p e r n i c i o u s a n e m ia f a c t o r p r e s u m a b ly r e a c h e d t h e f e t a l
l i v e r b y p a s s i n g fro m t h e c i r c u l a t i o n o f t h e m o th e r t h r o u g h t h e p l a c e n t a
in to th e f e t a l c ir c u la tio n .
L T h e s e f i n d i n g s s u p p o r t t h e v ie w t h a t w i t h d r a w l o f t h e a n t i p e r n i c i o u s
n e m ia p r i n c i p l e b y t h e f e t u s may b e a n e t i o l o g i c f a c t o r i n t h e p r o d u c t i o n
S
[
f a m a c r o c y t i c a n e m ia o f p r e g n a n c y .
.E v id e n c e i s
d is c u s s e d r e l a t i v e to th e n e c e s s ity o f th e a n tip e r n ic io u s
n e m ia p r i n c i p l e f o r n o rm a l e r y t h r o p o i e s i s i n t h e f e t u s .
The p o s s i b i l i t y
I s c o n s i d e r e d t h a t t h e a n t i p e r n i c i o u s a n e m ia p r i n c i p l e i s n o t n e c e s s a r y
jpor n o rm a l e r y t h r o p o i e s i s i n t h e f e t u s .
72
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B ib lio g ra p h y
(1 )B e n c e , J . ,
Z t s c h r . P . K l i n . M e d ., 1 2 6 ; 1 2 7 , 1 9 3 3 .
( 2 ) B e r g l u n d , H . , W a tk in s , C .H ., a n d J o h n s o n , R . , P r o c . S o c . E x p . B i o l ,
a n d M e d ., 2 5 : 8 3 4 , 1 9 2 7 .
( 3 ) C a s t l e , W .B ., Am. J . M ed. S c i . , 1 7 8 : 7 4 8 , 1 9 2 9 .
( 4 ) C a s t l e , W .B ., a n d T o w n sen d ,
W .C ., I b i d . , 1 8 2 : 7 4 1 , 1 9 3 1 .
( 5 ) C a s t l e , W .B ., H e a th , C .W ., a n d S t r a u s s , M .B ., I b i d . ,
( 6 ) C a s t l e , W .B ., T ow nsend, W .C ., a n d H e a th , C .W ., I b i d . ,
182:
7 4 1 , .1931.
1 8 0 : 305, 1930.
( 7 )G o ld h a m e r , S .M ., C y c lo p e d ia o f M e d ic in e (R e v . V o l . ) , P h i l a d e l p h i a ,
P .A . D a v is & C o ., P . 1 3 7 , 1 9 3 6 .
(8 )G o ld h a m e s S .M ., I s a a c s , P . ,
188: 193, 1934.
a n d S t u r g i s , C .C - , Am. J . M e d ., S c o .,
(9 ) G r e e n J * , K l i n i s c h en e x p e r i m e n t e e l o n d e r z o e k o v e r a n e m ia p e r n i c i e s a
e n v o o r w a a r d e l i j k e d e f i e i e n t e , A m sterdam , S e h e lte m a a n d K o lk e ra a ’ s
B o e k h a n d e l, 1 9 3 5 .
( 1 0 ) H a r t f a l l , S . J . , and W itts ,
L . J . , Guy’ s H o s p . R e p ., 8 3 : 2 4 , 1 9 3 3 .
(1 1 )H e lm e r, O .M ., P o u t s , P . J . ,
184, 1934.
a n d Z e r f a s , L . C . , Am. J . M ed.
S c i., 188:
( 1 2 ) I s r a e l s , M .C .G ., a n d W il k in s o n , F * J - , Q u a r t . J . M e d ., _5: 6 9 , 1 9 3 6 .
( 1 3 ) M id d le to n , W .S ., a n d S tie h m , R .H ., Am. J . M ed. S c i . ,
180: 80 9 , 1930.
|1 4 ) M i l l e r , ^ . K . , a n d R h o a d s , C . p . , New E n g . J . M e d ., 2 1 1 : 9 2 1 , 1 9 3 4 .
'1 5 )R e im a n n , P . , K l i n W c h n sc h r* , 1 3 ; 4 1 3 , 1 9 3 4 .
|1 6 ) R i c h e t e r , 0 . , Iv y , , A .C ., a n d K im , M .S ., P r o c . S o c . E x p . B i o l , a n d Med.
29: 1 0 9 3 , 1932.
; i 7 ) S i n g e r , K . , W ien. k l i n . W c h n s c h r ., 4 5 : 1 0 6 3 , 1 9 3 2 .
;i8 ) bta s n e y , J . ,
490, 1937.
a n d H i g g i n s , G .M ., S t a f f M e e tin g Mayo C l i n i c , 1 2 :
1 9 ) S t a s n e y , J . , H i g g i n s , G .M ., a n d M ann, P . O ., I b i d . , P . 6 9 9 .
2 0 ) U n g le y , C .C ., a n d J a m e s , G .V ., Q u a r t. J . M ed. 27_: 5 23, 1 9 3 4 .
2 1 )W ig o d s k y , H . S . , a n d I v y , A .G -, P r o c . S o c . E x p . B i o l , a n d M e d ., 38:
787, 1938.
( 2 2 ) W il k in s o n , E . J . , a n d K l e i n , L . , L a n c e t , 1 : 7 1 9 , 1 9 3 2 .
( 2 3 ) W in t r o b e , M .M ., a n d S h u m a c k e r, H . S . ; ( a ) B u l l . J o h n s H o p k in s I io s p .
52: 3 8 7 , 1933. (b ) J . C lin . I n v e s t . , 14: 837, 1 9 3 5 .
( 2 4 ) W in tr o b e , M .M ., K i n s e y , R . E . , B l o u n t , R .E ., a n d T r a g e r , W ., Am. J
M ed. S c i . , 1 9 3 : 4 4 9 , 1 9 3 7 .
75
P a r t IV
T he E f f e c t o f I n d o l e o n t h e Red B lo o d C e l l s
o f N o rm al a n d G a s t r e c to m iz e d D ogs
76
T he u s e o f i n d o l e t o p r o d u c e a n a n e m ia i n d o g s w h ic h w as a f f e c t e d
by l i v e r e x tr a c t (v id e i n f r a
) p ro m p te d t h i s e x p e r i m e n t .
T he p u r p o s e
o f t h e e x p e r i m e n t w as t o d e t e r m i n e t h e e f f e c t o f i n d o l e on t h e r e d b lo o d
c e l l s o f n o r m a l a n d g a s t r e c t o m i z e d d o g s i n o r d e r t o d e te r m i n e i f t h e
r e s u l t a n t a n e m ia c o u ld b e u s e d f o r t h e a s s a y i n g o f a n t i - p e r n i c i o u s a n e m ia
s u b s ta n c e s .
R e v ie w o f t h e L i t e r a t u r e
The e a r l y l i t e r a t u r e
c o n c e r n in g i n d o l e , i n d i g o , e t c . fro m t h e tim e
o f H i p p o c r a t e s w as c o m p ile d by M a r t i n (1 ) i n 1 8 4 6 .
The l i t e r a t u r e fro m
1825 t o 1 8 9 5 w as r e v ie w e d by Sim on ( 2 ) .
T h at a b lu e s u b s ta n c e o c c u rre d o c c a s io n a lly i n u r in e had b een
m ow n f o r c e n t u r i e s , b u t t h e i d e n t i f i c a t i o n o f t h i s s u b s t a n c e a s i n d i g o
aras f i r s t m ade b y P r o u t i n 1840 ( 3 ) .
T hat th e p r e c u rs o r o f in d ig o , i . e .
in d o le , i s a n o rm a l p ro d u c t o f i n t e s t i n a l d i g e s t io n a s a r e s u l t o f b a c ­
t e r i a l a c t i v i t y w as s t a t e d b y N e n c k i i n 187 4 ( 4 ) .
Sim on s t a t e s t h a t
Baumann b y d i s i n f e c t i n g t h e g a s t r o - i n t e s t i n a l t r a c t o f a dog w i t h c a lo m e l
cau sed a l l t r a c e s o f in d o le to d is a p p e a r .
th a t th e
I t w as S im on1s b e l i e v e ( 2 )
f r e e h y d ro c h lo ric a c id o f g a s tr ic ju ic e i s th e a c tiv e a g e n t i n
c o n tr o llin g th e d e g re e o f i n t e s t i n a l p u tr e f a c tio n .
"T h e i n d i c a n e l i m i -
a a t i o n may h e n c e b e r e g a r d e d a s r u n n in g a c o u r s e p a r a l l e l t o t h e
ie g re e o f a n t i s e p t i c — v iz . g e rm ic id a l— a c t i v i t y o f th e g a s t r i c j u ic e , i . e . ,
th e a m o u n t o f f r e e h y d r o c h l o r i c a c i d a p p e a r s t o s u g g e s t i t s e l f . "
T h is
s t a t e m e n t w as b a s e d o n h i s own o b s e r v a t i o n s a n d t h o s e c o l l e c t e d fro m t h e
L ite ra tu re .
I n n i n e t y o n e c a s e s o f a c h l o r h y d r i a due t o c a rc in o m a o f t h e
3to m a c h , t h e am ount o f i n d i c a n i n t h e u r i n e w as i n c r e a s e d i n s e v e n t y a n d
77
i n f i f t e e n i t w as a b s e n t o r n o t i n c r e a s e d .
I n t w e n ty o t h e r c a s e s o f
a c h lo r h y d r ia due to h y s t e r i a , a c u te g a s t r i t i s , an d g a s t r o e n t e r i t i s ,
h ad in c r e a s e d u r in a r y in d ic a n e x c re tio n .
I n p e rs o n s w ith u l c e r and
in c r e a s e d h y d r o c h lo r ic a c id c o n te n t o f th e g a s t r i c
i n d i c a n e x c r e t i o n w as u s u a l l y f o u n d .
a ll
ju ic e , in c re a s e d
I n o th e r h y p e rc h lo rh y d ria s e i t h e r
n o r m a l o r d e c r e a s e d v a l u e s w e re f o u n d .
T he p o s s i b l e r e l a t i o n b e tw e e n i n d o l e a n d p e r n i c i o u s a n e m ia
was p o i n t e d o u t b y I ’o n t e s a n d T h i v o l l e some t h i r t y y e a r s a f t e r S im o n ’ s
s ta te m e n t.
In a s e r ie s o f f iv e p a p e rs ( 5 ,6 ,7 ,8 ,9 )
t h e s e men i n v e s t i g a t e d
t h e r o l e o f h i s t i d i n e a n d t r y p t o p h a n e i n r e d b lo o d c e l l p r o d u c t i o n .
T hey
p o i n t e d o u t t h a t t h e s e tw o am ino a c i d s c o u ld n o t b e s y n t h e s i z e d i n th e ,
Dody.
When g i v e n t o r a b b i t s a n d d o g s , h i s t i d i n e a n d t r y p t o p h a n e ,
e ith e r
s i n g l y o r i n c o m b i n a ti o n , i n c r e a s e d t h e r e d b lo o d c e l l c o u n ts a n d hemo­
g lo b in v a lu e s .
When r a t s w e re p l a c e d on a lo w t r y p t o p h a n e an d h i s t i d i n e
l i e t t h e y l o s t w e ig h t a n d lo w e r e d b lo o d v a l u e s w e re o b s e r v e d .
A d d in g
t h e s e tw o am ino a c i d s t o t h e d i e t r a i s e d t h e l e v e l s b a c k t o n o r m a l,
l a b b i t s on a n o r m a l d i e t w e re g i v e n l y s i n e , l e u c i n e , p h e n y l a l a n i n e a n d a
l i x t u r e o f try p to p h a n e and h i s t i d i n e .
T h o se a n im a ls on l e u c i n e , l y s i n e ,
).r p h e n y l a l a n i n e d i d n o t h a v e i n c r e a s e d b lo o d v a l u e s , b u t t h o s e a n im a ls
'e c e i v i n g t h e m i x t u r e o f t r y p t o p h a n e a n d h i s t i d i n e
b l o o d c e l l c o u n ts a n d h e m o g lo b in v a l u e s .
show ed i n c r e a s e d r e d
A lth o u g h t h e i r d a t a i s n o t
m t i r e l y c o n v i n c i n g , t h e y i n v e s t i g a t e d t h e e f f e c t o f t h e s e tw o am in o a c i d s
>n p e r n i c i o u s a n e m ia p a t i e n t s
(1 0 ).
I ’o n t e s a n d T h i v o l l e b e l i e v e t h a t i t w as n o t i m p o s s i b l e t h a t
> e r n i c io u s a n e m ia i s
due to a l a c k o f try p to p h a n e and h i s t i d i n e due to
>oor d i g e s t i o n a n d j u i c e s w i t h r e s u l t i n g
d e s t r u c t i o n o f t h e s e am ino a c i d s
by b a c te r ia .
T hey b e l i e v e d t h a t l i v e r t h e r a p y i s o f v a l u e b e c a u s e i t
c o n t a i n s t h e s e tw o e s s e n t i a l am ino a c i d s .
One p e r n i c i o u s a n e m ia p a t i e n t
w as g i v e n d a i l y i n j e c t i o n s o f 100 mgm. o f t r y p t o p h a n e a n d 200 mgm. o f
h is tid in e .
In t h ir ty
d a y s t h e r e d b l o o d c e l l s h a d i n c r e a s e d fro m 1 .7 0
m i l l i o n t o 5 .1 0 m i l l i o n a n d t h e h e m o g lo b in fro m 4 5 $ t o 9 0 $ .
The p a t i e n t
g a in e d 20 k i l o s i n s i x m o n th s o f t h e r a p y a n d t h e b lo o d v a l u e s w e re m a in ­
t a i n e d w i t h o u t a n y o t h e r ty p e o f t r e a t m e n t .
so r a p i d l y n o r f o r so l o n g .
O t h e r c a s e s d i d n o t im p ro v e
T hey m e t w i t h c o m p le te i n s u c c e s s i n o l d e r
p e r s o n s who h a d h a d n u m e ro u s r e l a p s e s a n d h a d b e e n t r e a t e d w i t h a r s e n i c a l s .
R i e s e , h o w e v e r, h a d h y p e r p l a s t i c m arro w s w h ic h w e re i n t e r p r e t e d a s p e r h a p s
2o n t a i n i n g t h e c e l l s b u t no e m p ty in g th e m i n t o t h e b lo o d s tr e a m .
I n c o n n e c t i o n w i t h t h i s w o rk , i t
[ll)
s h o u ld b e m e n tio n e d t h a t B a a r
h a d p r e v i o u s l y r e p o r t e d i n 19 1 2 t h a t i n d i c a n e x c r e t i o n w as i n c r e a s e d
In p e r n i c i o u s a n e m ia .
E x p e r im e n ta l s t u d i e s
on t h e e f f e c t o f i n d o l e i n t h e o r g a n is m
e x te n d b a c k t o H e n c k i (1 2 ) who i n 187 6 d e m o n s tr a te d t h a t o n e gram o f
th e s u b s t a n c e a d m i n i s t e r e d t o a dog show ed no i n t o x i c a t i o n w h i l e two
gram s c a u s e d d i a r r h e a a n d h e m a t u r i a .
T he t o x i c p r o p e r t i e s o f i n d o l e w e re i n v e s t i g a t e d b y H e r t e r i n
L898 ( 1 3 , 1 4 ) .
I n t r a v e n o u s i n j e c t i o n o f i n d o l e i n d o g s an d r a b b i t s
r e s u lte d i n d e p re s s e d r e s p i r a t i o n and h e a r t r a t e , p r o s t r a t i o n ,
p u p ils ,
c o lo n ic sp asm s, and in c r e a s e d r e f l e x e x c i t a b i l i t y .
c o n tra c te d
H e r te r and
iiVakeman (1 5 ) m e a s u re d t h e c h a n g e o f i n d o l e t o i n d i c a n i n m i x t u r e s o f
L i v e r , k i d n e y , m u s c le , b r a i n , a n d b lo o d a n d fo u n d t h e i r o r d e r o f a c t i v i t y
fro m h i g h e s t t o l o w e s t t o b e i n t h e o r d e r n am ed.
T hey a l s o
in fu s e d
L n im a ls i n t r a v e n o u s l y w i t h i n d o l e u n t i l n e r v i o u s sym ptom s a p p e a r e d .
The
79
a n i m a l s w e re t h e n k i l l e d a n d t h e o r g a n s a n a l y z e d .
M u s c le c o n t a i n e d t h e
s m a l l e s t am o u n t a n d l i v e r t h e g r e a t e s t am o u n t o f t h e s u b s t a n c e .
The
a u t h o r s s t a t e t h a t t h e y b e l i e v e t h e l i v e r c o n t a i n s t h e l a r g e s t am ount
because i t
re m o v e s t h e m o s t fro m t h e b lo o d s t r e a m , n o t b e c a u s e i t i s t h e
l e a s t a c ti v e in d e to x if y in g i t .
P r o lo n g e d e t h e r o r c h lo r o f o r m a n e s t h e s i a
w as show n t o r e d u c e t h e a b i l i t y o f t h e l i v e r t o o x i d i z e i n d o l e a n d c o n j u g a t e
p h e n o l.
When 0 .1 t o 2 g ra m s o f i n d o l e was g i v e n d a i l y t o t h r e e human
s u b j e c t s f o r f o u r t o s e v e n d a y s t h e o n l y r e s u l t s n o t e d w e re f r o n t a l
h e a d a c h e s , a b n o rm a l c e p h a l i c s e n s a t i o n s , a n d a n i n d i s p o s i t i o n f o r m e n ta l
o r p h y s ic a l e x e rtio n .
L o n g c o n t i n u e d i n g e s t i o n o f s m a ll a m o u n ts o f i n d o l e o r p a r a c r e s o l
( 0 . 1 t o 0 . 2 g ram s f o r s i x t y t h r e e d $ y s ) w as w i t h o u t a f f e c t on t h e h e a l t h
o f r a b b i t s a n d g u i n e a p i g s , a c c o r d i n g t o W lad y czk o ( 1 6 ) , b u t p r o d u c e d
re g re s s iv e a lt e r a t io n s in th e v e s s e ls o f th e b r a in .
S m a ll a m o u n ts o f
i e s t r u c t i v e c h a n g e s i n t h e c e l l s o f t h e n e r v o u s s y s te m a s w e ll a s a p r o . i f e r a t i o n o f n e u r o g l i a w e re o b s e r v e d .
If,
a s H e r te r (v id e s u p r a ) s t a t e d , i n t e r f e r e n c e w ith l i v e r fu n c -
;io n re d u c e s th e a b i l i t y o f th e l i v e r to o x id iz e in d o le , th e a p p e a ra n c e
p f f r e e i n d o l e i n t h e u r i n e m ig h t b e u s e d a s a n i n d i c a t i o n o f l i v e r
fa ilu re .
V au g h an (1 7 )
e x a m in ed t h e u r i n e s o f p a t i e n t s w i t h l i v e r a n d
p u t r e f a c t iv e and i n t e s t i n a l s t a s i s d is e a s e s b u t f a i l e d to n o te f r e e in d o le
In t h e u r i n e .
On t h e o t h e r h a n d , F o r b e s an d N e a le (1 8 ) fo u n d no i n d o l e
Ln n o r m a l hum an u r i n e , b u t t h e
s u b s t a n c e w as p r e s e n t i n t h e u r i n e s o f
p a t i e n t s w ith c h ro n ic a r t h r i t i s ,
p e l l a g r a , d i a b e t e s , t u b e r c u l o s i s , to x e m ia
pf p r e g n a n c y , c o n g e s t i v e h e a r t f a i l u r e ,
an d l o b a r p n e u m o n ia .
T h e s e men
l o n s i d e r e d t h e p r e s e n c e o f 0 .0 5 mgm. p e r l i t e r o f u r i n e a s s i g n i f i c a n t
Ln a m o u n t.
V au g h an (1 9 ) s t a t e d t h a t h e b e l i e v e d t h i s am ount o f i n d o l e
80
w as due to b a c t e r i a l c o n ta m in a tio n .
P r o t e u s v u l g a r i s , p s e u d o d y s e n t e r y , a n d t r u e v i b r i o s w e re l i s t e d
b y C a r n e s a n d L e w is (2 0 ) a s some o f t h e o r g a n is m s c a p a b l e o f f o rm in g
in d o le .
T hey s t a t e d t h a t E . c o l i c a n p r o d u c e i n d o l e i n t h e p r e s e n c e o f
tr y p to p h a n e , in d o l p y ru v ic a c i d , o r d - in d o le l a c t i c
a c id .
In o r d e r to
d e te r m i n e i f o r g a n is m s i n u r i n e c o u ld b e r e s p o n s i b l e f o r t h e p r e s e n c e
o f f r e e i n d o l e , t h e y p e r f o r m e d t h e f o l l o w i n g e x p e r im e n t.
w as o b t a i n e d b y p a s s i n g u r i n e th r o u g h s t e r i l e
filtra tio n
S t e r i l e u rin e
s y s te m s .
A
p o r t i o n o f t h e s t e r i l e u r i n e w as s e e d e d w i t h E. C o l i , a n d t h e r e s t was
n o t seeded.
T he s e e d e d u r i n e s showred i n c r e a s e d i n d o l e c o n t e n t , w h ile t h e
c o n t r o l s c o n t a i n e d ‘t h e sam e am o u n t a s b e f o r e .
The i n o c u l a t e d f i l t r a t e s
show ed a p o s i t i v e i n c r e a s e i n f i v e t o s i x h o u r s ; f u r t h e r i n c u b a t i o n
p ro d u c e d n o m o re i n d o l e .
fo rm ed i n t h e u r i n e .
I n t h e p r e s e n c e o f 0 .2 $ g l u c o s e , n o i n d o l e w as
T hey s t a t e d t h a t t h e i n d o l e t h u s fo rm e d d o e s n o t
a r i s e fro m t h e o r g a n is m s t h e m s e lv e s b u t t h a t u r i n e c o n t a i n s a s u b s t r a t e
fo r in d o le p ro d u c tio n .
T ftis s u b s t r a t e i s a s m a ll c o n s t a n t am ount a n d i s
i r o b a b l y t r y p t o p h a n e o r some c l o s e l y a l l i e d i n d o l e d e r i v a t i v e .
A n a l y s e s o f t i s s u e s f o r i n d i c a n w e re m ade by O l i v e t ( 2 1 ) .
th e o r d e r o f t h e i r c o n t e n t ,
In
t h e a n a ly s e s r e v e a le d t h a t k id n e y s con­
t a i n e d t h e g r e a t e s t am ount f o ll o w e d b y l i v e r ,
s m a ll i n t e s t i n e , s k i n ,
D ra in , a n d p a n c r e a s ; t h e s p l e e n c o n ta i n e d o n l y a t r a c e .
No f r e e i n d o l e
vas fo u n d i n t h e b lo o d s tr e a m f i f t e e n m in u t e s f o llo ^ 'i- n g t h e i n t r a v e n o u s
: n j e c t i o n o f 50 rngju. o f i n d o l e .
O n ly 1 0 # t o 1 5 # o f t h e s u b s t a n c e w as c o n -
^ e r t e d t o in d o x y l g ly c u r o n ic a c id .
The t h e o r e t i c a l y i e l d o f i n d i c a n
, ih o u ld b e n e a r l y 100 mgm. b u t o n l y 15 t o 16 mgm. c o u ld b e f o u n d m t h e
a n im a l.
A f t e r t h e i n j e c t i o n o f i n d o l e , t h e l a r g e s t a m o u n ts w e re fo u n d i n
;he b r a i n a n d l i v e r .
E s s e n t i a l l y t h e same r e s u l t s w e re r e p o r t e d by
81
d e G a e ta n i ( 3 2 ).
T he a p p a r e n t d i s c r e p a n c y b e tw e e n t h e t h e o r e t i c a l a n d a c t u a l
r e c o v e r y o f i n d i c a n f o l l o w i n g i n d o l e i n j e c t i o n w as d i s c u s s e d by Bohm
(2 2 )
i n a l e n g t h y a r t i c l e on i n d o l e m e ta b o lis m .
T h is a u th o r s t a t e d t h a t
t h e r e w as a n u m b er o f i n t e r m e d i a t e s u b s t a n c e s i n t h e c o n v e r s i o n o f i n d o l e
t o i n d i c a n w h ic h a c c o u n t f o r t h e d i s c r e p a n c y .
T h ese s u b s ta n c e s a p p e a r
w i t h s u b s t i t u t i o n on t h e A lp h a a n d B e ta p o s i t i o n s on t h e p y r r o l r i n g .
I n t h e s t u d y o f t h e o r g a n s w h ic h r e g u l a t e i n d o le m i a a n d in d o x y le m ia
f o l l o w i n g t h e i n j e c t i o n o f i n d o l e , H o u s s a y (2 3 ) m ade t h e f o l l o w i n g o b s e r ­
v a tio n s .
I n d o l e i n t r o d u c e d i n t o t h e i n t e s t i n a l lu m en
i s r a p id ly a b so rb e d .
I f t h e d o s e g i v e n i s 20 mgm. o r m o re p e r k i l o , t h e l i v e r c a n n o t a r r e s t
a l l o f t h e i n d o l e w i t h t h e r e s u l t t h a t t h e r e may be 10 t o 20 mgm. p e r
l i t e r i n t h e p o r t a l b l o o d a n d 5 t o 12 mgm. p e r l i t e r i n t h e a r t e r i a l
b lo o d .
T he am ount i n t h e p o r t a l b lo o d i s r e d u c e d t o t r a c e s i n two h o u r s .
I f l e s s t h a n 20 mgm. p e r k i l o i s g i v e n , i n d o l e i t s e l f d o e s n o t a p p e a r
Ln t h e b l o o d s tr e a m , i n s t e a d i n d o x y l i s f o u n d .
Dlood s t r e a m , b u t i n d o l e i s
I n d o x y l r e m a in s i n t h e
r a p i d l y t a k e n up b y t h e t i s s u e s ,
D eing m o s t c a p a b l e o f f i x i n g i t .
I f i n d o l e (1 0 m gm .) i s
th e l i v e r
in je c te d in to
t h e j u g u l a r v e i n o f a n e p h r e c to m iz e d d o g , i t s l e v e l i n t h e b lo o d q u i c k l y
lim in is h e s .
T h i s o c c u r s e v e n when t h e d i g e s t i v e t r a c t i s
e x tirp a te d .
H ow ever, i f t h e l i v e r i s re m o v e d , b lo o d i n d o x y l r i s e s o n l y a v e r y s l i g h t
a m o u n t.
H o u s s a y , l i k e O l i v e t a n d Bohm, fo u n d t h a t i n d o l e w as n o t q u a n ­
tita tiv e ly
c o n v e r t e d t o i n d o x y l i n t h e o r g a n is m .
He r e c o v e r e d g O -3 3
ngm. o f i n d i c a n f o r 100 mgm. o f i n d o l e .
D i s c u s s i n g t h e m e c h a n ism s f o r t h e t r a n s f o r m a t i o n o f s u b s t a n c e s t o
in d o x y l i n t h e o r g a n is m , H o u s s a y e t a l . ( 2 4 )
l i s t t h r e e m e th o d s .
88
1 . G e n e ral tr a n s fo r m a tio n ,
e x tr a h e p a tic and e x tr a i n t e n t i n a l .
O n ly o r t h o - n i t r o - p h e n y ^ - p r o p i o n i c a c i d c a u s e s a r i s e i n
in d o x y l a f t e r th e l i v e r and k id n ej^s, o r g a s t r o - i n t e s t i n a l t r a c t
and k id n e y s a r e e x t i r p a t e d i n th e dog.
8 . H e p a t ic t r a n s f o r m a t i o n — o n l y i n d o l e c o n v e r t e d to i n d o x y l .
3 .T ra n s fo rm a tio n w ith in te r v e n t io n o f g a s t r o - i n t e s t i n a l a p p a ra tu s
and l i v e r ,
An e l e v a t i o n o f in d o x y l i n t h e b lo o d s tr e a m may
a r i s e fro m t h e f e e d i n g o f B e t a - i n d o l e a l d e h y d e , e - n i t r o p h e n y la c e ta ld e h y d e , B e ta - in d o le p y ru v ic a c id , i s o t i n e ,
m e th y l i n d o l e .
I f th e
sto m a c h a n d i n t e s t i n e a r e e x t i r p a t e d and
th e s u b s ta n c e s i n j e c t e d ,
i s e x tirp a te d .
B e ta -
th e r i s e
i s l e s s t h a n when t h e l i v e r
H o u ssay th e o r i z e s t h a t t h e i n t e s t i n e s p ro b a b ly
c h a n g e t h e s e s u b s t a n c e s to a p r e c u r s o r (w h ic h may b e i n d o l e )
v% 4ch i s t h e n t r a n s f o r m e d by t h e l i v e r t o i n d o x y l .
-Ln m an, H o u s s a y (2 5 )
s t a t e s t h a t i n d o l e fo rm e d by t h e b rea k d o w n
o f t r y p t o p h a n e by b a c t e r i a i n t h e gut i s a b s o rb e d i n t o t h e b lo o d stre a m .
I t p a s s e s t o t h e l i v e r w here i t
i s c o n v e rted in to in d o x y l.
The k i d n e y
e l i m i n a t e s t h e i n d o x y l.— 0 . 9 t o 36 mgm. p e r d a y i n man b e i n g e l i m i n a t e d
a s in d o x y l o r p o ta ssiu m in d o x y l s u lp h a te .
Four f a c to rs in th e r e g u la tio n
o f in d o x y le m ia a r e :
1 . Amount o f i n d o l e p r o d u c e d a n d a b s o r b e d i n t h e i n t e s t i n e .
2 . C o n v e r s io n o f i n d o l e t o
in d o x y l i n t h e l i v e r a n d i t s
d e s tru c tio n .
3 . S to ra g e in th e b lo o d .
4 . E lim in a tio n by th e k id n e y s .
The r o l e o f t h e l i v e r i n c o n v e r t i n g i n d o l e t o i n d o x y l was c o n f i r m e d
b y L a r o c h e a n d D e s b o r d e s ( 2 6 ) who i n j e c t e d i n d o l e i n t r a v e n o u s l y i n t o
83
r a b b i t s , d o g s , a n d g u in e a p i g s .
U s in g n o r m a l, e v i s c e r a t e d ,
s p le n e e to m iz e d d o g s , a n d d o g s w i t h t h e
d i g e s t i v e t r a c t e x c lu d e d , a n d d o g s w i t h t h e p o r t a l v e i n a n d h e p a t i c
a r t e r i e s l i g a t e d , B a ra c,
(2 7 ) a g a i n show ed t h e im p o r ta n c e o f t h e l i v e r
in c o n v e rtin g in d o le to in d ic a n .
A b s o rp tio n s tu d i e s u s in g th e r a t ,
a s r e p o r t e d b y N i c o l a i (2 8 )
d e m o n s t r a t e d t h a t 50% o f t h e i n d o l e was a b s o r b e d i n t h i r t y
s e c o n d s when
1 c c . o f a s o l u t i o n c o n t a i n i n g 2 .3 0 m g m ./c c . w as p l a c e d i n a 15 gm.
l e n g t h o f je ju n u m .
A l l o f t h e i n d o l e w as a b s o r b e d i n s i x m i n u t e s .
Some
o f t h e i n d o l e r e a p p e a r e d i n t h e g u t i n o n e h o u r , an d l a r g e q u a n t i t i e s
in f o u r h o u rs.
He c o n c lu d e d t h a t t h e g u t i s b o t h a r e s o r p t i v e a n d
e x c re to ry a g e n t o f in d o le .
S t u d i e s by I n o u e (2 9 )
d e m o n s tr a te d t h a t m o re a b s o r p t i o n o f i n d o l e
b a k e s p l a c e i n t h e s m a ll bow el t h a n i n t h e c o lo n .
in d o le p a s s e s in to
Ln t h e l u n g s .
t h e lym ph s t r e a m .
P a r t o f th e a b so rb e d
T h is p o r t i o n i s c o n v e r t e d t o
in d ic a n
He c o n f ir m e d t h e im p o r ta n c e o f t h e l i v e r i n i n d i c a n sy n ­
t h e s i s , a n d w as a b l e t o
d e m o n s tr a te t h a t l u n g a n d l i v e r e m u ls io n s w e re
s a p a b le o f t r a n s f o r m i n g i n d o l e i n t o i n d i c a n .
Lf t h e p H i s b e lo w 6 .2 o r a b o v e 9 i 6 .
a n u ls io n i s h e a te d t o
T h is d o e s n o t o c c u r , h o w e v e r,
N e ith e r w i l l i t o c c u r i f th e
60 ° f o r t h i r t y m i n u t e s .
I n r e g a r d t o t h e r e l a t i o n s h i p o f i n d o l e t o b l o o d , b lo o d f o r m in g
a le m e n t s , an d t h e f o r m e d e le m e n ts o f t h e b lo o d , t h e r e h a v e b e e n re m a rk ­
a b ly fe w i n v e s t i g a t i o n s .
B u n g e le r (3 0 ) r e p o r t e d t h a t d e g e n e r a t i o n a n d r e g e n e r a t i o n o f t h e
Done m arro w i n a n e m ia s w as c o r r e l a t e d w ith t h e am ount o f a r o m a tic s u b 3t a n c e s i n t h e b l o o d s tr e a m a n d t h e l e n g t h o f s i c k n e s s .
and i n d o l e a s a r o m a t i c s u b s t a n c e s i n t h e b l o o d .
He d e f i n e d p h e n o l
84
When i n d o l e i s a d d e d t o
a s o l u t i o n o f g u in e a p i g c o r p u s c l e s i n
a m o u n ts a m l l en o u g h so a s n o t t o c a u s e h e m o l y s i s , d e G a e t a n i (3 1 )
r e p o r t e d t h a t t h e o s m o tic r e s i s t a n c e o f t h e r e d b lo o d c o r p u s c l e s was
i n c r e a s e d , w h i l e t h e p e r m e a b i l i t y an d s e d i m e n t a t i o n r a t e w e re lo w e r e d .
I n i n v e s t i g a t i n g t h e r o l e o f i n d o l e i n t h e a n e m ia f o u n d i n
u r e m i a , d e G a e ta n i (3 3 )
p ig s .
i n j e c t e d i n d o l e i n t r a p e r i t o n e a l l y i n t o g u in e a
I n o n e g ro u p w h ic h r e c e i v e d 30 mgm. p e r k i l o , t h e b lo o d was
e x a m in e d o n e h o u r f o l l o w i n g t h e i n j e c t i o n .
The a v e r a g e s o f t e n a n im a ls
w e re :
R ed b lo o d c e l l s
P re -in je c tio n
P o s t-in je c tio n
5 .8 9
S .2 1
H e m o g lo b in % W h ite b lo o d c e l l s
8 7 .9
7 8 .8
A n o th e r g ro u p r e c e i v e d 15 mgm. p e r k i l o
t w e n ty d a y s .
8670
6500
f o r f o r t y t o o n e h u n d re d
A m i l d a n e m ia w h ic h h a d a t e n d e n c y to w a rd h y p e rc h r o m ia
a n d m i c r o c y t o s i s t o g e t h e r w i t h a le u k o p e n i a a n d n e u t r o p h i l i a d e v e lo p e d .
The a u t h o r s t a t e s t h a t t h e d a t a i s i n s u f f i c i e n t t o s a y w h e th e r t h e i n d o l e
a c t e d a s a h e m o l y ti c a g e n t o r a b o n e m arrow d e p r e s s a n t , b u t h e d o u b t i f
i n d o l e i s r e s p o n s i b l e f o r t h e a n e m ia o f u r e m ia .
T he e f f e c t s o f i n d o l e o n t h e r e d b lo o d c e l l s o f d o g s r e c e i v i n g a
n o rm a l a n d a d e f i c i e n t d i e t h a v e b e e n r e p o r t e d b y R hoads ( 3 4 ) , R h o ad s
and B a r k e r ( 3 5 ) , R h o ad s and M i l l e r (3 6 ) an d R h o a d s , B a r k e r an d M i l l e r ( 3 7 ) .
When s t o m a t i t i s a p p e a r e d i n d o g s f e d a b l a c k to n g u e d i e t , t h e y w e re g iv e n
L gm. o f i n d o l e p e r d a y i n a d d i t i o n t o t h e d i e t .
The r e d b lo o d c e l l
v a l u e s d r o p p e d s h a r p l y , o c c a s i o n a l l y a s lo w a s 1 .0 0 m i l l i o n p e r c u . mm.
The sam e am ount o f i n d o l e f e d t o n o rm a l d o g s on a n o rm a l d i e t h a d no
s ffe c t.
i n a n o t h e r s e t o f d o g s , w hen t h e a n im a ls h a d becom e s t a b i l i s e d
to i n d o l e f e e d i n g , t h e b l a c k to n g u e d i e t w as g i v e n c a u s i n g a s e v e r e
a n e m ia w h ic h r e s p o n d e d t o l i v e r e x t r a c t i n
a d m in is tra tio n .
s p i t e o f c o n ti n u e d i n d o l e
T he m arro w o f t h e s e a n im a ls show ed i n c r e a s e d e e l l u l a r i t y
a n d p r e d o m in a n c e o f im m a tu re f o r m s .
B i l i r u b i n e x c r e t i o n w as m e a s u re d i n b i l e f i s t u l a
r e c e i v e d 1 gm. i n d o l e i n o n e s i n g l e d o s e .
d o g s w h ic h
The b i l i r u b i n e x c r e t i o n w as
m e a s u re d f o r s e v e r a l d a y s f o l l o w i n g t h e i n d o l e .
T h e r e w as a s h a r p r i s e
i n b i l i r u b i n e x c r e tio n th e d ay f o llo w in g in d o le a d m in is tr a tio n , r e tu r n in g
t o n o r m a l l e v e l s i n tw o t o f i v e d a y s .
A s e v e r e a n e m ia c o u ld b e in d u c e d i n d o g s w hose b a s i c d i e t c o n s i s t e d
o f m i l k w h ic h w as s u p p le m e n te d w i t h i n d o l e .
I n a l l o f t h e s e a n im a ls i n w h ic h a s e v e r e a n e m ia w as in d u c e d by
i i e t a n d i n d o l e , f e e d i n g l i v e r e x t r a c t , y e a s t , o r a n o r m a l d i e t w ould
r e s t o r e t h e r e d b lo o d c e l l v a l u e s t o n o rm a l l e v e l s .
T he a u t h o r s c o n c lu d e d fro m b i l e
f i s t u l a d o g s w h ic h r e c e i v e d
i n d o l e a n d t h e b l a c k to n g u e d i e t a n d h a d l i v e r e x t r a c t f o r t r e a t m e n t
b h a t t h e a n e m ia i s a r e s u l t o f t h e v i t a m i n d e f i c i e n c y a n d i n d o l e was
d ue t o i n c r e a s e d l y s i s o f a l l h e m o g lo b in c o n t a i n i n g c e l l s .
The r i s e i n
t h e b l o o d l e v e l s f o l l o w i n g l i v e r t h e r a p y t e n d s to show d e c r e a s e d h e m o ly s is
and d e c r e a s e d c e l l p r o d u c t i o n .
T h ese e x p e rim e n ts s tim u la te d i n t e r e s t i n t h i s c o u n try in th e
h e m o ly tic p r o p e r t i e s o f i n d o l e .
P o n d e r (3 8 ) fo u n d t h a t c e r t a i n c o n ­
c e n t r a t i o n s o f i n d o l e c o u ld b e u s e d t o c a u s e l y s i s i n a r a b b i t c e l l s o d iu m c h l o r i d e - i n d o l e s y s te m .
T h i s l y s i s c o u ld b e i n h i b i t e d by seru m
Dr p la s m a a n d w as p r e c e d e d b y d i s c s p h e r e t r a n s f o r m a t i o n .
fo u n d t o b e a p o w e r f u l a c c e l e r a t o r o f l y s i s b y s a p o n in .
to h a v e tw o a c t i o n s , l i k e b e n z e n e ,
L y s in s .
(l)
ly tic
I n d o l e was
In d o le a p p e a rs
a n d (2 ) a c c e l e r a t i o n t o
On t h e o t h e r h a n d , i n d i c a n w as fo u n d t o be n e i t h e r l y t i c
nor
36
a c c e le ra to r;^
in f a c t i t
h a d a n i n h i b i t o r y a c t i o n on s a p o n in l y s i s .
-P onder s u g g e s t e d t h a t f l o o d i n g i n d o l e i n t o
t h e b lo o d o f d e f i c i e n t d i e t
d o g s a l l o w s t h e e n t r a n c e o f a w eak l y s i n o r a c c e l e r a t o r f o r l y s i n s a l r e a d y
p re s e n t.
T a y l o r (3 9 ) i n v e s t i g a t e d t h e b l o o d c h a n g e s p r o d u c e d i n r a b b i t s
r e c e iv i n g in d o le s u b c u ta n e o u s ly i n 40$ a lc o h o l .
g iv e n s u b c u t a n e o u s l y e a c h d a y t o
E i g h ty mgm. o f i n d o l e
e a c h o f t h r e e r a b b i t s c a u s e d a m ark ed
f a l l i n r e d b lo o d c e l l v a lu e s i n t h i r t y
days.
When t h e d o s e w as r e d u c e d
t o 56 mgm. p e r w eek , t h e r e d c e l l v a l u e s im p r o v e d .
R e tu rn in g to th e
f i r s t d o sa g e th e v a lu e s a g a in f e l l b u t a f t e r f i f t e e n d ay s began to r i s e
s lo w ly , r e t u r n i n g t o n o r m a l i n s p i t e o f t h e c o n ti n u e d a d m i n i s t r a t i o n o f
in d o le .
T h e h e m o g lo b in c o n t e n t o f t h e c e l l s d i d n o t r i s e a s r a p i d l y a s
th e c e l l c o u n t.
I n a d i s c u s s i o n o f t h e e t i o l o g y o f p e r n i c i o u s a n e m ia a t C o ld
S p r in g s H a r b o r ( 4 0 ) , R h o a d s p o i n t e d o u t t h a t t h e i n a b i l i t y t o p r o d u c e
r e g u l a r l y a n e x p e r i m e n t a l m a c r o c y t i c a n e m ia
u s in g j u s t a d i e t a r y d e f i -
s ie n c y m a k e s t h i s h y p o t h e s i s a s t o t h e e t i o l o g y o f p e r n i c i o u s anem ia
m te n a b le .
Some h e m o l y ti c f a c t o r m u st b e t a k e n i n t o a c c o u n t .
T h is
h e m o l y ti c t h e o r y i s b a s e d o n a n i n c r e a s e d o u t p u t o f b i l e p ig m e n t d u r in g
s x a c e rb a tio n s o f th e d is e a s e .
T h i s i n c r e a s e m u st be d u e t o
(l)
in c re a s e d
le r a o ly s i s o r (2 ) f o r m a t i o n o f b i l e p ig m e n t w i t h o u t t h e b re a k d o w n o f hemo­
g lo b in e .g .
fro m p r o t o p o r p h y r i n ,
^he l a t t e r is n o t lik e ly .
R h o a d s w e n t o n t o p o i n t o u t t h a t h e m o ly s is p r e s u p p o s e s i n c r e a s e d
; e l l p r o d u c t i o n , h e n c e t h e h y p e r p l a s t i c b o n e m arrow i n p e r n i c i o u s a n e m ia ,
t o b r i n e r h a s -shown (4 1 ) t h a t t h e r e i s a n i n c r e a s e d e x c r e t i o n o f ty p e I
m d t y p e I I p o r p h y r i n s i n p e r n i c i o u s a n e m ia w h ic h d i s a p p e a r s d u r in g
87
re m is s io n s o f th e d is e a s e ,
(v id e
R h o ad s r e p o r t e d h i s e x p e r i e n c e v.dth i n d o l e
s u p r a ) a n d a l s o e x p e r i m e n t s w i t h a t r a n s p l a n t e d l o o p o f ile u m .
If
t h e t e r m i n a l t h i r d o f t h e i l e u m i n d o g s w e re s h o r t c i r c u i t e d by an
a n a s t o m o s i n g p r o c e s s a n d t h e d i s t a l fe w c e n t i m e t e r s e x p l a n t e d and e n c l o s e d
i n t h e s k i n , p r e s s u r e o n t h e e x p l a n t e d se g m e n t w ould c a u s e s t a g n a t i o n a t
w ill.
d ie t i t
T h is c a u se d i n in d ie a n e m ia .
''“h e n t h e a n im a ls w e re on a r e s t r i c t e d
w o u ld c a u s e a s e v e r e a n e m ia , m i l d m a c r o c y t o s i s ,
s i d e r o s i s , and
i n c r e a s e d num ber o f im m a tu re h e m o p o ie tic e le m e n ts i n t h e b o n e m arro w .
1/tfhen t h e t r a n s p l a n t e d lo o p o p e r a t i o n w as p e r f o r m e d i n b i l e f i s ­
t u l a d o g s , p r e s s u r e o n t h e e x p l a n t e d lo o p c a u s e d a n i n c r e a s e i n t h e b l o o d
i n d i c a n , a d e c r e a s e i n r e t i c u l o c y t e n u m b e r s , a n i n c r e a s e i n b i l e p ig m e n t
o u t p u t , a n d a m o d e r a te a n e m ia .
N orm al v a l u e s r e t u r n e d vrtien t h e n o rm a l
c o u rs e o f t h e i n t e s t i n a l c o n t e n t s was r e s t o r e d .
No r e v ie w o f t h e i n d o l e l i t e r a t u r e
w ould b e c o m p le te w ith o u t
a e n t i o n i n g t h e w ork o f B i e b l (4 2 ) who i n v e s t i g a t e d t h e e f f e c t s o f
p h e n o l i c s u b s t a n c e s a n d i n d o l e on r a b b i t s b u t f a i l e d t o r e c o r d t h e
e f f e c t s on th e b lo o d .
A lth o u g h t h e w ork w as a l a r g e u n d e r t a k i n g , t h e
i a t a an d c o n c lu s io n s a r e c o n fu s in g .
M e th o d s
T he d o g s u s e d i n t h i s e x p e r im e n t w e re s e l e c t e d fro m a n im a ls
r e c e i v e d i n t h e l a b o r a t o r y fro m t h e p o u n d .
They r e c e i v e d t h e r e g u l a r
s to c k l a b o r a t o r y d i e t t h r o u g h o u t t h e e x p e r i m e n t , a n d r e c e i v e d no l i v e r
o r o rg a n m e a ts .
T h c e e s e r i e s o f e x p e r i m e n t s w e re p e rf o r m e d w i t h t h e f o ll o w i n g
groups o f a n im a ls I
88
G ro u p I — F o u r g a s t r e c t o m i z e d d o g s a n d tw o n o rm a l d o g s r e c e i v e d 1
gram o f i n d o l e e a c h p e r d a y f o r 224 a n d 63 d a y s r e s p e c t i v e l y .
The ty p e
o f o p e r a t i o n s on t h e g a s t r e c t o m i z e d d o g s w e re :
G a s t r e c to m iz e d Dog # 1 — O p e r a te d tw o m o n th s b e f o r e i n d o l e
g iv e n .
G a s t r e e to m iz e d Dog # 2 — O p e r a te d o n e m o n th b e f o r e i n d o l e
g iv e n .
G a s t r e c to m iz e d Dog G .E .P .— G a s t r e c to m iz e d f i v e y e a r s b e f o r e
in d o le g iv e n , a ls o
s u b to ta l
■i
p a n c r e a te c to m y a n d e n te r e c to m y
tw o y e a r s p r i o r t o i n d o l e .
G a s t r e c to m iz e d Dog G .E .P .X .— G a s t r e c to m iz e d s i x y e a r s b e f o r e
in d o le g iv e n .
S u b to ta l pan­
c r e a t e c t o m y and e n te r e c t o m y two
y e a rs p r io r to in d o le .
G roup I I — Two g a s t r e c t o m i z e d d o g s (N o s . 1 andi 8) and tw o n o rm a l
lo g s r e c e i v e d 1 g ram o f i n d o l e e a c h d a i l y f o r 210 d a y s .
B egan a f t e r f o u r
n o n th s l a y o f f fro m G roup I .
G roup I I I — Same a n im a ls a s G roup I I ,
ia c h d o g .
2 . 5 g ram s i n d o l e p e r d a y t o
T h i s w as b e g u n im m e d i a te l y a f t e r G roup I I w as e n d e d .
T he i n d o l e w as g iv e n i n g e l a t i n c a p s u l e s .
The a n im a ls w e re o b s e r v e d
c a r e f u l l y a f t e r e a c h d o s e t o m ake c e r t a i n t h a t t h e i n d o l e w as t a k e n .
B lo o d f o r c o u n t s ,
e t c . w as o b t a i n e d fro m t h e a n t e c u b i t a l v e i n
an d p l a c e d i n a d r y v i a l c o n t a i n i n g h e p a r i n .
i n t h e u s u a l m an n e r.
B lo o d c o u n ts w e re p e r f o r m e d
H e m o g lo b in d e t e r m i n a t i o n s w e re m ade a c c o r d i n g t o t h e
Newcomer m e th o d .
One m o n th f o l l o w i n g t h e l a s t i n d o l e p e r i o d , g a s t r e c t o m i z e d d o g s
89
N o s . 1 a n d 2 , a n d b o t h n o rm a l a n i m a l s w e re g iv e n 2 . 5 gram s i n d o l e e a c h
a n d b l o o d i n d o l e d e t e r m i n a t i o n s w e re m ade a t o n e h o u r a n d t h r e e h o u r s
fo llo w in g th e a d m in is tr a tio n o f th e in d o le .
The b lo o d i n d o l e v a l u e s w e re
d e te r m i n e d a c c o r d i n g t o t h e m e th o d o f M azzocco ( 4 3 ) .
B lo o d i n d o l e m e th o d :
1 .P o u r , d r o p b y d r o p , 1 c c . o f o x a l a t e d b l o o d i n t o 3 c c . o f e t h e r
i n a 10 c c . t a l l g l a s s — s t o p p e r e d v i a l ,
^ h a k e m o d e r a t e l y tw o m i n u t e s i n a
v e r tic a l p o s itio n .
2 . D e c a n t e t h e r w ith a r e c u r v e d p o i n t e d P a s t e u r p i p e t .
3 . Add 2 c c . m o re e t h e r a n d a g a i n s h a k e a n d d e c a n t .
4 . R e p e a t w i t h 2 c c . m o re e t h e r .
5 . R e p e a t w i t h 1 c c . m o re e t h e r .
6 . C o l l e c t t h e d e c a n t e d e t h e r i n a c l e a n a n d d r y 30 c c . t e s t
t u b e a n d a d d e n o u g h 9 5 $ a l c o h o l t o b r i n g t h e v o lu m e t o 12 c c .
7 . Add 10 d r o p s o f a 5$ a l c o h o l i c s o l u t i o n o f p - d im e th y l a m id o b e n z a ld e h y d e .
8 . Add c a r e f u l l y 2 c c . h y d r o c h l o r i c a c i d
9 . P r e p a r e know n q u a n t i t i e s o f i n d o l e
(D 1 . 1 9 ) .
i n e t h e r fro m 10 Gamma t o
L Gamma a n d c o m p a re i n c o l o r i m e t e r .
T h i s m e th o d w i l l d e te r m i n e a b o u t 8 3 -9 3 $ o f t h e i n d o l e i n t h e b l o o d .
The c o l o r d e v e lo p e d i s a t a maximum i n 5 -7 m i n u t e s , an d a t t h e e n d o f
t h i r t y m in u t e s t h e r e i s 9 2 -9 4 $ o f t h e o r i g i n a l m a x im a l c o l o r .
U ro b ili­
n o g e n i n t h e b l o o d w i l l g i v e t h e sam e c o l o r .
R e s u lts
The a v e r a g e r e s u l t s f o r G roup I a r e show n i n T a b le I a n d P i g u e s
I and I I .
T he a v e r a g e r e s u l t s f o r G roup I I a r e show n i n T a b le I I a n d
f i g u r e s I and I I .
T he r e s u l t s o f G roup I I I a r e show n i n T a b l e . I I .
G ro u p I —-The r e d b lo o d c e l l , h e m o g lo b in a n d h e m a t o c r i t v a l u e s o f
t h e g a s t r e c t o m i z e d d o g s f e l l o f f v e r y m a r k e d ly i n t h e f i r s t week
e x p e rim e n t.
o f th e
T h e re w as a s lo w , e r r a t i c r e t u r n o f t h e r e d b l o o d c e l l an d
h e m o g lo b in v a l u e s to w a rd t h e p r e - e x p e r i m e n t a l l e v e l s , b u t t h e h e m a to ­
c r i t v a l u e s r e t u r n e d m o re r a p i d l y .
W ith t h e su d d e n d ro p i n b lo o d c o u n t
t h e r e w as a s im u l ta n e o u s i n c r e a s e i n t h e m ean c o r p u s c u l a r v o lu m e ,
The
n e a n c o r p u s c u l a r v o lu m e r e m a in e d a b o v e t h e p r e - e x p e r i m e n t a l l e v e l 120
d ay s a f t e r t h e i n d o l e w as s t o p p e d .
T he n o rm a l d o g s r e a c h e d m o re s lo w ly , t h e r e d b lo o d c e l l a n d hemo­
g l o b in v a l u e s d e c l i n i n g s lo w ly b u t s t e a d i l y d u r in g t h e f i r s t tw e n ty e i g h t
la y s o f t h e e x p e r im e n t a n d t h e n g r a d u a l l y r e t u r n i n g to w a rd n o r m a l.
T h e re
was n o t a s m ark e d a n i n c r e a s e i n t h e m ean c o r p u s c u l a r vo lu m e o f t h e r e d
c e l l s i n t h i s g ro u p a s i n t h e g a s t r e c t o m i z e d g r o u p .
By tw e n ty e i g h t d a y s
f o l l o w i n g t h e c e s s a t i o n o f t h e i n d o l e f e e d i n g s , t h e b l o o d v a l u e s w e re a t
a p p r o x i m a t e l y n o rm a l l e v e l s .
G roup I I — I n b o t h t h e n o rm a l a n d g a s t r e c t o m i z e d d o g s o f t h i s
g ro u p t h e r e a c t i o n w as v e r y s i m i l a r t o t h a t o f t h e n o rm a l d o g s i n G roup I
and t o e a c h o t h e r .
G roup I I I
The g a s t r e c t o m i z e d d o g s w e re n o t a f f e c t e d d i f f e r e n t l y
on t h e l a r g e r d o s a g e o f i n d o l e t h a n on t h e
s m a lle r d o s e .
T h e re w as a g a i n
a s m a l l d e c l i n e i n r e d b lo o d c e l l a n d h e m o g lo b in v a l u e s w i t h a n i n c ­
r e a s e d m ean c o r p u s c u l a r v o lu m e b u t t h e s e t e n d e d t o r e t u r n t o n o rm a l l e v e l s
in s p i t e o f t h e c o n t i n u e d i n d o l e a d m i n i s t r a t i o n .
T he n o rm a l d o g s w e re m o re s t r i k i n g l y a f f e c t e d w i t h t h e l a r g e r
l o s e , t h e r e d c e l l a n d h e m o g lo b in v a l u e s f a l l i n g o f f p r o g r e s s i v e l y w i t h
i n c r e a s e s i n t h e m ean c o r p u s c u l a r v o lu m e .
T h ir ty th re e days a f t e r th e
c e s s a t i o n o f t h e i n d o l e , t h e r e d b lo o d c e l l v a l u e s w e re a g a i n n o r m a l.
T he b l o o d i n d o l e d e t e r m i n a t i o n s b e f o r e a n d a f t e r a s i n g l e o r a l
d o s e o f S . 5 g ra m s o f i n d o l e w e re :
Dog N um ber
G a s tre c to m iz e d
G a s t r e c to m iz e d
N o rm al 10
N o rm al 11
1
2
0 h o u rs
1 hour
3 h o u rs
O y /lO O c c .
0
0
0
l O O O y /l O O e e .
630
0
857
600 ^ 1 0 0
600
600
948
T h e s e r e s u l t s i n d i c a t e t h a t t h e i n d o l e g iv e n o r a l l y i s a b s o r b e d
r e a d i l y a n d i s p r e s e n t i n t h e b lo o d s tr e a m i n l a r g e a m o u n ts a t t h e end o f
th re e h o u rs.
D is c u s s io n
A s R h o a d s h a s p o i n t e d o u t , t h e i n c r e a s e d h e m o ly s is i n p e r n i c i o u s
a n e m ia d o e s n o t l e n d i t s e l f w e l l t o a t h e o r y o f d i e t a r y d e f i c i e n c y .
H ow ever, i t
i s n o t i m p o s s i b l e t h a t su c h a d e f i c i e n c y m ig h t l e a d t o
d e f e c t i v e c e l l s w h ic h a r e m o re r e a d i l y rem o v ed b y t h e b lo o d d e s t r o y i n g
o r g a n s t h a n a r e n o rm a l c e l l s .
I t i s n o t p o s s i b l e to s t a t e d e f i n i t e l y
w h e th e r o r n o t a n a c t u a l h e m o l y ti c a g e n t i s a t w o rk .
C e r ta i n ly w ith th e
i c h l o r h y d r i a p r e s e n t i n p e r n i c i o u s a n e m ia we m ig h t e x p e c t a n i n c r e a s e d
I n d o le p r o d u c t i o n i n t h e g a s t r o - i n t e s t i n a l t r a c t w h ic h m ig h t l e a d t o
In cre a se d in d o le a b s o rp tio n .
I f Ponder i s
c o r r e c t, th e in d o le i t s e l f
v h i l e n o t b e i n g a s t r o n g l y h e m o l y ti c a g e n t m ig h t a c c e l e r a t e a h e m o ly s in
a l r e a d y p r e s e n t , o r a t l e a s t r e n d e r t h e c e l l s m ore s u s c e p t i b l e t o d e s ­
t r u c t i o n b y t h e b lo o d d e s t r o y i n g o r g a n s .
T J h lik e R h o ad s a n d M i l l e r , we h a v e f o u n d t h a t d o g s g i v e n o n e gram
92
o f i n d o l e p e r d a y w i l l d e v e lo p an a n e m ia w h ic h g r a d u a l l y r e c e d e s an d n o rm a l
v a l u e s a r e a g a in f o u n d i n s p i t e o f t h e c o n t i n u e d i n d o l e a d m i n i s t r a t i o n .
When t h e d o s a g e i s
s t e p p e d u p , h o w e v e r, n o rm a l d o g s a p p e a r t o d e v e lo p a n
i n c r e a s i n g a n e m ia w h ic h i s p r o m p tly r e l i e v e d b y d i s c o n t i n u i n g th e i n d o l e .
G a s tre c to m iz e d dogs on t h e
o th e r hand a r e b u t l i t t l e
d o s e , r e a c t i n g i n m uch t h e
same m a n n e r a s t o t h e s m a l l e r d o s e .
Do t h e s e r e s u l t s
i s a c c e l e r a t e d by i n d o l e ?
su g g est
a f f e c t e d by t h e l a r g e r
a n o rm a l h e m o ly s in i n t h e b lo o d w h ic h
T h e se d a t a a r e to o few t o a n s w e r t h i s q u e s ­
tio n .
I n f a c t t h e q u e s t i o n c a n n o t b e a n s w e re d u n t i l c o r r e l a t e d s t u d i e s
o f th e
s i z e o f t h e d o s e , t h e a p p e a r a n c e o f t h e b o n e m a rro w , an d b lo o d
I n d o l e l e v e l s a r e m ad e.
W i l l i n d o l e i n s u f f i c i e n t a m o u n ts c a u s e m a c r o c y t o s i s ?
d a t a i t w o u ld a p p e a r t h a t t h i s i s v a r i a b l e .
From o u r
C e r t a i n l y no s e v e r e m a c ro c y ­
t o s i s a p p e a re d i n o u r a n im a ls .
C o n c lu s io n s
One gram o f i n d o l e g i v e n o r a l l y p e r d a y t o n o rm a l a n d g a s t r e c ­
to m iz e d d o g s o n a d o r m a l d i e t w i l l c a u s e a m il d a n e m ia w h ic h t e n d s t o
r e c e d e t o w a r d n o rm a l v a l u e s i n s p i t e o f c o n ti n u e d i n d o l e a d m i n i s t r a t i o n .
Two a n d o n e - h a l f g ram s o f i n d o l e g iv e n t o n o rm a l an d g a s t r e c t o m i z e d
lo g s o n a n o rm a l d i e t w i l l c a u s e a m i l d a n e m ia i n t h e g a s t r e c t o m i z e d
lo g s w h ic h t e n d s t o r e c e d e t o n o r m a l, a n d a s lo w ly i n c r e a s i n g anem ia i n
lo rm a l d o g s .
I n b o t h i n s t a n c e s t h e a n e m ia i s r e l i e v e d by d i s c o n t i n u i n g
bhe i n d o l e .
^ n d o le i n s u f f i c i e n t d o s a g e w i l l c a u s e a m il d m a c r o c y t o s i s i n
lo r m a l a n d g a s t r e c t o m i z e d d o g s .
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B ib lio g ra p h y
( 1 ) M a r t i n , L . , H e l l e r s A r c h iv . p . 1 9 1 , 1 8 4 6 .
( 2 ) S im o n , C . S . , Am J . M ed. S c i . 1 1 0 : 4 8 , 1 8 9 5 .
( 3 ) P r o u t , P . , N a t u r e a n d T r e a tm e n t o f Stom ach an d U r i n a r y A f f e c t i o n s ,
L ondon, 1846, 3 rd e d . , p . 96.
( 4 ) N e n c k i , K ., B e r d e r D e u ts c h . Chem.G e s e l l p s c h . 9_:1 5 9 3 , 1 8 7 4 .
( 5 ) F o n t e s , G ., a n d
T h i v o l l e , L . , C om pt.
( 6 ) i b i d . 105: 969,
1930.
( 7 ) ib id . 105: 965,
1930.
( 8 ) i b i d . 106: 217,
1931.
( 9 ) i b i d .. 1 0 6 : 2 1 5 ,
1931.
(1 0 ) i b i d .l0 6 :
1931.
219,
( 1 1 ) B a a r , G . , W ie n e r A r c h iv . f .
R end. S o c . de B i o l . 1 0 5 : 9 6 7 , 1930
i n n . M ed. _5: 3 53, 1 9 2 3 .
( 1 2 ) N e n c k i , Y . , B e r d e r D e u ts c h . Chem. G e s e l l s c h . 1875 £ .2 9 8 .
( l 3 } H e r t e r , C .A ., Med. R e c . 5 3 : 5 69, 1 8 9 8 .
( 1 4 ) H e r t e r , C .A ., N .Y . M ed. -J. 68: 89 a n d 1 1 6 , 1 8 9 8 .
( 1 5 ) H e r t e r , C .A ., a n d Wakeman,
A .J . , J .
E xp. Med. 4: 3 0 7 , 1 8 9 9 .
(1 6 )W la d y c z k o , S . , A nn. de l ’ I n s t . P a s t e u r 7: 336, 1 9 1 3 .
(17)V aughan, S.
L . , P r o c . S o c . E x p e r. B i o l , an d Med. 29_:
(1 8 )E o i? b e s , J . C . a n d N e a l e , R .C ., J . L a b . C l i n . Med.
20:
523, 1932.
1017, 1935.
( 1 9 ) V a u g h a n , S .L . , J . ^ a b . C l i n . Med. 22: 39 9 , 1936.
(2 0 ) C a r n e s , H .E . , a n d L e m s , G .T ., J . La b . C l i n . Me d . 23: 4 5 9 , 1 9 3 8 .
( 21)O liv e t, J . ,
Z e i t f u r K l i n . Me d . 1 1 0 : 1 9 5 , 1929.
( 22)B ohm , P . , B io ch em . Z e i t s c h r . 2 9 0 : 1 3 7 , 19o7.
( 2 3 ) H o u s s a y , B .A ., D e u l o f e n , V ., an d M azzo cco , P . , C om pt. r e d d . S o c . de
B i o l 11 9 : 8 7 5 , 1935 and S o c . de B i o l , de B uenos A i r e s , A p r i l 4 , 1 9 3 5 ,
p .877.
$7
( 2 4 ) H o u s s a y , B .A ., D e u lo f e v , V ., a n d M azzo cco , P . , C om pt. r e d d . S o c . de
B i o l . 1 1 9 1 8 7 7 , 1 9 3 5 an d S o c . d e B i o l , de B uenos A i r e s , -^-pril 4 1935
9 .8 7 5 .
( 2 5 ) H o u s s a y , B .A ., Am. J . Me d . S c i , 1 9 2 : 6 15, 1 9 3 6 .
( 2 6 ) L a r o c h e , G ., a n d D e s b o r d e s , J . A nn. de M ed. 52: 2 2 1 , 1 9 3 2 .
( 2 7 ) B a r a c , G . , C om pt. r e d d . S o c . de B i o l . 1 3 2 : 4 1 , 1 9 3 9 .
( 2 8 ) N i c o l a i , E . , K l i n , K c h n s c h r. 1 8 : 1 3 3 8 , 1 9 3 9 ,
( 2 9 ) l n o u e , H .,, J .
C h o sen Me d . A s s . 24: 1 0 2 , 1 9 5 4 .
( 3 0 ) B u n g e le r , W ., Z e n t r a l b l . f .
a l l g . P a t h , und P a t h . A n a t. 60: 3 4 9 , 1 9 3 4 .
( 3 1 ) d e G a e t a n i , G .E ., M o rg a g n i 7 5 : 679, 1 9 3 3 .
( 3 2 ) d e G a e t a n i , G .E ., B iochem e T e r a p i a S p e r im e n t. 20: 3 8 2 , 1 9 3 3 .
( 3 3 ) d e G a e t a n i , G .E ., an d C o n s o l i , A ., H e m a to lo g ic a 17 : 7 7 , 1 9 3 6 .
( 3 4 ) E h o a d s , C . P , , P r o c . S o c . E x p e r . B i o l , a n d Med. 5 6 ; 6 52, 1 9 3 7 .
( 3 5 )R h o a d s , C . P . , a n d B a r k e r lAi.H., J. Exp. M ed. 6 7 : 26 7 , 1 9 3 8 .
(3 6 ) E h o a d s , C . P . , a n d M i l l e r , D .K ., J . Exp. M ed. 6 7 : 2 7 3 , 1 9 3 8 .
( 3 7 ) P h o a d s , C . P . , B a r k e r , lAi.H., a n d M i l l e r , D .K ., J . E xp. M ed. 6 7 :
299, 1938.
( 3 8 ) P o n d e r , E . , P r o c . S o c . E x p . B i o l , an d Med. 3 8 : 237, 1 9 3 8 .
( 3 9 ) T a y l o r , W .T ., Med. A nn. D i s t . o f C olu m b ia 8_: 3 6 3 , 1 9 5 9 .
( 4 0 ) P h o a d s , C .P * , C o ld S p r in g H a r b o r S ym posia o n q u a n t i t a t i v e B io lo g y
V o l. V, 1 9 3 7 , p . 4 1 0 .
( 4 1 ) D o b r i n e r , K. , a n d R h o a d s , C .P - , J . C l i n . In x re s. 17: 9 5 , 1 9 3 8 .
(4 2 )B i& l, M ., B e i t r .
B i e b l , M ., Z e i t .
z u r p a t h . a n a t . 84: 2 5 7 , 1 9 3 0 .
f .d .
i b i d . 87: 436, 1933.
i b i d . 88: 1 ,
1933.
i b i d . 88: 370, 1933.
i b i d . 891 5 2 9 , 1 9 3 3 .
g e s . e x p . M ed. 87: 4 1 6 , 1 9 3 3 .
98
i b i d * 91: 622, 1 9 3 3 .
i b i d . 93:
5 1 5 , -1934.
(4 3 )M a z z o c c o , P . ,
Comp. r e n d . S o c . de B i o l . 1 1 9 : 8 7 9 , 1935 a n d So:c. de
B i o l , de B u e n o s A i r e s , A p r i l 4 , 1 9 3 5 , p . 8 7 9 .
N orth w estern
University
Library
P a rt V
T he E f f e c t o f a G o ld b e r g e r D i e t on t h e G row th
a n d R ed B lo o d C e l l s o f R a ts
T h is e x p e r im e n t w as b e g u n i n c o n n e c t io n w i t h t h e e f f e c t o f i n d o l e
on t h e r e d b lo o d c e l l s o f d o g s .
T he o r i g i n a l i d e a was t o t e s t t h e e f f e c t
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tin ic
a c id d e f ic ie n t d i e t .
b e p e rfo rm e d on r a t s ,
T he a s s a y o f t h e d i e t f o r V ita m in B was t o
u s i n g t h e i r g ro w th c u r v e s .
I t w as th o u g h t
a d v i s a b l e t o d e t e r m i n e a t t h e sam e tim e t h e e f f e c t o f a n i c o t i n i c a c i d
d e f i c i e n t d i e t o n t h e r e d b lo o d c e l l s o f a d u l t a n im a ls t o s e e i f a
m a c r o c y t o s i s m ig h t d e v e lo p w h ic h w ould re s p o n d t o l i v e r e x t r a c t .
R e v ie w o f t h e L i t e r a t u r e
The n e c e s s i t y o f n i c o t i n i c a c i d i n th e d i e t o f t h e r a t f o r n o rm a l
g r o w th h a s r e c e i v e d m uch c o n s i d e r a t i o n .
R hoads a n d M i l l e r ( l )
f e d s i x r a t s a m o d if ie d G o ld b e rg e r d i e t
w h ic h c a u s e d a n e m ia , a c h l o r h y d r i a ,
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th is d ie t.
and l o s s o f h e m o p o ie tic e le m e n ts
A l l s i x r a t s g rew i n a nozm al m a n n e r on
I n a n o t h e r s e r i e s o f e x p e r im e n ts ( 2 ) , y o u n g r a t s f e d a d i e t
w h ic h c a u s e d c a n i n e b l a c k to n g u e m a i n ta i n e d a n o rm a l g ro w th r a t e .
U s in g G o ld b e r g e r d i e t s 123 a n d 3 2 4 , B o o k e r an d H ausm ann (3 )
f o u n d t h a t t h e a d d i t i o n o f 40 mgm. p e r d a y o f a V ita m in G c o n c e n t r a t e
im p ro v e d t h e g r o w th o f t w e n t y - e i g h t d ay o l d r a t s on t h e d i e t s .
B i r c h , C h ic k , a n d M a r t i n (4 ) m o d if ie d t h e G o ld b e r g e r d i e t 123
b y o m i t t i n g t h e s u c r o s e a n d c o t t o n s e e d o i l an d r a i s i n g t h e c o rn c o n t e n t
t o 80 % o f t h e d i e t .
T h r e e g r o u p s o f r a t s w e re f e d t h e d i e t .
One g ro u p
r e c e i v e d t h e d i e t a n d l a c t o f l a v i n , o n e g ro u p t h e d i e t a n d a u t o c l a v e d
y e a s t,
and t h e t h i r d g ro u p j u s t t h e d i e t .
e q u a lly a s w e ll.
P .P .
d e fic ie n c y .
A l l t h r e e g r o u p s g rew
The a u t h o r s c o n c lu d e d t h a t r a t s w ere n o t s e n s i t i v e t o a
101
L a r g e r f o o d i n t a k e s a n d b e t t e r w e ig h ts t h a n t h e c o n t r o l s w e re
o b t a i n e d by Funk a n d F u n k (5 ) when n i c o t i n i c a c i d o r n i c o t i n a m i d e w e re
a d d e d t o e i t h e r o f tw o d i e t s .
The d i e t s w e re ( l )
p u r i f i e d fo o d s and
co d l i v e r o i l a n d ( 2 ) a r t i f i c i a l d i e t a n d v ita m i n c o m p le x .
N o rm al g r o w th i n r a t s on a d e f i c i e n t d i e t r e s e m b li n g a b l a c k
t o n g u e d i e t w as in d u c e d b y am yl a l c o h o l e x t r a c t s o f l i v e r e x t r a c t s .
T h i s w as shown b y M a c ra e and E d g a r (6 ) t o b e some s u b s ta n c e o t h e r t h a n
n ic o tin ic a c id .
II
N e i t h e r n i c o t i n i c a c i d , n i c o t i n a m i d e , o r c o h y d ro g e n a s e
c o u ld r e p l a c e y e a s t f i l t r a t e o r e l u a t e f a c t o r s i n t h e d i e t o f r a t s .
T he same o b s e r v a t i o n s w e re m ade by Cook, C l a r k e , a n d L i g h t ( 7 ) .
T hey a l s o fo u n d t h a t t h e r e was no s u p p le m e n ta r y e f f e c t on g ro w th when
n i c o t i n i c a c i d w as a d d e d t o c o n c e n t r a t e s fro m r i c e p o l i s h i n g s o r y e a s t
a d e n y lic o r n u c le ic a c id s .
B a e r so n a n d E v a n s (8 ) fo u n d t h a t r a t s r e c e i v i n g a d i e t i n w h ic h
a l l o f t h e known f a c t o r s o f t h e V ita m in Bg com plex w e re s u p p li e d e x c e p t
t h a t o f t h e m u ltip ly a b so rb e d l i v e r f i l t r a t e
f a i l e d t o g row .
G row th
w as n o t r e s t o r e d b y n i c o t i n i c a c i d o r y e a s t n u c l e i c a c i d , o r a com­
b i n a t i o n o f t h e tw o .
T h e s e f i n d i n g s a r e c o n fir m e d by 1 1 - S a d r a t a l . ( 9 ) .
T he c o n c l u s i o n s r e a c h e d by C h ic k , M a c ra e , an d M a r tin (1 0 ) when
r a t s w e re f e d a d i e t c a u s a t i v e o f sw in e p e l l a g r a vfere:
( 1 ) N ic o t in a m i d e o r n i c o t i n i c a c i d
c o n s titu e n ts f o r th e r a t ,
s re n o t e s s e n tia l d ie ta ry
or
(2 )T h e r e q u i r e m e n t o f t h e r a t f o r t h e s e two i s v e r y s m a ll i n
c o m p a r is o n w i t h t h e p i g .
C o m p le te n i c o t i n i c a c i d d e f i c i e n c y f o r 150 d a y s was a c c o m p a n ie d
b y no s p e c i f i c p h y s i o l o g i c a l d i s f u n c t i o n i n r a t s on a s y n t h e t i c d i e t .
T h o s e a n im a ls on t h e d i e t who r e c e i v e d n i c o t i n i c a c i d h a d g ro w th c u r v e s
102
s i m i l a r to th e -u n tre a te d r a t s ( B ir s c h , 1 1 ).
H e lm e r a n d F o u t s (1 2 ) a n d M a r g o l i s , M a r g o l i s , an d S m ith (1 3 )
h a v e r e p o r t e d t h a t t h e G o ld b e r g e r d i e t i s d e f i c i e n t i n r i b o f l a v i n a n d
V ita m in B-j_ a s w e l l a s som e s u b s t a n c e c o n t a i n e d i n l i v e r e x t r a c t w h ic h
p r o m o te s t h e g r o w th o f r a t s .
H e lm e r an d F o u ts fo u n d t h a t t h e a d d i t i o n
o f n i c o t i n i c a c i d d i d n o t c a u s e a n i n c r e a s e i n w e ig h t p e r week a s g r e a t
a s t h a t c a u s e d b y a d d in g l i v e r e x t r a c t t o t h e d i e t .
R ib o f la v in a lo n e
was n o t s u f f i c i e n t t o r e s t o r e n o rm a l g ro w th , b u t r i b o f l a v i n a n d n i c o ­
t i n i c a c i d t o g e t h e r w e re b e t t e r t h a n e i t h e r a l o n e .
Maximum g ro w th w as
o b t a i n e d w i t h l i v e r e x t r a c t , r i b o f l a v i n a n d an d n i c o t i n i c a c i d c o m b in e d .
M a rg o lis e t a l
fo u n d t h a t dogs s u b je c te d to r e p e a te d a tta c k s o f b la c k
to n g u e e v e n t u a l l y f a i l e d t o r e s p o n d t o n i c o t i n i c a c i d , b u t w ould r e s p o n d
bo l i v e r e x t r a c t .
A com bined s u p p le m e n t o f n i c o t i n i c a c i d , t h i a m i n and
r ib o f la v i n gave r e s u l t s
s im ila r to l i v e r e x tr a c t.
U n l i k e o t h e r a n im a ls w h ic h t h e y i n v e s t i g a t e d , H a r r i s an d Raymond
(1 4 )
r e p o r t e d t h a t e v e n a f t e r lo n g p e r i o d s on a n i c o t i n i c a c i d f r e e d i e t ,
ra ts
c o n ti n u e d t o e x c r e t e n i c o t i n i c a c i d i n t h e u r i n e .
3U g g est t h a t t h e r a t
T h is w ould
s y n t h e s i z e s n i c o t i n i c a c i d o r o r g a n is m s i n t h e g a stro <
I n t e s t i n a l c a n a l o f t h e r a t s y n t h e s i z e t h e s u b s ta n c e a n d make i t a v a i l a b l e
;o t h e r a t .
G u in e a p i g s , on t h e o t h e r h a n d , w ere f o u n d by H a r r i s a n d Raymond
t o e x c r e t e d e c r e s i n g a m o u n ts o f n i c o t i n i c a c i d i n t h e u r i n e u n t i l when
d e f i c i e n c y sym ptom s s e t i n t h e r e i s no n i c o t i n i c a c i d s e c r e t e d i n t h e
u rin e .
T h is i s i n l i n e w i t h t h e e a r l i e r w ork o f M i l l e r an d R hoads (1 5 )
who r e p o r t e d t h a t g u i n e a p i g s on a m o d if ie d G o ld b e r g e r d i e t d i e d u n l e s s
g i v e n a s u p p le m e n t o f l i v e r e x t r a c t o r a n a u t o l y s e d y e a s t p r e p a r a t i o n
(V e g e x ).
S in c e l i v e r e x t r a c t p r e v e n t e d o r c u re d t h e d i s e a s e w h ile
103
v e n t r i c u l i n w as w i t h o u t a f f e c t ,
is
t h e y c o n c lu d e d t h a t " t h e d e f i c i e n c y w h ic h
c a u s a tiv e o f c a n in e b la c k to n g u e i s c lo s e l y a l l i e d t o th e d e f ic ie n c y
w h ic h i s e t i o l o g i c i n p e r n i c i o u s a n e m ia " .
T hey s u g g e s te d t h a t t h e d e a th
o r s u r v i v a l o f g u i n e a p i g s f e d a b l a c k to n g u e d i e t m ig h t s e r v e a s a t e s t
f o r t h e p o t e n c y o f s u b s t a n c e s a c t i v e i n t h e t r e a t m e n t o f p e r n i c i o u s a n e m ia .
N ot o n ly i s th e t i t l e
o f t h e a r t i c l e " A b se n c e o f d i e t a r y A n ti-A n e m ia
s u b s t a n c e s i n d i e t c a u s a t i v e o f b l a c k to n g u e " m i s l e a d i n g , b u t a l s o no
d a t a c o n c e r n i n g a n e m ia a r e m e n tio n e d i n t h i s p a p e r .
T h e r e h a v e b e e n , h o w e v e r, some i n t e r e s t i n g o b s e r v a t i o n o f t h e
e f f e c t s o f V ita m in B d e f i c i e n t d i e t s on t h e b lo o d o f r a t s .
G y o rg y e t a l .
(1 6 ) o b s e r v e d t h a t r a t s k e p t on a s y n t h e t i c d i e t c o n t a i n i n g V ita m in B^,
la c to fla v in ,
a n d V ita m in B g , i n a d d i t i o n t o t h e o t h e r e s s e n t i a l v i t a m i n s ,
d e v e lo p e d n a s a l h e m o r r h a g e , t e a r h e m o rrh a g e , a n d p u r p u r a t o g e t h e r w i t h a
p a n m y e lo th is is .
T h is c o n d i t i o n c o u ld b e c u r e d o r p r e v e n t e d b y s u p p le ­
m e n tin g t h e d i e t w i t h P e t e r s e l u a t e — a n e l u t i o n o f a c h a r c o a l a d s o r b a t e
3f a n a q u e o u s y e a s t e x t r a c t w i t h a c i d a l c o h o l .
L a t e r (1 7 ) t h e s e o b s e r ­
v e rs r e p o r t e d t h a t t h e a d d i t i o n o f n i c o t i n i c a c i d t o t h e d e f i c i e n t
l i e t w o u ld c a u s e a c o m p le te s u p p r e s s i o n o f t h e p a n m y e l o p h t h i s i s .
I f th e
r a t s w e re a n e m ic , t h e n i c o t i n i c a c i d w o u ld c a u s e r e c o v e r y .
R a t s f e d a d i e t c o n t a i n i n g o n l y t h i a m i n a n d l a c t o f l a v i n fro m t h e
fB" g ro u p w e re r e p o r t e d b y O e s t r e i c h e r a n d O v e rb e e k ( 1 8 ) t o h a v e a
p r o l o n g a t i o n o f t h e p h y s i o l o g i c a l l e u k o p e n i a fo u n d i n y o u t h .
) lo o d c e l l p i c t u r e w as h a r d l y a f f e c t e d .
The r e d
N i c o t i n i c a c i d h a d no e f f e c t
m t h i s le u k o p e n ia .
One o t h e r r e p o r t s h o u ld b e m e n tio n e d o n l y t o p o i n t o u t t h a t
r e p e t i t i o n o f t h e same e x p e r im e n t i n c a s e o f d o u b t i s n o t o n l y d e s i r a b l e
b u t n e c e ssa ry .
W i l l s a n d M e h ta (1 9 ) f o u n d t h a t r a t s f e d a d i e t r e s e m b l i n g
104
t h a t o f Mohammedan a n d H in d u women i n I n i d a d e v e lo p e d B a r t o n e l l a a n e m ia .
T h i s d i e t i s lo w i n b o t h V i t a m in s A an d B.
The s t r i k i n g t h i n g a b o u t t h i s
r e p o r t i s t h a t t h e r a t s a l l d e v e lo p e d t h e a n e m ia w i t h i n t e n d a y s o f one
a n o t h e r a l t h o u g h t h e y w e re a t v a r i o u s p e r i o d s i n t h e d i e t fro m 44 t o 104
days.
T h i s w o u ld a p p e a r t o h a v e e v e r y s i g n o f b e in g an e p id e m ic .
w ork w as n o t r e p e a t e d t o s e e i f
The
t h e phenom enon c o u ld b e o b s e r v e d a g a i n .
B a r t o n e l l a b o d i e s w e re fo u n d i n t h e b lo o d s tr e a m a n d t h e a n im a ls a l l h a d
t h e u s u a l sym ptom s a n d s i g n s o f a b a r t o n e l l a i n f e c t i o n .
E x p e r im e n t a l M eth o d s
Two g r o u p s o f r a t s fro m s t o c k a l b i n o r a t s w ere s e l e c t e d f o r t h i s
e x p e rim e n t.
One g ro u p o f tw e n ty o n e r a t s (fro m tw o l i t t e r s )
th a t l i t t e r
w e re d i v i d e d so
m a te s w e re p l a c e d i n t h e c o n t r o l and t r e a t e d g r o u p s — t e n
t r e a t e d and e le v e n c o n tr o l a n im a ls .
T h e se a n im a ls w ere w eaned a t tw e n ty
o n e d a y s o f a g e a n d p l a c e d on t h e i r r e s p e c t i v e d i e t s .
The c o n t r o l a n im a ls
r e c e i v e d P u r i n a dog c h e c k e r s a n d t h e t r e a t e d a n im a ls r e c e i v e d t h e m o d if ie d
G o ld b e r g e r d i e t ( v i d e i n f r a ) .
E ach a n im a l r e c e i v e d a s a s u p p le m e n t t o t h e
d i e t 20 Gammas e a c h o f l a c t o f l a v i n an d t h ia m i n e c h l o r i d e d a i l y .
The
a n im a ls w e re w e ig h e d d a i l y f o r tw o m o n th s an d w e re c o n tin u e d on t h e d i e t s
Tor o n e y e a r .
A t t h e e n d o f o n e y e a r c o m p le te a u t o p s i e s w e re p e rf o r m e d on
a l l o f t h e r a t s a n d m i c r o s c o p i c s e c t i o n s w e re m ade o f t h e b o n e m arrow
fro m v e r t e b r a e , u p p e r , m id d l e , a n d lo w e r p o r t i o n s o f t h e f e m u r , a s w e ll
a s s e c tio n s o f th e l i v e r ,
s to m a c h , s m a ll and l a r g e i n t e s t i n e s , k i d n e y s ,
s p le e n , lu n g s , and h e a r t.
T he o t h e r g r o u p o f r a t s ,
o n e l i t t e r o f f o u r t e e n r a t s , w e re
d i v i d e d i n t o t e n t r e a t e d a n d f o u r c o n t r o l a n im a ls .
They r e c e i v e d t h e
same
105
d i e t s a s th e f i r s t g ro u p .
T he a n im a ls w e re p l a c e d on t h e d i e t s a t 158
d ay s o f a g e and k e p t on t h e d i e t u n t i l d e a th o r te r m in a tio n o f th e
e x p e r im e n t a f t e r 575 d a y s ( 7 3 3 d a y s o f a g e ) .
Red b lo o d c e l l c o u n ts an d
h e m o g lo b in d e t e r m i n a t i o n s w e re m ade a t i r r e g u l a r i n t e r v a l s th r o u g h o u t t h e
e x p e rim e n ta l p e r io d .
T he d i e t u s e d w as a m o d i f i c a t i o n o f G o ld b e r g e r 1 s d i e t num ber 2 6 8 . {19)
M o d if i e d G o ld b e r g e r d i e t
C o rn m e a l
1600 gram s
C a l if o r n ia b la c k eyed p e a s
185
C o tto n se e d o i l
125
Cod l i v e r o i l
100
C a s e in
225
C a lc iu m c a r b o n a t e
15
Sodium c h l o r i d e
15
The p e a s w e re c o o k e d i n w a t e r , t o w hidh t h e s a l t h ad b e e n a d d e d ,
u n t i l a m ush w as o b t a i n e d , t h e n t h e c o r n m ea l an d c a s e i n w e re a d d e d
t o g e t h e r w i t h e n o u g h w a t e r t o fo rm a s t i f f m ush, an d co o k e d u n t i l s o f t .
The r e s t o f t h e i n g r e d i e n t s were, a d d e d and t h e d i e t m ix e d c o m p l e te l y ,
c o o le d , a n d s to r e d i n
th e ic e box.
T he d i e t was p r e p a r e d f r e s h e a c h w eek,
i r o n w as n o t a d d e d t o t h e d i e t b e c a u s e i t w as b e l i e v e d t h a t enough w as
p r e s e n t and a v a il a b l e i n th e in g r e d ie n ts u se d .
The c o m p le te a v a i l a b i l i t y
o f t h e i r o n i n th e b la c k eyed p e a s h a s been d e m o n s tra te d (2 0 ).
R e s u lts
T he a v e r a g e w e i g h ts o f t h e c o n t r o l and t r e a t e d a n im a ls a r e g i v e n
in T a b le I .
I t w i l l b e s e e n t h a t a l t h o u g h t h e a n im a ls r e c e i v i n g t h e
G o l d b e r g e r d i e t c o n t i n u e d t o g ro w a f t e r b e i n g p l a c e d on t h e d i e t , t h e i r
g r o w th w as much s l o w e r t h a n t h a t o f t h e c o n t r o l s .
A t t h e end o f two
m o n th s , t h e t r e a t e d a n im a ls w e ig h t a p p r o x i m a t e ly o n e - h a l f a s much a s t h e
c o n tro ls .
A t t h e en d o f a y e a r t h e y w e ig h e d a b o u t t w o - t h i r d s a s m uch a s
th e c o n tr o ls .
A t t h e en d o f a y e a r on t h e d i e t ,
A n im a ls
N um ber
C o n tro l
7
Red b lo o d
c e lls
8 .7 8
T re a te d
8
8 .5 4
t h e b lo o d v a l u e s w e re :
H em oglobin
1 2 .7 1
1 1 .9 8
T h e s e f i g u r e s a r e v e r y s i m i l a r a n d w e l l w i t h i n t h e n o rm a l r a n g e
f o r r a t s i n o u r c o lo n y .
A t n o tim e d i d t h e r a t s on t h e G o ld b e r g e r d i e t show a n y
u n u s u a l sym ptom s.
th e y e a r .
One d i e d o f m id d le e a r d i s e a s e and one e s c a p e d d u r in g
Of t h e t r e a t e d a n im a ls , o n e d i e d o f m id d le e a r d i s e a s e and
t h r e e e s c a p e d d u r in g t h e y e a r .
A t a u t o p s y , t h e r e w e re no g r o s s d i f f e r e n c e s b e tw e e n t h e t r e a t e d
and c o n t r o l a n im a ls e x c e p t f o r t h e d i f f e r e n c e s i n w e i g h t.
E x a m in a tio n
o f t h e m ic r o s c o p i c s e c t i o n s a l s o r e v e a l e d no d i f f e r e n c e i n t h e two g r o u p s
T he d a t a c o n c e r n in g t h e s e c o n d g r o u p o f r a t s a r e g iv e n i n T a b le
[I.
The m o s t s t r i k i n g t h i n g i s t h e p l a t e a u i n w e ig h t i n t h e a n im a ls
r e c e i v i n g t h e G o ld b e r g e r d i e t .
A f t e r a n a v e r a g e g a in o f tw e n ty gram s t h e
F i r s t w e e k , t h e r e w as v e r y l i t t l e
f l u c t u a t i o n i n t h e w e ig h t o f t h e a n im a l
l u r i n g t h e r e m a in d e r o f t h e e x p e r im e n t.
The r e d b lo o d c e l l num bers a n d
h e m o g lo b in c o n t e n t r e m a in e d w e ll w i t h i n t h e n o rm a l l i m i t s t h r o u g h o u t t h e
e n t i r e e x p e rim e n t.
A l l o f t h e r a t s l o s t fro m t h e e x p e r im e n t e i t h e r
e s c a p e d ( 4 ) o r d ie d o f m id d le e a r d i s e a s e ( 3 ) .
107
Table I
H a ts o n G o l d b e r g e r D ie t
A ll s t a r t e d on 2 1 s t day o f age
T re a te d
A n im als
10
10
10
10
10
10
10
10
10
10
10
10
8
8
A v e ra g e
w e ig h t
21
23
26
27
31
37
40
44
51
56
62
74
138
176
C o n tro ls
A v e ra g e
g a in
0
M2
n
*6
/3
"f4
■hi
+5
/6
te
Days on
d ie t
0
3
7
11
16
21
26
31
36
40
45
52
228
365
^ A n im als
11
11
11
11
11
11
11
11
11
11
11
11
6
7
A v e ra g e
w e ig h t
Aver?
g a in
21
29
36
47
62
73
94
105
}.24
136
144
154
225
255
0
TQ
+1
/-ll
M15
-M l
>-81
-+11
ML2
+ 8
■y-10
108
T a b le I I
R a t s o n G o ld b e r g e r D i e t
15 8 d a y s o l d when p l a c e d on d i e t
733 d a y s o l d a t end o f p e r i o d
T re a te d
^ R a ts
10
10
10
9
9
' 9
9
9
9
9
9
9
9
9
9
9
9
6
5
A v e ra g e
w e ig h t
gm s.
179
184
204
205
209
214
184
200
200
202
195
200
204
218
248
224
C o n t r o ls
R .B .C .
X 106/
Cu.Mm.
Hemo­
Days
g lo b in
d ie t
Gms/lOOc c .
9 .3 2
9 .3 0
9 .7 5
9 .7 1
9.2.0
1 0 .4 2
9 .0 0
9 .4 8
9 .6 5
9 .7 3
9 .5 0
9 .5 4
9 .2 1
9 .5 0
9 .0 9
8 .9 8
9 .4 4
9 .5 2
8 .0 0
1 2 .1 1
1 3 .5 0
1 3 .7 7
1 3 .1 7
1 3 .0 7
1 5 .7 5
1 3 .6 4
1 6 .5 0
1 5 .5 4
1 6 .1 4
1 5 .1 0
1 4 .0 7
1 5 .7 5
1 3 .0 1
1 4 .7 5
1 3 .5 0
1 5 .4 4
1 6 .4 4
1 3 .7 0
0
0
7
21
84
114
128
142
156
170
184
198
216
250
283
334
395
494
575
# R a ts
A v e ra g e
w e ig h t
gm s.
R .B .C . HemoX 1 0 6/
g lo b in
Cu. Mm. G m s/100 cc*
180
4
4
4
4
4
4
4
4
4
3
3
2
2
220
233
237
242
234
234
246
254
317
288
9 .5 3
9 .5 8
9 .3 7
9 .0 3
9 .4 0
9 .3 4
8 .7 0
9 .8 0
9 .8 2
8 .1 3
9 .1 3
8 .4 5
7 .9 1
1 6 .1 4
1 7 .3 6
1 6 .6 6
1 5 .5 7
1 5 .5 9
1 4 .9 5
1 7 .0 0
1 2 .9 2
1 6 .4 3
1 3 .2 6
1 5 .9 2
1 5 .7 3
1 4 .5 1
109
D is c u s s io n
From t h e d a t a g i v e n , i t
i s o b v io u s t h a t t h e m o d if ie d G o ld b e rg e r
d i e t i s d e f i c i e n t i n some s u b s t a n c e n e c e s s a r y t o p ro m o te g ro w th i n y oung
and a d u l t r a t s .
e v id e n t.
T hat t h i s
s u b s t a n c e i s n o t r i b o f l a v i n o r t h ia m in e i s a l s o
S i n c e , h o w e v e r, n i c o t i n i c a c i d w as n o t g iv e n t o s e e i f i t w ould
p ro m o te t h e g ro w th o f t h e a n i m a l s , we c a n n o t s a y w h e th e r n i c o t i n i c a c i d i s
th e la c k i n g s u b s ta n c e o r n o t .
From t h e o b s e r v a t i o n s o f o t h e r s , i t w ould
a p p e a r t h a t i t i s n o t n i c o t i n i c a c i d w h ic h i s l a c k i n g b u t some o t h e r su b ­
s ta n c e p r e s e n t i n l i v e r e x t r a c t s .
C e r t a i n l y t h e d e f i c i e n c y i n no way im p a ir s t h e b lo o d fo rm in g
riech an ism i n t h e r a t ,
e v e n a f t e r p r o lo n g e d f e e d i n g o f t h e d i e t .
We a r e a t a l o s s t o e x p l a i n why t h e G o ld b e rg e r d i e t u s e d i n
; h i s e x p e r im e n t f a i l e d t o p ro m o te g ro w th n o r m a l ly , w h ile o t h e r e x p e r a e n t e r s h a v e fo u n d t h a t t h e d i e t w i l l p ro m o te g ro w th .
T h e re a r e no
g r o s s o r m ic r o s c o p i c o b s e r v a t i o n s w h ic h w i l l a c o u n t f o r t h i s d i f f e r e n c e .
We a r e a l s o a t a l o s s t o e x p l a i n o u r i n a b i l i t y t o p r o d u c e a p a n n y e l o p h t h i s i s i n t h e r a t s a s d i d G y o rg y .
A t no tim e d u r i n g t h e e x p e rim e n t
l i d we s e e a n y o f t h e sym ptom s d e s c r i b e d by G yorgy.
T he o b s e r v a t i o n s o f V a i l s and M ehta c o n c e r n in g t h e o c c u r r e n c e o f
3 a r t o n e l l a a n e m ia i n r a t s on a d e f i c i e n t d i e t s i m i l a r t o o u r s i s i n t e r ­
e s t i n g i n t h a t a l t h o u g h o u r c o lo n y i s known to be i n f e s t e d w ith B a r to n ­
e l l a , we d i d n o t s e e a n y e v id e n c e o f B a r t o n e l l a anem ia i n a n y o f t h e r a t s
ise d .
It
i s v e r y p r o b a b l e t h a t t h e an e m ia w h ich w i l l s o b s e r v e d was i n t h e
a a t u r e o f a n e p id e m ic .
We h a v e n e v e r s e e n an e p id e m ic o f B a r t o n e l l a
i n f e c t i o n s i n n o rm a l r a t s .
I t m ig h t b e o o s s i b l e t h a t i n t h e f a c e o f a
s e v e r e d e f i c i e n c y , b o d i l y d e f e n s e s m ig h t b e lo w e re d enough t o p e r m it a n
110
o v e rw h e lm in g B a r t o n e l l a i n v a s i o n , h u t b e c a u s e o f t h e w id e ly s e p a r a t e d p e r i o d s
o n t h e d i e t a n d t h e o c c u r r e n c e o f t h e a n e m ia i n a l l o f t h e r a t s w i t h i n t e n
d a y s o f o n e a n o t h e r , t h e e v id e n c e w ould seem t o p o i n t m ore t o an e p id e m ic
t h a n t o a d i s e a s e a r i s i n g fro m t h e d e f i c i e n t d i e t .
C o n c lu s io n s
T he m o d if ie d G o l d b e r g e r d i e t 268 u s e d i n t h i s e x p e rim e n t i s
d e fic ie n t in
some s u b s t a n c e n e c e s s a r y t o p ro m o te g r o w th i n y oung and a d u l t
p a ts .
P r o lo n g e d f e e d i n g o f a m o d if ie d G o ld b e r g e r d i e t i s w ith o u t e f f e c t
lp o n t h e r e d b lo o d c e l l s o f r a t s .
A d m i n i s t r a t i o n o f t h e m o d if ie d G o ld b e r g e r d i e t t o y o u n g r a t s
from w e a n in g t o o v e r o n e y e a r o f a g e , a l t h o u g h n o t p r o m o tin g g ro w th i n a
lo rm a l m a n n e r , h a d no e f f e c t s w h ic h c o u ld b e d e m o n stra te d a t a u to p s y
e i t h e r t h r o u g h g r o s s o r m ic r o s c o p i c i n s p e c t i o n .
Ill
B ib lio g ra p h y
( 1 ) R h o a d s , C . P . , a n d M i l l e r , D .K ., J . C l i n . I m r e s . 1 4 : 1 5 3 , 1 9 3 5 .
( 2 )R h o a d s , C . P . , a n d M i l l e r , D .K ., S c ie n c e 8 1 : 1 5 9 , 1 9 3 5 .
( 3 ) B o o k e r, L . E . , a n d H ausm ann, G .H ., Am. J . P h y s i o l . 1 1 4 : 4 2 9 , 1 9 3 5 .
( 4 } B i r c h , T .W ., C h ic k , H . , a n d M a r t i n , C .T ., B io ch em . J . 31: 2 0 6 5 , 1 9 3 7 .
( 5 ) P u n k , C ., a n d P u n k , I . C . , J . B i o l . Chem 1 1 9 : XXXV, 1 9 3 7 .
( 6 )M a c ra e , T . F . , a n d E d g a r, C . E ., B iochem J .
31: 2 2 2 5 , 1 9 3 7 .
{ 7 )C o o k , C .A ., C l a r k e , M .E. a n d L i g h t , A . E . , P r o c . S o c . E x p e r . B i o l ,
a n d Me. 3 7 : 5 1 4 , 1 9 3 7 .
(8 ) E m e rso n , G .A ., a n d E v a n s , H .M ., P r o c . S o c . E x p e r. B i o l , and Med. 3 8 :
195, 1938.
( 9 ) E 1 - S a d r , M .M ., H in d , H .G ., M a c ra e , T . E . , an d W ork, C .E «, N a t u r e 144:
73, 1939.
( 1 0 ) ^ h i c k , H . , M a c ra e , T . E . , M a r t i n , A * T .P ., and M a r tin , C . J . ,
J . 32: 8 4 4 , 1 9 3 8 .
B iochem .
(1 1 )B lirc h , T .W ., J . N u t r i t i o n 1 7 : 2 3 1 , 1 9 3 9 .
( 1 2 )H e lm e r, O .M ., a n d F o u t s , P . J . , J . N u t r i t i o n 1 6 : 2 71, 1 9 3 8 .
( 1 3 ) M a r g o l i s , L .H ., M a r g o l i s , G e o ., and .Sm ith, S . G . , J . N u t r i t i o n 17:
63, 1939.
( 1 4 ) H a r r i s , L . J . , a n d Raym ond, _W .D ., B iochem . J .
33: 2036, 1 9 3 9 .
( 1 5 ) M i l l e r , D.K . a n d R h o a d s, C . P . , P r o c . S o c . E x p e r. B i o l . Med. 32:
419, 1934.
( 1 6 ) G y o rg y , P . , G o l d b l a t t , H ., M i l l e r , E .R * , a n d F u l t o n , R . P . , J . Exp.
Med. 66: 5 7 9 , 1 9 3 7 .
( 1 7 ) G y o rg y , P . , P r o c . S o c . E x p . B i o l , an d M ed. 37* 732, 1 9 o 8 .
(1 8 )G e s t r e i c h e r , F . , and O v e rb e e k , G .A ., A c ta . B re n . N e e r la n d . 9: 2 4 9 , 1 9 3 9 .
1 9 ) W i l l s , L . , a n d M e h ta , M.M. , I n d . J . Med. R e s . L3. 665, 19e>0.
(2 0 ) G o ld b e r g e r , I . , an d S c h r e l l , W .A ., W eekly U .S . P u b l i c H e a l t h R e p t s .
4 5 : 3064, 1 9 3 0 .
[ 2 1 ) A seham , L . , S p e i r s , M ., an d M addox, D ., S c ie n c e 9 0 : 5 9 6 , 1 9 3 9 .
112
E p i lo g u e
We h a v e f a i l e d t o i n d u c e a m a c r o c y t i c a n e m ia , r e s e m b lin g p e r n i c i o u s
a n e m ia , i n m o n k e y s ,
s w in e , d o g s , an d r a t s th r o u g h t h e a d m i n i s t r a t i o n o f
d e f i c i e n t d i e t s o r a h e m o ly tic s u b s t a n c e .
We h a v e f a i l e d t o a l t e r a
n a t u r a l o c c u r r i n g m a c r o c y t o s i s i n n ew born r a t s t h r o u g h t h e a d m i n i s t r a t i o n
o f s u b s t a n c e s know n t o b e e f f e c t i v e i n p e r n i c i o u s a n e m ia ,
We h a v e demon­
s t r a t e d t h a t t h e f e t a l c a l f p o s s e s s e s i n i t s l i v e r an a n t i - p e r n i c i o u s
a n e m ia s u b s t a n c e .
T h e s e e x p e r im e n ts o n l y a d d t o a n e v e r i n c r e a s i n g l i s t
t o p r o d u c e a n a n e m ia i n a n im a ls s i m i l a r t o p e r n i c i o u s a n e m ia .
Df t h e e t i o l o g y o f p e r n i c i o u s a n e m ia i s
of fa ilu re s
The s o l u t i o n
so i n t i m a t e l y l i n k e d w i t h t h e
l e c e s s i t y o f o b t a i n i n g a n im a ls w h ic h c a n be u s e d t o a s s a y a n t i - p e r n i c i o u s
a n e m ia s u b s t a n c e s a s t o c a u s e a v i c i o u s c i r c l e .
A n im a ls w i t h t h e a n em ia
iTOuld l e a d r e a d i l y t o t h e s u b s t a n c e r e s p o n s i b l e f o r r e m i s s i o n s w h ic h w ould
.e a d o u i c k l y t o t h e e t i o l o g i c f a c t o r s .
On t h e o t h e r hand i f we knew t h e
s t i o l o g i e f a c t o r s , t h e n e x p e r i m e n t a l a n im a ls c o u ld p r o b a b ly b e f o u n d ,
a n d a m ore c o n v e n ie n t a s s a y m e th o d w ould b e e v o lv e d .
The p r e s e n t l i m i ­
t a t i o n s o f i n v e s t i g a t i o n s on p e r n i c i o u s a n em ia p a t i e n t s h a s h i n d e r e d p r o g r e s s
] L a rk e d ly .
T he i n a b i l i t y t o r e p r o d u c e t h e d i s e a s e i n a n im a ls s u g g e s t s two
]lo s s ib ilitie s .
l . T h e a n im a ls so f a r i n v e s t i g a t e d do n o t r e q u i r e t h e a n t i - p e r n i c i o u s
a n e m ia s u b s t a n c e .
2 . I f t h e s u b s ta n c e i s r e q u i r e d , t h e a n im a ls a r e
b le to
s y n th e s iz e
t h e s u b s t a n c e o r h a v e i n t h e i r g a s t r o - i n t e s t i n a l c o n t e n t s o rg a n is m s
c a p a b l e o f p r o d u c in g t h e s u b s ta n c e o r i t s p r e c u r s o r w h ic h t h e
a n im a l a b s o r b s and u t i l i z e s .
113
T he l a t t e r t h e o r y i s n o t a n i m p o s s i b l e o n e .
It
i s known t h a t
a n im a ls o r o r g a n is m s t h e y c o n t a i n a r e a b l e t o s y n t h e s i z e c e r t a i n o f t h e
v ita m in s .
I t i s n o t i m p o s s i b l e t h a t t h e p e r n i c i o u s a n e m ia p a t i e n t f a i l s
to
p r o v id e t h e s e o r g a n is m s w i t h t h e p r o p e r s u b s t r a t e f o r p r o d u c in g t h e
a n t i - p e r n i c i o u s a n e m ia s u b s t a n c e o r i t s p r e c u r s o r .
Df
T h i s may b e t h e r e s u l t
t h e a c h l o r y h y d r i a a n d a c h y l i a s e e n i n p e r n i c i o u s a n e m ia .
De a n enzym e i n t h e g a s t r i c j u i c e w h ic h p r o v i d e s t h i s
In g e s te d fo o d .
s u b s t r a t e fro m
T h ere h a v e been i n d ic a ti o n s o f t h e p o s s i b i l i t y o f o th e r
g a s t r i c enzym es o t h e r t h a n t h o s e a l r e a d y d e s c r i b e d .
Df
T h e re may
in c u b a tin g g a s tr ic
C e r ta in ly th e e f f e c t
j u i c e a n d m e a t, f e e d i n g v e n t r i c u l i n , o r d u o d e n a l
lu c o s a m ig h t e q u a l l y a s w e l l b e u s e d a s a n a rg u m e n t f o r t h e p r e s e n c e o f a
3 p e c i f i c enzym e a s f o r t h e p r e s e n c e o f an " i n t r i n s i c ” an d " e x t r i n s i c " f a c t o r .
P e r n i c i o u s a n e m ia r e m a in s a n o d d i t y i n t h e m e d ic a l w o r ld .
;he d i s e a s e c a n n o t b e c u r e d , r e m i s s i o n s c a n b e o b t a i n e d .
A lth o u g h
By w hat H u b s ta n c e
; h i s i s b r o u g h t a b o u t , o r how t h e l a c k o f t h i s s u b s ta n c e o r i g i n a t e s , we
ire s t i l l a t a l o s s to d e s c r ib e .
A ck n ow ledgem ent s
I w ish, t o a c k n o w le d g e t h e i n v a l u a b l e a i d o f a l l o f t h e m em bers
Df t h e d e p a r tm e n t a n d e s p e c i a l l y t h a t o f D r. I v y u n d e r w hose g u id a n c e a n d
d i r e c t i o n a l l o f t h e s e p r o b le m s w e re c o m p le te d .
To D r. R .A . B u s s a b a r g e r
m d D r.
W illia m H . B a c h r a c h I am e s p e c i a l l y i n d e b t e d f o r s u g g e s t s
le lp i n
s o l v i n g m any o f t h e p ro b le m s w h ic h a r o s e .
and
F o r h i s v e ry v a lu a b le a id i n s tu d y in g th e p a th o lo g ic a ls e c t i o n s and h i s
le lp in
m a t t e r s o f t e c h n i q u e , I am i n d e b t e d t o D r. F .D . Gunn.
V ita
B o rn S io u x C i t y , I o w a , J u n e 1 2 , 1 9 1 5 .
A tte n d e d g r a d e s c h o o l s M o v il l e a n d S io u x C i t y , Io w a , h i g h s c h o o l
S io u x
C ity , Iowa.
A tte n d e d M o r n in g s id e C o l l e g e 1 9 3 2 - 3 3 , Y a n k to n C o l l e g e 1 9 3 3 -3 4 a n d sum m ers,
L934, 1 9 3 5 .
E n t e r e d m e d ic a l s c h o o l U. o f S o . D a k ., V e r m i l l i o n , S o u th D a k o ta 193 5 a n d
a t t e n d e d 1 9 3 5 -3 7 .
A s s i s t a n t i n p h y s i o l o g y 1 9 3 6 -7 .
B . S c . d e g re e g r a n t e d
L937.
A tte n d e d N o r t h w e s te r n U n i v e r s i t y M e d ic a l S c h o o l a n d G r a d u a te S c h o o l J u n e ,
.937 t o A u g u s t, 1 9 4 0 .
R e s e a r c h a s s i s t a n t i n p h y s io l o g y 1 9 3 7 -4 0 .
I .S c . 1 9 3 8 , a n d B.M. 1 9 4 0 .
G r a n te d
116
P u b lic a tio n s
A l l a r t i c l e s m a rk e d w i t h a s t e r i s k w e re p u b l i s h e d w i t h c o l l a b o r a t o r s .
L .* A ss a y o f a V ita m in K p r e p a r a t i o n f o r V ita m in D, P r o c . S o c . E x p . ^ i o l .
a n d M ed. 38: 7 8 5 , 1 9 3 8 .
2 . * E f f e c t o f L i v e r E x t r a c t , I n j e c t e d P a r e n t e r a l l y i n t o t h e P r e g n a n t M o th e r,
o n t h e E r y t h r o c y t e s o f N ew born R a t s . P r o c . S o c . E x p e r . B i o l , a n d Med.
38: 787, 1938.
3 . The A n t i - P e r n i c i o u s A nem ia E a c t o r i s P r e s e n t i n P e t a l C a lv e s ’ L i v e r s ,
Am. J . P h y s i o l . 1 2 3 : 2 1 5 , 1 9 3 8 .
* T h e P r e s e n c e o f t h e A n t i - P e r n i c i o u s A nem ia E a c t o r i n a n E x t r a c t o f
F e t a l B o v in e L i v e r s , Am. J . M ed. S c i . 197: 7 5 0 , 1 9 3 9 .
l .* T h e E f f e c t o f L o c a l A p p l i c a t i o n o f T e s t o s t e r o n e i n a n O in tm e n t on t h e
G ro w th o f t h e P e n i s i n t h e R a t . P r o c . S o c . E xp. B i o l , an d Me. 5 9 :
307, 1938.
5 .* T h e E f f e c t o f G a s t r i c M ucin on t h e H e m o g lo b in R e g e n e r a t io n i n A nem ic
D o g s. J . L a b . a n d C l i n . M ed. P5: 1 3 , 1 9 3 9 .
3 .* T h e E f f e c t o f N o rm al Human G a s t r i c j u i c e , i n j e c t e d P a r e n t e r a l l y a n d
G iv e n O r a l l y t o t h e P r e g n a n t M o th e r , on t h e E r y t h r o c y t e s o f N ew born
R a t s . Am. J . P h y s i o l . 1 2 6 : 6 5 5 , 1 9 3 9 .
7 .* T h e E f f e c t o f
G a s tr e c to m y on t h e M onkey.
A nn. I n t . M ed. 1 5 : 1 0 2 8 , 1 9 3 9 .
3 ,T h e E f f e c t o f I n d o l e on t h e R ed B lo o d C e l l s o f N o rm al an d G a s tr e c to m iz e d
D o g s. Am. J . P h y s i o l . 1 2 9 : 4 9 7 , 1 9 4 0 .
3 .* T h e E f f e c t o f T e s t o s t e r o n e , E s t r o n e , a n d E s t r a d i o l A p p lie d l o c a l l y t o
t h e P e n i s o f t h e R a t . E n d o c r in o lo g y 26: 1 0 7 8 , 1 9 4 0 .
L 0.*A lum inum P h o s p h a te i n t h e T h e ra p y o f P e p t i c U l c e r : E f f e c t o f Alum inum
H y d r o x id e on P h o s p h a te A b s o r p t i o n .
In P re ss.
M anual
KA M a n u a l on P r e s c r i p t i o n W r i t in g
N o r t h w e s te r n U. M ed. B o o k s to r e , C h ic a g o , 1 9 4 0 .
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