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1939

Antuitrin-S Injections in Pregnant Rats. Antuitrin-S Injections in Pregnant Rats.

Russell Mark Coco Louisiana State University and Agricultural & Mechanical College

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Recommended Citation Recommended Citation Coco, Russell Mark, "Antuitrin-S Injections in Pregnant Rats." (1939). LSU Historical Dissertations and Theses. 7805. https://digitalcommons.lsu.edu/gradschool_disstheses/7805

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MANUSCRIPT THESES

Unpublished theses subm itted fo r the m as te r^ and doctor*s

degrees and deposited in the Louisiana S ta te U niversity L ibrary

are a v a ilab le fo r in sp ec tio n . Use of any th e s is i s lim ited by the

r ig h ts of the author* B ib liograph ical references may be noted, but

passages may not be copied unless the author has given perm ission.

C red it must be given in subsequent w ritte n or published work.

A l ib r a ry which borrows th is th e s is fo r use by i t s c lie n te le

i s expected to make sure th a t the borrower is aware of the above

re s tr ic tio n s*

LOUISIANA STATE UNIVERSITY LIBRARY

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UM1 Number: DP69183

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ACEHOWLEDCaSEKT

flte author wishes to eatress M e appreciation for the kind

advise and assistan ce of Dr* S« L. M illar o f tha Department of

Zoology, Physiology and Entomology, under whose d irection th is mark

m s done* and to Dr* W* H* dates* Head of the Department o f Zoology,

Physiology and Entomology.

TABLE OF contents

pagei . u s t o f t a b l s s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ▼

XI. LIST OF FIGURES . . f . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v l

r a . a b s m j t . . . . . . . . . . . . . . . . . . . . . . . ...........

IV. INTRODUCTION MB DISCUSSION OF 'LTSMAWBM . . . . . . . . . . . .......... 1

v . m aterials m> methods .......... m

VI. ME NATURE* BIOLOGICAL SOTOTS* ORIGIN MR CLINICAL USm

ox.......................... ..................................................... . . . . . . . . . . . ........... s i

VH . THE EFFECT OF INJECTING MTOTRIN-S INTO PREGNANT ANIMALS . 34

The E ffect on G estation and P arturition . . . . . . . . . . . . . . . . . 34

The Condition and Size o f the Fetuses • . . . . • • • « . . « • • • • • . • 43

The Condition o f the U teri 33

The Condition o f the Ovaries .......... 34

The Postahortive H istories o f Injected Animals • • • « . . . • • • 61

The Condition o f the Placentae . . . . . . . . . . . . . . . . . . . . . . . . . . 68

T i l l . THE EFFECT OF PR0GE3TM AND AMUITRIN-S PIUS PRGGSSTIH

ON PREGNANCY « . . . ......................... . ............. 70

IX. THE EFFBST OF BIMIL BHJE AND BXABZL BINE PLUS

m m m tM -s on pregnancy . . . . .......... 72

X. THE EFFECT OF 0IM IL BLUE* DIMIL M PLUS MTtJITRIN-S,

AND TRYPAN BLUE ON NOH-PRSGNANT MBSAL3 .............. 83

XI. THE PERSmBILOT OF THE PLACENTA ............. 86

X U . THE EFFECT OF LACTATION ON GESTATION ............ 88

X III. DISCUSSION.................. 90

i l l

TABLE OF CONTENTS (C o a t 'd )

pag9xrv. smmARr /m cmcwsxm . .................... . . . . . . . 96

XV. U«WT0KB CrM) .................................... . . . . . . . . . . . . . . . . . . . . 99

XVI. SXPLANATIOH OF FIGURES . ................................ . . . . 10?

XVII. BIOG&APHY ............................................................. 115

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LIST OF FIGURES

pageFlgur® I , A Graph Showing the E ffect o f One Preparation o f

Prolan When Injected in to Immature Rats .............. 107

Figure 2 . A Photograph o f Three Dead Premature Fetuses from

Animal No. 38003(a) ........... . . . .................... 107

Figure 3 . A Photograph o f Three Dead and One Living Premature

Fetuses from Animal No. 38003(d) .......... 108

figu re 4* A Microphotograph o f a Section Through the Left

U terine Cornu o f Multiparous Animal No* 38003(a) . . . 108

Figure 5* A Mlcrophotograph o f a Section Through the Left

Ovary o f Animal No* 38003(d) ................* 109

Figure 6* A Microphotograph o f a Section Through the Right

Ovary o f Animal No. 38007(e) .......................... 109

Figure 7* A Microphotograph o f a Section Through the Right

Ovary o f Animal No. 38009(c) . . . . . . . . . . . . . . . . . . . . . . . 110

Figure 3 . A Microphotograph o f a Section Through the Right

Ovary o f Animal No. 38001(a) .......................................... * 110

Figure 9* A Mlcrophotograph o f a Section Through the Right

Ovary o f Animal No. 38008(h) .......................................... I l l

Figure ID* A Microphotograph o f a Section Through a Corpus

Luteua o f Pregnancy o f Animal No. 38006(d) * 111

Figure 11. A Microphotograph o f a Section Through a Corpus

Luteum o f Pregnancy o f Animal No. 38007(c) . . . » . • • • • 112

v i

LIST OF PIOOHES (Cont'd)

pageFigure 12. A Mierophotograph o f a Section Through a Corpus

Luteua o f Pregnancy o f Animal No, 38009(d) 112

Figure 13. A Microphotograph o f a Section Through a Corpus

Luteum o f Pregnancy o f Animal No. 38006(b) . . . . . . . . . . 113

Figure 14, A Microphotograph o f a Section ‘Through a Corpus

Luteum o f Lactation o f Animal No, 38006(b) . . . . . . . . . . 113

Figure 15, A Microphotograph o f a Section Through a Placenta o f

Animal No. 38008(F) .............. 114

v i l

ABSTRACT

The d ifferen t opinions o f in vestigators using crude pregnancy

urine ex tra ctsv the sca rc ity o f reported stu d ies concerning the e ffe c t

o f A ntuitrin-S (Parke, Davis and Company’s Pregnancy Urine Extract) on

pregnancy, and the desire fo r a mors complete understanding o f the

hormonal in terrela tion sh ip s concerned with pregnancy and parturition

led the author to undertake th is study. The e ffe c ts o f A ntuitrin-S,

progestin (Parke, Davis and Company’s Lipo-Lutln), and the v ita l dye,

D lan il B lue, on la te pregnancy In the rat were in vestigated .

Except for a few animals o f the hooded v a r ie ty , the animals used

in th is in v estig a tio n were o f albino stock Mua norvegicus albinos

Hatai 1907* A to ta l o f 158 animals were employed.

A ntuitrin-S , when in jected in dosages of 50-75 R. U* on the 18th

day o f pregnancy {gestation 21-82 days) in the r a t, resu lted in b ir th ,

prematurely or at tern , to liv in g , or dead, or liv in g and dead fetu ses

in 93.9 per cent o f the animals in jected . The liv in g fetu ses were

u sually son -v iab le, and parturition was not postponed nor prolonged.

These re su lts are believed to be due to the action o f the A ntuitrin-S

acting on the uteroplacental complex e ith er in d irectly through the ovary,

or d ir ec tly on the uterus and placenta, or both; th is consequently

resu lted in a decreased blood supply to th e fe tu s and p lacental detach­

ment. These re su lts are in contrast to those of Hoopes (1934) and Hoopes

and King (1935) who obtained prolonged gestation and postmature fetu ses

with sim ilar dosages.

v i i i

Dosages Of 150-200 R. IT. usually resu lted in parturition being

delayed and in sane eases prolonged from 1-5 days. The fetu ses died

In utero before or a t tern j l except ion) and in some eases even death

o f the mother resu lted . I t Is thought that the b irth mechanism was so

deranged by th e a u to ly sis and necrosis of dead fetu ses that the female

was unable to give b irth to her young. I t might a lso be that uterine

contractions were inh ib ited to some degree , but th is i s s t i l l

problem atic.

In jection s made on the 19th, 20th and 21st days usually resu lted

in th e b irth o f v iab le fetu ses at term. I t would seem th at the 18th

day i s the c r it ic a l point so far as the tolerance o f the fetu ses to

A ntuitrin-S la concerned, and that in jectio n s made la te r had l i t t l e

o ffs e t on the subsequent growth o f the fe tu se s# and on the b irth

mechanism.

A 1 per cent aqueous so lu tion o f the v ita l dye. D ianil B lue, when

in jected in trap eriton eally in dosages o f 4 c c . on the 14th, 15th,

16th, and 17th days, resp ectiv e ly , resu lted , in the m ajority o f cases,

in fe ta l death and an impairment o f the physical conditions o f the

mother, sometimes even resu ltin g in her death. These r esu lts are not

In accord with those o f Long and Evans (1922) who were unable to detect

any serious to x le e ffe c ts from the use o f D ianil Blue in pregnant

r a ts , but they are in agreement with those o f Meyer who was able to

in h ib it the estrous cycles o f rats by the us© of th is dye. These

e ffe c ts were shown to be due to the to x ic ity of the dye since i t was

to x ic to normal, non-pregnant animals as w e ll.

ix

Comparative h is to lo g ic a l stu d ies o f the ovaries, u te r i, sad

placentae ware made and these were correlated as far as possib le with

the condition o f the mother and that o f the fetuses* The ovaries were

found to contain large c y stic and a tr e tic f o l l ic le s , hemorrhagic

f o l l i c l e s , lu te a l cy sts (some with trapped ova), corpora lu tea o f

pregnancy, and normal-looking induced corpora a fter the 2nd-3rd day

fo llow in g in jectio n s o f Antuitrin-S*

I t i s the author9 s opinion that the corpora lu tea induced by the

extract were not normally functioning ones sin ce (1) o f 4 animals

which aborted dead young, 2 copulated and became pregnant on the same

day that b irth occurred, and the other 2 conceived on the 3rd day post­

partum; (2) the fetu ses did not liv e past term (1 exception out o f 63

ca ses); (3) some animals with well-formed induced corpora aborted th e ir

young prematurely; (4 ) progestin in jected in 5 animals in la te preg­

nancy in doses as high as 3 X* U* (3 Com er-Alien Rb, tF*), had no

e ffe c t on gestation* This i s in agreement with th e work o f Bnnde {1938)

who was unable to a ffec t gesta tion in the rat by the use o f 3 Rb. TJ.

o f h igh ly active progestin per day vfcen in jected in la te pregnancy*

The r e su lts obtained in th is study, therefore, show that: (1)

A ntuitrin-S , when in jected in la te pregnancy* may act as an abort len t

or i t may act in postponing parturition depending upon the dosage and

time o f In jection , but i t may not resu lt in prolonging gestation ;

(2) the v ita l dye. B ian il B lue, is able a lso by means o f i t s to x ic ity

to the animal to cause abortion, fe ta l death, and even death o f the

mother; (3) progestin in dosages as high as 5 I* B. did not prolong

x

gesta tion ; (4) the uteroplacental complex plays an important role i s

m aintaining pregnancy and in determining the time o f parturition;

(5) the need for b etter pu rified ex tra cts, and Improved methods o f

assay for preparation used for experimental and c lin ic a l purposes i s

c lea r ly shown.

x i

m m m n m m> m m m tm m Lim & rtm s

Httch wwfe has been don* in an attempt to solve the hormonal in ter­

rela tion sh ip s aaseeiated w ith pregnancy and p artu rition , fair in v e sti­

gators, howevor, have reported on th e e ffe c t e f A »tu itrt»-S in the rat

as related to m e phase o f the nosh# Hoopoe (1934), using A atuitrin - 8

from Parhe9 Davis and Ofiflapsny, regu larly obtained prolongation e f

pregnancy w ith the development of fe tu se s o f excessive e ls e in m e

follow ing a sin g le subcutaneous in jectio n of 75 Bat U nits adm inistered

four days before tom # fh s normal length of gestation for her animals

was given as 23 days# Hoopoe end Bing (1936), a lso using r a ts , reported?

**1 * Pregnancy can regu larly he prolonged for 9 or more days by cue in jectio n of 75 Rat Unite o f human pregnancy urine extract (A n tu itrin -S ), given on the 19th or 2<tth day o f gestation# 1

2# P arturition sue delayed u n til the 25th , 26th or 27th day and was prolonged over 2 or more days#

2 , fe tu se s remained d iv e and continued to grew fo r 2 or 3 days beyond term*

4« H ecroeis o f retained fetu ses began a fte r th e 26th day and i s indicated by a mueoldal, blood-stained discharge from the vagina#

5# H isto log ica l examination o f the ovaries revealedinduced corpora lu tea by the S lat day ©f pregnancy* but sub­sequent retrogression of these could not be d e fin ite ly estab­lish ed by the method used# fhe induced corpora are regarded as talcing over the function of those of pregnancy and th eir Secretion as delaying the onset o f labor,

6 , Attempts ut estzu s, as indicated by vaginal smears, were observed as early as the 27th day and are not incompatible with fe tu ses in utero#*

rin g (1938), studying the e ffe c t of A ntuitrin-S adm inistered ia tra -

psrltoneaH y during the f i r s t h a lf o f pregnancy, found that 50 B . H#

in jected on or before th e day o f im plantation (im plantation occurs on

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adult male and female fa te whan in jected subeutanecfusiy. Be i s o f the

opinion that the action o f human P . H. on the hypophysis in the rabbit

I# p artly in d irec tly through the gonad® and p artly d irec tly upon the

hypophysis it s e lf# In case o f the rat the e ffe c t Is an in d irect one

mly through the gonad, and he sta te s that th is Is presumably due to

the inh ib itory act 1cm o f ea trln or male ear hormone# Ihraas and Simpson

(19S0) s ta te that the ch aracteristic prolongation o f g esta tion eaa be

secured by the adm inistration once d a ily of a h igh ly concentrated fra ctio n

Obtained from the human placenta or P. tJ# ex tracts made in th e ir labora­

to ry . they obtained sim ilar r e su lts by the im plantation o f l - g glands

Of the anterior hypophysis d a lly for the la s t $ days o f gestation*

P rietem (1 9 3 2 ) in jected A ntuitrin-3 d ir ec tly into mature f o l l ic le s

Of rabbit ovaries and obtained corpora lu tea a fter 4 days* No changes

rare noted in the contralateral ovary* The induced corpora degenerated

early and he mas unable to produce a decidual reaction which indicated

that these corpora lu tea did not function normally* Levin, Katsman* and

Beiey (1931), by subcutaneous in jectio n o f 1500*7000 mouse u n its o f P . U#

extract administered from the 14th-21st days* always delayed or prevented

parturition in th e pregnant rat* fhey sta te that th eir re su lts mere not

due to to x ic ity *

Thus the litera tu re concerning the p h ysiological action o f P* tr*

ex tra cts, including A ntuitrin-S , i s not in agreement* the consensus o f

opinion, however, i s that these extracts prolong gesta tion and in h ib it

parturition when given in la te pregnancy*

She term s, prolonged gesta tion and postpone^ p artu rition , are

rather lo o se ly used In the literatu re* The w riter Is o f the opinion

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gestation depending upon the dosage end time of adm inistration

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of pseudopregnanoy. They state that the corpora lutea appeared to he

9fu n ctio n a l. The pseadopregnant condition in the rabbit u su ally la s t s 16

Aays (Allen** Sen and Internal S ecretion s, p .505)* In two la te r papers

Hackel and A llen (193S, 1939) repeated that d aily In jection s of

estra d io l monobenaoate (Progynoa B) varying from 50*500 international

unit* per k ilo adm inistered at anytime from the £7th»51st day, postponed

delivery in most cases in rabbits* la 14 cases parturition was delayed

longer than 55 days* A ll fetu ses bora or obtained by operation or a t

autopsy were dead* In no case were postmatur© fetu ses Obtained as they

were k ille d w ithin 5 days and died in utero* They were dead and found

to be necrotic by the 52ad day* In the la s t th ird of pregnancy* Injee*

tio n s for only a day or two produced premature delivery of dead fe tu ses

w ithin 3*4 days* When the In jections were continuous* the placentae and

fetu ses were injured but the corpora lu tea were maintained by estrogen

and the fetu ses were retained in utero* They further sta te that tbs

ovaries must be present i f delay la parturition occurs a fte r the in jec ­

tio n o f the estrogenic hormone as shown by control experiments with

castrated rats* They concluded, frcsa these experiments* that regression

o f th e corpora lu tea was prevented by increasing the estrogenic lev e l*

This i s in contrast to Bunde and Creep’s work on hypophyseet omired

rats*

The Corpus Luteum and I t s R elation to Other Endocrine Organs

A ssociated w ith Reproduction a Numerous researches have been reported

on th e corpus luteum and i t s relation to other endocrine organs associated

with reproduction* The corpora lu tea are necessary for im plantation and

the maintenance of embryos la the uterus* and th e ir destruction daring

result# in reeorptlon or abortieiu This

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generally known that hypophy sec t oeny results In the persistence of

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ctomy so early as tbe IStli day of gestation was M to kayo

no affect os subsequent development of tbe corpora lutes of p

maintained by soois extrahypophyseal influence* They aafce no suggestion

12

«* to what th is in iluaas* ®i«&t he* Aatwood and Greep (1938} hare

demonstrated a corpus lutoam ^ebiaulatliig substance in tb s n t placenta.

This fcoftt#a*t 1fcey s ta te , secreted by the developing ehorlm a fte r the

N r Of pregnancy se ts to maintain and enlarge the corpora o f preg-

steady whose continued secretion prolongs gestation to tern* f ly e r (1983)

abate# that the hypophysis la e sse n tia l tea? ovulation and the l i f e of

the oorpue la teen and the maintenance o f pregnancy In th e rabbit*

fhao fron a review o f the lite r a tu r e , i t would a®m th a t th e o ffse t

Of hypophyaectomy in la te pregnancy in th e rat 1# s t i l l open to

In vestigation .

H etffio g io a l Observations on Corpora Patent long and Staan# (1928)

were able to determine the age and fun ctional a c tiv ity o f the corpora

in tea b f a study o f the lip o id granules. Degenerate corpora always

contained large lip o id granules * The technique need was that o f Benda

given by Sieves and Bueefeerg (1908). Warbrition (1934) in studying the

cytology o f the corpora lu tea o f the ewe found that the reappearance o f

fa t droplets was th e f ir s t ind ication o f retrogression .

Purther R elation of Gonadotropic Hormones to the Gonads; .In 1931,

reveld , Hisaw and leonard reported a separation o f P. S . B. and &. H.

by pyridine extraction of anterior lo b es. The f * S . H, fra ctio n was

found to be water so lu b le and the L. H. water insoluble* These washers

have pointed out that the action of the L* S . appeals to depend upon

the prior action of the P, S. H* They a lso state that the fom ation o f

hemorrhagic f o l l ic le s (Blutpnnfcte) i s due to 1 , H. acting on the

developing f o l l i c le s , S elye, C ollip and Thcaascm (1933) sta te that the

13

corpora lu tea which p ersisted a fte r hypophyoeotoBy in the rat inh ib ited

lu te in isa tio a o f the thecal c e lls* They farth er s ta te that the A* P . L.

homaoae (I*. H*) prolonged the l i f e o f the corpora lu tea present a t

hypophyaeotoay and that th is hormone did net lu ts in isa theeal c e lls i f

I t had true lu te a l c e l ls to act upon* In a la te r paper by C ollip* Selye

and Williamson (1938), they found that chronic treatment o f adult hype*

phyeectaraized r a ts with p ig anterior p itu itary extract resu lted in

ovarian sin e regression and severe atrophy* The ovaries war© resista n t

to the gonadotropic action o f th e extract used, but also* at le a s t to

some degree to implants of rat hypophyses* Massive doses of A* P* 1*

prepared from urine fa ile d to cause fo llic u la r growth or corpus luteum

form ation, but led ex clu sive ly to pronounced theea lu te in lza tio su

Contrariwise, th is never happens in normal adult rats*

Poster and Hiaaw (1939) wore ab le to produce, by the use of F. 3* K*

in immature ra b b its, only pure fo llic u la r growth without a tr e s ia , cy stic

degeneration m ovulation* Intravenous in jectio n s o f an ovulating

mixture (F . 8 * 1 . and 1 * B*| in animals previously treated with IV 3* H.

suboutaneously fo r 9 days, resu lted in ovulation o f 40-60 f o l l ic le s

with subsequent lu te ia ia a tio n and psendopregnancy* F* S. B« and higher

concentrations o f 1 * H* resu lted in a tresia , cy stic degeneration and

lu tein isatiem * Experimental ovulation under these conditions always

fa ile d even i f they used 1000 mgnu Of $** H* or more* Bunde and Oreep

(1936) using young adult hypophyseetomized r a ts , obtained almost to ta l

regression of the p ersistin g corpora lu tea w ith marked dim inution of

ovarian weight by the use of 1* H. The character of the corpora lu te a ,

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lith * 1 4 tb day of gesta tion and Abortions occurred w ithin 36-48 fare*

A fter tb s la s t in jection# Giving ©etrin a l l in one dose did not canse

Abortion* | f *23-4.0 u n its o f oxytocin mm $ivea immediately a fte r

260 u n its o f e str la had been Injected over a Id hr* period , th e abortion

follow ed with fa ir regu larity in 1-6 hre* Iteri& s and Newton (1935),

aleo working with m ice, reported that th e In jection of 400-500 sense

h a lts o f e s tr ia adm inistered on the 15th day of pregaaaey increased the

se n s it iv ity of the uterus to oxytocin so th at one un it o f p ito c in ,

otherwise without e ffe c t * now caused uterine cont raction which was

su ffic ie n t to h i l l or d e liv er ease o f the lit te r s * The sane e ffe c t was

obtained i f a phenolic eth er-solub le fraction of pregnancy urine mo

used*

P. B* Smith (1932), by removal o f the p osterior lobe o f the

hypophysis* showed that in the ra t the p osterior lobe m e not necessary

eith er fo r th e genesis or maintenance of u terine contractions at parturi­

tion* fh le has more recen tly been v e r ifie d by Peaeharss and Long (1935)

and several others* Reynolds (1932) sta te s that as l i t t l e a s *05 cc* of

ovma^-stimalabing substance o f human P . V* (Parke, Davie and Co*)

induced a tran sitory decrease in the m o tility o f the uterus o f the

unanesthetized rabbit in 6 -8 hrs* large amounts {*4-1*0 4 0 *) in the

presence o f the ovary resu lted in ovulation and u terin e quiescence. He

further sta tes that In h ib ition of m o tility is independent of th e ovaries

but the ovaries contribute to the cause i f they are present* In a second

paper (1938) Reynolds, using the f is tu la method, concluded that e str in

i s sp ec ific and i s the causative agent for the in it ia tio n and maintenance

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eat of 8 rabbits injected for 5 days with ©etrin during the later stages

13

Of g esta tio n , ons aborted and the remaining 4 retained th e ir fetu ses

with consequent resorption . Hypophyseotoray did not in terfere with

responses o f the uterus to ex tra cts o f the corpus la teu a .

&tat» i t seems w ell established that in the ra t the corpus luteum

dees not in h ib it uterine m o tility nor does e str in se n s itiz e the uterus

to p lto e in , but in the rabbit i t would seem that progestin does in h ib it

u terin e m o tility and e str in a lso se n s itise s the uterus to p ito c in .

T heB ole o fth e Placenta in Reproduction; Researches conducted

during the past 7 years or so have served to reemphasize the role o f the

placenta as am snAoorih* organ in regard to reproduction. C olllp and

h is associa tes (1990~31, 1954) have announced patent preparations

containing a . f * L. substances obtained from the human placenta. K linger,

Burch and Cunningham (1795) Injected an alcoh olic ©attract of human

placenta in spayed r a ts . twenty-one t 9 days a fte r spaying, th e hypophyses

wore removed from the in jected animals and transplanted in to subcutaneous

t is su e s of in fa n tile m ice. A study of the mice ovaries showed hemorrhagic

f o l l ic le s and corpora lu tea to be present* They concluded that the

placen tal extract stim ulated the A. P . and that the e str in decreases the

ovary-stim ulating power o f the hypophysis, thus verify in g the contentions

Of Leonard, Meyer and Bisaw (1791). B est on (1935), studying the ro le o f

the placenta in the mouse, concluded that the placenta plays an important

part in the in h ib ition of estru s during the la tte r h a lf o f pregnancy and

probably a lso in determining the date o f parturition . The fetu ses were

k ille d at varying in terva ls up to the 15th day of gestation but the

placentae were l e f t in ta c t. The lo s s o f the placentae was the sign al

19

f m th e conseneement o f © stras, and a fte r th is , abortion. The placentae

TO# delivered at term. Eetrue occurred X*£ days a fter uterine

evacuation* Boston termed the evacuation o f these placentae a t term

"paeudo-parturltlon^ Hateriua {195$} reported that 9 oat o f 3d pregnant

female rata b ila te r a lly ©varieetoaiizea during the la t te r part o f preg­

nancy reached term or proceeded beyond provided cmly one fe tu s mas

Carried in u tero . So k ille d the fetu ses in utero by means o f a cautery

needle but the placentae were l e f t in tact* The b irth mechanism appeared

d e fin ite ly to break down* He suggested, without much evidence, that the

placenta secretes progestin or a p rogestin -lik e substance* Rirseh (1933),

in stu d ies conducted on th e control o f the length o f gestation in the

r a t, concludedi (1 ) The fe tu s played no important part in governing the

length o f pregnancy in the r a t; K linger, Burch and Cunningham (1933)

had reached sim ilar conclusions* (ft.) normal pregnancy in the rat i s

invariably terminated by ovariectomy, u sually w ithin 48 hours; (3 ) The

placenta was necessary for m aintaining u terine quiescence ch aracteristic

o f pregnancy; (4 ) The placento~uterine complex was important in main­

ta in in g gesta tion and probably in con tro llin g i t s length* He sta te s

that i t secretes progesterone or a sim ilarly actin g principle*

C oilip (1934), Bain (1933), and Astwood and Greep (1938) were

unable to produce any detectable a ffec ts by the im plantation of rat

placentae. Her could C oilip ©t a l secure resu lts from the us© of

placentae from dogs, rab b its, p igs or cows, m ss Hain (1985) implanted

8 ra ts w ith 1 , ft or 3 rat placentae apiece between the 1 1 th and 17th

days of gestation and the length o f gestation of the treated ra ts was

20

unaffected . Astwood and Oreep (1988) reported that the in jectio n of

large amounts o f fresh rat placental suspension in to 4 ra ts a t frequent

in terv a ls from the 19th 4ay o f pregnancy onward did not delay parturi­

tio n . l iv e fe tu ses o f normal b irth weight were delivered on th e Slat*

Sind day.

Holland (1928}, in a study on the cause o f fe ta l death found that

ten of seventeen dead macerated human fetu ses had unusually sm all

placentae. He could fin d no other apparent cause o f death.

Studies on the perm eability of the placenta have been le w numerous.

Ccurrier (1931). studying the e ffe c ts ©f the female sen hormone (e str in )

in pregnant guinea p igs (p . 1339). sta tes;

"Cett® hormone fo llie u la ir e in jectee a la fem elle do cobaye geetante, traverse 1© placenta e t a g it sur I ’appareil g en ita l dec foetu s fem elles dent d i e provoque le developpement

He further sta tes (p . 1303); *Le foetu s lui-meme eat capable de reagir in utero.^ 8© l fon in je c ts do la fo llle u lln e a fo rtes doses a uae fsm ells p lein e dans la m ise-bas, e l le traverse 1© placenta et vient se n s ib ilise r le traotus gonito-maraaair© des foetu s fem elles e t la am cH e dee foetu s maXes.*

L e ll, lib e r and Snyder (1938), using a non-toxic dye, phenol-

sulphonphthalein, on stu d ies in placental tranaaaiseion and p em ea b ility

during idie la s t th ird of gestation in the rabbit, found that: (1) The

in jectio n o f each fetu s with 3 mgm* of dye resulted in an output during

a 6 h r . period o f ©ore than 30 per cent of the dye frcaa the fe tu s at

the 22nd day o f pregnancy, while le s s than IS per cent of th© dye was

excreted at the 29th day. I f the gestation sac (fe ta l membranes) were

in jected with 3 mm* Of dye, on output of 4 per cent resu lted in three

and two-third hours at 21 days and 40 per oent at 30 days of pregnancy.

21they coneluded, therefore, th a t there was a decrease in p lacental trans-

o f dy© as tern approaches and In Increase in perm eability of

the to ta l membranes. Huggett (1927) measured the fe ta l and maternal

blood-gas ten sio n s la the goat and concluded that the d ifferen ce between

the oxygen and carbon dioxide pressures on the maternal and f e ta l sidee

was su ffic ie n tly great to explain the d iffu sio n of gases from the

maternal to the fe ta l blood through the placenta. He asphyxiated the

mother and ra ised the Carbon dioxide ten sion in the maternal blood so

that the fe ta l riotous blood absorbed carbon dioxide from th e mother.

He a lso obtained marked in h ib itio n of oxygen, passage to the fe tu s .

Shimldau (1922), studying the perm eability of the placenta of ra ts

and mice to certain dyes* found that the perm eability of the placenta

fo r each dye investigated (8 basic dyes and 15 a d d ones) was the same

for the rat and mouse. He a lso concludedJ

*We are ju s tif ie d la saying that the fe tu s i s free from the dye when no color i s detected in the fetu s by the same method of detection that reveals the sta in in th e mother, fo r th is reason X excluded the dyes which do not deeply sta in the maternal body.*

A ll the basic dyes that ShimidaGU tested passed through the placenta,

and 6 o f the 13 sulphonic acid dyes (E-SQ^S) passed through but 7 o f

them did n ot. D ian il Blue £E* which the w riter used in th is study was

not In vestigated , however. H is r esu lts ind icate a lso th a t any lip o id -

Soluble dye w ill pass through the p lacen ta. Shialdatt further sta te s:

*fh© perm eability o f the placenta i s to be considered as a phenomenon o f u ltra f iltr a t io n in Beehhold’s sons© (*Q7-f08); the placenta here corresponds to an u ltr a - f ilte r .

Xhe power o f the dyes to pass through the placenta runs p a ra lle l to the co llo id a l sta te o f th e ir so lu tion in the serum, esp ecia lly to th e ir a b i l i ty to spread in a gel of high percentage."

32

h m g and IIvans (1928), in th e ir monograph (p . 6 0 ), sta te concerning

the ace o f D ian il Blue m that:

* fh is dye, in aqueous so lu tion a deep reddish-blue, or b lu ish -v io le t can b© adm inistered at any time in the sexual h istory o f eh animal in doses so large as to f i l l the peri­toneal cav ity and on d or 6 su ccessive days without serious to x ic e ffe c t and u su a lly , Indeed, without any detectable toad# o ffse t m the animal.*

»# X* Meyer (personal eesjnnnicatlcn}, however, was able to in h ib it

the e str e ss cycle o f ra ts when th is particu lar dye was in jected in

amounts coaparable to those o f long and Ivans*

Hamilton and Wolfe (1908) in jected testosteron e propionate

(Perandrexi from Ciba Co*) in d a ily subcutaneous doses o f 500 gamma from

the 10th-17th day of pregnancy in the rat* $be young showed a masculine

appearance as evidenced by a large c lito r is and sometimes the lade o f

a vaginal o r ifice* I t is evident in these experiments that the placenta

was permeable to the bexmono injected*

the fa c t th a t n u trition and metabolism play an important ro le in

reproduction was reemphasised by the work o f Mason in 1935, and others*

Mason sta te s that vitamin A deficiency could f ir s t be detected by the

presence o f a continuous estrcus smear* In r a ts d efic ien t in v ltea in

A, and in A and X, abneim&l cern iflca tio n p ersisted for some time prior

to mating and throughout pregnancy* Dnder such conditions fe ta l death

and resorption , or e ls e b irth or reten tion of dead fetu ses in utero

a fter prolonged gestation and d if f ic u lt parturition , resulted* mien

Complete resorption did not take p lace, pregnancies often were prolonged

to the 84th, 85th and 86th days and were follow ed by a marked disturbance

o f the b irth mechanism* P arturition often extended over 3 days; the

23

tm eXes m m X ugly were in great d istress# Often the fe tu se s , only

p a r tia lly extruded, were decapitated or badly macerated by the mother

In an attempt to remove them f rcsa the vagina, Dead and liv in g fetu ses

were often eaten by the mother* Placentae were sometimes found expelled

from the uterus# Death of the mother occasionally occurred during

parturition or a few days later* F ifty -e ig h t per cent of th e fetu ses

were delivered dead or found dead in u tero, and about 70 per cent o f

the liv in g fe tu ses delivered died w ithin 5 days* He sta te s that mor­

t a l it y was due to the inherent weakness o f the young and impaired

la c ta tio n in the mother# The s iz e o f the fe tu s had no sig n ifica n t bearing

upon d if f ic u lt p artu rition . The ovary and hypophysis were not s ig n if i­

cantly impaired* prolongation of pregnancy in A -deficlent ra ts was

d e fin ite ly not due to hyperfunction o f the corpora lutea* so Mason

sta te s .

la c ta tio n as le la te d to Reproduction! sin ce the mammary apparatus

i s c lo se ly associated with reproduction, i t may be b r ie fly stated th at

estr in seems necessary to cause the development o f the duct system of

the gland, progestin for the growth of the lobules and prolactin fo r

the actual In itia tio n of secretion (A llen 's Sex and Internal Secretion s,

Chap. 211). Smith and Smith (1933) were able to in h ib it la cta tio n by

the in jection of large doses of e str in in pregnant rab b its. They are

of the opinion th at retained placentae, when abortion occurred, inh ib ited

la c ta tio n due to the th ee lin present in the placentae. Lafcr and Kiddle

(1936) temporarily suppressed the estrous cycles In ra ts by the d aily

In jection of 15-30 u n its of p ro lactin . Sim ilar in jectio n s of progesterone

24

(proluton) were without e f fe c t . Gcmnon (1937), la Evan’s laboratory,

rap ortad p a r tia l or complete cessa tion o f la cta tio n in the ra t by the

adm inistration o f large amounts o f A ntuitrin-3 beginning at b irth and

continuing for 3-5 days.

The E ffect o f Lactation on the Length o f G estation: Braabell

(1237) , review ing tb s lite r a tu r e , sta tes that Xataste as early as 1882

recognised that in a number o f mammal a the period o f gesta tion was pro­

longed during la c ta tio n . In mice suckling 3 or more young, gestation

mss prolonged and th is prolongation was due to an arrest in the develop­

ment o f the embryos a t an early stage. Daniel (1910) reported tea

instances o f prolongation of gesta tion from 2-10 days in mice suckling

3-10 young. He concluded that the period of gesta tion varied d ir ec tly

w ith the number o f young suckled. He went so fa r as to say that gesta tion

was prolonged approximately 1 day fo r each young suckled. H is data were

meager, however, since he only had 5 fem ales. In non-suckling female

mice the s iz e o f the l i t t e r bora had no e ffe c t on the length o f g esta tion .

Kirkhera (1916), c ited by Brambell (1937), investigated the stage o f

development attained by the embryos in 21 mice suckling from 3-8 young

and k ille d at d a ily in tervals from 6-24 days postpartum. He compared

these w ith a standard se r ie s o f embryos from non-suckling fem ales that

had become pregnant at the postpartum estru s. He concluded that in the

non-suckling mouse, implantation occurred at the c lo se o f the 5th day

and in the suckling mouse i t occurred at the end o f the 13th day of

pregnancy. This delay he stated was due to the lo s s o f surplus

nourishment through the asrassary glands, preventing the uterine mucosa

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26Im plantation in la c ta tin g shrews* Ha a lso found that the growth of the

corpora o f the second pregnancy waa arrested by la cta tio n in the bank

t e le (C leithrioaonye glareolus britanaieua, M iller ), the number of

embryos in utero had no influence m the duration o f gestation*

Estrua reappears in the suckling rat from 35-40 days a fte r the

postpartum estru s (Long and Brans, 1930)* They attrib uted th is retarda­

tio n to corpus luieum inhib ition*

la the rat the postpartum ovulation may taka place anytime between

the 16th-B4th hour a fter lit te r in g (long and Brans, 1933)* The resu ltin g

corpora lu tea p ersist for a much longer time i f the animal i s In sta tin g ,

not infrequently fo r 30 days, during which time ovulation does not

occur* B&rkee (1936) had described a sim ilar condition fo r the mouse

and had pointed out that persistence o f the corpora lu tea without

ovulation i s serrulated with the number o f suckling young, eetrua

appearing sooner when there are 1-2 In the l i t t e r than when there are

several* Hlsaw (unpublished work) sta te s that th is i s true for ra ts

a lso ; l i t t e r s o f 1-3 postpone ovulation from 12-20 days; these of 4 -7 ,

17-24 days; and thos o f 8 -11 , 23-39 days (A lienas Sex end Internal

S ecretion s, p* 804)*

Thus, summarising, i t would seem that prolongation o f gestation

occurs in the rat when 6 or more are suckled, and that there i s a

correlation between the number suckled and the duration of gestation*

Several attem pts have been made to explain th is phenomenon, but we are

forced to admit that the answer i s s t i l l problematic*

Thus, from a review o f the lite r a tu r e , i t i s evident that in the

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AatuitriiH S (fsrfce, Davie and Company** Anterior P itu itary-L ike

Sex Hormone) i s a standardised, h igh ly p u rified extract o f pregnancy

urin e. I t contains the anterior p itu itary-lik © sex hoaeaion®, which

con sist* o f th e lu te in iz in g hormone end a sm all quantity o f the f o l l i e le -

stim ulating hormone# Concerning the chemical properties a? Prolan

(P* IT* extracto) H. B. fan hyke la h is beefc on m e Physiology and

l l i a i w a i s l ^ states*

"Properties which prolan does net p ossess can he given With seme assurance# B n til i t has hem iso la ted a s a pur© substance* however* le s s that w ill u ltim ately he recognized as accurate can he said about i t s chemical nature. Prolan in aqueous so lu tion i s rather rapidly inactivated hy heating (above 60%)* esp ecia lly I f the so lu tion is b o iled . Askew and P aries (1933) considered th is t o h e due to hydrolysis inasmuch as dry prolan read ily withstood an hour’ s heating at 100® in the presence o f oxygen. According to von Kuler and Zondek (1934) prolan resembled enzymes as to the conditions (temperature and pH) under which i t was inactivated* I t was a lso inactivated by u ltr a v io le t rays end by hydrogen peroxide, but not by a d ipeptldase. fa ir ly pure preparations o f prolan do not contain protein hut they have u su ally been considered to resemble a derived protein ( e .g . , a polypeptide). Prolan i s said to be inactivated by trypsin but not by pepsin (Raise and Haurowitss, 1989; Wiesner and M arshall» 1961$ and O thers), good preparations o f prolan contain no amines, tyrosin e (negative M illoa reaction ), tryptophane (negative Adamklewiez rea ctio n ), adenine, phloreglueinol, halogen, sulphur or phosphorus* the most potent preparations are said to contain 1-2 per cent h is tid in e , 6 per cent argin ine, and carbohydrate equivalent to about 7 per cent glucose (C, 43 per cen t, K, 6 per cen t, 0* 39 per cen t, and u , 18 per cen t)."

$h© gonad-stimnlat ing sub stance found in pregnancy urine does not

exactly resemble any chemical fra c tio n secured from anterior lobe

substance. I t i s le s s e ffica c io u s than p itu itary implants or extracts

31

38

Of whole anterior lob© substances* H. IS* Evans, in h is chapter on

Dysfunction of th e P itu ita ry in GlanduXax Physiology and therapy

summarizes the d ifferen ces hetman anterior Xcfee substances end the

ganad~stljmil&t ing substance o f pregnancy urine as follovrs:

*1* I t cannot stim ulate the growth o f f o l l ic le s and corpora in ovaries o f hypophyseetoaized anim als.

8* I t does not a ffe c t th e development o f the immature avian testes* This i s read ily produced by anterior lobe substance*

$* I t has coaly s lig h t e ffe c ts cm tb s monkey © vary-- e ffe c ts th a t do not lead to reddening o f th e sex skin*This i s produced by anterior lefee substance*

4* When administered to normal immature r a ts , no m atter how frequently or in chat doses* i t rarely produces ovarian weights in excess of from 40 to 70 mg* in n inety- s in hours* whereas with anterior lobe substance increased e ffe c ts are secured with increased dosage, the ovaries atta in in g from two to three tim es the weight stim ulated by pregnancy u rin e•* (See fig * 1#)

0* W* Corner, in h is chapter on Corpus Intern Therapy in glandular

Physiology and Therapy sta te s that iin tu itrin -3 i s h igh ly potent in

producing corpora lu tea in r a ts , but that evidence i s to ta lly lacking

that the human ovary can be so lu tein ized* Monkey ovaries certa in ly

cannot be so affected* Corner further sta tes that i f favorable

therapeutic r e su lts fo llow the use o f pregnancy urine ex tra cts, the

r e l ie f o f symptoms can therefore scarcely be considered to be due to

the corpus luteurn*

There are reasons fo r believ ing that the gonadotropic factor o f

the urine of pregnancy a r ises through the function of chorionic tissu e

and i s not a product of the anterior hypophysis. The evidence as stated

by Evans is*

33

*1* The chorionic t is su e o f the placenta M e large asssuats o f gonadotropic substance in i t f as demonstrated by im plantation o r in jectio n o f ex tra c ts , P e r e a s the an terior lob© o f wG&am dying in pre^ianey has l i t t l e o r no gonadotropin hormone in i t (B iilip p )*

2* 1&« sura# im m & m i s present in h i^ i aaounts in a l l types o f ehorioatc tissu e growth, e sp e c ia lly hyd atid ifom stole and chorionepitheliom * A eth er in women or in

5 . 4 lo r e case o f hydaiiflform m l# has been described in which the body flu id s were sin gu larly lacking in th is substance. Study o f the uterine m ss showed that the m l# had been delim ited from the maternal circu latory system by a m ss iv e fib r in boundary m m 9 conceivably preventing leakage o f the horcs>ne shoxm to be actu a lly abundant in the mole t is s u e ,

4 . 'fhe hormone content i s high in the chorionic v lH i young ova as w ell as in the body flu id s before

Bark©, Davis and Company sta te that A ntuitrin-S has been found to

be o f c lin ic a l value in the following, conditions* functional uterin©

ajssnorrfrsa and ollgasisn&rxhea , habitual or threatened abortion.

dystrophy), and m m v u lg a r is . Exactly how Autuitrin^S brings

about b en efic ia l e ffe c ts in cases o f habitual or threatened abortion

in worsen i s not d e fin ite ly known.

THE EFFECT OF XHJECTIEG AHTOITRIN-S IHTO PREGHANT ANIMALS

The E ffect m G estation and P arturition

A to ta l o f 15$ animals were employed in th is study* Of th is to ta l

55 were in jected with A ntuitrln-S in sin g le doses o f 25-200 H* T7. ranging

from the 15th~2Xst daya, in c lu siv e . These data are summarized in

Table I and more b r ie fly as follow s:

Ho* o f Animals Days Injected Amount Injected

0 18 50 R .8.1 18 651 17 708 18 701 15 754 16 784 17 75

14 1© 756 I f 755 80 781 21 758 18 100Z 16 1502 18 1508 18 2001 19 2002 15,16 30,303 15,16,17,18 25,25,85,25

For the sake o f c la r ity i t m y be restated here that the gestation

period for the animals in our colony was usually between 21 and 22 days.

Of the 2 animals receiving 80 R, 8 . on the 18th day, 1 gave b irth

to dead fetu ses on th e 20th and the the other gave b irth at normal tim e

on the 22nd to v iab le fe tu se s . The 2 animals in jected with 65 R. D. on

the 18th and 70 R. 8 . on the 17th, resp ectiv ely , gave b irth to dead

34

35

m m x* m m m m w m m m m m m m w m mm movmm *s

Animalnumber

-f

30 30

Beys o f Pregnancy

1 » 30 81 88 88 84 35 36 37

»S

M .

78

m m i s l

370oa(e) 78

BA

BBSCBimoN m TABUS I .

Sh# numbers abenre the horizontal lin e s under the days o f pregnancy ind icated represent the amount o f Antuitrin*® in jected In ret u n ite . e-Thla e«lrwi gave b irth to one fetu s on each of days S I, 22, 23, i t and

28. On the 28th d&y an autopsy revealed 8 dead fetu ses in u tero . b«tPerlod o f gestation usually £1*8$ days.t«Abbreviatt0U0 used are as follow s? B» birth of one o r more fe tu ses;

B» Death o f mother; a , autopsy performed; 1 , liv in g fe tu s e s ; d, dead fe tu ses; r , fe tu se s being resorbed*

♦•M sltiparous animals; a i l other® were primiparous.***4jgad in two se r ie s of in jec tio n s.

m u s t* )

m

j m a u i n ma70 814

aaaa**) ?o M88013(1) m MS®

70

70 Bd

70

« — ,..— ..........,.... rT 75 - A

78 84

w e p ta ).............. , 78 BA

a p u f c ? .... ............ 76 Bd

Z 78 Bd

8 78 Bd

i ________ ________ 75

37

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SS0 0 9 t f J TO m

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200

200

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36

lui&be#' ...If , !« T 80 2i _ j * _

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n ___BX45 ,0 0 5 ,., n Bd

W00B(*1 75 " Bd

SOO BI

(8 ■ n _ ^ . Tm

87003(a) 78

37002(b) 78 B1

37 ■f* T

39

fetu ses on the 22nd day* Of the 9 receiving 70 R, TX. on the 18th, 2

gave b irth to dead fe tu ses and 3 gave b irth to liv in g and dead fetu ses

on the 21st; another autopsled on the 21st had dead and p a r tia lly resorbed

fe tu ses in utero; a 7th had liv in g and dead and the other 2 had dead

fe tu ses on the 22nd day# The one animal which received 75 R* U* on th e

15th had liv in g and dead fetu ses on the 82nd* Three fem ales receiving

78 R* tT* cm the 16th aborted on the 20th; 2 of these had dead fetu ses

while the 3rd had liv in g and dead fe tu ses (Rigs* XX and I I I ) . The 4th

female of th is se r ie s gave b irth to 1 dead fe tu s on the 2 1 st« Autopsy

Of th is animal revealed 2 liv in g and 1 dead p a r tia lly resorbed fetus*

Of the se r ie s o f 4 animals to which were administered 75 R* IT* on the

17th, 2 gave b irth to dead fetu ses on the 21st and the remaining 2

autopsled on days 18 and 19, resp ectiv e ly , had liv in g ones in utero*

14 animals in jected w ith 78 R* tf* on the 18th were e sp ec ia lly in te re st­

ing* 1 autopsled on th e 19th had liv in g fetu ses In utero; 3 aborted

dead fe tu ses on the 20th; 6 gave b irth to dead, and 1 to liv in g fetu ses

on the 21st; 1 had liv in g and dead fetu ses on the 21st and autopsy on

the 22nd showed her to have dead and p a r tia lly re sorbed fetu ses in u tero .

The remaining 8 animals o f th is ser ie s gave b irth on the 22nd to liv in g ,

and liv in g and dead fe tu ses, resp ectively . 1 of the la tte r anim als,

Rfo. 37004(c), and a lso Ho. 30009(f) were the only animals of the group

receivin g 75 R. XI* on the 18th day which gave b irth to viab le young.

The 6 animals injected with 75 R. t?* on the 19th a l l gave b irth on the

28nd day 8 had dead, 3 liv in g and dead, and 1 had liv in g fe tu se s . 3

animals adm inistered 75 R, XX. on the 20th had viab le young on the 22nd

40fand X receiv in g 78 R» TJ* on the 21st gave b irth to v iab le fe tu ses on

the 23rd. 5 animal a were in jected with 100 R. XX, on the 18th; 2 gave

b irth on the 20th and 1 on the 81st to dead fetu ses; a 4th gave b irth to

1 dead fe tu s on each or the g is t , 22nd, 23rd, 24th and 25th days, and

autopsy on the 26th revealed 8 more dead fetu ses in utero which had

evid en tly been dead for 4-8 days a s indicated by th e degree of a u to ly sis ,

n ecrosis, and resorption that had t&kan p lace. The dead fe tu ses and

th e ir membranes were much decolorised and em itted a most o ffen sive odor.

The 5th animal gave b irth to 1 dead fetu s on the 22nd and then d ied on

the 24th day. autopsy revealed 8 dead fetu ses in utero which had

undoubtedly died before or a t term. I t seems probable that the female

was too weak to give b irth to them. 2 animals receiving 180 R. U. on

the 16th gave b irth to dead fetu ses on the 21st and 24th days,

resp ectiv ely . In the ease of the g animals receiving 180 R* U. on the

18th , 1 had liv in g and dead fetu ses on the 2 1 st, and the other died on

the 23rd. Read fetu ses were found in utero at autopsy. 1 of 2 fem ales

which received 200 I* XJ. on the 18th gave b irth to 1 dead fetu s on each

Of days 84, 28, 26 and 27; the other had not yet given b irth on the 25th

when an autopsy revealed 4 dead fe tu se s , 1 placenta s t i l l attached, and

7 liv in g young, some of which were possib ly a l i t t l e heavier than tern

fe tu se s, sin ce they averaged 5 .5 ©as. in m ig h t. This i s the only case

in the whole se r ie s of animals in which gesta tion was markedly prolonged

sin es an animal which gave birth during the e a r lie r part of the 23rd

day cannot be considered a case of prolonged gestation as th is i s w ithin

the lim its o f normal varia tio n . The one animal receiving 800 R. TJ. on

the 19th gave b irth to liv in g fetu ses on the 22nd day.

41

The 2 an imal s in jected with 30 It* D* per day each on the 15th and

16th days were autopsled on the 19th* 1 had liv in g , and the other

liv in g and dead fetu ses in utero* Of 3 additional fem ales in jected

with 85 R. U* each per day on the 15th# 16th# 17th, and 18th days, 1

died on th e 80th and autopsy of the 2nd on the same day# and of the 3rd

on the 81st revealed that a l l 3 had dead fetu ses In utero*

I t I s evident from Table I that animals in jected on the 15th-18th

day with dosages of 50-75 R# 0* aborted dead# or liv in g , or dead and

liv in g fetu ses before or at term* liv in g young were usually not v iab le

at b irth (fab le II}* In no case w ithin th is ser ie s was postponed or

prolonged parturition detected even a fte r hourly and in many cases

continuous observations* Of 6 animals Injected with 75 R* U. on the

19th only 1 gave b irth to viab le fetu ses; and 5 receiving 75 R* XT* on

the 80th day, a l l gave b irth to v iab le young* In no case when in jectio n s

were made on the 19th or 20th was gestation prolonged# since In a l l

instances th e period of gestation was 22 days* These r e su lts can hardly

be compared to those of le v in , Katsman and Daisy (1931) who in jected

1500-7000 Mouse Units* They agree in part with those of Hain (1935)

who had only 4 anim als, but they are in contrast to those of Hoopea

(1934), and Hoopes and King (1938) who obtained a regular prolongation

Of pregnancy in the rat of 3 or more days with 75 R* IT* of A ntuitrln-S

administered on the 19th or SOfch day of gestation* They a lso obtained

a delay in parturition u n til the 25th, 26th, or 27th day with parturi-

tio n extending over 8 or more days. They further reported that the

fe tu ses remained a liv e in utero and continued to grow for £ or 3 days

42beyond term*. I t i s important to note that in the present experiments

a l l animals in jected on the 20th and Blot day gate b irth at term to

viab le fe tu se s .

Dosages o f 100 R# U* administered on the ISth day gave varying

resu lts* 5 eases resu lted in the birth of dead fetuses-*—- 2 on the 20th

and 1 on the 21st day} in 2 other animals under sim ilar treatment

parturition teas postponed in both eases as evidenced by th e fa c t that 1

gave b irth to I dead fe tu s on the 22nd and died on the 24th day a fter

which autopsy revealed dead fetu ses in utero* la the other case 1 dead

fe tu s was bom cm each o f the 21 st, 22ndf 23rd, 24th and 25th days}

autopsy on the 26th revealed 5 dead fetu ses in utero which evidently

had died 4-5 days previously*

I t was thought that i f the doses were graded or extended over a

longer period o f tim e that perhaps the animal might be able to excrete

the hormone fa s t enough so as to prevent or modify i t s e ffects* With

th is in mind 3 animals were in jected each w ith 25 E. TJ. per day on days

15# 16, 17 and 1$ making a to ta l o f 100 R*tf. In 8 of these cases

subjected to autopsy on the 20th and 21st days# respectively# dead

fetu ses were present in utero* The 3rd animal died on the 21st day and

a lso had dead fetu ses in utero a t autopsy* Evidently in these 3 cases

c ited the homone could not be excreted fa ct enough to prevent i t from

having i t s e ffe c ts (See a lso animal Wo* 5 in Table I)*

When doses o f 160-200 R. TJ* were adm inistered, 4 out of 7 animals

showed parturition to be postponed and prolonged for as long as 1—5 days*

These were d e fin ite ly not cases of prolonged gestation but cases of

postponed parturition with the p ossib le exception of 1 case# No* 38009(c),

43

which had liv in g and dead fetu ses in utero on the 25th day. The fa ct

that the fetu ses died before or at tern (exception e ited above) was

ascertained by exploratory laparotomies or autopsies or by th e ir being

bom* Further evidence to substantiate these observations was supplied

by the fact that necrosis and au to lysis of the fe tu ses had proceeded

so far that an exceedingly offensive odor was nearly always present when

autopsy was performed or birth occurred* The extract in conjunction

with the putrefaction of dead fetu ses in utero resulted in the birth

mechanism being upset and eventually in most cases caused the mother’s

death*

I t i s believed that the induced corpora lutea were not responsible

for these eases of postponed parturition since the fetu ses died before

or at term (exception previously c ite d ) , and some animals with w ell-

formed induced corpora gave birth prematurely or at term to dead ar nan-

viable fetuses* Also 2 animals copulated and became pregnant on the

same day that abortion occurred. Further evidence w ill be c ited under

the o ffse ts of v ita l dyes in a la te r section of th is paper*

The Condition and Size of the Fetuses

Bata obtained on the condition and s ize of the fetu ses of 63

animals injected with 50-200 E* tF. of Antuitrin-S on the 15th-21st day,

in c lu sive , (Table® II ana I I I ) showed that of a to ta l of 506 fetu ses

accounted for by the presence of placental s i te s (which indicated that

the mothers had eaten th e ir young), by exploratory laparotomy, autopsy

or b ir th , only 74 were liv in g 72 hours postpartum, the remaining 432

44

x a b u s n . cmnnm os rsmrn mm m m s m n o i s s i m mmmn^a

inject"

II

ed Inratunit*

M » o rofyoung

and

I?

Condition of young when

COP SAP* lop* foraed1

f

Condition of young at birth®

n

Ho* o f young dead 72 hrs* post* p ar tun

Ho* ofyoungliv in gn hr a*o f morspost*partum

e&

1 5 0

16i f

16

ijyeBD^ 3222222; 11 0Te

a 0 nr& 75ISO

1L5Dmm

0SKa 16 0

6

Description of Table XX

a-Number of young bom and eaten as indicated by the presence o f p lacental Alton in the uterus when autopsy or exploratory laparotomy was perforated* No young could be found in the cages* * lso lo s s Of weight w ithin IE hrs* previous to examination indicated that the animals had given birth*

b*Bxp. la p ., exploratory laparotomy* B, dead; Lf liv in g ; R, being resorbed* c-Numbsrs in parentheses indicate the umber of days since conception

occurred*JHSnltiparous animal; had been injected in a previous s e r ie s . i4ll other

animals wore priaiparous.♦-Only case In Which l iv e fetu ses wore found in utero past term*

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47

being, o f course, non—v ia b le« Thus 14*6 per e@Qt of the fetu ses were

viab le and 85,4 per cent were non-viable. Of 270 fetu ses from 32

animals injected on the 18th day with dosages of 50-200 R. 17., 97.6

per cent o f these were non-viable with 2 .4 per cent v iab le . Viable

fetu ses were bom to 1 animal receiving 150 B. U. on the 16th, to 1

receiving 50 R. 0 . on the 18th, to 1 receiving 75 R. V, on the 18thf to

% receiving 75 R. TT. on the 19th, to 1 receiving 200 R. TJ. on the 19th,

to 3 receiving 75 R. t7. on the 20th, and to 1 receiving 75 R. T7. on the

81st day. In case of the remaining 52 animals a l l the fetu ses were non-

viable* th is means that 82.6 per cent of the females injected had non-

viable young, and only 17.4 per cent had viable young and these were for

the most part animals Injected on the 20th and 21st days. No difference

was detected between primiparous and raultiparous animals.

Thus i t would seem that on the 18th day of pregnancy the tolerance

of the fetu ses to the e ffe c ts of Antuitrin-S i s considerably le s s than

i t i s on the 19th, 20th and 21st days, ©ale i s exactly what one would

expect since as pregnancy advances the fetus i s more fu lly developed and

approaching parturition time while the function of the corpora lu tea i s

beginning to wan®. According to long and Ivans (1922) the corpora lutea

of pregnancy in the rat ©ease functioning about 12 hours before parturi­

t io n . In addition the placenta© certain ly must ba in the early stages

of separation from the uterine wall by the 20th or 21st day since birth

in some cases did occur as early as the 21st in non-injoctod animals.

Day 18 i s esp ecia lly important since i t corresponds more or le s s

to day 19 of Hoopes and King’s experiments.

48

In th is aeries of experiments only 1 animal. No. 38009(c), injected

with $00 R. H* m the 18th had liv in g fetu ses in utero past tern . An

autopsy on the 25th day revealed 1 dead and 7 liv in g fetu ses in the l e f t

uterine cornu, 3 dead p a rtia lly resorbed fetu ses and one almsot com­

p le te ly resorbed fe tu s with i t s placenta s t i l l present in the right

cornu* the liv in g ones averaged 5*5 gms* at autopsy but even though

they were the heaviest fetu ses recorded, they showed no evidences of

postmaturlty such as the v ib r issa s , the shin and the extrem ities recorded

by Hoopes (1934) and Hoopoe and King (1935)*

fe tu ses bom past term from animals Injected with the larger doses

(150-800 R* ?*) not infrequently appeared compressed as though they had

been subjected to unusual pressure* At tim es, but not always, parturi­

tio n was so prolonged that several hours were required for the expulsion

of a s in g le fetus* la two animals at lea st i t took 8-6 hours to expel

one fetus* I t should be mentioned in these cases o f postponed parturi­

tion that the mother® usually got weaker and weaker as pregnancy con­

tinued, refused to e a t , and f in a lly died* With the smaller doses

resorption was rather the exception while with the larger doses i t was

usually the rule* I t seemed that in acme few cases the fetu ses with the

larger placentae usually survived longer than those possessing smaller

one®* Hoad fetu ses removed by exploratory laparotomy or autopsy possessed

decolorized membranes. Out of 506 fe tu ses, 185 (36.5%) were eaten by

the mothers a® evidenced by the presence of fresh placental s i t e s or

placentae found when an exploratory laparotomy or autopsy was performed*

Whether those were stillb o rn or not i s unknown* At the other extreme,

49

for example, were two females injected with 75 R. IF* on the 16th day*

Mo. 38008(a) aborted 3 dead premature fetu ses on the 20th day. These

weighed 1*7 gms. eeeh (F ig. 2 ) . The other. Ho, 88003(d), had at lea st

9 fetuses* Two of these were partly eaten, 3 weighed 1 ,7 , 1*6 and .9

gwSf, resp ectively (Fig* 3 ) , Another fetu s apparently ju st born was

s t i l l l iv in g la the enveloping f e ta l membranes. This young with i t s

membranes and attached placenta weighed only 2*3 gms. (F ig, S ). The

other 4 young had been eaten a s indicated by the placental s i t e s present.

These were both, of course, oases of abortion of premature fe tu ses.

Undoubtedly the placentas had pulled loose and the growth of these

fetu ses had been arrested early due to the in jection s since normal 80

day old fetu ses were much heavier than th ese .

During the ea r lier part of th is study, 12 pregnant animals were

in jected subcutaneouely with 40-100 R. U* of Antuitrin-S varying from

the Sth-l&th day of pregnancy* In an attempt to get a more gradual

e ffe c t than when only one dose was given, the in jection s were made on

each animal on 2 consecutive days* For example, an animal was given

85 S* U. on the 8th day and 25 S . U* on the 9th day thus making a to ta l

dosage of 50 R. U. Autopsies of a l l 18 animals on the 18th day revealed

l iv e , normal-sised fetu ses in ©very case.

King (1938) was able to prolong pregnancy with the consequent

birth of postmatur© fetu ses by the intraperitoneal in jection of 50 R. U.

of Antuitrin-S provided the extract was administered within 4 days a fter

implantation* The resu lts given above do not necessarily contradict

those Of King since the method of in jection was not the same, but

neither do they substantiate them.

so

Further proof that AatuitriiHS resulted in injury to the fetu ses

i s home out by the fact that some of those bom a liv e were placed with

fo ste r mothers which had young of comparable age* but they fa ile d to

survive* In cases in which experimental animals had viable young,

usually these young were not up to par as shown by their growth curves

even though the l i t t e r was a small one. There might be two possible

explanation to t h is . On© i s that the young were bom inherently weak

and were consequently slow to begin growth; the other i s that lacta tion

was inh ib ited , at le a s t p a r tia lly , or possibly i t was a ocrabination of

these two fa c to r s . In % eases young from uninjected mothers placed with

in jected ones soon a fte r they had given birth did not survive. This

would agree with the findings of Connon (195?) who was able to in h ib it

la c ta tio n in mice and r a ts , completely or p a r tia lly , by the in jection of

A. P . L. hormones given at birth and continuing for 5*5 days. I t i s

a lso In accord with the findings of &sith and Smith (1955) who were able

to in h ib it lacta tion in rabbits with large amounts of e s tr in . I t i s

possib le that in the experiments c ited here that the estrin produced by

the stimulated f o l l i c le s in animals injected with Antuitrin~S was

responsible for some inh ib ition o f la cta tio n .

Since the Injection of 75 R» on the 18th day regularly (2 excep­

tion s) caused the birth of non-viable fetuses before or at term, and

since the average number of young per l i t t e r in our colony i s between

7 and 8 , i t was thought that the injection of 10 R. per fetu s inside

the fe ta l membranes might possibly throw lig h t on how the Antultrin-S

had i t s e ffeo ta . A ccordingly some of the fetuses of 4 animals were

treated as fo llo w sj a multiparous female, No. 38014(a), was found by

exploratory laparotomy on the 19th day, to have 5 fetu ses in the right

oorau and S in the le f t* The membranes of the 5 fetu ses In the right

eonai were injected by means of a fine needle and a 1 ce. tuberculin

syringe with 10 B* U* of Ant u itr in-8 each* Special care was taken not

to injur© the fetu ses with the needle, and a lso to prevent as l i t t l e of

the enclosed flu id from escaping a s was possible* 37 hours a fter being

in jected the fetuses were removed by a Caesarian operation* One of the

5 injected fe tu ses was dead and almost completely resorbed* The

remaining 4 were smaller than the uninjected ones on the l e f t s id e .

They were s t i l l a liv e but le s s active than the uninjected ones as they

could not be made to move by probing them lig h t ly through the fe ta l

membranes with a pair of forceps* The ones on the l e f t side moved

vigorously under the seme stimulus* The placenta© of the injected fetu ses

were almost completely detached from the uterine wall as they could be

l i f t e d out without the appearance of any fresh blood| those of the unin­

jected ones were s t i l l firmly attached, hard to remove, and upon removal

profuse bleeding was in itia ted*

By means o f an exploratory laparotomy i t was found that female No*

38015(h) carried 8 fetu ses in utero on the 18th day, 4 in each cornu*

The fe ta l membranes of the 4 on the right side were injected with 10

B. TJ. of Antuitrin-S each while the membranes of the ones on the l e f t

side were punctured in a manner sim ilar to the injected ones but no

hormone was given. Autopsy a fter 48 hours showed that 2 of the 4

injected fetu ses had died and the other 2 were liv in g but seemed anemic

52

and were smaller than the uninjected ones* Also the placentae of the

Injected ones pulled loose e a s ily and showed no m acular connections,

while those of the uninjected had a narrow, whitish ring around the

outer edge but they s t i l l retained a central vascular connection*

A third animal, Ho* 38015(1), had 3 fetu ses in the l e f t cornu* The

2 nearest the vagina were injected with 10 R. U. each and the membranes

of the one proximal to the uterine tubes were punctured only. After 48

hours, the uninjected fe tu s was liv in g and i t s placenta was s t i l l firm ly

attached and vascular* The injected fetus adjacent to the vagina was

s t i l l l iv in g but i t s placenta was loose ly attached and non-vascular.

The 3rd fetu s had died and was being resorbed* I t s placenta was a lso

loose ly attached and non-vascular.

your of the 7 fetu ses of a 4th animal, Ho* 38015(1), received 10

1* TJ. each on the 18th day and the membranes of the remaining 3 were

punctured only* On the 19th day the mother was quite sick and a blood­

stained ssicoidal f lu id was seen to ooze from the vagina* One dead young

of about normal s iz e was bora on the 21st day* Autopsy on the 22nd

revealed that the 4 injected fetu ses had died and were d e fin ite ly tinder-

going au to lysis and necrosis and their placentae were completely detached

and avascular. One of the uninjected fetu ses had been born but the

Other 2 were s t i l l l iv in g with firm placental attachments*

Thus from th e evidence at hand i t would seem that the in jec tio n

of 10 R. TJ. of A ntultrin-S into the f e t a l membranes of rat fe tu ses on

the 18th or l§ th day of pregnancy resu lted in a detachment of the

placenta© and fe ta l death.

The Condition of the Uteri

53

Almost Invariably on the End day a fter in jection , a yellow ish ,

mucoidal f lu id was seen to oosse from the vagina. As pregnancy proceeded

the co lor changed to a reddish hue, and la te r to a dark red due to the

leakage o f blood from the placentae which were losing th e ir uterine

connections and la ter to the autolyals o f some of the necrotic fe tu ses.

In the case o f animals in which parturition was postponed th is bleeding

became profuse and i t was not uncommon to be able to cause the exudation

of blood from the vagina by gently handling the animal. At autopsy an

appreciable quantity o f dark-colored blood was usually found to be

present In the uterus. In cases where the fetuses were not v iab le , the

vascular attachments o f the placentae were loose and sometimes en tire ly

lacking. The uterus quite often was hard and fibrous-like but contracted

readily when stimulated with a pair o f small forceps.

H isto log ica l examinations o f the u teri Indicated that in seme few

cases a sloughing o f f of the epithelium of the uterine lumen had occurred

at places (F ig . 4 ) , but th is was by no means the ru le. That the injury

was not o f a permanent nature was indicated by the fa ct that 2 females

receiving 75 R. TJ. each on the 18th aborted on the 20th and 21st day,

resp ectively , and Immediately became pregnant. Right others became

pregnant within 3—31 days postpartum (Table VI).

The oviducts were found to have developed thick w alls a fter the

in jection o f Antuitrin-S.

The Condition o f the Ovaries34

In contrast to the corpora lutea of most mammals, those of the mouse

and rat remain as su bstantia l, fa ir ly well preserved structures so that

4 or 5 se ts may be present in the ovaries o f a non-pregnant animal*

Pregnancy not only in h ib its ovulation but by the 20th day has caused

resolution of the other corpora present in the ovary previous to th is

pregnancy (Long and Evans, 1922). I t is important to determine i f

possib le how c lo se ly the h isto lo g ica l ovarian changes can be correlated

with the physiological phenomena of abortion, b ir th , postponed parturi­

tion and prolonged gestation . With th is in view the ovaries o f 24

injected pregnant animals, 4 injected non-pregnant, and 2 uninjected non-

pregnant ones were studied. These data are summarized in Tables I I I ,

IV and V.

In order to p o s itiv e ly identify the corpora o f pregnancy from those

induced by AntultrixHd, a v i t a l dye, D ianil Blue 2R, was used* D ianil

Blue SR is a combination of orthotolidine diazotized and coupled in

alkaline solu tion with one molecule of 1,8 diozyaaphthaline 3.6

disu lfon ic acid (Chromotrop© acid) and one molecule of alpha-naphtol

4 monosulfonic acid (Neville-Winther acid ).

The structural formula for D ianil Blue 2R is:

OH OH OH

SO3 N2

SOjNa

55

This dye enables one to d istinguish p o s itiv e ly the corpora lutea

of pregnancy from those induced by Antultrin-S in jectio n s, since those

of pregnancy take up the dye while the Induced ones (those formed after

dye in jection ) do not. Also the older the corpora, the more intensely

they s ta in . Thus the ovary usually showed blue and flesh-colored ,

unstained corpora when i t was removed 48 hours or longer a fter in

in jection o f Antultrin-S; when hemorrhagic f o l l i c le s due to the extract

were present, they were bright red (color of blood) in co lor . A second

criter ion which was also used to d istinguish these corpora is the size

and number of the lip o id granules when Benda's flu id had been used as

a fixative* The corpora of pregnancy contained larger but fewer

granules while the induced ones contained smaller but more numerous

granules, Thus the s ize and number of the granules can be used as a

measure o f the amount o f regression that has taken place (Long and

Evans, 1922), The macroscopical observations were thus confirmed micro­

sco p ica lly , The corpora formed after the f ir s t postpartum estrus and

those of lacta tion could be distinguished from those of the la st

previous pregnancy by the same method (Gompare H g s . 13 and 14 ),

The usual procedure was as follows: On the day the placental

sign was present (usually the 14th) the pregnant animal was injeeted,

at the beginning of the experiments with 4 or 5 c c . , but la ter with

3 c c . , of a 1 per cant aqueous solu tion of the dye on each of 4 con­

secutive days (14th-17th, in c lu siv e ). The in jections were made not

sooner than 3-4 hours after the animals had been watered and fed since

the introduction of such large quantities of flu id into the abdominal

cav ity , no natter how much care one night take in i t s adm inistration

in most oases so disturbed the physiology of the animals that they

refused to eat and became inactive and remained crouched in a corner of

th e ir cages. Usually, however, they seemed to have recovered within

4-5 hours a fter the in jection s. It w ill only be mentioned here, but

elaborated on la ter In the section dealing with the e ffe c ts o f dyes on

pregnancy, that th is dye, as injected in these experiments, often

caused death o f the fe tu s , a u to ly sis , resorption, abortion and even

death of the mother. Long and Evans (1922), using the same dye and pro­

cedure, state that the dye was not seriously to x ic . R, 1C. Meyer

(personal communication), however, was able to inh ib it the estrous

cycle o f rats by the use of Dlanil Blue, I t i s the author’s under*

standing that these dyes (Dianil Blue and Trypan Blue) are d if f ic u lt

to duplicate in manufacture, and it may be possib le that the Dlanil Blue

used here was not actu ally of the same constitution as that of Long and

Evans or that i t contained impurities which Long and Evans' did not

have.

Antuitrin-S was generally administered the day following the la s t

Injection of D lanil Blue, and the condition of the animal was elways

carefu lly recorded before in jectin g the hormone. The ovaries were care­

fu lly examined when an exploratory laparotomy or autopsy was performed

and in some cases they were removed at intervals of 24, 36, 48, 96 and

ISO hours to determine the time required for various dosages injected

on d ifferent days to cause lu te in iza tion and to correlate these changes

with the condition of the fetu ses. Complete data are given in

57

Table 111, but It may be stated here that the in jection o f 75 R* U. of

Antultrin-S on the 18th day caused lu te ln lza tion of mature f o l l i c le s

within 5-5 days which did not become functional enough to prolong

gesta tion . Hot uncommonly i t was found that large lu tea l cy sts , as

pictured by Sngle and Smith (1929), were present (F igs. 5 and 7); the

antra of some of these were f i l l e d with blood (F ig, 6 ). These cysts

were e sse n tia lly large fo l l ic le s in which the granulosa c e l ls had

lu te in ized , but for some reason the fo llic u la r antra had not been

invaded by lu te a l ce lls* In other cases the theca interna c e l ls were

lu tein ized but the granulosa c e l ls had not done so . In some ovaries

excessively large f o l l i c l e s were present but these were for the greater

part cystic and a tre tic (F ig, 6); when they were present, they were

often the predominant structures in the ovary. In certain other cases

the f o l l i c l e s became hemorrhagic (F ig. 9 ) , often showing the presence

of 6-10 of the blutpunkte of Zondek and Ascheim in one ovary. Since

these animals were not k illed with ether, the hemorrhage cannot be

ascribed to this*

Dosages o f 150-200 R* D, apparently caused a more rapid lu te in iz a -

tion but not to a marked degree*.

Two animals were injected with 0.3 mgm* of colchicine per kgm* of

body weight and 9.5 and 11.5 hours a fter , respectively , they were

autopsled. No mitoses could be detected h is to lo g ica lly in the ovaries,

u teri or placentae.

Ovulation was not found to occur during pregnancy, but followed

i t s termination. At no time were ova detected in the oviducts* In 4

s a

TABLE l i t * 0BOTVA2I0HS <# IHJ10TED PEBGHANT AJ&H&S

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Ato!8W5lt iUitultrln-S in jected in

u n its

Days a fter in jection tissu e rerc&cved

Histology of the ovaries*

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Condition at time of in jection

Amount of Antuitrin^S injected la pat u n its

tetya a fter aJaction dasues svsioved

Histology o f tiie ovaries

Remarks

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38007(d)** 6 bears postpartum...... ... laetating& < to ib r* 5 days postpartum ...........k$& & A_____ - ...Aa°ta&a&..

61

cases o f Injected females ova were seen trapped in the newly foimed

corpora lu tea , indicating that ovulation had at le a s t not taken place

in these f o l l i c l e s .

The Postabortive H istories of Injected Animals

A summary of the postabortive h is to r ies of 10 animals is given in

Table YI « Two of these animals must have ovulated within 24 hours after

abortion since they were seen in copulation and became pregnant from

th is co itu s . Two others conceived on the 3rd day postpartum. In these

cases i t i s evident that the induced corpora did not inh ib it ©Btrus.

This is In direct contrast to the work o f Hoopes (1934) and Hoopes and

King (1935) who obtained what they believed to be functional corpora

lu tea by the use of 75 R* XT. of Antuitrin-S administered on the 19th

and 20th days o f gestation (gestation 23 days) in the ra t, but th is

agrees in part with the resu lts o f Bain (1935) who injected 50 R. U. o f

Antuitrin-S into 4 rats on the 14th-l?th day of gestation . However, the

number of animals she used was too small to be of any sign ifican ce.

These resu lts also support those of D#Amour and D*Amour (1933) who

found that P. U. extracts when injected with large amounts of estr in in

la te pregnancy in rats did not increase the number of completed preg­

nancies over that obtained when the estrin alone was used. It is

evident that in th e ir experiments the estr in in some way inhibited the

L. H. present in th e ir extract from forming functional corpora lu tea .

The calculated number of days postpartum the remaining 5 animals

conceived were 1 on the 15th, 1 on the 17th, 2 on the 20th, and 1 on

62

t m t i x . mxtBOBirra atsK ffiiis o f m s o r m m u a a *

sNVMnmikMbWWwMPir Ant***© lt t j . inMF*.

Itay o f1 BTeftEUJ&J*

days a fter la j*

Jtaya a fter abortion next l i t t e r b«wm

Calculated no. o f days conception occurred a fter abortion*

86003(a) n 16 4 84 . m

88003(e) n 17 8 m ■ *&

98011(b) n 17 6 u 80

asooefg) m i f S m 17

38009(f) n m 8 m 8

aso is(b ) n 1 0 8 83

38105(6} 7 0 u 8 4£ SO

36Ca3(f) n : iB Z 88 b•erne day

88015(a) 78 IB 3 $9 7

38015(e) 78 IS 8 88 same day*

*-Penalea wa» kept with Hale* constantly after abortion) net laotating exeept No. 36011(b) ibioh suckled four young,

a-Tsronty-two daps taken as the length of gestation. b-Antaale seen in copulation) keens pregnant.

S3

the 3 1 st, I f the induced corpora were responsible for th is delay in

conception In these 5 casesp they must hare functioned as long as those

o f pseudopregnaney (2 cases) and longer (3 cases) . I f th is i s true

then th is would agree with the findings o f Shelesnyak (1933) who was

able to produce declduomata in immature rats by pregnancy urine in*

jeo tion s, (Fig* 1 illu s tr a te s the e ffe c t of prolan on immature ra ts .)

I t is p o ss ib le , of course, that in the la s t 5 cases mentioned above

these animals copulated soon a fter abortion but became pseudopregnant

when th is copulation was in fe r tile * The pseudopregnant condition after

a s te r i le co ition Is also known for the rat (hong and Ivans, 1922; and

others)* Although ovulation occurs spontaneously in these animals,

sexual stim ulation prolongs the l i f e of the resu lting corpora lutea and

the next estrus i s inhibited for about 12*15 days during which, the

physiological and anatomical changes o f the reproductive tract are lik e

those o f early pregnancy. Long and Ivans found that th is condition

could also b© induced in rats by mechanical stimulation of the cervix

u te r i . The exact nature o f th is stimulus is not known but available

evidence points toward I t s being a nervous action on the anterior lobe

o f tha hypophysis which in turn secretes a hormone or hormones which

a ffec t the ovary, e l ic it in g ovulation and lu te in development. That the

nervous stimulus does not act d irectly upon the ovary i s shown by the

fact that the stimulus produced by coitus induces ovulation in ovarian

g ra fts . In case of the 5 animals mentioned above i t i s possible then

that the corpora induced by copulation and not those due to the extract

were responsible for delaying estrus 29-31 days. This, however, could

64

not have happened In the remaining 5 animals where copulation occurred

within 1-7 days postpartum (Table VI),

I t Is the author's b e lie f that in most cases these induced corpora

did not function to any appreciable extent, i f in some cases indeed

they functioned at a l l . Support for th is opinion is found In the follow­

ing facts: (1) gestation was not prolonged (1 possible exception out

of 63 oases) but in the majority o f oases birth was premature even

though well-formed induced corpora lutea with small antra were present

In the ovaries; (8) 8 females were seen in copulation and became preg­

nant on the same day they aborted, and 8 additional animals copulated

and became pregnant on the 3rd day postabortion; (3) in 1 animal, No.

38009(h), injected with 150 R. 0 . of Antuitrin-S on the 16th day, the

corpora lu tea of pregnancy were removed on the 18th day and by the 80th

day au to lysis was taking place, The fetuses died in utero and were

resorbed which indicated that the induced corpora lutea did not maintain

the pregnancy. Friedman (1932) was unable to produce a decidual reaction

In rabbits when corpora lutea induced by the in tra fo llicu la r injection

of Antuitrin-S were present Indicating that these corpora were non­

functional.

In the experiment of Hoopoe and King (1935), they sta te that a

study of typ ica l cases showed injected animal Ho. 5 to have c e l ls o f an

estrous type on the 27th day and that she litte re d on the 27th and 28th

days. She mated on the 39th day but did not become pregnant; Ho. 169

had a few small masses of cornified c e l ls on the 30th day; and Ho. 27

showed cornified c e l ls on the 29th day though she died on the 32nd with

65

4 fetu ses in utero# They further sta te that:

"Attempts at estru s, as indicated by vaginal smears, were observed as early as the 27th day and are not incompet- ib le with fetu ses in utero,"

I t i s possib le that since c e l l s of the estrous type were present

as early as the 27th day that the induced corpora lutea were e ither not

functioning or functioning at a low le v e l and that the toxic condition

of the animals due to dead fetuses in utero was responsible for in­

h ib itin g estru s. Certainly an animal in a toxic condition due to dead

fetu ses undergoing autolysia and resorption in utero, could not be

expected to run nomal estrous cyc le . This i s further supported by the

fact that D ianil Blue, testicu la r and muscle t is su e s , and other sub­

stances which are non-hormonal in nature, w i l l Inhibit the estrous

cycles of otherwise normal rats (Meyer, 1933, personal communication}#

The corpora of pregnancy in animals injected with Antuitrin-S

possib ly showed some signs of retrogression as judged by the s ize and

number of the lip o id granules, ce llu lar vacuolation and leueocytic

in f iltr a t io n (Compare F igs. 10, 11 and 12),

I t has been known for some time that ovariectomy in rats in the

la tte r third of pregnancy resu lts in abortion or/and resorption of the

fetu ses i f these are 2 or more in number (Johnson and Ghallans, 1931;

H aterius, 1936; Kirsch, 1938; and others), idealizing the p o ss ib ility

that the estr in produced by the large f o l l ic le s in animals receiving

Antuitrin-S might be instrumental in causing abortion, and since i t

was reported that th eelin in la te pregnancy would resu lt in a derange­

ment of the birth mechanism (D*Amour and D*Amour, 1933), 2 animals

66

were injected with 200 I* ft, o f th ee lin (Parke, Davis and Co.) on days

17 end 18, resp ectively . The former had liv in g young on the 22nd end

the la t te r had 2 liv in g and 1 dead fetu s on the S lat day, 5 days hence

a l l o f her young were dead. I t la inconceivable that the ovaries

could produce a quantity o f th eelin comparable to that Injected in a

span of 2-4 days, and consequently th is does not seem to explain the

resu lts obtained in these experiments. This statement Is further

supported by the findings o f Levin, Katsman and Solsy (1931) who were

unable to a ffe c t gestation or survival of the l i t t e r in la te pregnancy

In rats with pu rified , cry sta llin e th eelin and th ee lo l In dosages as

high as 600 R. U.

In another part o f th is study, the corpora lu tea o f pregnancy were

carefu lly "picked out" o f the ovaries of 6 animals, 2 on the 16th, 1 on

the 18th, 1 on the 19th and 1 on the 81st day of pregnancy by means of

& very f in e pair o f forceps. As shown in Table VII, the 8 animals in

which the corpora were removed on the 16th day had aborted by the 18th,

A third animal In which removal o f the corpora occurred on the 18th

showed auto lysls to be taking place on the 80th, and she resorbed her

young instead of giving b irth to them. The 4th animal in which the

corpora were removed on the 19th gave birth to 2 dead fetu ses on the

81st, and to 4 more dead ones on the 23rd; she died, a fter giving birth

to the 4 fetu ses just mentioned, on the 83rd, Autopsy revealed 4 other

deed fetu ses In utero. The 5th animal with i t s corpora removed on the

21st day at 10580 A. K, was dead at 8:00 F.M, of the seme day. Autopsy

67

mm nu am mm® m mmsmm m& omtm imu m mmmt

Animalnumber

: fixso of piragnansy eoxpora m m rmoVed Bay out t$£t

t History ; Observations

saoio la) 16th-fi;45 P.II* 17th-et00 P,H,- IStlHBsOO A4I* 46th

S t i l l pregnant* Had aborted, l i t t e r of 3 ,

16th~3t45 F.M* IF1MMQ& P*M* ldth-atoo a , m, 44th

S t i l l pregnant* Had aborted, l i t t e r o f 4*

88009(h)* 18th-ll;0O iUM, SOth-9400 Ail*

25rd-6i00 A*M, 26th-7*00 J» JU

28th70th8©th

Autolysis as la d i- eated by blood­stained raacoidalflu id from vagina* S t i l l pregnant» S t i l l pregnant; reeorbing.Placed with male* l i t t e r o f 2* Pregnant again- palpated.

88009(e) 19th-*lS?16 ?«M. 20thglst-l*© 0 F.M*

EStrd-5:30 P*M,

Autolysis*& dead fetuses bora,4 more dead fetu ses bora*Animal dead; autopsy revealed dead fetu ses in utero*

seoio(o) E lat-10100 iuM* £lst-5:Q0 F4{* 81at-8*00 P*M,

Hot doing w e ll, Dead; autopsy revealed dead fetu ses in utero*

a-Had previously resolved 160 B.H. of iintuitrin-3 chi the 16th day of pregnancy*

68

revealed dead fetu ses in utero a lso . After a l i t t l e experience,

these corpora could he restored whole with very l i t t l e a t no bleeding.

I t w il l be noticed that 1 of these animals was previously injected with

150 R. 0 . o f Antuitrin-S on the 16th day of pregnancy and the corpora

were removed on the 18th* I t was hoped that the induced corpora would

maintain pregnancy but th is did not occur* That the ovaries were not

injured to any extent by th is operation was shown by the f e e t that 3

of these animals became pregnant soon afterwards (Table VII).

The Condition o f the Placentae

Regardless o f the dosage or day o f In jection , in no case were

fetu ses bam or found dead In utero previous to the 2nd day follow ing

the administration o f Antuitrin-S, Autopsies 1 day after in jection of

animals Ho. 37001(b) and Ho. 3 , which had received 75 R. U. on the 17th

and 18th days, resp ectively , revealed liv in g fetuses in utero. Additional

cases examined by means o f exploratory laparotomies a lso revealed th is .

However, many dead fetu ses wore found on the 2nd day post in ject Ion, and

consequently the placentae were not found to be loosened from th e ir

uterine attachments previous to th is time. The placenta© f ir s t lo s t

th e ir attachments along the periphery as indicated by the fact that not

uncommonly a w hitish , anemic ring was present around the outer edge o f

the placenta# When the placenta© wore pulled loos® with a pair of

forceps, i t could be seen that th® central part of th© attachment surface

In some cases was s t i l l intact and vascular. Placentae from injected

69

animate ware fixed and prepared for comparative stu d ies . H isto logical

examination o f these revealed in many cases, esp ecia lly those with

higher dosages, hemostasis, Infarct formation and necrosis (Fig. 15)*

The maternal side o f the fe ta l placenta was more affected .

the w m n o f m tm m m m A m sv m w s m m progestin on pregkmgy

I t the induced corpora lutea were responsible for prolongation o t

pregnancy in the experiments of Hoopea (1954), Hoopes and King (1935)

and Bunde (1938), then i t saeas lo g ica l that large amounts o t progestin

should accomplish the same resu lts . For th is reason progestin was in­

jected in 3 animals as indicated in fab le F i l l . I t can be seen that

progestin in the amounts used had no appreciable e ffe c t on gestation*

Bunde was unable to prolong gestation In la te pregnancy in rats by the

use of highly purified and active progestin in doses as large as 3 Rb*

0 . per day# He further reported that progestin plus th ee lin gave the

same e ffe c ts as progestin alone# fhua the resu lts obtained her® sub­

stan tia te the findings of Bunde, and suggest that the Induced corpora

lu tea obtained in th is study were not functional to any sign ifican t

degree. I t i s hard to understand how the corpora lu tea , both those of

pregnancy and the induced ones combined, could possibly secrete an

amount of progestin equal to or more than the 3 Rb# IT, per day which

Bunde injected#

Parke, Davis and Company sta te that Antuitrin-S administered in

connection with progestin is sometimes more efficacious than progestin

alone# With t h i s in mind 2 animals, Nos# 38013(a) and 38018(b) were

injected with Antuitrin-S and progestin as indicated in fable VIII#

In these 2 cases no sign ifican t prolongation of pregnancy resulted#

Of a to ta l of 19 fetu ses born to these S animals, 1? were viable and

2 non-viable#

70

71

tms Ttxtf m m m m mmrnm m m® p ro g b stik0r WPMhRGT

AniiwlnoBtber

Day o fpregnancyin jected

M tu itrtn -S in jected in fiUtf*

Aisottat Progestin* in jected in St*t*

len gth of gestation

He . o f young bora and triable

88018(f) id 119 120 *5Cl 1M 1 22 9(9)

88018(d) m10

1im>w

m<*

*921 122 1 22 13(13)

38018(a) 14 72 .8 29 12(10)

ssoisu) IS 112 120 1sa . 2 83* »(d)

38018(b) IS 72 119 '*•20 .421 1 83 7(9)

*~T&& progestin used was Vtu&&9 Davis and CoBjpany’s Lipo-Lutin* tHTefflftle bleeding p r o n e ly fro® vagina m 22nd day* S4 hrs* a fter the

f i r s t injection* a co lor less uaieoidal secretion was seen to ooae fromthe vagina*

e-8<i*ibers ta parentheses Indicate the asafcer of Tiable young! included la auabere on the l e f t .

THE EFFECT 0 ? HIAJJIL BUTE AND D IM IL BLUE PUTS

mrnvmms otr pregnancy

Five pregnant animals wars injected with D ianil Blue only and 15

with B ianil Blue followed by Antuitrin-S. The resu lts are given in

Tables IX and X. In 2 o f the 5 in jested with the dye alone, no serious

tox ic e ffe c ts were noted, but In the other 2 animals dead fetuses and

in 1 ease death o f the mother resu lted .

When the dye and hormone were both injected, the dye was considered

to be tox ic when fresh blood was found on the vulva before injection o f

the hormone was begun, since th is was known to indicate that autolytle

processes were taking place* When auto lysis was detected a fter the

in jection of Antuitrin-S, It must be attributed to the action of

Antultrln-S alone or to the combined action of both*

It can be seen from Table X that o f these 15 eases o f animals

receiving B ian il Blue and Antuitrin-S, 7 cases o f autolysis and 1 ease

o f f e ta l resorption were found to occur before th© in it ia tio n of

A ntultfin-3 injections* The e ffe c t on th© fetu ses is also given in

the ta b le .

Thus i t i s concluded that B ianil Blue alone, in the amounts in­

jected , Is capable of producing abortion, death o f the fe tu ses, and in

some eases, death o f the mother (Table IX). These resu lts are not in

accord with those o f Long and Evans (1932) who could detect no serious

e ffe c ts due to the in jection of D ianll Blu© in pregnant ra ts , but they

are In agreement with those of Meyer (personal communication) who was

78

73

mi xx* m wm of zwmnm mm* mm mm mmm Msmm*

Animalsomber -

P arity F or B

Dayinjected

jjjaount injected in fli*

Observations*and remarks

8M0#{b) * m

j&Oth

gJS&d

4

4

4

f i t t e r of 7*

S800?(«) » 0

: 5

Ho au to lysis detected | U tte r o f U *

86009(b) P ie th 4

19th 4 AUtolysis.

80th 4 Auteiysls*

AutoXysisj suckling 2 young from another mother*

£8nd Autopsy revealed 2 uonaal-si^ed dead fetu ses and 3 placental sites*

*~Xajected 1$ aqueous solution o f D ianll Blue 2R intr& peritoneally.

wEhe dve m i considered to be tox ic when a blood-stained, raucoidal f lu id m e seen to exude from tbs vagina*

74

*MM IX , (O oat’4 )

numberP arity P COPM

PayInjected

Amount in jected

in ee*.

Gbeervationsa and remarks

S8009{ft) P 80th 4

S la t 4

88n& 8 dead and 8 liv in gfetu ses bom* th eseyoung war© placed witha fe s te r wether hutwere nen-'Viabl©.

38013(1) P 13th 4

10th 4

17th 4

X8th 4

23nd A utolysis} 8 dead younghern* Animal died;autopsy repealed nofetu ses In u tero .

75able to Inhibit the estrous cycles of rats with D ianll Blue. As to

ho* th is Is accomplished la s t i l l problematic but from the evidence at

hand i t i s the author’s b e l ie f that the dye "poisons” the en tire system

o f the mother, and so disturbs her physiology end consequently her birth

mechanism to such an extent that she i s unable to give birth to the

fetu ses which have died in utero* Indeed in some instances the dye

become so tox le that the mother died. In most ca ses$ D ianll Blue plus

Antuitrin-S gave sim ilar resu lts (table X)«

76

tiusax. wt m m or mmnm wum. mm m asmma-& m o m u m m m sm je P

Lnfvi*]smarter

i t o t t r : l o r d

Bayin jected

m ow* iajaatad

in e<u

Observations*and ipeoHStiflta

88006(e) it 14th

16tfc

16th

i m

18th

80th

I ta t

8

8

4

4

180 R.tr. Aat,*6 in j .

A ttta lysis.

Animal' died o*j S la t dayf autopsy showed resorptloa o f dead fetu ses aeeanpenied by vary offen sive ©dor*

88006*d) M 14th

10th

16th

4

4

4

1n k 4 A u tolysis.

18th A utolyois; ?§ &#tf# Ant*-S inj*

19th Animal not doing n a il; autopsied 36 hrs* a fter iint .-S in j .

*-XnJect«i X* aqueous eoXutian o f EianU Blue 2R lntraperitoneally .

Sat Units <vivt■ -8 , Hjituitsln-Ss 1&J*» in jection .

iiii

^1*4 «

4 Is k 31 , *? 13ffsl

£

i 1 1 5? &*§*,-I t I ti UI-*js1«1 1 * * ijlm

t i 11 c im

aSi

« 1

I 2 *

1 f s I

* ♦ * a®

3 1 « 1*

1 1 # 3 s*f 3 1

4 If3

© pi

e ©« S i o § *-t at as «t

a»%

. iss^

R1*!

^

^

^•*

*> ■*

<#

9m**t

5

+* S

5

4»

4**

ife

<& ft*

#

O* #4

Hi fl

rl »4

H-

$

$

d

d

fl s

a a a a a I3 3 3 3

fc*f»ft A.

St»

a

H-r*g

*!•**

f

9

vmx x,7a

■ F erity '. f m m ■ ' ia jeeted

■ Aaount inJedteO

la ee*

Observations*

le th 150 R.U. Ant.-S In j.

i m AUtolysls*

80th Autopsied as not doing troll; autopsy revealed 6 dead and 5 liv in g fetu ses o f aameal s ir e .

38001(e) F 15th 4

10th 4 Autolysis*

17th 4 Autolysls* .

v m 4 Aut©lysis*

19th 75 Ant.~S in j .

S l i t Autopsy 48 hrs? a fter Aat«*fi inj* revealed 4 resorbing fetuses*

38001(a) P 15th

16th

4

4

17th 4 **ut © lysis.

18th 50 B*U« Mt»^S Inj •

20th 4 AUt©lysis*

2 M AutopBy 96 has. a fter iait*~S inj* revealed 2 liv in g and 3 dead nom al~sized fetuses*

79

CAKUC X, (Cant'd)

AnimalywfKay

Bayin jected

Mount in jected

in c e .

Observations* and re©&T)cs

38003(c) u 14th 4

18th 4 AUtolysls.*

16th

81st

4

78 jU9* M ti,’# |hj*

Autoiysls# \

A u to lysis5 autopsy revealed 5 dead, 8 liv in g , 1 p a r tia lly re~ sorbed fe tu s , and 4 placentas which indicated 4 young had been bora and eaten*

w 15th

16th,

8

3

17th 3 Antolysis*

18th Hot doing w ell; 73 R.O. Aat.-S in j .

......

81st Autopsy 48 hrs. a fter Aiit*~S inj* revealed 9 liv in g normal-sized and 4 dead p a rtia lly resorbed fetuses* Placentae were ea s ily detached from uterine w a ll.

80

ZABUt X . {O O Et'dJ

Via/toy P «rX

Boxin jected

Amount

in sc .

Observations*eyd

WMWtj.} 9 le th

16 th

n

3

m fe 3 ^ u to ly sis.

m ufin a le not doing w ell at St 30 A«2S« Female almost desdatSiO O B.M. | autopsy revealed 3 dead fe tu ses , 3 of which wore p artia lly resorted.

S8008<c) 9 U tli

15th

16th

17th

80th

3

3

3

103 E#f. 4 a t ^ injk

Autopsy 86 hrs* a fter iiUt.-S lnj« revealed 16 normal-s i zed liv in g fe tu ses.

360U{ls) M 14th

13th

16th

18th

3

3

78 R.U. Ant4*8 inj*

6 liv in g and 3 dead fetu ses horn. Another l i t t e r of 3 bom 62 days a fter abortion* Young net v iab le.

TABLE X. (ContfdJ

81

58988(6)

Parity? « p K

14th

ISth

i m

I8th

S2n&

Dayinjected

Amomit injected

in cc*

Observations* and remarks

100 K«0« Ant**S Inj*

Female almost dead; autopsy revealed 10 dead fetu ses shieh were accompanied by a very o ffen sive odor* Evidently a ease of postponed parturition*

38009(a) P U th

IStfc

16th

18«i

19th

75 B.O. Ant.-S in j .

Autolysis*

Autopsy 72 h rs. a fter Ant.«S in j . revealed 1 l iv e and 7 dead fe tu se s , 2 o f which were p artia lly resorbed*

teas, x. (cont* a)

Aniasalmwaher

Parity 3? ©Jp H

Bay Amount in jected

1b cc.

Observai i ons* and reiaarks

F 3

16th 5

17th 3

18th 78 E .0 . Ant.-S in j .

£Snd autopsy revealed 9liv in g normal-sizedfetuses*

a m EFFECT OF IHJEGT2N0 DIANIL BLUE, DIANIL BLUB PLUS ANTUITRIN-S,

and m&w m m into non-priotant animals

I t now became desirable to te s t the to x ic ity o f v ita l dyes on

normal, non-pregnant animals. Accordingly, 1 male and 3 females were

injected with tbs D ianll Blue as shown In Table XI. (In addition 1

female received 76 ft. U» of Antuitrin-S the day following the la s t in­

jection. of dye.) Of the 4 animals used only the male survived*

In a 2nd ser ies 1 male and 3 females were injected with Trypan

Blue. Of th is group (Table H I) the male and 2 females died and the

other female was doing w ell when she was ©utopsied on the 17th day

after receiving 8 cc , of dye.

From these resu lts I t i s evident that in the amounts injected

these dyes were seriou sly tox ic in most cases* It Is apparent that

Trypan Blue when given in the same concentration and amounts i s more

tox ic than D ianll Blue since the animals injected with Trypan Blue

became sluggish and died sooner then those receiving D ianll Blue. It

i s in order to s ta te here that these dyes were kept on ice in the same

b o ttles in which they were received and were diluted with s te r i le d is­

t i l l e d water so that to x ie ity cannot be attributed to technique or

contamination a fter the dye was received.

63

84

im b u e » . t s b tiw m m tm e m v ix tm t m m jx b m m m mv m x m w m m m . m m s *

imaibey'00%

f or HPate .

in jectedAmount

injected la cc .

Observations* and remarks

S6G16{a} iff

Beo*14th Dee*18th Dec.l9th

4444

Doing w ell; dye not serious­ly to x ic «

MttU(fc) f Deo#lSthDecdSthDee*l4thB eeasthDe0«18th

4444

trot doing w ell at IOiOO *«1M dead a t 3x80 F»M« Dye toxic*

38016(c) * Bee*18th Deo.13th Pee.14th Bee.15th B eeasth P©C.20t tk

4444

Being well* Bead; dye toxic*

88003(a) f m y n t h m y uath m y 13thm y 14th July 13th

m y 18th

4444

Ket doing w ell; 75 R.B* *jit.-S in i*Bead; death evidently due to to x ic ity of dye*

a-ln jeeted a 1# aqueous solution of D ianil Blue intraperitaneally . *-ftw poor condition and death o f the animals following in jections

were considered d efin ite signs of t o i ic i t y .

85

tm x xxx, ms mrm nr xmonm m a sunt vm ms-mamm m m a

dXtilininnnihor

00*I «r it

Datein jected

amount* Injected 1a ee*

Observations and ranarka

38017(a) H DeCwX2,»38Dce*15tfcB®e*14th

Bec*X5th

444

Set doing w e ll.Hat doing m l l i abdominal muscles have lo s t tono. Dead; dye toxic*

S8Gi?(b) fBeo.XStfcD©C*X4th

Bee,15th

44 • 4

Hot doing ve il*Hot doing v e ils abdominal muscles have loot tone* Dead;dye toxic*

3803.7(c) f Dec .18tbIleO*10t&B©8

44

iliumn . i . i f . ii . i . |j tit t . t t ! nf ij i , , . v i ii i

Hot doing v e il*Deads dye toxic*

380X£(a) * Hot* 8th Hot. $th H a v .im

44

Doing v e i l ; dye not seriQusly toxic* Sacrificed to obtain tissu es for h ie to log iea l studies*

a-laja«t«4 a 1)5 a®»#«M aolatlas ot trypan Bine intraperltonaelly.

* m m s m m i u n Of th e m a

The question concerning the permeability of the placenta i s an

import ant one hut as f e t explanations are problematic. I t i s known

that certain hormones are capable o f passing through the placenta and

affectin g the fetu ses (Courrier, 1031; L s ll , Liber and Snyder, 1932;

Hamilton and Wolfe, 193B; and others), Shimidzu (1922), investigating

the permeability of v i ta l dyes in mice and ra ts found that 6 out o f

13 su lfon ic acid dyes investigated passed through the placenta while

7 o f them did not, The action of B ianil Blue was not investigated .

According to Shimidzu:

"W© are ju s tif ie d in saying that the fetus i s free fromthe dye when no color is detected in the fe tu s by the samemethod of detection that reveals the sta in in the mother,for th is reason I excluded the dyes Which do not sta in the maternal body,**

In no case in th is study was the dye detected in the fetuses o f the

animals receiving B ianil Blue,

In addition, the fe ta l membranes o f 6 fetuses of an animal carry­

ing 13 fetu ses in utero wore injected with B ianil Blue as follows:

3 received 0,15 co. and 3 others received 0,1 ce . each on the 16th day.

On the 17th (23 hrs. a fter) the mother was sick and therefore autopsied.

A ll 6 of the Injected fetu ses were dead end almost completely re sorbed,

8 of the uninjected ones were recently dead and 8 were s t i l l liv in g

and of normal sin e. A ll placenta© were s t i l l attached and vascular.

Bo dye could b© detected in the maternal t is su e s , from these stu d ies,

86

8?

It Is concluded that the rat placenta i s impermeable to D ianil Blue,

and th is dye may he added to Shimidasu's l i s t of 7 impermeable ones,

the gonads o f a few o f the fetuses o f animals injected with

Antuitrin-S ware examined under the d issecting microscope, but no

e ffe c t due to Aatuitrin-S could he detected.

th e m m n oar l a c ta t io n oh g e s ta t io n

I t has been known for sons time that la cta tio n in certain mammals

resu lts la prolonged gesta tion , hat the explanation for th is phenomenon

1« s t i l l unsolved, Various theories hare been promulgated, however*

Daniel (1910) was o f the opinion that the period of gestation varied

d irec tly with the number o f young suckled* Kirkham (1916) found

delayed implantation due to lactation to be the cause of prolongation*

King (1915) thought that a lessening of the food available for the fe ta l

young in rats was the explanation, Mlrskala and Grew (1915), working

with mice, concluded that the length o f prolongation showed no correla­

tion with the number o f fetu ses in utero or the number of young being

suckled. B ain's studies on rats (1954) agreed with these o f Minskaia

and Grew, except that when the number o f young in utero exceeded 5 ,

gestation was prolonged*

The data c ited in Table XIII is hard to interpret* Of the 3

animals In which gestation was prolonged, i t would seem that there is

l i t t l e I f any correlation between the number o f young suckled or those

carried in utero* Fosaibly the number of fetu ses in utero and the

number of young being suckled might both be responsible* I t was not

uncommon to find the suckling young of females carrying fetuses in

utero to be in poor physical condition, as i f they were undernourished,

quite o ften the mothers, when they were carrying young in utero, at©

th o ir young, and esp ec ia lly so when they were suckling large lit te r s *

More data w il l have to be obtained before any defin ite conclusions can

be drawn*88

89

wsm MUf tm immt m ummm m wmmwP

A&lffialraypftmiy

Humber young cackled

Period «f gesta tion - :

toys

Number youngborn

S8009(h) 3 S3 S5-2 5 «L 8<& 4 S3 3t 9 S i 9A t m . 1034-3

”38

-------— ;-------6

a-3?emales eoneetvea. a t f i r s t poetpaxtum sstras*

DISCUSSION

I t i s d if f ic u lt to reconclie the r e su lts o f these experiments with

those o f Scopes (1934) end Hoopes end King (1935), I t i s possible that

the Aatuitrin-S used by them was o f a d ifferent potency than the one

used here. However, sines they do not state whether or not they re­

assayed th e ir ext root on th eir own animals It Is impossible to make any

comparison on th is b asis . I t la possib le a lso that in th e ir animals

the extract caused more rapid lu te in izetlon with a subsequent prolonga­

tion of gesta tion , i f the induced corpora were functional, than was the

case in th is study. I t i s w ell to note that in th is work no difference

In potency could be detected between the Antuitrin-S obtained from

Parke, Daw Is and Company o f New Orleans and that so kindly supplied by

Dr# Oliver Kamm of Parke, Davis and Company, D etroit. I t would have

been interesting to know the postpartum h isto r ies o f th e ir experimental

animals more fu l ly than i s given la th e ir paper. They sta te , however,

that attempts at estrus occurred as early as the 27th day and were not

incompatible with fetuses In utero# I t might be possible that the

physiological condition of their animals du© to a u to ly s is , necrosis,

and resorption of dead fetuses would be so affected that they would not

be able to run normal estrous cycles after parturition even though pro­

g estin were lacking. Gertainly other substances besides progestin are

known to in h ib it the estrous cycles of ra ts . Some of these are

extracts o f te s tic u la r and muscle tissu es and certain v ita l dyes

90

91

(Meyer, 195$, personal communication). Hoopes (1954) and Hoopes and

King (1955) did not te s t the e ffe c t o f progestin on pregnancy.

I t Is the author’s opinion that the corpora lu tea formed from the!

Injection of Aatultrin-S In th is study were not normally functioning

Corpora lu tea . An evidence for t h is opinion may be c ited the follow­

ing facts: (1) 4 out o f 10 observed animals copulated and became

pregnant within 1-3 days postabortion, 2 on the same day and 2 on the

3rd day. I t i s evident that in these eases the induced corpora were

unable to in h ib it eetru s. (2) A ll animals injected with dosages o f

BO-95 R. U. gave b irth prematurely or at term to l iv in g , or dead, or

l iv in g and dead fetu ses even though induced lu tea l tissu e was present

in some eases* Friedman (1932) injected Antuitrin-S d irectly into

mature f o l l i c l e s o f rabbit ovaries and obtained corpora lutea after 4

days. These induced corpora degenerated early , and he was unable to

produce a decidual reaction , which indicated that these corpora did

not function normally*

The presence o f extremely large and cystic f o l l i c le s in ovaries

of injected animals indicated that in these cases the corpora did not

in h ib it th e ir development. Also the fact that Leathern and Morrell

(1839) were able to induce estrus and mating in anestrous dogs by the

use of pregnancy urine extract, fo llu te in , indicates th at, in some

animals at le a s t , pregnancy urine extract can cause the growth of

f o l l i c l e s with consequent secretion of estr in in quantities large

enough to cause ©strus and that th© induced corpora lutea i f present

may not be able to inh ib it the action o f the estr in .

9 2

I f in th e experiments o f Hoopes (1934), Hoopoe and King (1935)

and Baade (1938) the induoed corpora la tea were responsible for the

prolongation o f gesta tion which they reported, then i t would seen

lo g ic a l that the corpus luteum homone (progestin) should accomplish

the same resu lts when injected alone in pregnant animals at the time

that the induced corpora lu tea were thought to be functional enough

to prolong gestation in th e ir animals* However, Bund© (1938) was

unable to o ffse t gestation in the rat with dosages of progestin as

large as 3 Rb* tf. a day; progestin plus t heel in gave the same resu lts

as progestin alone. The r e su lts obtained in th is study substantiate

those o f Bunde with progestin, since pregnancy could not be prolonged

with dosages as large as 5 Bb* V. Hoopes (1934) and Hoopes and King

(1935) fa ile d to t e s t the a ffec t o f progestin on gestation .

I t i s possible that the X*« H. (in Antuitrin-S) was injurious to

the already regressing corpora lutea of pregnancy since i t i s known

that in jections o f 1 . H. into hypophysestomized rats w ill cause

regression o f the corpora lutea which p ersist after hypophyseetomy

(Bund© and Greep; 1936; Greep, 1938), and consequently caused them to

cease functioning before they noimally would. This plus the increased

amount of th ee lin from the stimulated f o l l i c le s might have been

responsible for detachment of th© placentae. It i s a lso possible that

the Antuitrin-S acted d irectly on the placentae and caused them to

become detached.

Th© resu lts obtained with th® higher dosages (150-200 R. U.) which

d iffe r from those with the smaller doses (50-75 R* U*) reemphasize the

fa c t that th e acne preparation nay produce d ir ec tly opposing r e s ta ts

e lm adm inistered in amount* which vary to a considerable degree, The

resu lts here obtained cannot he attributed to impurities in th© extract

since th is p o s s ib i l ity was amply tested on non-pregnant animals o f

varying ages (immature and adult) that were maintained under conditions

sim ilar to those o f the experimental ones.

I t i s believed that the larger doses t i l l e d the fetu ses fa ster

than the sm aller doses by causing a more rapid decrease of the blood

supply through the placentae and that the subsequent necrosis and autoly­

s i s o f dead fetu ses resulted in the mother becoming so affected by the

toxins which then disturbed the b irth mechanism to such an extent that

she could not g ive b irth to her young, whether they were dead or a liv e ,

la these oases o f postponed parturition the female often died.

The e ffe c t o f v ita l dyes on pregnancy as obtained in th is study

would possib ly explain in a way the apparently co n flic tin g resu lts

reported by various investigators who supposedly used sim ilar unpttrlfled

preparations or made th e ir own ex tracts . I t fu r th e r points oat the

need for better purified ex tracts , and improved methods o f assay for

preparations used for experimental and c l in ic a l purposes sine© i t i s

seen that tox ic substances (non-hormonal in nature) may cans© resu lts

sim ilar to those produced by certain extracts of a hormonal nature.

The action o f Antuitrin-S her© seemed to be in the manner o f an

abort le n t , rather than playing a ro le in prolonging g e s ta tio n . As

Corner s ta te s (Glandular Physiology and Therapy) i t i s hard to assign

the reported b en efic ia l e ffe c ts obtained by the use o f A n tu itrin —S, in

94

the lig h t o f these experiments, as applied to eases o f habitual abortion

in women, to the induced corpora lu tea , end esp ecia lly so when there i s

no concrete evidence that Antultrin-S causes lutein izaticm in the human

ovary# I t further points out a need for more extensive researches on

the e ffe c ts of Antuitrin-3 on the human ovary#

It i s strongly believed from the resu lts o f these studies that the

uteroplacental complex in the rat plays an important ro le la the main­

tenance of pregnancy and in detexmining the time o f parturition , and

that parturition is not due alone to the degeneration o f the corpora

lu tea o f pregnancy* Astwood and Creep (1958) demonstrated the presence

o f a progestin -lik e substance in rat placentae#

The theory o f parturition as given by Reynolds {1957} i s , at the

le a s t , an in terestin g one# He proposes that in la ta pregnancy the d is ­

proportion between growth of the uterus and that of the fe tu s w ill

resu lt in a gradual but ever increasing intauterine tension# This

condition plus the fa ct that estr in increases uterine co n tra c tility

would resu lt in a condition incompatible with the retention o f the

uterine contents# Thus birth would occur# He further sta tes that since

the uterus has reached the lim it o f i t s capacity to grow at th© end of

gesta tion , that e ith er abortion or prolongation could occur depending

upon th® thae i t takes the uterus to reach i t s growth capacity due to

estr in and the ^distention mass” (sum of weights o f fetu ses, fe ta l

f lu id , placentae and membranes, and maternal placentae)# Reynolds, in

a paper previous to the one just cited {1932), also surmises that the

hypophysis might be one o f the factors (besides corpus luteum)

S5

responsible for tbs in h ib ition of the uterine contraction in the rat

in the in tereatra l periods since the estrone cycles ere so rapidly

recurrent that the corpus luteum i s said to he non-functional and i s

aerely a "h isto log ica l ornament.* He was able to inh ib it these con­

tractions with whole urine of pregnancy. I f th is i s the case In the

experiments here reported, i t would have to be that the smaller doses

{50-75 8 . U .) increased uterine co n tra c tility and that the larger doses

(150-800 H. U.) decreased i t . Ho d efin ite statement can be made

concerning th is view.

SHUKAHT f m Q Q m W Bim

The e ffe c t Of Antuitrin-S, progestin , and o f a v i ta l dye (D ianil

Blue SB) o& pregnancy wee investigated . The following conclusions

have been drawn:

1 . Antuitrin-S, when injected in dosages o f 50-76 R, IT. on the

18th day of pregnancy (gestation 81-2£ days) In the r a t , resulted in

b irth , prematurely or at term, to living* or dead, or liv in g and dead

fetu ses in 93.9 per cent o f the animals in jected . The liv in g fetu ses

were usually non-viable* and parturition was not postponed nor pro­

longed* These resu lts are believed to be due to the action o f the

Antuitrin-S acting on the uteroplacental complex either in d irectly

through the ovary* or d irectly on the uterus and placenta* or both;

th is consequently resulted la a decreased blood supply to the fetu s

and p lacen ta l detachment* These resu lts are in contrast to those o f

Hoopes (1934) and Hoopes and JCiag (1935) who obtained prolonged

gestation and postmature fetu ses with sim ilar dosages.

2 . Dosages o f 150-200 R. IT. usually resu lted in parturition being

delayed and in some cases prolonged from 1-5 days. Th® fetu ses died

in utero before or at term (1 exception) and in some cases even death

o f the mother resu lted . It i s thought that the birth mechanism was so

deranged by the auto lysis and necrosis o f dead fetu ses that the female

was unable to g ive b irth to her young. It might £lso be that uterine

contractions were inhibited to some degree, but th is Is s t i l l

problematic.96

9?

3» In jection s made on th© 19th, 20th and g is t days usually

resu lted in the b ir th Of v iable fetu ses at tezm. I t would seen that

the 18th day i s the c r i t ic a l point so far as the tolerance o f the

fetu ses to Antuitrin-S Is concerned, and that in jections node la ter

had l i t t l e e ffe c t on th e subsequent growth o f the fe tu se s , and on the

b irth mechanism*

4 , Progestin in doses as high as 8 1. (5 Corner-Alien Rb, tf.)

when injected in la te pregnancy was found to have no appreciable e ffec t

on gesta tion , These resu lts v er ify those o f Btrnde (1938),

5 , The v i t a l dye, D ianil Blue 2R, when injected in doses o f 4 cc,

d a lly from the 14th-X?th days in c lu sive , resulted in the majority o f

oases in death o f the fetu ses and even of the mother, showing the

e ffe c t o f a tox ic substance in pregnant ra ts . When Dianil Blue and

Trypan Blue were injected in the same amounts in to non-pregnant, adult

animals, the Trypan Blue was found to be the more toxic of the two,

6 , The physiological phenomena of premature abortion, and of

postponed and prolonged parturition were correlated as far as possib le

with h is to lo g ica l stud ies o f the ovaries, u te r i, and placentae. I t i s

the author’s opinion, from the resu lts o f these stu d ies, that the utero­

placental complex plays an important rol© in maintaining pregnancy and

in detexmining th© time of parturition, and that birth i s not due

so le ly to the regression o f the corpora lutea of pregnancy with the

resu ltant decrease in progestin secretion . It Is strongly believed

that the corpora induced by the administration of Antuitrin-S in

dosages o f 150-200 R, U. were not responsible for the cases o f

98

postponed parturition* Abortion occurred in animals with well-formed

induced corpora Indicating that these corpora were not normally

functioning ones* Certainly In the la t te r case, they did not prolong

gestation nor did they in h ib it or prolong parturition* This i s in

contrast to th e resu lts o f Hoopes (1934) and Hoopes and King (1935)•

7* Attention i s ca lled to the need for b etter purified extracts

and better methods o f assaying preparations used for experimental and

c lin ic a l purposes, since to x ic ity might lead to resu lts comparable to

that obtained by certain supposedly purified extracts*

LITERATOHI QVm

A llen , I# 1994* Sex and Internal accretions* The Williams and W ilkins Co** Baltimore*

A llen , W* H. 1932* Physiology o f the Corpus lutaum.In terrelation sh ip o f O estrin and th© Corpus Luteum as Beten&ned by th e ir E ffects in the Adult Babbits Am* X- Physiol* 100:650- 468*

A llen , W* K. and 9* W* Comer# 1929* Physiology of the Corpus Luteum. 111* Normal Growth and Implantation o f Embryos a fter Tory Early Ablation of tho O varies, under tho Influence o f Extracts o f tho Corpus Luteum. dm* X* Physiol* 88:340-546*

* 1930* Physiology o f tho Corpus Luteum.:* Maintenance o f Pregnancy in Babbits a ftsr Early Castration

by Corpus Luteum E xtracts. Pros* Soe. Exper. Biol* and Hod* 87:403-405*

Allan* W, M. and G* P* Seefcel* 1926* Prolongation of the Corpus Luteum in tho Pseudopregnant Babbit* Science 84:161-168.

1939* fa ilu r e of Progesterone to PreventResorption of Embryos in Babbits Castrated In Tery Early Pregnancy. Science 86:409-410.

., . * 1939* Baiat ©nance of Pregnaney bySogQSterone in feabbits Castrated on th© H th Bay. Am* «T* P hysiol. 123:31-35*

Allen* W* M* and B* K. Meyer. 1935* Physiology of th© Corpus Luteum.IX* The In h ib ition o f ©estrin by Progestin-containing Extracts of the Corpus luteum* Aaat* Bee. 61:487-439.

Astwood, E* B« and E. 0* Creep* 1938* A Corpus Luteum-StimulatingSubstance in the Bat Placenta* Pros. See. Exper. B io l, and Med. 38:7X3-716*

B ellerby, C. W. 1935. Termination o f Pregnancy in the Rabbit by Intra­venous In jection of Anterior lobe P itu itary E xtract. J . Exper. B id * 18:886-295*

Bronhell. P . W* R* 1937. Th© Influence of la cta tio n on th© Implantation o f the Mammalian Itabryo# Am* J* ©bst. and Gynec. 33:948-953.

99

100ABEde* 0* A* 1998* She E ffects o f Certain Gonadal end Gonadotrepie

Rermottee on the G estation Period of the Hat. Endocrinol. 23:545- 38$.

Bond#* 8* A# end JU 0* Creep. 1936* Suppression o f P ersistin g Corpora lu te a in Bypephyeootomized Bate* Pros. S ee. Bsper. B io l, end Med. 35*235-237,

G ollip , f* B* 1937. She Standard!zati on of .anterior P itu itary Hoxaonee. An. F* Chat, and Gynee, 33*1010-1016.

G ollip , J* B ., B . Sely© and y . E. W illiamson. 1938. Changes in the Hypophysis and the Ovaries o f Bate Chronically treated with an Anterior P itu itary E xtract. Endocrinol. 23!879-284*

G ollip , J . B*t B» 1 . 'Thomson, y . S . 1 . Brown, II. K. McPhail and I . E* Williamson* 1934. P lacental Homaaaes. Endocrinol. 15$315-323.

Cornea, P . E* 1937, E ffect o f Anterior P itu itary-L ike Homans* on L actation in the Albino Hat* Bros. Boo, Bxper* B io l, and Med. 37*58*38,

C carrier, R, 1931* Lee Hormones SoxneUes Fam elles. Ccmpt. Bond.See. m ol* 107:1367-141£.

Courrler, H. and R* Kohl. 19 SI* Bourellee Recherche* ear L1 Act ion do la F o lllc u lia e chess la Lupine on Phase Lutenlnique. Compt* Hand. 8 0 0 . B id , 107*1847*1848.

BtAmour, F* E* and M* 0* D^jaour. 1933. Studies in the Action of E strin in Pregnancy* Am, 1 . P h ysio l, 105$26-87*

D aniel, 1 , F . 1910* Observations on the Period o f G estation In White Hide. J . Hiper. Bool. 9*865-870.

Donaldson, H. E. 1924, the Bat* Bata and Reference Tables. Memoirs W lstar In stitu te of Anatomy and Physiology, Philadelphia.

Emery, F , E . 1937* A Study o f the Augmentation of Ovarian Weighte as E ffected by 2in© S u lfa te , A ntuitrin-S and Thyroid Im plants. ^&*J . Physiol* 118$316-320*

Engle, g . f , and 0 . Memod. 1928, The E ffect of B ally Transplantation o f the Anterior Lobe on the Course ©f Pregnancy in the Sat and Mouse. Am. 1 . P hysiol. 85:518-526.

finale B, TV and P . I . Smith. 1929. The Origin of the Corpus Luteum #ln th e Hat as Indicated by Studies upon the L utein izatioa o f the C ystic F o llic le . Anat. Res. 43:239-849.

101

IVWRft* H. M*. M* *Rd 8# I . Simpson. 1981* R elation of Prolanto .in terior Hypophyseal Hormones. Pros. Sec. Eaper. B io l, and Mod* 88*845-847.

Evans. H. H* and M. B. Simpson. 1989* Impairment o f the BirthMechanism Boo to Hormones from the Anterior Hypophysis. Pros.Boo. Sagpor. B io l, and Mod. 86*595*897.

f w ld * H# L ., ?# U Hisaw and 3 . 1 . Leonard. 1931. the Gonad8t herniating and the Lutelnlfcing Koraones o f the Anterior lobe o f the Hypophesis* An. 7 . Phjrelol. 97*891-301.

HfSor* W. X* 1938. Hypophyaeeteaay in Pregnant R abbits. Am. 7 .P h ysio l. 104*204-015.

Poet or, M# A* 1938. The D ifferen tia l Action o f P itu itary Gonadotropic Hormones upon the Secretory Capacity o f the Graafian F o llic le and Corpus luteum. Am. 7 . P hysiol. 101*633-639.

F oster, II. A. and H. 1# Fevold. 1938. The Ia t errel& tlonship o f the P itu itary Gonadotropic Hormones in fo llic u la r Development and O tulation cm the Xuvenila Babbit. Am# IT. P h ysio l. 1815625-832.

Friedman, X« H. 1938. On the Mechanism of O tulation in the Babbit.V. The $ fte e t o f D irect Intxufollicul& r In jection s o f Extract a o f tr in e o f Pregnancy* Am. 7 , P h ysio l. 99j332-337.

Glandular Physiology and Therapy. 1935. m * Med. A ssn ., Chicago.

Goodman. L . 1935. The E ffect of tr in e from Pregnant Wctaen on theOvary-Stimolating Potency of the Hypophyses of Babbits and B ats.Am. Jr. Physiol# 1X1*3X2-320.

Creep, R. 0 . 1938. Structural and functional Ovarian Response o fEypophyaeetomis&ed Adult Rets Treated with Gonadotropic Hormones. Anat. R es. 70s02-33.

. 1938. The E ffect of Gonadotropic Hormones on the ' J 1 1 P ersistin g Oorpora Lutea in Hypophysectomiaed R ats. Endocrinol.

23:154-163.

Haim. Annie M. 1938. The Physiology of Pregnancy in the B at. I . TheProlongation and Interruption of Pregnancy. Quar. 7# Bxper. P hysio l. 22*249-267.

. 1934# The E ffect o f Sucfcling on the Duration of ''-ifregiisEu cy in the Rat (Wlstar ^libino). 7 . Exper. B io l. 11*279-288.

102

Physiology of Fregnancy in the Rat: nnHOMhal In vestigation Inte th e Mechanism o f P artu rition . E ffeet « » th e fem ale Set o f the Ante-ft&tal Adm inistration o f Gastrin to the Mbther* Qnar. I* Bxper* Physiol* 85il&L«a43*

Hamilton, J# b„ and J* M, Wolfe. 1938# The E ffect o f Male HomoneSubstances upon Birth ana Prenatal Development in th e Bat* Anat.Bee* 7 0 *455*4 4 0 *

Harne, 8# ft* 1952* A Study o f the Excised Bterufl o f th© Rat, i t stlStxm M eplacm ent and Ir r ita b ility to F ltu itr in , with Reference to the Oeetroae Cycle* Am. I* P h ysio l. 1001531*338*

H arris, It. 0* and I* I . P fiffner* 1939* Extracts o f Corpora Lutea in H alation to Pregnancy* Anat* Bsc* 44:205-206.

B aterlus, H* 0* 1958# Redaction o f l i t t e r Sine and Maintenance o f Pregnancy in the Oophorectomissod Rat: Evidence Concerning the Endocrine Role o f the P lacenta, m # I* P hysio l. 114:599-406.

H eekel, G* P . mod W* H* Allen* 1958* Inhib ition o f P arturition in the Rabbit by the In jection o f Estrogenic Hormone# Science 87:308* 503*

: . , .. . , . * 1939, Maintenance o f the Corpus luteum~~" aid 'in h lb itioa o f P arturition in th© Babbit by In jection of

Estrogenic Hormone. Endocrinol* 84:137*148*

Holland, B* 1988* Th© Causation o f fo e ta l Death. Reports on Public Health and Medical Subjects. Bo* 7:118-113.

Hoopes, Edith 0 . 1954* Prolonged Pregnancy in Albino Rat follow ing in jec tio n o f Pregnancy Brine E xtract. Proc. Soc. Eaper. B id . and Med. 5H 1U 6-1U 7.

Hoopes, Edith € . and la s s ie L* King. 1933* Prolongation o f Pregnancy la the Rat by the In jection o f Human Pregnancy Brine Extract.Am* I* Physiol# 111:507-514.

Raggett, A. St 0* 1987. fo e ta l ELood-Cfos Tensions and Gas Transfusion through the Placenta o f the Goat. I . o f Physiol* 68:373-384.

Johnson, 0* I . and Joanna Challans# 1932. Ovariectomy and Corpus Luteum E x tra ct Studies on Rats and Ground Squirrels* Endocrinol. 16s 876-284,

K elly , 0 . L* 1931* The E ffect of In jection s o f Female Sex Hormone (O eatrin) on Conception and Pregnancy in the Guinea P ig . Surg. Gyaee* and Obst. 52:713-788.

105

^ftg* E den B* 1913. Bm» Anomalies in th e G estation of th# albino Rat (Mb# norvoglcus a lb iim s). B io l. B a ll. 24:877-591.

King, Icsst© I*. 1938. Influence on the Duration o f G estation on the In jection o f Pregnancy Grin# Extract in th# Hat before end a fter Im plantation. ssu f . P h ysio l. 122*458-459.

Klzrse}i,R# E. 1988* A Study # f the Control of Length of G estation la l e t with lo to s on Maintenance end Termination of Pregnancy.

m* Ji P h ysio l. IBS 886-93.

Klinger* H .# f . C* Burch end R. g . tamineM m. 1933. the Role o f th# Placenta la the Maintenance of Hypophyseal A ctiv ity Baring Pregnancy. Dorg. Gynec. and Obst. 8$*137-14®.

Lahr, M* L. and o . R iddle. 1938. Temporary Suppression « f EstrousCycles la the Rat by P rolactin . Proc. Soc. Exper. B io l, and Med. 341880-883.

Leathern. I . 1 . and 3T. A. M orrell. 1989. Induction of Mating ia the Bog with Pregnancy Grin® E xtract. Badocrinol. 24:149-196.

L o ll, W* A#., K# B* lib e r and f* f « Snyder. 1988. A Q uantitative Study o f P lacental franaRiccion and th# Perm eability o f f e ta l Membrane# at Various Stages of Pregnancy. Am. J . P hysio l. 100:21-23.

Leonard, S* L . and P. 1 . Smith* 1984* Responses o f the Reproductive System of Rypopfaysectoiaized Rats to In jection s of Pregnancy-Brine d etra cts. I I . The fem ale. Anat. Bee. 5 8 *175 - 2 0 3 .

L«vin, L* , P . A. Katsman and l . a . Daisy# 1951. Effect© o f Estrogenic Substances and the B utcinising Factor m Pregnancy in the JLbino Rat. Endocrinol* 15:207-213.

Long. J . A. sad H* M. Evans. 1922* The Ocstrous Cycle in the Rat and i t s Associated Phenomena. Mam# Daiv* C a lif. 6:1-148.

Marrian, G. F . sad W. E. Newton. 1988* The Synergism between Oestrin and Oxytocin. J . Physiol* 84s 133-147*

Mason, E. B. 1135. fo e ta l Death, Prolonged G estation, and D iffic u lt p artu rition in the Rat as a Result of Vitamin ^ -d eficien cy . Am.J . A»at* 57 s 305-349.

Kaser, G* and B. R. K ats. 1953. C lin ica l Evaluation o f Combined Prolan and Anterior P itu itary Therapy. E n d ocrin o l. 17 s709-722.

Moves, P* and I . Duesberg. 1908* Die S p em a to zy ten tc ilu n g en be! der H em isse (Vespa crabro L. )• Arch. f . Mikr. A nat. 71*38.

104

® W « B# 2C* 1908« Inhib ition o f the Oeetroaa Cycle (Personal ooamnnloatioii)*

Rewton, W* H* 1988# ^seudo-Parturi tion" in the House, and theR elation cf the Placenta to Post-partum Oestrus# f * Fhyaiol.84 s 196-207#

Meleon* W# ©*# y* J# p flffn e r and B* 0# B&terlus. 1980# TheProlongation o f Pregnancy by Extracts o f Corpus Luteum. Am. y* Physiol# 91s690-695.

Darke* Davis wad Co# 1988# C lin ica l Bees o f ^atuitrin-S* Therapeutic Botes# Bo# 4*97-102#

Parhe4 Davis and Co* 1989# Therapeutic Botes* Hoe* 7 - 6 *2 2 1 *

Parks, DaviS and Co* 1938# Progestin; Functions and Therapy# Therapeutic B otes. Bo# 10*271*876#

Parkes, 4# 8 # 1926# Observations on the Oestrcus Cycle o f the Albino Mouse# Proa. Bey# See# of London# ser ie s B* 100*151*170.

Badges, A# 8 * 1930* On the Synergism between O estria and Oxytocin#I# Physiol* 69s463-472*

Parkes, A# S . and 0# WV B ellerby. 1986. Studies on th e InternalSecretions of the Ovary* II* The E ffects o f In jection o f the Oestrus Producing Bomono during Pregnancy. J# Physiol* 62*145- 155*

Peneharz, R. I# and I . A. Long* 1938* Hypophysectoaay in th e PregnantBat* mm* J# Anat* 53S117-139*

Reynolds, S. B« M. 1982. The Action o f Ovary-stim ulating Substance o fHuman Brine of Pregnancy on U terine M otility in th© tTnanesthetised Rabbit# ksu I* Physiol* 100*545-552.

* 1958* Humoral facto rs A ffecting Uterine M otility#' S d ^ S n c i # 16*19$-198*

, # 1987* Mature of Uterine C o n tractility , a Survey~9TleesmF'feendfl* P h ysio l. Rev. 17?304*384.

» 1987# Chronic Uterine D istention and i t s R elationT"nrrnWTnr to '^ h i SfiT of gestation* Am. J . Obst. and Gynec. 33*968-978.

Robson, y* M. 1935* The E ffect o f O estrin or the Uterine ReactiTity and i t s R elation to Experimental Abortion and P arturition . I . Physiol* 84?121-132.

_-r- -r 1938* Th© Response o f the Uterus o f BypophysectoalredRabbits to Irbraets of the Corpus Luteum* J* Physiol * 643896- 301*

S elye, | t4 J* B* C ollip and 0* t>* Thomson* 1933* On the E ffect o f the A nterior IHtnitary-LIJce Hormone on the (hrajy of the Hypephyseeto- mi**d Bat* Endocrinol* 175494-000*

- - .............. . . 1933* E ffect OfHypophysectca^f upon Pregnancy and la c ta t ion . Proc. Soe* Exper* B iol* and Bad* 30*589-890.

. .. ... . . , ■ * 1934* Studies on. the E ffectof Pregnancy m the w r y T S a f * mSec• 385139*14$*

Skelesnyale, M. 0* 1933* The Production of Deeiduom&ta in Immature Bats by Pregnancy Urine Treatment, m* J* Physiol* 1045693*499*

Shiraidzu, tm 1988* On the Perm eability to ly en tu ffs o f the Placenta o f the Albino Rut and the Ih ite Mouse* Am* 1 * Physiol* 62:£02- 884*

Smith, Margaret 0 . 1926* On th© Interruption o f Pregnancy in the Bat by the In jection o f Ovarian F o llicu la r E xtract. Bull* Johns Hspfcta Hasp * 39:2Q3-214.

Smith, P* E# 1930* Hypophyseetomy and a Replacement Therapy In the Bat* Am* I* Anat* 435803—373*

, 1938* Th© Ib n -lsso n tia llty of the P osterior Hypophysis In P artu rition . Am* 1* Physiol* 995343-348*

Smith, C* tan S* and Q. W* Smith* 1933, The In h ib ition o f Lactation in Babbits with Large ^mounts o f Oestrin* Am* I . Physiol* 1035 336-361*

Snyder, F* F* 1934* Prolongation of Pregnancy and Complications o f P arturition in the Babbit Following Induction of Ovulation near Term* Bull* Johns Hophins Hosp* 5451—83*

Snyder, F* F* and Rosemfeld. 1937. D irect Observation of Intra­u terin e Respiratory Movements of the Fetus and the Bole of Carbon Dioxide In th e ir R egulation, m * J . P hysiol. 119 *153-166,

106

f* and 6* B. W islocki. 1931* further Observation* upon tits Experimental Production o f Ovulation in the Rabbit* Boll* Johns Hopkins Hoep* 49*106*020.

fee l* H* If* ,. 1926* The I f f s o t s o f In jectin g Anterior Hypophysial f lu id ©n the Course of G estation in the Sat* Am. J# Physiol*nwiwM*

Tan Pyhe, H. B* IS36* fho Physiology and phaisaacology of the P itu itary Body# The Univ. o f Chicago Press* Chicago*

Barbflttem* Tirgene* 1934* 2fce Cytology of the Corpora lu tea of the Ewe* J .& orp h el. 56il8I*202*

H aloek ly 0* B* and 1* Goodman. 1934* $he E ffect o f in te r io r lobe .^street or Concentrated Susan Urine o f Pregnancy upon the Early fa r t of G estation In the Babbit. S9*35?S*381.

EXPLAHmGH OF FIGURES

107

F ig . X* Graph showing the e ffe c t of one p reparation of prolan

when in jec ted in to immature r a t s , Ten anim als were need fo r each

dose. From Coester (1938), Arch. exp. P a th , P h a m .> GLXVXXI: 745-752

Copied from The Physiology and Pharnmcology of the P i tu i ta ry Body by

H. B. Tan Bylce* p . 219*

F ig . 2. Three dead premature fe tu ses aborted on th e 20th day by

prireiparous animal No. 58003 {a} which had received 75 R, U. of

A ntuitrin-S on the 16th day of g e s ta tio n . X2. The fe tu se s weighed

1.7 gms. each. They were preserved and photographed in 95 p e r cent

a lcoho l.

Figure 2

Oe

st

ru

s—

or

c

or

po

ra

lu

te

a i

n

ov

ar

ies

-PE

RC

EN

TA

GE

OF

AN

IMA

LS

M

03

to

o

o

o

o

oo

l/l

o

Fig* 3* Three dead aad on© Hiring premature fe tu se s aborted

on the 20th day by animal Ho* 38003 (d) which had received 75 R* V*

o f A n tu itrin -S on th© 16th day of gestation* X2* The th ree dead

fe tu se s weighed 1*7, 1*6, and ,9 gras*, respective ly* The 4 th young

was s t i l l l iv in g w ith in i t s f e t a l membranes, and to g e th e r with i t s

membranes and p lacen ta weighed 2*3 gras* The fe tu se s were preserved

and photographed in 95 per cent alcohol* The th ree dark sp o ts on

the p lacenta are blood spo ts in d ica tin g where the p lacen ta m s

attached*

Fig* 4 . A sec tio n through the l e f t u te rin e cornu o f raultiparous

animal Mo* 38003 (a) taken on th© 25th day showing the amount o f

in ju ry to the l in in g o f the lumen* X2I. This animal had received

150 R. U* of A n tu itrin -3 on the 16th day o f gestation* She gave b i r th

to four dead fe tu ses on the 24th and to two a d d itio n a l ones on th©

25th day* Four fe tu ses (not eaten) weighed 4 , 3*7, 4*3, and 3*4 gms*,

re sp ec tiv e ly . P a r tu r it io n required not over te n minutes fo r one

fetus* A most o ffensive odor accompanied the b ir th o f each fe tu s due

to the fa c t th a t they had undoubtedly died before o r a t term* This

i s ev iden tly a case o f postponed p a rtu ritio n *

Figure 4*

Fig* 5 . A sec tio n through the l e f t ovary o f animal Ho.

38008 (d) which had received 100 a* B* o f A n tu itrin -S on th e 18th

day o f gestation* X17. This animal had been prev iously in jec ted

w ith 3 ee« o f D ian il Blue on the 14th , 15th , and 16th days, respective ly*

She was k i l le d on th e 26th day a s she had not given b i r th to h e r

young y e t , and was almost dead* At autopsy ten dead fe tu se s were in

utero* The only induced lu te a l t is s u e was th a t represen ted by the

two la rg e lu te a l cy s ts shown with la rg e a n tra . The so lid corpora

lu te a a re those of pregnancy* This i s a ty p ic a l case o f postponed

p a r tu r i t io n . The r ig h t ovary of th is animal contained two hemorrhagic

f o l l i c l e s .

Fig* 6* A sec tio n through the r ig h t ovary of animal No*

38007 (c) which had received 150 R. U* of A n tu itrin -S on the 18th

day of gestation* X17. This a n im l had previously been in jec ted

with 4 cc* of D ianil Blue on the 14th , 15th , 16 th , and 17th days,

respectively* She was k il le d on the 20th day as she m s not doing

w ell and had been bleeding from the vagina since the morning of the

19th. Autopsy revealed four norm al-sized dead fe tu se s which ev i­

den tly were not long dead as they were not yet deco lo rized , and th ree

liv in g ones. Note the th ree la rg e a t r e t i c f o l l i c l e s w ith convoluted

m i l s , and two hemorrhagic lu te a l cy sts in one o f which the lu te a l

t is s u e is invading the an trun a t one s id e .

Figure 6.

F ig . '7, A sec tio n through the r ig h t ovary o f primiparous

animal Ho* 3800$ (e) which had received 200 B* II. o f A n tu itrin -S

on the 18th day o f g e s ta tio n . X17. She m s autopsied on the 25th

day since she had net yet given h ir th to he r young* Autopsy re ­

vealed fou r dead and seven l iv in g fe tu se s , and one p lacen ta w ith

no fetus* The liv in g fe tu ses averaged 5 .5 gms* in m igh t* The

ovary contained on© lu te a l cyst with a trapped ovum (upper l e f t ) *

two la rge a t r e t i c f o l l i c l e s , corpora of pregnancy ( la rg e r ones)

and young induced corpora (sm aller ones). This m s the only case

where liv in g fe tu ses were found past term .

F ig . 8 . A sec tio n through the r ig h t ovary of animal Ho.

38001 (a) which received 75 B. U. of A ntu itrin -S on the 18th day o f

g e s ta tio n . X17. She had previously received 4 cc . of B ian il Blue

on the 15th , 16th, and 17th days, resp ec tiv e ly ; and she a lso received

an a d d itio n a l 4 cc. on the 20th. On th e 17th and 20th days a dark ly-

sta ined mucoidal f lu id was seen to ooze from the vag ina . This sec tion

shows two hemorrhagic lu te a l c y s ts , a non-hemorrhagic one, four

c y s tic and a t r e t i c f o l l i c l e s , and corpora lu te a o f pregnancy.

Figure 8

Fig* 9* A sec tio n through th e r ig h t ovary of animal He* 28008(h)

which had received 85 B. U* o f A n tu itrin -S on each o f days 15, 1$, 17,

and 18* XI?. On th e 19th day she was in jec ted w ith .2 ingot* of

c o lch ic in e , and was autopsied 9*5 h r s . a f t e r th is* Autopsy revealed

one dead fe tu s in u te ro . Section shows one heiaonhagic f o l l i c l e , c y s tic

and a t r e t i c ones, induced corpora w ith a n tra , and corpora o f pregnancy*

Fig* 10* A section through a corpus luteum of pregnancy of

animal No. 38006(d) which had received 75 It* U* of A n tu itrin -S on

the 18th day. X43G. She had previously been in jec ted w ith 4 cc*

o f D ian il Blue on the 14th , 15th , 16th, and 17th days, respective ly*

An autopsy 36 hrs* a f t e r the in je c tio n o f A n tu itrin -S revealed four

dead fe tu se s th a t were being resorbed* She c e l l w alls of the lu te a l

c e l l s were in d is t in c t and leucocytic i n f i l t r a t io n m s evident

in d ica tin g regression* Compare with Figures 11 and 12 of the same

m agnificat ion .

Figure X©*

Fig* XI* A sectio n through a corpus lutetsa o f pregnancy o f

aniiaal Ho* 38G07(c} which had received 150 E. U* o f A ntuitrin-S

on the 18th day o f gestation* X430. She had previously horn in­

jected with 4 co . o f D ianil Blue on the 14th, 15th, 16th, and 17th

days, respectively* Autopsy on the 20th day repealed s i s dead and

three liv in g fe tu ses in u tero . Xhe cytoplasm o f the lu te a l c e l ls

was highly vacuolated and the c e l l m i ls were in d istin c t in d icatin g

regression* Compare with Figures 10 and 1Z o f the same m agnification*

Fig* 12* A section through a corpus luteum o f pregnancy of

animal He* 28000(d) i& lch had received 25 B* XI* per day on the 15th,

16th, 17th, and 18th days* 2430. Animal had ju st died a t 11:00

P . M. on the 19th day. Autopsy revealed ten dead fetu ses in utero*

Compare with Figures 10 and 11 of the same m agnification.

3

*

‘f

J

Fig* 15* A sectio n through a corpus luteum o f pregnancy o f

a n im ! Ho. 38006(b) suhieii m& autopsied on the Sth day postpartum*

X430* She m s suckling seven o f her young. Hote the large and le s s

numerous lip o id granules m compared to those in Figure 14 o f th e same

m agnificat ion and from the m m ovary*

Fig* 14* A sectio n throu^ a corpus luteum o f la cta tio n o f

animal Ho. 38006(b) \3hich m s autopsiod on the 5th day postpartum*

X43Q* Hote the sm all and more numerous lip o id granules as compared

to those in Figure 13 o f the same m agnificat ion and from the same

ovary* 5&e granules in th is sectio n coapar© in s iz e and number to

those o f the corpora o f a comparable age induced by the in jectio n

o f A ntu itrin -S .

Figure 14,

whieli had received 100 R. if* o f A ntuiirin-S o s th© 18th day o f

gestation* X13* She m s autopsied o s the 88th day s iso e She had

not yet g ives b irth to her young. Autopsy revealed e ig h t fetus©*

tjhieh had evid en tly bees dead fo r f iv e o s s ix days, H isto lo g ica l

essar&nations revealed n ecro sis, hesiostasis and in farct form ations.

BIOGRAPHY

B u ssell Mark Coco was bora in B ordelonville, Louisiana, Avoyelles

P arish , on June 82, 1909, He attended the Voorhies Grammar School o f

that parish , and graduated from the Bordelonville High School in 1926,

In June 1928, he entered the Louisiana State Normal College at

N atchitoches, Louisiana, During h is junior and senior years, ho was

laboratory a ssista n t in b io logy. He was awarded the A, B. degree from

that co llege in 1951, at which time he was facu lty representative »nd

V aledictorian o f h is c la s s .

During the session 1951*52, Mr. Coeo was a graduate research

a ssista n t in zoology at the Kansas State College o f Agriculture and

Applied Science at Manhattan, Kansas, at which in stitu tio n he was

awarded the M. S . degree in August, 1952. He was e lected a member

o f Phi Kappa Phi and Gamma Signs D elta , two national honorary

fr a te r n itie s , at th is school.

Mr. Coco taught high school biology and mathematics in Avoyelles

Parish fo r three years (1952-35} and was Instructor in Biology at the

Louisiana State Normal College during the summer session o f 1955.

In 1936 he entered the Louisiana State U niversity to do graduate

work in zoology, where he held teaching fellow ships for the session s

1936-3? and 1957-58. During the summer of 1937, he attended school at

th© U niversity of W isconsin.

At present Mr. Coco is a graduate teaching fellow in Zoology at

the Louisiana State U niversity and is a candidate for the Fh. D. degree

in Zoology at that in s titu tio n .

115

EXAMINATION a n d t h e s is r e p o r t

Candidate: R u sse ll Mark Coco

Major Field: Zoology

Title of Thesis: -Antuitrin-S Inj e c t io n s in Pregnant Rats

Approved:

Major Professor and Chairman

D ean of the Graduate School

EXAMINING COMMITTEE:

tr f k s iA a jA A a ^ d ^ j

Date of Examination: