THE EFF3CT OF TWO TRANQUILIZERS AND A RZWTED
COYPOUFD ON THE FiAT3 OF FISSIOPI AKD THE
SURVIVAL OF PARAMECIUh4 CAUDATUM
A Thesis
Presented to
The Graduate Division
Drake University
In Partial Fulfillment
o f the Requirements far t h e gegre?
K a s t z r of A r t s
THZ EFFZCT OF TWO TRAKQUILIZERS AIID A k!ZLAT?D
COil4POIP?D O?: THE RATE OF FISSIOH Al:D THE
SURVIVAL OF PARABECITJM CAUDATUM
by
Emerson G. Smith
Approved by Committee:
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Dean of t h e Graduat~/Divisi'.on
TABLS OF COPI'rENTS
PAGE
Ci-IAPTFR
.
...................
I1 . RISTORY . . . . . . . . . . . . . . . . . . . . .
I11 . MATTRIALS AND ivIZTHODS . . . . . . . . . . . . . .
I V . RKSULTS AND DIS'JUSSIOT\J . . . . . . . . . . . . . .
V . SUMMARY . . . . . . . . . . . . . . . . . . . . .
15
......................
33
I
INTRODUCTICN
BIBLIOGRAPHY
1
3
20
31
LIST OF T A B L E
I,
11.
The E f f e c t f S e v 5 r a l C o n c e n t r a t i o n s o f
on t h e S u r v i v a l and T i s s i o n
Thorazin
o f Paramecium Caudatum
22
The E f f e c t s of S e v e r a l C o n c e n t r a t i o n s o f
phenergar@ on t h e S u r v i v a l and F i s s i o n of
Paramecium C a u d a t u m .
24
dh
............
.............
111.
43
The E f f e c t s o f S e v e r a l S o n c e n t r a t i o n s of -4tara
o n t h e S u r v i v a l a n d F i s s i o n o f Paramecium
Caudatum
...................
26
CHAPTER I
INTRODUCTION
T r a n q u i l i z e r s have been w i d e l y used i n I n d i a f o r a t
least f i v e c e n t u r i e s f o r a v a r i e t y of ailments including
s n a k e b i t e , d y s e n t e r v , c h o l e r a , f e v e r s , i n s o m n i a and i n s a n i t y . 1
I n c o n t r a s t t o t h i s , t r a n q u i l i z e r s o f known p h a r m a c o l o g i c a l
s t r u c t u r e , h a v e b e e n u s e d i n t h e U n i t e d S t a t e s s i n c e 1954.
D u r i n c t h i s l a s t d e c a d e h o w e v e r , t h e u s e and u n d e r s t a n d i n g
of t r a n q u i l i z e r s has increased rapidly u n t i l a t present t h i s
class o f d r u g s o c c u p y a p r o m i ~ e n tp o s i t i o n i n t h e f i e l d s of
m e d i c i n e , p h a r m a c o l o g y and b i o l o g y .
I t a w D e a r s t h a t most i n v e s t i g a t i o n s w i t h t h e s e c3m~ o u n d s? a v e b e e n c o n d u c t e d on h i g h e r a n i m a l s , i n which t h e s e
a-ent,s
a r e c z ~ a b l eo f e x e r t i n g a u n i q u e t y p e o f s e l e c t i v e
c e n t r a l n e r v o u s s y s t e m d e p r e s s i o n , d i f f e r i n g from m a s t s e d a t i v e s w:'lich a c t b y o r o d u c i n ? a s e n e r a l c e n t r a l n e r v c u s systern
.
: : , .
L i t t l e i s knov:n o f t h e i r a c t i o n o n 3 i c r 0 0 r g a n -
isms.
It, wns t h e
F U ~ ~ O S 3f
S
this invzstigati2n t c s t ~ d y
th+? & ? f f e c t of two w a t e r s o l u b l ? , o r ~ 2 r i . zconpouncis known a s
1~h:)i-l.?:; 0. N i l s o n
0 1 s S r i s v a l 3 , T e s s b z o k cf O r -
-
Lrnn i. r: ! ~ , ~ i t :-. i i c i n nand
l
F h n r n s c e u t i c a l C h ~ r n i a , -,-Lr .~ - -( - E 1i &
1
n
I :
J. 3. L i o p l ~ c o t t , P o . 7 j y7.-. -?;--a_.
,
+
~ : l Z ,
@ ~ t a r a a and
,
tranquilizers (Thorazin
compound ( p h e n e r g a &
a c l o s e l y related
on t h e r a t e of f i s s i o n and t h e survival
o f t h e ~ r o t o z o a nParamecium caudatum.
The e f f e c t o f t r a n q u i l i z e r s and r e l a t e d compounds on
p r o t o z o a n s h a s r e c e i v e d l i t t l e a t t e n t i o n , a l t h o u g h t h e effect,
of s u c h compounds o n v e r t e b r a t e s , e s p e c i a l l y mammals, i s
under active investigation.
P a r a m e c i a h a v e l o n g been one o f t h e f a v o r e d protozoans
f o r r e s e a r c h work.
They a r e v i s i b l e u n d e r a low powered
m i c r o s c o p e , h a v e a r a p i d r a t e o f r e p r o d u c t i o n , t h r i v e 111a
great v a r i e t y o f rnodia, a r e e a s y t o c u l t u r e ar,d p o s s e s s t o
some extent g r e a t e r c g x p l e x i c i t y t h a n many o t h e r u n i c e l l u l a r
o r q e n i s m s , which undoubtedly p a r t i a l l y aczaunts f o r t h e i r
U s P u y t o r a c , A n d r i v o n a n d ~ e r r e , ls t u d y i n g t h e c y t o -..
. , . : I . I.T.,
-:tic
~ c t i o no f n i c k e l s a l t s ( n i c k e l c h l o r i d e , s u l f a s s a n d
n i tr:it*3) on P a r a m e c i u n cauciatun found t h a t t h e t i n ? necesszr:;
;r
r-.r,-:.:r c i l i a t r - s m o t i c n l e s s i s n l r c p o r t i o n a l c o t n e
nickel-
b u t th2.t
..
;,,a : : i i r , i ~ ~ i r . ti i m e
j
for r - ~ a r c o s i s ;t h s t . rnztln5 t y ? e s h s v e v e r y
.
i ;- . ; r . ? n t s e r , s i t i v i t y t o n i c k e l s a l ~ s ;a n d tnat s_':sr c & r c c r i s
ha
l t ) . De?tlytorac,
i. . ; n 3 r i v r n ,
aria
9. S a r r e , f 1 : y ~ 2 n a r ;,-ur-r-ZL
r n t i c . \ c t i o n o f f\ i c k e l Salt? o n P a r a m e z l u : ~caud:.t';nl,"
~
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-
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,
4
a b o u t t w e n t y g e n e r a t i o n s must p a s s b e f o r e t h e r a t e o f m u l t i p l i c a t i o n i s a.gain normal.
~ e t e r s ,i n~ 1 9 0 4 , made a n e x t e n s i v e s t u d y o f t h e e f f e c t
o f c a l c i u m c h l o r i d e , p o t a s s i u m c h l o r i d e and c h l o r o f o r m on t h e
d i v i s i o n r a t e o f s e v e r a l p r o t o z o a n s i n c l u d i n g P. caudatum,
-
a n d f o u n d t h a t t h e r a t e o f f i s s i o n c a n be c o n t r o l l e d t o a
g r e a t extent b y v a r y i n g t h e c o n c e n t r a t i o n and t h e time o f
e x p o s u r e o f t h e o r g a n i s m t o t h e compounds used.
P.
-
c a u d a t u m showed t y p i c a l " a v o i d i n g r e a c t i o n f 1 toward
pH c h a n g e s , a n d t o many o t h e r c h e m o t a c t i c s t i m u l i , s u c h a s
s o d i u m c h l o r i d e , c a l c i u m c h l o r i d e and magnesium c ' h l o r i d e
according t o ~
r
d a p e c u l i a r motor r e s p o n s e which
~ Hel n.o t e~
l a s t e d f o u r t o f i v e s e c o n d s and sometimes l o n g e r and c o n s i s t e d
m a i n l y o f r o t a t i o n s around t h e t r a n s v e r s e a x i s of t h e body,
instead o f t h e ' ' a v o i d i n g r e a c t i o n , " when o r g a n i s m s w s r e s u b j e c t e d t o a 40 m i l l i r n o l a r p o t a s s i u m c h l o r i d e s o l u t i o n .
O b s e r v i n 5 parrnscia with a n e l e c t r o n m i c r o s c g ~ e , P i t e l k a
a n ~ a r d u c z ,a ~t t h e U n i v e r s i t y o f C a l i f o r n i a , noted t h a t
1 ~ h.
. P e t e r s , " 3 i v i s i o n i n P r o t o z o a , ' l J o u r n a l of thP
l i r n e r i c a n !\czdenv of Arts
-a n d S c i e n c e s , SXXIX ( A p r i l , l i : 3 1 ) ,
45.1.
2 ~ D
. ryl,
" C o n t r i b u t i o n s t o 2diechanism o f : h e n o t a c t i c
Kes p o n s e i n Paramecium caudatum, 'f Animal S e h a v i o r , li ( A p r i l A u l v , 1363), 3Q5.
3 ~ o r o ~P iht e~l k a a n d Eiala Parciucs, n 3 1 e c t r o n i ~ ~ i c r o s c o p e O b s e r v a t i o n s on P a r a l y z e d P a r a m e c i u x , " J o u r n a l cf
IJ r n t , o z o o l o ~ y ,I X ( ~ u ~ u Ssutp p l e m e n t , IQo?), 4.
5
n i c k e l s u l f a t e p a r a l y z e s t h e s o m a t i c c i l i a and t h e c o n t r a c t i l e
vacuoles.
in 0.005
I m m e r s i o n o f t h e o r g a n i s m s f o r two t o f i v e m i n u t e s
-
0.01 p e r c e n t n i c k e l s u l f a t e , f o l l o w e d b y washing
a n d s u s p e n s i o n i n t a p water o r c u l t u r e medium, l e a v e s some
i n d i v i d u a l s u n a f f e c t e d , o t h e r s s u f f e r temporary p a r a l y s i s
o n l y , and some s u f f e r p a r a l y s i s accompanied by l o s s o f c i l i a
o v e r t h e a n t e r i o r o n e - f o u r t h t o o n e - t h i r d o f t h e body w i t h
s u b s e q u e n t f u l l r e c o v e r y , o r more e x t e n s i v e d e c i l i a t i o n , b l e b
f o r m a t i o n and d e a t h .
~ u s a , a l s o u s i n g a n e l e c t r o n microscope, observed t h a t
P. c a u d a t u m , a f t e r b e i n g i n d u c e d t o d i s c h a r g e t h e i r t r i c h o -
c y s t s by e l e c t r i c s h o c k , a r e a b l e t o r e g e n e r a t e f i f t y p e r
c e n t o f t h e i r n o r m a l complement o f t r i c h o c y s t s w i t h i n f o u r t o
s i x hours.
discuss in^ t h e r e c o v e r y o f p a r a m e c i a a f t e r i m m o b i l i z a t i o n b y X - i r r a d i a t i o n , # i c h t e r m a n 2 s t a t e s t h a t many imrriobil i z e d s n e c i m e n s may r e m a i n s 2 e m i n g l y dead t h r o u g h a t w e n t y -
f n l l r :?cur p e r i o d o n l y t o r e c o v e r l a t e r .
Immobilized p 2 r a n 3 c i z
s e t t l ~t o the b o t t o m o f t h e c u l t u r e medium t h e s a n e as t h e
d e a d ones.
They d o n o t i n g e s t f o o d and show a c t i v e c y c l o s i s
' k y r a s Yusa, " U e c ~ r o o n Piicroscope O b s e r v a t i o n s cn
U e v e l o n i n f f I ' r i c h o c v s t s i n Paramecium ~ a u d a t u r n , ' ;~o u r n a l of
I ) r o t , o z o o l o ~ y , IX ( j i u S u s t t
, 1 ? 6 2 ) , b.
2iialr~ilWichterrnan, * S u r v i v a l a n d R e p r o d u c t i v d A + b i l i ~ : :
:,fl;cr X - i r r a d i a t i o n i n f o u r s o e c i e s o r Paramecium," i c u r n a l
o!.
, 159-53.
- i ' r o t o z o o l o q v , V I I I (Mav, 1
6
a n d c i l i a r y movements a r e n o t a p p a r e n t .
Recovery b e g i n s b y
c i l i a r y a c t i v i t y , and t h e n t h e r e o c c u r s a g r a d u a l and s l o w
g l i d i n g o f t h e o r g a n i s m on t h e b o t t o m o f t h e d i s h w i t h o u t t h e
c h a r a c t e r i s t i c s p i r a l swimming which o c c u r s n o r m a l l y and l a t e r
a f t e r major recovery.
A m o t i l i t y i n h i b i t o r s h o u l d b e r e v e r s i b l e and non-
l e t h a l , e v e n when i n c o n t a c t w i t h t h e o r g a n i s m f o r p r o l o n g e d
p e r i o d s of t i m e .
E y s and ~ a r n o c k ,u ~s i n q t e n q u a t e r n a r y
ammonium c o m ~ o u n d sa s m o t i l i t y i n h i b i t o r s on
P.
caudatum
c o n c l u d e d t h a t t h e t i m e b e t w e e n a p p l i c a t i o n and e f f e c t v a r i e s
w i t h t h e concentration of t h e solution.
Using a l o w power
m i c r o s c o p e t h e y o b s e r v e d t h a t m o t i l i t y i n one g i v e n o r g a n i s n
i s n o t a n a l l o r n o n e phenomenon, b u t t h a t o n e c a n a p p r o x i -
m z t e b e t w e e n n o r m a l movem2nt and i m m o b i l i t y i n d e g r e e s o f
75, 5'3 a n d 2 5 p e r c e n t o f n o r m a l .
The e f f e c t o f t h e s e met'lon-
i u m d r u ~ sz z v e a n e x t r e m e l y s h a r p end p o i n t .
of
2.7
A f t e r six d a y s
m i t i l i t v , d i l u t i o n w i l l r e s u l t ir! a c t i v e g r o w t h f o r
TJ. c a u , i ? t u m .
-
A n e s t 5 e C i c s a r e known t o d e c r e a s e n e t e b o l i s m i n t h s
h i rhpr a n i m a l s .
Pjurnerous w o r k e r s h a v e i n v e s t i g a t e d $ h e sf-
f r c t s o f v a r i o u s a n e s t h e t i c s on
.
P.
caudatum.
~eichsenrinq~
1 Van ' 7 ~ sa n d L. 2 . ;$arnocli, l f I n h i b i L i o n o f ~ . . o t i i i t y
-,
9 '
X (r\
a
: :?rp11-:'1 ; r A ; 3 t 5 ~ y i D
~ nr i
o v z r n h a r , lQs3), 465-Sc.
2 'I. -
l ~ ~ ,L
"ournal
of
?ro5@232lcq;.,
L e i c h s e n r i n ~ , ' l I n f l u e n c i n z t h z h a t ? of Oxyssz
'.c7urn.l
of
Cclnsr1;n:~ti o n Ln U n i c e l l u l a r O r g a n i s m s , " .:tm?r7iean
h
.i P 1
L,Y \.TI ( 1 ) 6 3 ~ 3 n t \ 3 r ,11325 ) , ? L - h - .
-----9
.;1.
7
s t u d i e d t h e o x y g e n c o n s u m p t i o n ( a s t a n d a r d measure o f metabo-
l i s m rate) of P. c a u d a t u m i n v a r y i n g c o n c e n t r a t i o n s o f s e v e r a l
a n e s t h e t i c s i n c l u d i n g e t h e r , chloroform, n i t r o u s oxide and
ethylene.
E t h e r and c h l o r o f o r m d e c r e a s e d t h e oxygen consump-
t i o n , while n i t r o u s o x i d e and e t h y l e n e had l i t t l e o r no e f f e c t
o n t h e amount o f oxygen used.
k t t h e end o f one h o u r o f
c h l o r o f o r m a n e s t h e s i a , a n d two h o u r s o f e t h e r a n e s t h e s i a , t n e
p a r a m e c i a w e r e r e t u r n e d t o f r e s h medium i n wnich, a f t e r a
t w e n t y - f o u r h o u r t i m e l a p s e , most o f t h e o r g a n i s m s from
c h l o r o f o r m were dead.
The o r g a n i s m s from e t h e r , a f t e r a
t w e n t y - f o u r h o u r p e r i o d , had r e t u r n e d t o a l m o s t normal oxygen
consumption.
?hen t h e o r g a n i s m s were removed from c h l o r o f o r m
a n e s t h e s i a , a f t e r t h i r t y minutes, they a l s o returned t o almost
n o r m a l oxyqen conslmption i n twenty-four hours,
r C a r l ; ~work i n 1 d 6 9 by R i c h a r d s o n , as c i t e d by B i l l s , 1
yenersli
72d
t h a t t h e t c x i c i t y of t h e a l c o h o l s i n c r e a s e s r o u z n l y
w i t h t h e i n c r e a s e i n m o l e c u l a r w e i g h t and t h a t t h e n a r c o t i c
.?ffect i n c r e ~ s e s? o r e ra:>i31y t h a n t h e t o x i c i t y .
f i r n e d by.' zxrnsrous l z t e r i r ? v e s : i s ? t c r s .
b-e::
:;::lt,
: !
n;jrzmt2cia,
!
T n i s nas
3 i l l s found
I n a s o l u t i z r , w i t h o u t f ~ o z ,l i v e I s n g e r wit3
:73.9 ~ :t ij c - l t i t ,
3rj that
stzrvinq c l i l t u r e s ccr. sver,
5 e r??s:,oreil tc ~ r ~ s p e r i tb y t h e a i d i t i o n o f s u i t a b l e axz~r,:s
8
of any a l c o h o l .
Exposure o f para:n-ci.~ t o rsak e t h y l ~ I c o h o l
i n c r e a s e s t h e i r s u s c e p t i b i l i t y t o a s t r o n g e r dose of e t h y l
a l c o h o l , a n d t o five o t h e r a l c o h o l s .
D e p e n d i n g on t h e c o n c e n t r a t i o n , a l c o h o l s have v h r i e d
effects o n t h e b i o l o g i c a l p r o c e s s e s o f p r o t o z o a . 1
Con-toxic
c o n c e n t r a t i o n s o f e t h y l a l c o h o l a c c e l e r a t e d t h e r a t e of d i v i s i o n of p r o t i s t s .
-P.
I n media devoid o f n u t r i t i o n , s u r v i v a l of
c a u d a t u m was p r o l o n g e d by a d d i n 5 e t h y l a l c o h o l , t h u s s u b -
s t a n t i a t i n g p r e v i o u s work.
Higher c o n c e n t r a t i o n s of a l c o h o l
a n d of o t h e r n a r c o t i c s g a v e r i s e t o conspicuous t o x i c synptoms,
s u c h a s s l o w i n g c f movement, r e d a c e d r a t e o f f o r m a t i o n o f f o o d
v a c u o l a s , a n d s l o w e r p u l s a t i o n o f c o n t r a c t i l e vaciloles.
o f Poland, l e f t p a r a m e c i a
a l c o h o l f o r twenty-four
a t t;e
,,
, .-,
-.. 1
:c;.:ir:
end o f
t:?5
:.r:d
t h e s e v e r a l c o n c e n t r z t i o n s cf
n o u r s , c o m t i n g t h e d e a d and t h e l i v i n g
period.
~ t . 23 i c i r . t ~ - r ? : i o n
ir!
X i s c c n s e n t r ~ t i ~ n3si . i
cf the c e l l s .
32 c c ~ c l u d o d :hat
552
$53
o f ;ke cor.!:ound.
" l ~ s c ~ ~ i ea ndd j e r ~ a n , c i t e d b p 3 r y l , '
I
-:
~ 2 %
czase
c h e r ~ ~ t a e~f ft ei c~t s o f a l c o h o l s I ~ c r s a s e d7iti-i
n 1 , ~ 1 e c : ! l , iwr ; i r i t
9ryl,3
i n 1, .? o r
ncsej
2.
p e r c e n t s o l u t i o n s of e t h y l a l c o n o l
l > . 3ry1, 1 1 2 h e a o t a c t i c and T o x i c 3 f f e c t s of L O W ? ~
..,l c o t i c l ~ :cri 1';rarneciua
c a u j n t j i n , '1 Acta E i c l o g i z e Y x o s r i x i ? n t ~ l i s ,
:(I;: ( t ' t n : : : i ~ l , 1 0 5 ~ P) dq-log,
9
q a t h e r , a f t e r a f e w m i n u t e s , on t h e p e r i p h e r y o f t h e d r o p and
t h e y h y p o t h e s i z e d t h a t t h i s a c t i o n was d u e t o t h e e v a p o r z t i o n
o f e t h o n o l o n t h e p e r i p h e r y and c o n s e q u e n t r e l a t i v e d i m i n u t i o n
o f c o n c e n t r a t i o n a s corripared t o t h e c e n t r a l a r e a o f t h e drop.
This h y ~ o t h e s i swas l a t e r c o n f i r m e d b y D r y l .
Many i d e a s h a v e b e e n a d v z n c e d and l a t e r d i s c a r d e d on
exactly how n a r c o t i c s p r o d u c e t h e i r e f f e c t .
R e g a r d l e s s of
how t h e e f f e c t i s d e r i v e d , i t i s g e n e r a l l y a g r e e d t h a t , n a r c o t i c s are g e n e r a l c e n t r z l nervous system depressants.
The
word, n a r c o t i c , i s d e r i v e d f r o m t h e Greek work, vapkau, meaning
"I a m
stupefied."
P a r a m e c i a h a v e f r e q u e n t l y beer, t h e s u b > e c t
o f e x p e r i r n e ~ t a t i o nt o d e t e r m i n e what e f f e c t n a r c o t i c s h a v e on
a sinsle c e l l .
It h a s been determined t h a t higher concentrations of
a n r ; r c o t i c a r e r e q u i r e d t o d e ~ r e s su n i c e l l u l a r o r g z n i s m s t n a n
are n e c e s s z r v t o 2 3 3 r e s s r n u l t i c e l l u l a r orgzinisms. 1
'.!sir,c t h e n z r c o t i c s , e t h y l c a r b a a a t e and c h l o r a l 3:,rc',r.t;
9
?
' r ( u r t L - i n 1 0 4 5 wzs a b l e t c i n h i b i t t h e d i v i s i o n r a t 3 cr' ? , o l ~ o 5 3
s t , l ?i n i i doh'?? t o z e r o .
-
':-ndrlc$ina
ar? i n v e s t i q a t i a n
t3
dotsraine the effezt
':i. Y. d a n d e r s o n , IfThe P r e s e n t S t e t i i s o f t n a ?''?ssri-s
o T ! ! s r c c e i e , lf F h y s i o l o q i c a l 3 e v i rw, X ( k r r i l , 1
, 171-73.
21t.
. .
. . i. ! o i ~c:il
TI
L. i]l~rt,I f > : a r c ~ s i and
s
C e l l 3 i v i s i o r i Ir, P r c ~ o z c a ,
i ; u l ? e t i n , LSILVIII ( F e b r u r - r : , ~ ,19i.+'.)
,
12-2P.
10
o f k i n e t i n ( 6 - f u r f u r y l Aminopurine) and i n d o l e - 3 - a c e t i c
acid
on t h e m u l t i p l i c a t i o n rate of P. c a u d a t u m , i4cManus and S u l l i -
van1 a n d o t h e r w o r k e r s c i t e d i n t h e i r r e p o r t , o b t a i n e d i n d e f i n i t e results.
All f o u n d a d a y t o d a y v a r i a t i o n o f i n c r e a s e
a n d d e c r e a s e i n t h e d i v i s i o n r a . t e o f p a r a m e c i a and were unc e r t a i n as t o t h e c a u s e .
Wing2 s t u d i e d t h e r e s p o n s e s o f p a r a m e c i a t o t h e t o x i n s
of cigarettes.
The f o l l o w i n g c i g a r e t t e components were bubbled
o r soaked i n twenty m l . of water:
c i g a r e t t e smoke, unburned
t o b a c c o , t h e a s h o f t h e e n t i r e c i g a r e t t e , and t h e a s h o f t h e
c i ~ a r e t t eD a p e r o n l y .
Each r e s u l t i n g s o l u t i o n i n h i z h e r c o r -
c e c t r a t i o n s p r o v e d t o b e l e t h a l and was accompanied by b i z a r e
t y p e s o f d e f o r m a t i o n and i r r e g u l a r p r o t r u s i o n s .
The w o r d , t r a n q u i l i z e r i s d e r i v e d f r o m t h e L a t i n word,
r ; r a . n q ~ ~ i l l a sn ,e a n i n : ; c 2 l r 1 , q x i e t , o r s t i l l .
Sone con;gounas
r e f ? r r r . d t o a s t r a n q u i l i z e r s a r e a l s o a t a r a c t i c drugs.
The
w o r 2 , a t a r a c t i c , comes f r o n t h e Greek word, Ataraxic, m?aninz
ccol, i n ~ a s s i v s , n o t m e n t a l l y d i s t u r b e d , w i t h o u t c o n ? ~ s i o n ,
lelusinns o r h a l l u c i r a t i o n s .
.,.
1?4ary
ib!c$,anus a n d K a t h l e e n S u l l i v a n , "Tffec~sof 0
a n d I n d o l e - 3 - A c e t i c Acid on J 5 u l t i p l i c s t . i ~ r ii i a ~ e sP A
Pararneci i l m , " ? , a t u r e , ICI,>;X:~I;I( A u ~ u s ,t 1 0 0 1 ) , 010.
k in.ti.9
:illre
..
a a n c , " D i f f e r e n t i a l X e s p o n s e s cf Parzn-cium
1ia t o ' : i ! ? ~ l - e t t e Cc,;i;ponents, X z t u r e , SiXLYST.'II (I:iarc+,
!
luc!?)
,
ff
OL+h-l,".
11
was t h e first of t h e a t a r a c t i c p h e n o t h i a z i n e d r u g s t o be synt h e s i z e d ( i n 1950).
T h e s y n t h e s i s was accomplished by t h e
R h o n e - P o u l e n c S p e c i a L a b o r a t o r i e s of France. 1
C h l o r p r o m a z i n e r e d u c e s s p o n t a n e o u s motor a c t i v i t y and
" d r i v e w a n d c a u s e s q u i e t n e s s , d e t a c h m e n t and i n d i f f e r e n c e t o
t h e e x t e r n a l e n v i r o n m e n t , s l e e p i n e s s and d r o w s i n e s s w i t h o u t
inducing hypnosis o r anesthesia.
Animals n o r m a l l y f i e r c e and
d i f f i c u l t t o h a n d l e become p e a c e f u l and e a s y t o h a n d l e when
t r e a t e d w i t h chlorpromazine.
I t h a s been shown t o i ~ c r e a s e
t h e h y p n o t i c e f f e c t o f b a r b i t u r a t e s and p r o l o n g s t h e d u r a t i o n
of n i t r o u s o x i d e a n d e t h e r a n e s t h e s i a i n e x p e r i m e n t a l a n i m a l s .
C h l o r p r o m a z i n e may b e u s e d t o l o w e r body temperature and i e c r e a s e oxyrren u p t a k e ,
I t i s w i d e l y used i:: p s y c h i a t r i c x e d i -
c i n e i n t h e t r e a t m e n t of h y p e r a c t i v e , d i s t u r b e d r n e ~ t a l
~ a t i e n 7 s 2.
Z h l o r n r o m a z i n e i s ~ l s e d :c, i e p r e s s v o m i t i n 3 a s s c c i a t e d
wi1!:
n u r n 3 r c u s c a u s e s sllc? a s p r e g n a n c y , r z c l i a t i o n s i c k n e s s ,
n a r c c t i c s , c a n c e r , lukemia, s t c .
I t h a s roved t o be a v a l u e
i n t n e t r e a t x e n t o f a c u t e a l c o h o l i s m , d e l i r i u m trernens, h i c -
.,
l 9 ~ ~ 2 1! ?dl a i r , ; ~ c d e r n D r u . : ~ for th2 Prea;?lent of l:ental
1 1Lness (
f
i I l l i n c i s : J h a r l - s 2. T h s g a s , l " ? ) ,
r?.
.3s.
.. :.
L?wis, & Introducti.-n
F h ~ r a a c c l c c ~( Y
. alti1 " ~1,j
'Ph?
\:i:?ians
a
n
d
;:'ilkins
3?:;a~!P
m o r ~ ,,V!~-vland
:
I-.
7 LO-1.'.
1
,-
.
The s e x u a l a c t i v i t y o f male r a t s wss reduced by Chlorpromazine.
M a l e r a t s r e c e i v i n g C h l o r p r o m a z i n e r e q u i r e d more
tima t o mount a n d t o c o p u l a t e .
They accomplished few mounts
a n d f e w e r c o ~ u l a t i o n sd u r i n g a g i v e n t i m e p e r i o d . 1
The o f f s p r i n g o f a l b i n o r a t e s , i n j e c t e d w i t h i l e s e r p i n @
C h l o r p r o m a z i n e , o r PIeprobamat@ d u r i n g t h e g e s t a t i o n p e r i o d ,
a r e s i g n i f i c a n t l y s l o w e r than c o n t r o l s t o reach the c r i t e r i o n
of l e a r n i n g . 2
I n a l l c a s e s , r a t s and mice i n j e c t e d w i t h Chlorpromaz i n e o n e h o u r p r i o r t o a l e t h a l i n j e c t i o n of several b a c t e r i a l
t o x i n s , i n c r e a s e d t h e s u r v i v a l t i m e o v e r t h o s e r e c e i v i n g none .3
I n j e c t i n g r h e s u s monkeys w i t h Chlorprornazine produc2d
a n i n c r e a s e i n t h e t i m e r e q u i r e d t o r e a c t t o a s t i m u l u s due
t o " d e c o n d i t i o n i n g p r o p e r t i e s " and i t s g e n e r a l d e p r e s s a n t
effects.4
P r o m e t h a z i n s ( ~ h e n e r ~ ai sd a p h e n o t h i a z i n e d e r i v z t i v e
l p h i l i n 2. Lirnbzrdc and " r b e r t
3 a r r y 111, " 5 f f e c t s of
2 a f f e - ; ? . a n d 2 h l o r r r o m a z i n ? n n the S e x u a l 3 e n a v i o r o f h a l e
-:.LA,
)= +
S c i e n c e , Z X X V I I ( L a n u a r y , 1958), 5'4.
>
Lc:ack
B'erhoff, l f y f ' e c t s cf P r e n a t a l ~ u n i c i s t r a t i o nof
r r a n q u - i l i x e r s on Xaze L e a r n i n ? i b i l i t y , . 4 x ' r i c e ~ P s v c h o l o ~ i s t ,
B - L ~ I( i~ u n e , l c 3 0 2 ) , 3 0 7 .
-
j ~ .; r c e n h e r q a n d J . X. I n g a l l s , T13f.?ect cr' T r a n q u i l L o u r n s l of P h a r ; n a c e u t i c a l
i z e r s c n : J a c t e r i a l l'cxemias,
. ; c i ~ 2 n c ~ - .L
, I ( i a n u a r v , l o o Z ) , 71.
3
L ~ : , r l L e w i s S c h e s k e l , w S f f d c t cf 2 5 l o r p r o m a z i n .e . ( T n o r 2 i nl: ) a n 3 C h l o r d i a z t ? ~ c x i d e ( i i b r i u m ) or, 3 e l a y e J liiaccnlP<
13
and u n t i l r e c e n t l y Was r e f e r r e d t o and u s e d as a t r a n q u i l i z e r .
I t i s a a n t i h i s t a m i n i c compound, h a s a h y p o n o t i c a c t i o n , i s
a n a n t i c o n v u l s a n t a n d i s u s e d f o r p r e a n e s t h e t i c , a s well a s
i n a n t i h i s t a m i n e t h e r a p y , w h e r e some d e g r e e o f s e d a t i o n i s
not desirable.
The i m p o r t a n t c l i n i c a l u s e s a r e preoperative
m e d i c a t i o n and a n t i h i s t a m i n e t h e r a p y .
I t i s no l o n g e r u s e d as
a t r a n q u i l i z e r , b u t by u s i n g Promethazine i n combination w i t h
C h l o r p r o m a z i n e t h e d o s e o f t h e l a t t e r c a n b e r e d u c s d , and
s a t i s f a c t o r y c l i n i c a l r e s u l t s can b e o b t a i n e d w i t h o u t g e t t i n g
some o f t h e u n d e s i r a b l e s i d e e f f e c t s o f C h l o r p r o m a z i n e .
The
m a i n d i s a d v a n t a g e of P r o r n e t h a z i n e i s t h a t i t p r o d u c e s d r o w s i -
ness. 1
A r t i f i c i a l h i b e r n a t i o n may b e i n d u c e d by usicg a comb i n a t i o n o f P r o m e t h a z i n e and C h l o r p r o m a z i n e t o g e t h e r w i t h
r n . ~ t . l o ~ ifs o r c c o l i n y the body. 2
P r c m e t h a z i n e 2 i v e s p r o t e c t i o n a g a i n s t s e a - s i c k n e s s and
I t a l s o i s used i n t h e treatment o f vomiting
air sickness.
31Ae 50 " ; ~ r s , v o m i t i n q o f p r e q a n c r , s t c .
n f n.
2
i
tl.3
--jrTUr
qoutq
t,'1+3r;.p;:.
The s i d e e f r " e c t s
a n d b l u r r i n e : of v i s i o n may accorn~an:.~P r o m e t h a -
3
:jvdrr.xyzine (6tara@
lJis:aril!,
a benzigdral iierivacive
l\q.'altnr :;, R o ~ ta n d F r e d e r i c k 2. Yofnanr., ?h::sioloqicsl
Academic P r e s s , 1353 ) , p m 4=.d.
i'har.n?acolo,qv ( K P w Y o r k :
'>
d-Lt.wi~i,
E. a.,
' ~ b i j . , n.
13.
i+5$.
31:.
a n d a rninor t r a n q u i l i z e r when compared t o t h e p h e n o t h i a z i n e
d e r i v a t i v e s , i s a p o t e n t a n t i h i s t a m i n e and h a s s e d a t i v e and
t r a n c l u i l i z i n g p r o p e r t i e s w i t h no accompanying h y p n o t i c e f f e c t .
I t h a s v a s o d i l a t o r p r o p e r t i e s , i s used t o c a u s e s k e l e t a l
m u s c l e r e l a x a t i o n , a s a l o c a l a n e s t h e t i c , t o t r e a t nervous
i r r i t a b i l i t y , emotional s t r e s s , agitztion, anxiety, tensicn
and c e r t a i n s k i n d i s e a s e s . 1
T r a n q u i l i z a t i o n o f d a i r y b u l l s , from which semen i s
c o l l e c t e d a r t i f i c i a l l y , r e s u l t e d i n e a s i e r h a n d l i n g and more
l i v e spermatozoa per ejaculate. 2
The u s e s o f t r a n q u i l i z e r s a r e many.
The immediate
e x i e r ~ a ie f f e c t s c a n r e a d i l y be o b s e r v e d , and ar, u n d 2 r s ~ s n a i n <
of t h e mechanism o f t h e i r p h y s i o l o g i c a l a c t i v i t y i s r a p i d l y
developing.
CHAPTER 111
MATERIALS AND MZTHODS
I n o r d e r t o d e t e r m i n e t h e e f f e c t o f two d i f f e r e n t
t r a n q u i l i z e r s a n d a r e l a t e d c o r n ~ o u n d o n t h e r a t e of f i s s i o n
and t h e s u r v i v a l of
P.
caudatum, an a q u a t i c protozoan, t h r e e
compounds w n i c h a r e w a t e r s o l u b l e were s e l e c t e d .
An e f f o r t
a l s o w a s made t o s e l e c t i n d i v i d u a l conpounds which a r e w i d e l y
u s e d a n d w h i c h would r e p r e s e n t t h e major t r a n q u i l i z e r groups. 1
The R a u w o l f i a a l k a l o i d s make up t h e f i r s t major g r o u p .
T
, h i s q r o u ~c o n s i s t s o f f o u r c l o s e l y r e l a t e d d e r i v s t i v e s , of
t h e Asian s h r u b R a u w o l f i a s u r p e n t i n a and o t h e r rauwolfia
s o e c i l c , w h i c h p o s s e s s s i m i l a r p r o p e r t i e s i n c l u d i n z that, of
.FP:. n.- : j n a r i n . ~ 1 : \ 1s o l u b l e i n w a t e r :
t h u s no r o , p r ? s e n t a t i v ? of
t n i s qroup could be u s e i i n t n i s investigatior?.
The F h e n ~ t h i a z i n e - d e r i v a t i v e s , t h e s e c o n d m a l o r s r o u p ,
are d i v i d e d i n t o twc s u b ~ r o u p s . The Amino-propyl s i d e c h a i n
s~i-~;.-rc/lir,
con
s i s t s o f s e v e n t e e n s y n t h e t i c deriv-;:ives
,hloropromnzinr
,7
9 y J r o x i d e ('?'horaziri&,
inzludins
~ a r < Z c a c ; i l Q iki"5a?azn@)
o r ' - c h l o ~ o - 1 3 - ( 3 - 3 i m e t h y 1 3 . m 1 ~ o ~ r o ? ~ r p1 h) 2 n o t h i a z i c ~h y d r o c 5 1 c r i , j ~ ?w, i l i - 5 i s var:. s o l u b l e i n w z t e r a n d w i s o n e c z :ne
16
tranquilizers used in t h i s i n v e s t i g a t i o n .
The Amino-ethyl s i d e c h a i n i s a n o t h e r s u t r o u : , of
s y n t h e t i c p h e n o t h i a z i n e d e r i v a t i v e s c o n s i s t i n g o f f i v e com-
pounds which a r e very s o l u b l e i n rater.
Promethazlne hydro-
c h l o r i d e o r P h e n e r g a n hydrochloride ( ~ h e n e r ~ a & ,was s e l e c t e - :
to r e p r e s e n t this subgroup o f c l o s e l y r e l z t e d c o a p o u ~ d s .
Tho D i p h e n y l n e t h a n e d e r i v a t i v e s make u? t3e t h i r d
rmjor g r o u p .
Consistin,< of seven synchetic derivatives, t h i s
sroup i n c l u d e s H y d r o x y z i n e h y d r o c h l o r i d e ( k t e r a q vvisfsrilQ,
w h i c h is v e r y s o l u b l e i n w a t e r a n d was t h o second : r a c q o l. i- i z e r
chosen.
The three compounds used w i l l siibsequenti:~be r e f z r r a ~ 3
tc
35
~ h o r a z i n @ , F'henergar@,
and .\~ars@.
A cul5ure c f P. eaudatum was o b t a i n e d frca The 5 s ~ t h 3 r r
! ? i c 1 ; 3 : ~ i c aSl u r n l y 7911rany, N c K e n z i e , Ten~esse?. S e v e r a l
c.lor,l?s ~ 9 - e ~ s t a b l i s . i . e dt o a l l e v i z t ~ ,i n s o f a r a s ?ossLS1?,
',:I.?
nre:!llen!
o f ' ; n d i v i d u a l v ? r l a t i o n o f parzmecia.
! i s : ? a c l o r , ~ ,a sir.r.le n a r ~ a e c i u nw s s iscl3:e3
rlirb?t:,e
. .;ti ;l.eri
-J
4 :
a n d wc.asheii throu::?
i,::-:'4~r.-,
"
cf' .-1:::;s t u b j r ; . : ,
TX.
a aicrc-
n i n e s ~ o ?t l a t e d e p r s s s i 2 r . s o f
7 1. l 1 2 ~ 0 ? i ? 2 t , 2
:':,v?
r.:i;k
'To ? s "' a- ~
v-
was n a j e b;:
isstir. a
3 L 3 ~ 2
i n c i i a x ? t e r , ';hen d r s w i n g x n t i l ';h3
C o n p a n y , M i n n e a p o l i s , M i n n e s o t a , was used.
When removed f r o m
t h e n i n t h d e p r e s s i o n , t h e paramecium was p l a c e d i n a depress i o n c o n t a i n i n g one m l .
sion.
of c u l t u r e medium t o f a c i l i t a t e f i s -
The younp; c l o n e was t r a n s f e r r e d t o a t e s t tube of
c u l t u r e medium when a p p r o x i i n a t e l y t w e n t y i n d i v i d u a l s wwe
p r e s e n t , and l a t e r t o a l a r g e r c o n t a i n e r .
A l l c l o n e s and c u l t u r e s of paramecia were c u l t u r e d i n
d a r k , m o i s t c h a m b e r s a t a c o n s t a n t t e m p e r a t u r e o f twenty-sever!
d e ~ r e e sz e n t i 5 r a d e a s recommended b y ~ e a l e , l~ o n n e b o r n , 2and
others,
P a r a m e c i a f r o m o n e c l o n e ( c l o n e " 2 " ) were used throughout t h e investiqation.
L i m i t e d s t u d i e s were made t o determine
t h e v a r i a t i o n o f t r a n q u i l i z e r and r e l a t e d compound e f f e c t s
on i n d i v i d u a l s o f d i f f e r e n t clones.
A hay i n f l l s i o n medium w 2 s o r ? p a r e d , i n a mannsr s i a i l z r
t- '.ie
rn'lnue,;.s
~ e t ' n o du s e d S y ?.'i:hternan,3
by b o i l i n g Tor tHrer,ty
s i x ~ r 3 m si"f j r y ti.ng-,hg hay i n one l i t z r of s-,rlng
w ~ s s _ r . This i a y S r ~ t hwzs a u t o c l a v e d f o r f i f t e e n minutes a t
f lf t . 3 1 . ~ ~ c u n , _ l so f c r ? ; s u r e ,
c o o l e d s o r o o 3 temperacure,
:t. ? ? a l e , T h s : e n s t i - s of'
- Pararneciur -:.srolia ( : a ~ .-.
i-,rirjtr*-,5 n ~ l s n r l : I l n i v - . r s i t ; : P r e s s , ~ Q s L ) ,p. 42.
l:.
i n o c u l a t e d with one c c . o f b r o t h c o n t a i n i n g Aerobacter
a e r o q e n e s and a l l o w e d t o r i p e n t w e n t y - f o u r h o u r s b e f o r e using.
F r e s h nedium was p r e p a r e d f o r e a c h s e r i e s of tests.
The bac-
terial c u l t u r e was k i n d l y f u r n i s h e d by t h e b i o l o ~ ydspartrnent
a t D r a k e U n i v e r s i t y , Des Moines, Iowa.
WichtermanL and o t h e r s
u s e d A e r o b a c t e r a e r o g e n e s as a food s o u r c e f o r paramecia,
a l t h o u g h o t h e r k i n d s of b a c t e r i a have been used?
Giaser
and ~ o r i a 3f o u n d t h a t P. caudatum w o u l d n o t grow under s r s r i l e
c o n d i t i o n s w i t h o u t t h e p r e s e n c e o f some o c h e r orsanism to
serve a s a f o o d s o u r c e .
P r e l i m i n a r y experirnentati3r-i was conducted with ntzmerorrs
c o n c e n t r a + , i o n s of t h e compounds ";o d e t e r n i n e a f s a s i b l e razge
o f c a n c e n t r a t i 2 n s o f t h e s e c h e m i c a l s t o use i n t h i s i n v e s t i -
:isin% h a y i n f u s i o n medium a s t h e d i l u t i ~ gs o l u t i o n ,
t h e t h r ; 9 3 comuounds were d i l u t e d f r o n an o r i g i n a l concen;r-at i o n o f 25 ne;./cc.
J.
3 .')dl
dl mil. / c c . ,
a n i I).OLIOUOl
t3
ng./cc.
t h e following csncentrations:
me. / c c . ,
0.0501 mg./cc.,
3.1 K J ~ . / ' z c . ,
0.000131 ng./cc.
D i l u t i o n was ac~anplishzd5 p m a n s of
1 3 a l o h Wic h z ~ r m a n , f t j u r v i v a l 2nd ite?roductive i j i l i i y
:'.. f t-:r ;(-i r r a J i a t i o n in F o u r S ~ a c i 2 so f ? a r ~ i m e c i u . ? 1 ", L3urnal
o f ' IJ r o t o z o o l o ~ * s VIII
,
():a;.,
1 0 5 1 ) ) 15.
Z i { a l n h Wichterman, The S i o l o s y of P a r a r n e c i u ~ , p.
10c?.
;;.
; L
c
--i n J - :
f c r t2S P u r e
3 ~ .
; l a s e r 2 3 5 ;(, t.\. : ? r i a , ?ls'et':~ods
-,
y r o t o z c n , " :curnal o f - x 3 d r i i n a n ~ a l~ 9 J i
, of ;
i,T (
y ,
'Ic>?J),
$131.
-
19
a two cc. hypodermic s y r i n q e am? one ml, p i p e t t e s .
Ten
m i l l i l i t e r s o f each c o n c e n t r a t i o n was p r e p a r e d .
To a v o i d a n y p o s s i b l e d e t e r i o r a t i o n of t h e c o m n o ~ q b s
d u e +,a aqe, the dilutions were made j u s t p r i o r t o each series
of
tests,
and were kept in t i g h i l y capped d r o p p i n g b c t t l e s .
A n i n e d e p r e s s i o n spot p l a t e was used f o r each concen-
t r a t i o n o f e a c h chemical,
Four d r o p s (0.2 ml.)
of c u l t u r e
medium with t h e a ~ p r o p r i a t econpound was p l a c e d i n each depres-
sion a n d o n e paramecium was a d d e d , u s i n s a ,nicro?ipette.
These depression p l a t e s c o n t a i n i n g t h e paramecimn were i n c s -
bated a s p r e v i o u s l y d e s c r i b l d .
A t t h e end o f each twentg-
f o u r h o u r p e r i o d , o n e p a r a m e c i ~ ~was
q ';ransferred fron e a c h
deo-es s i o n to c o r r e s ~ o n d i n gf r e s h m e d i m i n a n o 3 h e r p l a t e
reqardless o f whether any d i v i s i o n had occurred.
Cbservations
w e r e m a d l fr e 3 u o n t l y d u r i n s t h e f i r s t t w e n t y - f o u r h o u r
and dailv thsreaft2r.
The r a t e o f f i s s i o n , s u r v i v a l time,
a n n ; ; r i r a n c e o f b l e b f o r m a t i o n , u n u s u a l a c t i o ? s and g e n e r a l
: a n t i li:y
w?re noted and recorded.
The t o t a l p r o 2 e n y of i n d i -
v i l u . 3 1 n3r2mecia i n e a c h d e p r e s s i o n was conpu:ed
o .P
t h r e ? da;.rs,
tinul,j
t p
assuainz t h a t a l l pa.rane!cium would hav? con-
d i v i d e a t th;
?~.?rra:d d n i ly ti.
ac t'ne s ~ d
same
frssh m e d i u n .
rat,^
a s chs on2 w'nich
w2.5
trans-
CHAPTER
IV
RTSULTS AND DISCUSSION
P r e l i m i n a r y i n v e s t i g a t i o n revealed t h a t subjecting
p a r a m e c i a t o ~ h o r a z i n a ,phenerga@,
o r ~ t a r a B i nconcentra-
o r more, r e s u l t e d i n i n s t a n t a n e o u s death.
t i o n s o f 0.1 mg./cc.
T o d e t e r m i n e t h e e f f e c t on t h e s u r v i v a l and t h e r a t e of f i s s i o n
of P. caudatum, t h e s e compounds i n c o n c e n t r a t i o n s of 0.1, 0.01,
0.001,
0.0001,
0.00001,
and 0.000001 mg./cc.
were used.
Pre-
l i m i n a r y s t u d i e s w i t h ~ i o r a z i n @were r e p e a t e d t h r e e t i n e s
f o r t h e p u r p o s e o f developing techniques f o r t h e study of the
p r o b l e m , a n d t h e i n v e s t i g a t i o n was r e p e a t e d i n i t s e n t i r e t y
t h r e e tines.
The
r ~ ~ i lofl tt h
~e
investigation are illustrated
k:r c i t l n ~ d
, a t a f r o ? t h e last s s r i e s of s t u d i e s .
Perc~rn5ag~s
w e r ? co,n=\ute.2 f r o a t n e r e s u l t s a f t e r u s i n g s i x t y - t n r . t e i n d i v i -
d u a l ~ a r a n i e c i af o r e a c h o f t h e t h r e e c m ? o m d s .
f
C c
-4.
,er b e i n q f i r s t i n t , r o d u c e d :o
c u l t u r e rnedtum con-
t a i n i n ~a cc7x:~ourilll, e a c i ~ a r a m 2 c i uw2s
~ continually s~bj:2ct?d
t
7
t h n sa!:;.?s ? e c i f i c c ~ m ? o u n d u n t i l t n e organis!? s e t t l e d t o
+,?r!
'notten
~1of
=it s e v 2 n i v - t w o
the! c i e ~ r c s s i o no r d n t i l t h e s t u d y was c e r x i n a s e d
h.zurs.
5 1 l b j t ? c t l n ~p: a r a m e c i a to s 9 . 1 rns./cc.
l " ~ ~ ~ r ; : , ir z. s?l !l l~
t.,d
con:ezcr:;l?n
i n a n i n s t a n t a n e o u s , c a n v u l s i v e rs2:tion
?f
2 1.
s u b s e q u e n t l y r e f o r r e d to as t h e *hatori reaction."
a c t i c n l a s t e d only f o r a f e w secan.'a
m.
in
rOis
and was f o l l o w e d by
s e t t l i n i . to t h e botto.? of t h e depression in
~2
twen$;.-
f i v e seconds.
c c n c e n f r a t i o n of T%orazin,&Iproduc-d
The 0.91 mc./cc.
S D Y ~ " I T
rnovernents
~ ~
accompanied by +,he a v ~ i d i ~r ega c t i o n ey%n
khou$h nn s o l i d o b s t a s l e was present.
After s t b ~ u tfive
~ i n u L e san u n u s u a l rriuuenent, occurred wh.ifc3 consis$ed cf p j y ~ a l
.~yra+,ir;n
e x c e ? t thacs t h e p c s t 2 r i o r er=3 of zh;
moved in a yawing-f ishfuai'.in:g s p i r a l ,
~arasecis~
T h i s novewer,:
can best
b e d 2 s c r i b e b as a I&yge ~ : i r c u i a r r ~ L ; a b i a nof t%e aos:eri.cr
end of c h 3 pararr,eciun w h i l e
st.^^^^
Tii q
.
.Ivi-*
;
,
rsz
,
c'i,,,i,,,
, , .>,
q m
was aecnm?ar,i&
f
;
L I7
ar!?
S~QF~
for:q.2rd
'
~r0,grsssic.n.
will 52 r e f e r r e d z:? z s fhs va;+
suSssquent;ly
t 2 i l i n 5 rzsction.
.:
anterior ezl,d r25al3e.j fa5rl-t
-4 s r a d ~ a l? ? c r e a s e ir: saeed was sbserv?d
n r ~ t l j i t 7 . r c ? - ? s e i in a p ? r o x i m z e l : ; Len tz f i f z e e n x i z : . u t l ~
2562
crr.ar::sm
L ~ Pbol;:~?
tc
52t;tllj
3f
t;cie d e ~ r i 3 ~ ~ i o ~ .
. ? h o r a z i r ?8
, ecncentrzzicn c f -.-).;751,5
.
?raved
i'L,.
cnrn
5e l e t i - 3 , 1 t o 3araze~iaS e t w e e n s i x and zwe?';:;i-fctlr
,h K r a d u a l s l 0 w i r . g of mpti:Li::r
.
~ a c i - : . j$ v
,2fi
pc,z.isi;nsl
-
Jp;ne
j:-, .l.-+,z;l ,?f'
surr,m,3rj
-.
P
2nd 2.03,!3$1 mg. ,/c,:.
?;-iram.;.c i 3 sl1rv-i v2d for at leas:
t:?-.t h r e e w'?al.:+a?. c z n c e n t r a t l c ? ~ .
z y l acEc:;r)t
0:'
tne
cf f - 3 c !>f
t~
~f~
<
i
~
was o b s s r v a e a x m -
u b a t c n y e z : ' j i a ~ . ~';rior2zine
9.Jd1, 0 .f];!G;il
?,r:l=i
rns
+
,
n?..n:
i
~
z13
CC?CS--
TTC..'~~ 1359
t;we~:?-r"?2r
n..
7
*
1 13
CJ2-s
a
V ~ ~ ~ J I :Jc S' z z ? I I S ~ S T ; ~ ~ ~ S
23
of ~ h o r a z i n a o nt h e s u r v i v a l and r e p r o d u c t i o n o f P. caudatum
as compared t o t h e c o n t r o l .
The s e c o n d compound used i n this i n v e s t i g a t i o n was
A 0 . 1 rng./cc.
phenereaa
c o n c e n t r a t i o n of phenergar@ r e s u l t e d
i n a n i n s t a n t a n e o u s * b a t o n r e a c t i o n f r followed by s e t t l i n g t o
t h e bottom o f t h e d e p r e s s i o n i n approximately t e n t c f i f t e e n
seconds.
A Phenergar@ c o n c e n t r a t i o n o f O.C1 rng./cc.
produced
f r a n t i c a c t i v i t y accompanied b y a n a v o i d i n g r e a c t i o n .
This
f r a n t i c a c t i v i t y w a s c o n s i d e r a b l y s l o w e r a f t e r one minute, and
d u r i n g t h e s e c o n d m i n u t e , a n a b r u p t c e s s a t i o n of m o t i l i t y
o c c u r r e d a n d t h e o r g a n i s m s s e t t l e d t o t h e bottom o f t h e con-
tainer.
p h e n e r g a @ c o n c e n % r a t i o n s of 0,001 and 0.000l mg. /cc.
nrcl:lucod a ~ r a , 5 . ~ slol..ring
al
o f m t i l i t y ar,d an o c c a s i o z k l
'fyax-:ail't
'Ti5
r ? s c t v i o z and no appar;r;t
two w e a k e s t concensrasilns o f ?henerga@
n7.c d.iJ9OQ31 mq./cc.,
f ~ trh e
S P V ~ P ~ ~ : - ~ W Ci c, l i r
?
,. i i
t
i:
0
'
.
?-?
9.3GOJl
r s s - ~ l t e dir, t h e s u r v i v a l o f scne par2-
-;thci
~
mocility z i f t e r t h r $ e t~
p s r i ~ d , b u t f i s s i c n wzs z c z -
.A j:;n~:ir:..of t;?? e f f e c s s of
i ' i e ni z r y a &
c . ~ ~ d : i t ~
comp;ircd
; ~ ,
t
:
t ? ~ evsri,?:s
or, t h e s u r v i v a l 2r.d . - e ~ r o J ! i c t i - n
a ccn:rol,
i s preser,t?d Ir T i b l e
2 1.
,
the r
co:r.yc;lrJ
usad i n - , h i s L ~ - u ~ s s ~ ~ : ; ~ L ~ c : ' . .
T r a n q u i l i z e r s have been u s e d i n I n d i a f o r a t least
f i v e c e n t u r i e s f o r a variety of ailaents.
In c o n t r a s t t o ,-his,
t r a n q u i l i z e r s of known p h a r m a c o l o g i c a l s t r u c t u r e , have been
u s e d i n t h e U n i t e d S t a t e s s i n c e 1954.
This c l a s s of d r u , g
OCCUPY a p r o m i n e n t p o s i t i o n i n t h e f i e l d s of medicine, pharmacology and b i o l o g y .
A s f a r a s can be determined, no i n v e s t i -
g a t i o n s have b e e n c o n d u c t e d t o s t u d y t r a n q u i l i z e r e f "- e c t s on
p r o t o z o a n s , a l t h o u g h t h e e f f e c t o f such compounds on v e r t e b r a t e s , e s p e c i a l l y mammals, i s under a c t i v e study.
I t was
t h e p u r p o s e o f t h i s i n v e s t i g a t i o n t o s t u d y t h e e f f e c t of two
'n'zt?r ~ 3 ? 1 1 b l - ? , c r % a r , i c corngoilnds kncv~r! s s t r a n q u i l i z e r s
("nrr7int.-
0a n d
Atara@
and a r e l a t e d c o q ~ o m d ( ~ h e n e r p r O )
cln the r a c e of f i s s i c n end t h e s u r v i v a l of t h e protozoa^
? a r 3 r n e c i : ~ acaud3tum.
T a c j l o f t:?? t h r 3 e cor?lpounds were d i l u t e d t o ccncegcra-
, 2.21, g.301, 0.9001, 0.30231, 0.303COl % * / c c *
1
ti,,nc
:lsi niT :i!nociv
Aay ne,-ji um as t h ? d i l u t i n , ~s o l ~ t i ~ n lndividual
*
7):lrnmeeia f r o 3 on- c l c ~ e ,were subj?~t?d t o t h ?
,
,
ion:
e-.cn
coKtnini-nr:
c,-?nc.l,;r,,i
c:lemi c i l c~rnaoun5ic
3.1 ni
o.
f ,-ult-,l;ra
i i.r o f con:,su!lds
5
'On-
3 1 3 t 3 de?r''-
s?Oi
nedium.
Vzr12'E
T
bx?os'Jre
'""
t u d i s d was c9ctin3Qus :Or s'v'p''J-
32
two h o u r s o r until t h e o r g a n i s m s e t t l e d t o t h e b o t t o m of t h e
depressi3n.
P a r a m e c i a w e r e k e p t i n d a r k , m o i s t chambers a t
a c o n s t a n t t e m p e r a t u r e of t w e n t y - s e v e n d e g r e e s C e n t i g r a d e .
s u r v i v a l ti-oe v a r i e d from t e n seconds t o seventy-two hours,
d e p e n d i n g o n t h e compound and t h e c o n c e n t r a t i o n used.
The
f i s s i o n of P. c a u d a t u r n was c o m p l e t e l y i n h i b i t e d i n a l l e x c e p t
t h e 0 . 0 0 3 0 0 1 rng. / c c .
c o n c e n t r a t i o n o f ~ h o r a z i n @ and ~ t a r a @ ,
a n d was r e d u c e d e v e n i n s u c h a d i l u t e s o l u t i o n .
It was concluded t h a t t h e s u r v i v a l of P. caudaturn
v a r i e s i n v e r s e l y w i t h t h e c o n c e n t r a t i o n o f t h e t h r e e compounds
u s e d and t h a t f i s s i o n i s r e d u c e d by weak c o n c e n t r a t i o n s o f
t r a n q u i l i z e r s a n d r e l a t e d compounds and c o m p l e t e l y i n h i b i t e d
by s t r o n q e r n o n - t o x i c
concentrati9ns.
BISLIOGRAPHY
A.
BOOKS
B e a l e , G. H e The G e n e t i c s of Paramecium.
U n i v e r s i t y P r e s s , 1954.
Cambridge, England:
B l a i r , D o n a l d . Modern Drugs for t h e Treatment of Mental
1962.
Illness.
S p r i n g f i e l d , I l l i n o i s : C h a r l e s ~.Thomas,
*
R r a d l e y , P. B.
Ph s i o l o i c a l Pharmacology.
dernic P r e s s , I n c . , 1 9 3
New Ilork:
kca-
C a l k i n s , G a r y N . , a n d F r a n c i s M. Summers. P r o t o z o a i n Biologi c a l K e s e a r c h . ??ewYork: Columbia U n i v e r s i t y P r e s s , 1941.
-
--+
J e n n i n g s , H. S. L i f e and D e a t h q e r e d i t 4 Z v o l u t i o n
he Gorham P r e s s , 1920.
Unicellular 0rqanisms.Boston
I f l a t e r i a l s and Methods i n t h e Study of P r o t o z o a .
Kirby, Herald.
B e r k l e y a n d Los ~ n ~ e l e c ~ a l l f o r Xn iivze r r i t y of Calif o r n i a P r e s s , 1950.
An I n t r o d u c t i o n ,t o Pharmacolo
L e w i s , J. 2.
M a r y l a n d : --m
l h ? d i l l i a m s ana
Baltimore,
1963.
? c o t , Y a l t e r S . , and ? r e c i e r i c k G. gofmann.
P n n r m a c o l o q y . ?Jew Ycrk: Academic P r e s s
Xichterman, italoh.
The B i o l o
o f Paraaecium.
-+ , ~ g j j .
Thz 3 l a k i s t o n Company,
6ew York:
C h a r l e s O., a n d 013 O r i s v o l d . Textbook of Organic
id;adicinal
F h a r m a c e u t i c e l Chemistry. Philade:?lia,
P e n n s y l v a n i a : L i p g i n c o t t , 1962.
'':i
lson,
3ills C h & r l e s 7 . ? l i ~p i a r n a c o l o j i c a l C m p a r i s o n of s i x
~ l c o Sinql!'
h ~ ~ and
~ i n Admixture on paramecium," :ourna:
o f P h q r m a c o l o r v and S x p e r i m e n 5 a l T h e r a p e u s i c s , X X I I
r;u.ruz;*
G5?.
3 h e c k e l , Carl Lewis.
"The Sffe c t of Chlorpromazine (Thorazine)
a n d C h l o r d i a z e p o x i d e ( L i b r i u m ) on Delayed i h t c h i n g Responses
in t h e ii'lacaca r n ~ l a t t a . ~ ' Unpublished D o c t o r a t e d i s s e r t a t i o n , Fordham U n i v e r s i t y , New York, 1963.
~ ~ l l e n w e i d e rJ o, h n A l b e r t , " D i f f e r e n c e s i n C r i t i c a l F l i c k e r
F r e q u e n c y i n P r o c e s s and R e a c t i v e S c h i z o p h r e n i c s Treated
w i t h T h o r a z i n e and Cornpazine."
Unpublished Doctorate d i s s e r t a t i o n , Fordham U n i v ? r s i t y , New York, 1963.
" E f f e c t of Method of Semen C o l l e c t i o n
blells, Milton Srnest.
and T r a n q u i l i z a t i o n on Semen G u a l i t y . l1 Unpublished Doct o r a t e d i s s e r t a t i o n , Oklahoma S t a t e U n i v e r s i t y , Rorman,
Oklahoma, 1962.
Burt
R. L.
r T N a r c o s i s and C e l l D i v i s i o n i n P r o t o ~ o a ,&~
i o g i c a l Bull.tj.n,
LXXXVIII ( F e b r u a r y , 1945 ) , 12-29.
C o n o l l y , D o r o t h y , A. 2. Reisn.?r, and ld. R. Sobey. nA Growth
M e d i a for t h e Mass C u l t u r e of Paranecium * A u s t r a l i a n
of S c i e n c e , XXIV (September, 19611, 144-45.
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