University of Southern California Dissertations and Theses

Some metabolic effects of androgenic steroids on rats fed different inadequate diets

(USC Thesis Other)

Some metabolic effects of androgenic steroids on rats fed different inadequate diets

PDF

Download

Open document

Flip pages

Download a page range

Download transcript

Copy asset link

Request this asset

Transcript (if available)

Content SOME METABOLIC! EFFECTS OF ANDROGENIC STEROIDS ON RATS
FED DIFFERENT INADEQUATE DIETS
A Thesis
Presented to
the Faculty of the Department of Biochemistry and Nutrition
The University of Southern California
In Partial Fulfillment
of the Requirements for the Degree
Master of Science in Biochemistry
by
Marcelo E* Nimni
June 1957
U N IV E R S IT Y O F S O U T H E R N C A L IF O R N IA
G R A D U A T E S C H O O L
U N IV E R S IT Y P A R K
L O S A N G E L E S 7
B.'o '57 XT 13
This thesis, written by
M&reeXo. .EfJraim.Nironi.............
under the guidance of /zia....Faculty Committee,
and approved by all its members, has been pre
sented to and accepted by the Faculty of the
Graduate School, in partial fulfillment of the
requirements for the degree of
..........................Jfaster.of. Science...........................
JO H N D. COOKE
...............................Aotm£.D.ean..................
Dean
Date to... 1.9.57-.............
Faculty Commiijtee
Chairman
TABLE OF CONTENTS
HISTORICAL INTRODUCTION ..........
Effects of Androgenic Steroids on
Nitrogen Metabolism • ••••••••
Effects of Androgens on Body Weight
and Growth »•«•*••••*••••
Methods for Assaying Anabolic Activity
of Steroids •*•••••••••••
Effects of Diet on Nitrogen Metabolism
and Body and Organ Weight as Influenced
by Androgens • • • • • • • • « « • • •
Effect of Testosterone Propionate on
Liver Weight and on Regeneration
After Hepatectomy ••••«•••••
Mechanism of Nitrogen Storage « • * •
Relative Activity of Various Androgenic
Steroids on Different Organs » • • • •
Summary of the Review • •• •• •• •
STATEMENT OF THE PROBLEM AND PLAN OF ATTACK
MATERIALS AND METHODS ...... * . . *
Metabolic Studies •••••«•• ••
Organ and Muscle Weight and Composition
Experiments ••*»••«•••• «•
Diets ••••»•«•••••«• • •
PAGE
1
1
5
6
8
11
11
13
15
17
19
19
21
23
PAGE
EXPERIMENTAL RESULTS . ......... 26
\
Nitrogen Balance Experiments • • . . 26
Effects of Androgenic Steroids on
Muscle and Other Organs of Normal and
Castrated Rats on Protein Free Diets* 36
DISCUSSION ..................
SUMMARY . 50
BIBLIOGRAPHY ......... 5l
LIST OF FIGURES
FIGURE PAGE
1 Effects of subcutaneous administra
tion of testosterone propionate on
nitrogen balance of castrated male
rats fed zein diet* • ••««•••* 30
2 Nitrogen balance of castrated male
rats fed zein diet and injected
subcutaneously with sesame oil,
(control experiment) ...•••* • 31
3 Effects of subcutaneous administra
tion of testosterone propionate on
nitrogen balance of normal male
rats fed zein diet •••••••»• 32
1 | . Effects of subcutaneous administra
tion of testosterone propionate on
nitrogen balance of castrated male
rats fed a diet containing 15 % of
a mixture of glycine, alanine and
sodium glutamate as only source of
nitrogen • •••••••*•••• * 33
5 Effect of subcutaneous administra
tion of testosterone propionate on
nitrogen balance of castrated male
rats fed gelatine diet • • ......... 3b
6 Effect of continuous daily subcu
taneous administration of testoste
rone propionate over a period of 7
days, on nitrogen balance of
castrated male rats fed gelatin diet. 35
LIST OF TABLES
TABLE PAGE
I Effect of testosterone propionate
on organ and musele weight of
normal and castrated male rats on
protein-free diets •••••••••• 39
II Effect of growth hormone and pitui
tary gonadotrophin on normal and
castrated male rats on a protein-
free diet for eleven days •••••• 1^2
III Effect of testosterone propionate
and norethandr©lone on organ and
muscle weights of castrated protein
depleted male rats • « • • • ♦ ♦ • * • I|3
HISTORICAL INTRODUCTION
Effects of Androgenic Steroids on Nitrogen Metabolism |
Experiments on dogs . In 1935$ Kochakian and Murlin
(1), studying the effect of an extract prepared from men’s
urine on the nitrogen balance of castrated dogs, found
that the injection of this extract, in an oily vehicle,
produced a decrease in the urinary nitrogen excretion.
This nitrogen retention increased with the amount of
injected hormonal extract up to a certain point, beyond
which there was no greater response. After cessation of
the injections the dogs went into a negative balance
before returning to normal. All these changes were limited
to the urea nitrogen fraction. Other constituents showed
no significant changes. Creatine was not detected in the
urine of castrated male dogs if animals were kept on a
creatine free diet. A slight increase in body weight took
I
place after the injection but returned to normal there
after.
In 1936 the same workers repeated the experiment
with androstendion (2). The same decrease in urinary 1
nitrogen was observed in the castrated dogs, but a single
normal control animal showed no response. Studies of blood
non-protein nitrogen during the retention period showed
no Increase, so the retention was assumed not to be due to
a storage of non-protein nitrogen constituents in the
2
plasma* Testosterone and testosterone aeetate were
studied with similar results (3). The maximum nitrogen
retained was approximately the same for all substances
injected and varied from 50 to 60 mg.of N/Kg. of body
weight/day* Thorn and Engel ( I j . ) , after injecting single
doses of 25 and 125 mg.of testosterone propionate to
normal dogs, found no nitrogen retention* However, daily
doses during seven days resulted in a great nitrogen
retention at the end of the injection period* This is not
in accordance with other experiments (5) where a slight
nitrogen retention took place in a normal female dog
during the injection period, while another bitch showed
no significant decrease in the urinary nitrogen excretion*
Experiments on rats♦ Studies of nitrogen metabolism
and body weight of castratecl male rats (6), indicated that
the injection of testosterone propionate produced a
nitrogen retention which reached a maximum after 2 to I } ,
days. This rate was maintained for about 10 days after
which there was a decrease in the nitrogen retained until
equilibrium was reached. The body weight increased during
the retention period, but began to decrease during the
continued treatment, as the nitrogen excretion returned to
normal. Animals reached their original weight together
with the occurence of a negative nitrogen balance.
According to Braasch et al* (7) nitrogen retention
took place in normal male and female rats after the dally
*
Injection of testosterone propionate as compared with
controls injected with peanut oil. The maximum nitrogen
retained daily was approximately 70 mg.for the females
and 50 mg. for the males. These values are considerably
larger than those reported by Kochakian and Beall (8),
who found values ranging from 12 to 20 mg. of nitrogen
retained for normal females, while gonadeeternized females
showed a slightly greater retention.
Experiments on man. The effects of androgenic
substances on nitrogen metabolism and body weight of
human subjects has been widely, investigated. It is known
that the presence of the testes in the body is associated,
with certain changes in the muscular system. In eunuchoi
dism the muscles are underdeveloped and their contours
are soft compared to normal male individuals. Methyl
testosterone did not induce nitrogen retention in 2 normal
young men during strenuous work, although there was a rise
of creatine excretion (9) ♦ Hie creatinuria gradually
decreased after cessation of treatment with the drug.
The same compound produced a retention of urinary nitrogen
in an eunuchoid (10). A retention of potasium, water and
nitrogen was observed in 6 normal individuals after
receiving testosterone propionate. The retention took
place even in the absence of dietary sodium (11). Methyl
k
testosterone Increased body weight in two cases or
testicular insufficiency (12). Other studies performed
on immature boys (delayed adolescence, pituitary dwarfism)
showed a nitrogen retention following treatment with !
i
i
▼arious steroids. Greatest activity was shown by testoste-j
rone esters and less by several androstenediol deriva
tives (13)*
Kenyon et. al. (II4 - ) investigating the effects of
testosterone propionate on the nitrogen metabolism of
normal and eunuchoid individuals, found that the degree of
nitrogen retention induced in the eunuchoid was approxi
mately double as compared to the normals. Two normal
women were also studied, and in one nitrogen retention
was of the same degree as in the eunuchoid. The other
women studied showed no response to the hormone
administration.
i
Intravenous administration of testosterone propionate
in a solution of human serum albumin over a period of
30 minutes to 18 human subjects had no effect on nitrogen
balance. The same amount administered as an intravenous
infusion during a period of 2 l j . hours caused a slight
nitrogen retention (15) •
This comparative review of the experiments on normal
and gonadectomized animals, and normal and hypogonadal
human subjects, seems to demonstrate that a definite
5
decrease of urinary nitrogen excretion takes place in the
absence or atrophy of the male genitalia, but when the
i
gonads function normally little or no nitrogen retention
is induced by androgens*
Effects of Androgens on Body Weight and Growth
The effects of testosterone propionate on normal
rats recovering from acute fasting were reported by
Kochakian (16). Rats which had been fasted for seven
days were treated with testosterone propionate for 136
days* Noninjected controls were observed simultaneously.
Initially the weight recovery was equal for both groups,
but later the noninjected grew better*
When the recovery from fasting of castrated male rats
injected with testosterone propionate was investigated, it
was observed that initially the injected rats attained
better growth, but following the second week of injection
the non treated control animals were already showing
better growth, which persisted throughout the experiment
(17)• Experiments performed by Geiger and El Rawi (18),
during which normal rats were injected with testosterone
propionate during protein depletion and repletion, failed
to show any body weight sparing or growth stimulating
effect of androgens. When the growth of castrated male
rats receiving testosterone propionate was compared to
castrated controls receiving sesame oil (19), the controls
6
grew better, even though the food intake was greater for
the testosterone propionate injected animals.
In all these experiments comparatively large doses
of testosterone propionate were injected, ( 1 to 2.5 mg.
daily). Testosterone propionate administered intraperito-
neally to normal 26 day old rats over a Sh day period, in
doses of 0.05 mg.six times a week, led to a significant
increase in body weight and length (20). This growth
stimulating effect of small doses of testosterone propio
nate stands in contrast to the growth depressing effect
of the larger doses of androgens which are required to
stimulate a nitrogen retention.
Castration of immature or adult mammals is known
not only to arrest or regress development of the accessory
sexual organs, but also to produce changes In many other
organs. Korenchevsky (21) showed that after castration
there was a decrease in the size of the seminal vesicles,
prostate, penis, preputial gland, liver, kidney and heart,
and an increase in the weight of the adrenals, thymus and
hypophysis. Testosterone propionate was able to induce a
repletion of the decreased organs and an involution of the
adrenals and thymus•
Methods for Assaying Anabolic Activity of Steroids
Kochakian (22) utilized the kidney repletion phenome
non as a basis for an assay of the anabolic properties of
7
androgens. Shis renotrophic activity was compared with
the weight increase of the seminal vesicles, which was
designated as androgenic activity. For 39 steroids
measured, the so called renotrophic and androgenic acti
vities ran more or less paralel. Shis renotrophic activity
was proposed as a measure of the anabolic activity of
androgens. Search for a more appropriate assay pointed to
the stimulating effects shown by testosterone propionate
on muscles that atrophy after castration, such as the
temporal muscles of the guinea pig (23) 5 and the perineal
complex of the rat, including the bulbocavernosus«
ischiocavernosus and levator ani muscles ( 2 i j . ) • The latter
in particular, because of its readiness for dissection,
was studied with respect to changes in weight that take
place after castration. Subsequently the effectiveness
of various androgenic steroids in promoting growth of
the levator ani muscle was investigated (25)•
The ability of androgens to increase the weight of
this muscle after depletion by castration was assumed to
be a myotrophic activity, and was supposed to be different
from the androgenic and renotrophic activities previously
mentioned.
Many steroids have been compared for their various
activities, as it would be of great significancy to
obtain a substance which would be highly myotrophic in
normal Individuals or convalescents during the 1 1 catabolic
phase”, and at the same time devoid of androgenic effects
conducive to masculinization•
Effects of Diet on Nitrogen Metabolism and Body and Organ
Weight, as Influenced by Androgens
The influence of the dietary nitrogen and caloric
intake on the nitrogen retention induced by testosterone
propionate has been studied by several investigators.
Castrated male dogs which were placed on a high
protein diet (23.5 % casein) showed an increased nitrogen
retention after testosterone propionate (26). When the
same animals were equilibrated on a low nitrogen Intake
(11.8$ casein), testosterone propionate was not able to
induce nitrogen retention. On the contrary, the nitrogen
output seemed to increase as a consequence of the
androgen administration. The same effect was noticed by
these investigators In previous experiments on rats on a
low nitrogen intake.
The effect of testosterone propionate on castrated
male rats placed on a protein free diet was studied by
Wijnans and De Groot (27), who found a brief nitrogen
retention as compared to controls which received only oil
injections. After approximately one week, both groups
attained a constant level of nitrogen excretion at the
unsurpassable minimum limit.
9
Diets of high protein content were studied for their
effect on the nitrogen retention induced by testosterone
propionate (28)« Levels of 17 %9 28 % and l j . 6 % casein
were fed to castrated male rats* The maximum daily
nitrogen retention and the total nitrogen retained were
not altered by the different levels of casein in the diet*
Also increasing the dose of hormone did not change the
response* The maximum nitrogen retained ranged from i | > 8 to
62 mg. of nitrogen/day/rat*
The effects of general dietary deficiencies on the
response of the prostate of the rat to the administration
of testosterone propionate was investigated by Grayhack
and Scott (29)• They fed incomplete diets, (vitamin free
casein plus water, and glucose plus water) and also a
reduced intake of an adequate stock diet to castrated
male rats. At low dosage (0*05 mg*of testosterone propio
nate/day) there was no difference between the prostatic
growth of the deficient and normally fed controls. At
hig£i doses of hormone (2 mg/day) there was more growth
of the prostate in the normally fed rats, although if
prostate weight is related to final body weight, the
relative weight of prostate is much greater in the
deficient animals*
During excess protein feeding growth is retarded*
Leathern (30) feeding rats high protein diets (78 % casein
or lactoalbumin) and simultaneously administering testos
terone propionate, noted no effect of the hormone on the
growth retardation• Also the increase of blood non-protein
nitrogen that takes place after feeding high protein diets
was not altered by testosterone propionate•
Clinical studies on malnourished patients after
severe trauma showed that the degree of nitrogen reten
tion was not influenced by various adequate nitrogen and
caloric intakes (31)•
Studies were performed on the effect of testosterone
propionate on the body weight repletion at a low nitrogen
intake, after an..experimental protein deficiency, (32)•
During the recovery period,(on a 7 % casein diet) a
greater weight recovery was observed in the testosterone
propionate injected animals* These results are not in
agreement with previously reported experiments (18). In
general the composition of the various organs studied
showed no significant differences, except for the dry
weights of the accessory sexual organs, which were
increased by testosterone propionate, and the dry weights
of thymus and carcass which were decreased by testosterone
propionate. The facts that the dry weight of the carcass
\
was decreased in the testosterone injected animals, that
no significant difference is recorded for the major bulk
of the organs studied, and that a slightly greater body
11
weight was attained by the testosterone propionate
i
injected animals suggest an increase in body water content.
Effect of Testosterone Propionate on liver Weight and on
Regeneration After Hepatectomy
In castrated mice a 9 % decrease in liver weight
occured after testosterone propionate administration, as
compared to untreated controls (33)• In other experiments
liver weight was not significantly altered 25 days after
castration, but low doses of testosterone Increased it
to above normal (34) • When testosterone propionate was
injected to normal female rats with normal estrus cycles,
no change in liver weight occured if injected during
estrus, but a significant decrease took place when the
hormone was administered at diestrus (35)*
Injections of 2 mg.of testosterone propionate daily
during 21 days to normal male and female rats produced
a decrease in liver weight In both sexes, (36)*
Liver regeneration after partial hepatectomy was
studied for normal and castrated male rats, and for normal
rats which received 0,25 mg.of testosterone propionate per
100 g. of body weight daily. No significant differences were
noticeable among the three groups (37)•
Mechanism of Nitrogen Storage
Comparative studies of testosterone propionate and
growth hormone on body and organ weight of castrated male
mice showed that both hormones induced approximately the
i
same increase in body weight (38)# The androgen exhibited
greater growth stimulation for the kidneys and secondary
sexual organs# The simultaneous administration of testos
terone and growth hormone produced a summation of effects*
Nitrogen balance studies showed that both hormones
induced the same degree of nitrogen retention on castrated
rats (39)* On cessation of injections a rebound or loss
of nitrogen occured in both groups of rats*
Studies on hypophysestomized rats showed that growth
hormone produced a much greater nitrogen retention and
body weight increase than testosterone*
The mechanism by which the organism stores nitrogen
after testosterone administration has not been accounted
for as yet* Kenyon and coworkers (I 4 . 0) found in eunuchoids
that the nitrogen retained was greater than that which
could be attributed to an increase in the mass of genital
tissue* Calculations of the theoretical increase in body
weight of eastrated male rats after testosterone propiona
te, on the basis of the retained nitrogen, furnishes a
higher value than that which is observed by weighing the
animal (Ip) •
Studies of the composition of the carcass showed that
1
i
after testosterone propionate there was no significant
13
change in the total nitrogen. An increase in water
content and a decrease in fat was reported (19)• No
changes were recorded for the plasma total protein, non
protein nitrogen, or amino acid nitrogen. ]
The presence of the pituitary and adrenals is not
| required for the nitrogen retention to take place, as it
can be induced in hypophyseeternized (i+2), as well as in
adrenalectomized castrated rats (1+3) •
Based on the fact that growth hormone and Insulin,
both of which are considered to have a protein anabolic
activity, lessen the increase of the amino acid level ©f
the blood which occurs after evisceration, a study was
made to determine whether testosterone propionate would
show the same effect (27). Eviscerated rats injected with
testosterone propionate did not seem to differ from control
animals in their blood amino acid nitrogen. The author
concluded that the point of attack of testosterone, could
not be the equilibrium between amino acids and proteins,
thus leaving the mechanism of action of testosterone as
an anabolic hormone unknown.
Relative Activity of Various Androgenic Steroids on
Different Organs
The fact that the relative potency of different
androgenic steroids might be greater for some organs than
for others, could be atributed to their different require
ments as to the threshold level and conditions for optimal
effectiveness of the various compounds (I 4 I1 -).
i
Patty acid esters of testosterone are more active
because of their slower absorption from the site of
injection* Using different esters of testosterone it was
possible to produce in the castrate infantile rat seminal
vesicles which were heavier than the prostate (as in the
case of the normal adult rat)• Using free testosterone
the postate always remained heavier than the seminal
vesicles, (as in the case of the castrated rat) (I 4 . 5) •
All the esters showed the same activity when
administered by intravenous injection. The choice of
bioassay method for determining activity is of great
importance. Androsterone and testosterone will both cause
growth of the cap oris comb; but if doses of each steroid
which would promote equivalent growth of the comb were
injected into castrated male rats, the effect on the
seminal vesicles would be very different (I 4. 6) •
The dilution in which the hormone is injected, the
solvent used (peanut oil, castor oil, propylene glycol,
etc.), and the time between injections can influence
enormously the responses, even though the total quantity
of active substance injected remains constant, ( i j - 7).
The several methods that have been proposed to
measure tin various specific activities attributed to
15
androgens do not always show correlative results. This
makes understanding of the mechanisms involved more
difficult* Unesterified 19-nortestosterone did not
induce nitrogen retention in castrated male rats (I 4. 8), but
I
when its myotrophic activity was measured by the levator
ani muscle increase method, it showed an activity close
to that of testosterone propionate (I4 . 9) • Its androgenic
activity measured by the increase of weight of the seminal
vesicles was very slight.
Attempts to produce substances which showed greater
anabolic but less androgenic activity has led to the
study of 19-nortestesterone and its derivatives . 17-ethyl-
19-nortestosterone has been developed for clinical use as
an anabolic agent, on the basis of its growth stimulating
effect on the levator ani muscle (50)•
Summary of the Review
After reviewing the literature on the effect of
androgenic steroids on nitrogen metabolism and body and
organ weight, the following conclusions can be arrived at:
1. Nitrogen retention is induced by testosterone
propionate and other closely related steroids in castrated
male rats• Normal male and female rats show the same
effects to a much smaller degree.
2* Body weight of castrated or normal animals is not
significantly increased by testosterone. On the contrary
16
a growth depressing effect seems to occur at large doses.
3. Testosterone propionate stimulates growth of the
| accessory sexual organs in normal and castrated male
I rats, but other organs of the normal rat are not increased
■ ( ■ i
I by hormone administration. Some organs, like the liver,
j
are significantly decreased.
I j . . The most commonly used methods for assaying the
anabolic activity of steroids are based on the assumption
that the myotrophic effect on the levator ani muscle,
is completely independent from the androgenic effect. No
satisfactory evidence is advanced for this assumption.
STATEMENT CHP THE PROBLEM AND PLAN OF ATTACK
It was the purpose of this investigation to study
the effect of testosterone propionate and other androgenic
or allegedly anabolic substances on the nitrogen metabo
lism and the body and organ weight and composition of the
normal and castrate male rat*
The anabolic effect of these androgenic steroids is
being widely investigated but no really satisfactory
evidence exists, except that of an induced nitrogen
retention, to justify the assumption of anabolic activity*
The mechanism by which nitrogen is retained following
androgenic stimulation, and the fate of this retained
nitrogen remains unexplained* We therefore decided to
further investigate some aspects of this problem*
1* In order to observe if the nitrogen retention
induced by androgenic steroids should be necessarily
associated with the process ©f growth, metabolic nitrogen
balance experiments were performed during the feeding of
incomplete diets to normal and castrated male rats* The
dietary protein utilized was one of low biological value
(zein or gelatin), or a mixture of 3 non essential amino
acids* These proteins or the amino acid mixture cannot
support growth, even when consumed at an adequate caloric
level, and mixed with an otherwise satisfactory diet* It
was proposed to investigate if nitrogen retention could
j
18
still take place under these conditions of dietary amino
acid deficiency*
I 2* In order to investigate if the growth of the
i
(
! levator ani muscle stimulated by testosterone propionate
J and other steroids was a phenomenom independent of their
i androgenic activity, as suggested by previous investigators
the following experiments were planned:
Hats were placed on protein free diets, which lead
to a general catabolic state* Body weight and organs are
rapidly depleted, especially the accessory sex organs* A
diet of this nature does not support somatic growth, but
in spite of this the accessory sexual organs (seminal
vesicles, prostate, penis, etc.) increase their weight
after receiving testosterone. This can only be attributed
to the strong sparing effect stimulated by testosterone
to promote growth of its target organs, and are a direct
consequence of an androgenic activity*
It was the purpose of this part of the investigation
to see if the levator ani muscle behaved as the accessory
sexual organs tinder similar protein deficient conditions*
MATERIALS AND METHODS
Male Wistar rats were used In all experiments*
Bilateral castration was performed under light ether
anesthesia* All animals were kept in individual cages,
and fed Purina Chow, ad lib*« except during the
experimental periods, during which the specific diets
were supplied.
Metabolic Studies
For the purpose of these experiments animals were
placed on the specific amino acid deficient diet 3 to I j .
days previous to to the initiation of the balance studies,
in order to deplete the existing 1 1 nitrogen pool”. During
this initial period an extremely high loss of urinary
nitrogen occured.
Animals were then put in individual metabolic cages,
and a weighed amount of food was offered in cups with
metal tops, in which a 1*5 inch hole had been cut* These
cups were placed inside larger cups. This avoided spilling
of food, and permitted a quantitative determination of the
daily intake* Water was supplied ad lib* Body weight was
checked daily.
Daily collections of urine and feces were made
separately. For this purpose a collecting bottle was
placed beneath the metabolic cage* A light bulb was fixed
20.
\
at the opening of this bottle in such a way that the
\
urine would slide down the walls of the bulb into the
bottle, whereas the feces would fall into a Petri dish
placed below# This allowed a complete separation of urine
and feces•
The cages were washed with distilled water and the
collected liquids were immediately filtered. The urine
filtrate was acidified with 0.25 ml. of conc. HgSO^, and
stored in the cold until the time of analysis. Urine
nitrogen was determined using the Folin-Nessler method,
modified by Koch and McMeekin (5l)» The feces were placed
in an oven at 80°C overnight and then stored in a
dessicator over CaSOj^ until the end of the experiment#
They were then pooled for each rat and the nitrogen
determined by the same procedure, after digesting an
aliquot with conc# HgSOj^#
The nitrogen balance was calculated from the
following expression:
Nitrogen Balance - Nitrogen intake - (Urine N +Feces N.i
Positive values represent nitrogen retention
(positive nitrogen balance) and the negative values
represent nitrogen loss (negative nitrogen balance).
The body weight was recorded every other day and the
daily caloric intake was calculated from the quantity ©f
the diet consumed#
21
Organ and Muscle Weight and Compos it ion Experiments
Throughout these experiments the animals were kept
on a protein free diet.
The effects of the following substances were inves
tigated:
| Testosterone Propionate (Lilly) 25 mg/ml* Injected
| at different dose levels.
I
Antuitrin Growth (Parke Davis) 10 rat units/ml. in
normal saline•
Pituitary Gonadotropin (Squibb) 10 rat units/ml* in
normal saline.
Norethandrolone (17-ethyl-19-nortestosterone)
dissolved in sesame oil.
At the end of the experimental period the animals
were sacrificed by a blow on the head. The levator ani
muscle was dissected according to the technique described
by Hershberger, Shipley and Meyer (52). For this, the
skin in the scrotal area, between the base of the penis
and the anus was removed. The posterior aspect of the
perineal complex was cleared of fat and connective tissue,
particular care being taken to clean the constrictions
at either end of the levator ani where it joins the
bulbocavernosus muscle. The rectum was transected just
caudal to the point where the levator ani loops around it
dorsally. The body of the muscle levator ani was then
22
freed of the rectum, and removed by incisions at the
points of attachment t© the bulbocavernosus muscle. The
levator ani muscle was cleared of any connective tissue
and weighed immediately. The levator ani muscle, seminal
vesicles, kidneys and diaphragm muscle were weighed to the
nearest G.l mg.on a chainomatic balance. Other organs
were weighed on a Roller-Smith torsion balance to the
nearest $ mg. To determine the water content of the
levator ani muscle, it was placed for 2I 4 . hours in an
oven at 80°C, allowed to cool in a dessicator and
weighed* Ihis method gave constant dry weight values. For
determining the total nitrogen of the levator ani muscle
the dry muscle was predigested by heating lf> minutes with
10% An adequate aliquot was measured and the
nitrogen determined by the technique described above.
Diets
The diets used in the course of these experiments
were the following:
Protein free diet
Corn Starch 1,100.
g.
Ruffex 100
I I
USP salt mixture # 2 80
t t
Cod liver oil
65
M
Wheat germ oil
65
n
Lard 80
t t
Choline hydrochloride
'K-
.2 g.
Methyl llnoleate i
g
Vitamin A (*-)
« B
50,000
5,ooo
units
t t
w G
lj.00 mg.
!' B*
M - Q
Dg
30
25
t t
t t
" b6
5
8 1
b12
o.► 02 mg.
Calcium pantothenate
50 mg.
NiacJLn amide 200
t t
Folic acid
2.5 mg*
(*) Vitamin source was Nine-Vitamin, High Potency,
(Van Camp Laboratories)
Zein diet
Corn starch
Cane sugar
Butter fat (anhydrous)
Zein
USP salt mixture # 2
Vitamin A
Natural mixed toeopherols
equivalent in biological
activity to
Calcium pantbth enate
Niacin amide
Corn oil
Choline hydrochloride
1 gram of diet
62 g.
8 g«
10 "
15 "
5 n
1250 TOP units
250 1 1 n
125 " "
0.5 mg*
0,025 mg.
200 USP units
0,5 mg.alpha -
tocopherol
0,25 mg,
5 mg.
2 ml.
0.3 g«
calories
Glycine, alanine, sodium glutamate diet. An iso-
nitrogenous mixture of these 3 amino acids was used in
Sodium
glutamate 8I 4..5 w
This mixture was used as a nitrogen source for
1,020 g.of diet.
Gelatin diet. This diet was made up as the zein
diet, by replacing zein with gelatin (15> %).
1 gram of diet - calories
All diets were analyzed for their nitrogen content,
and this value was utilized in calculating the nitrogen
balance•
place of the 1$ % zein. The rest of the diet composition
Glycine 37*5 g.
Alanine 1 1
EXPERIMENTAL RESULTS
The studies reported cover two different aspects
of the problem of protein anabolism induced by androgenic
hormones•
The nitrogen metabolism experiments supply informa
tion regarding the net effect of testosterone on the
whole animal, but give no indication of the mechanism
by which nitrogen retention is stimulated, nor about the
fate of this extra stored nitrogen*
Studies at the organ level were intended to
investigate whether the anabolic effect of androgens
could be extended to the whole organism, as implied by
the term t f anabolic hormone11, frequently used in the
current literature, or should be restricted to organs
directly linked to the sexual stimulating property of
these hormones•
Nitrogen Balance Experiments
During the course of these experiments animals were
fed diets which, because of their lack of some of the
essential amino acids, could not support body growth. The
caloric intake was always adequate but, because of this
amino acid deficiency, animals lost weight throughout all
the experiments reported*
The first three experiments were performed with
animals fed a diet containing 1$% zein as the only
protein source* Zein is a protein of low biological
value and lacks two essential amino acids, lysine and
tryptophane *
Figure 1 shows the effect of testosterone propionate
! on the nitrogen balance of castrated male rats fed the
zein diet* On the l^th and 5th days of the experiment,
all rats were injected subcutaneously with 0*1 ml of a
25 mg/ml* solution of testosterone propionate in oil, thus
making the total dose received by each rat equal to 5 mg*
of testosterone propionate.
Following the last injection, the urinary nitrogen
excretion dropped, and the animals went into a positive
nitrogen balance which lasted 3 days* On the last day of
the experiment, animals returned to the negative nitrogen
balance* Body weight loss did not seem to be affected by
the hormone injection. Calorie intake was adequate
throughout the experiment.
Figure 2 represents the values obtained for a
control experiment in which castrated male rats fed the
zein diet were injected with 0.1 ml.of sesame oil on the
ij.th and 5th day of the experiment • As can be observed,
animals maintained a constant negative balance, which was
not affected in any degree by the oil injections•
Figure 3 shows the effect of injecting testosterone
28
propionate (total dose per rat, 5 mg. ) to normal rats
fed the zein diet* Animals were In negative balanee
throughout the experiment, and contrary to the castrated
rats receiving the same treatment (Figure 1), the
androgenic stimulation was not able to decrease the amount
' of nitrogen excreted in the urine, which was maintained
at a constant level*
The effect of testosterone propionate on castrated
male rats fed a diet which only source of nitrogen was
an isonitrogenous mixture of three non essential amino
acids (glycine, alanine and sodium glutamate) is shown
in Figure l | _ . The urinary nitrogen excretion was constant
throughout the experiment. On the 7th day the animals
went into a slight positive nitrogen balanee, which is
most likely a consequence of a simultaneous increase in
the food intake that took place that day, and not due to
a decrease in the nitrogen excretion, which as already
mentioned remained at a constant level.
Figures 5 and 6 show the results of androgenic stimu
lation on castrated male rats fed the 15 % gelatin diet*
Gelatin is also a protein of low biological value,
lacking the essential amino acid tryptophane*
In the first of these experiments (Figure 5)» rats
were injected on the ij-th and 5th day with 2*5 mg. daily of
testosterone propionate. The nitrogen balance, which was
29
positive during the control period, became more markedly
so as a consequence of the urinary nitrogen retention
induced by the hormone injection. The decreased food
intake during the last 2 days of the experiment decreased
the net balanee and finally brought the animals into
negative nitrogen balance.
The experiment of Figure 6 was run under the same
conditions, but the treatment period was extended to
7 days, during which the rats received 2 mg. of testoste
rone propionate daily (Total dose, 17*5 ♦ As can be
observed, a positive nitrogen balance was maintained over
all the experimental period, and was greatly increased by
the prolonged administration of testosterone.
It is interesting to note that in spite of the
positive nitrogen balance, animals showed the weight
decrease characteristic of all the experiments. It should
also be pointed out that the calorie intake was adequate
for body weight maintenance.
30
+40
+ 20
Nitrogen
Balance
—
mg.N/day
-20
- -40
320
Body
Uelght -300
(g.)
280
Caloric 60
Intake
30
cal./day
T.P. 2.5
n
10 days
Figure 1. Effects of subcutaneous administration of Testosterone
Propionate on Nitrogen balance of castrated male rats fed zein diet.
Total number of animalsx 11
999999999994
3 1
0.1 ml S.O.,
20
Nitrogen
Balance 0
mg.N/day
-20
-40
—60
260
Body
Weight 240
(g.)
220
Caloric
Intake
cal./day
60
30
0
V
r -
days
Figure 2. Nitrogen balance of castrated male rats fed zein diet
and Injected subcutaneously with sesame oil. (Control Experiment)
Total number of animals: 9
32
■ * • 2 0
Nitrogen q
Balance
mg.N/day
-20
-iO
-60
-80
300
Body
Weight
(g.) 280
260
Caloric 70
Intake
35
Cal./day
2.5 mg T.P.
n
days
Figure 3. Effects of subcutaneous administration of Testosterone
Propionate on nitrogen balance of normal male rats fed zein diet.
Total number of animalsi A
Nitrogen + ^
Balance
mg.N/day ®
-20
-40
220
Body
Weight 200
(g.)
180
Caloric
Intake
Cal./day
50.
25
0
2.5 mg T.P.
n
days
Figure 4. Effect of subcutaneous administration of Testosterone
Propionate' on nitrogen balance of castrated male rats fed.a diet containing
15 % of a mixture of glycine, alanine and sodium glutamate as. only source of
nitrogen.
Total number of animals* 4
3k
80-
60
40
Nitrogen
Balance 20
mg.N/day
0
-20
-40
240
Body
Weight 220.
(g.)
200
Caloric
Intake
cal./day
70
35
2.5 mg T.p.
m
days
Figure 5. Effect of subcutaneous administration of Testosterone
Propionate on nitrogen balance of castrated male rats fed gelatine diet.
Total number of animals: 4
67
35
2.5 mg. T.P. daily
120
+ 80
40
Nitrogen
Balance
mg.N/day
Body
Weight
(g.)
Caloric
Intake
Cal./day
260
240
220
60
30
10 days
Figure 6. Effect of continuous daily subcutaneous administration of
Testosterone Propionate over a period of 7 days,on nitrogen balance of
castrated male rats fed gelatine diet.
Total number of animals: 4
549927
Effects of Androgenic Steroids on Muscle and Other Organs
of Normal and Castrated Rats on Protein Free Diets
In the last few years there has been a great deal
of interest concerning the Anabolic properties of
testosterone and closely related steroids*
We proposed to investigate in the following
experiments the effect of some recently introduced
steroids with allegedly increased anabolic and decreased
androgenic properties* For the screening of such compounds
Eisenberg and (Jordan (25) suggested a bioassay method in
which the weight changes of the levator ani muscle of
male rats are determined after injection of androgens*
According to these authors the observed weight changes
indicate the ”myotropic”, anabolic, effect of the
compounds•
From our observations on the size and growth of this
muscle under various conditions it appeared that the
weight changes of the levator ani muscle might be a
sex linked function, the growth effect of the steroids
representing an androgenic rather than a general myotro-
phic effect* It was therefore questionable whether the
levator ani method could be used as an indicator of the
anabolic effect of steroid compounds* The present studies
deal, among other points, with the validity of this method.
|
37
In the first group of experiments the effect of a
protein-free diet was investigated# The results, conden
sed in Table I (Group I), show that the animals on
protein-free diets lost body weight , as expected. The
levator ani muscle, the seminal vesicles, and the kidneys
participated In this general loss in body weight and
were smaller than in animals on a complete diet.
In further experiments (Group II), the effect of
testosterone propionate on protein depleted animals was
investigated. Every other day, the animals received 2.5 mg-
of testosterone propionate in sesame oil, injected
subcutaneously. Rats maintained on stock diets and
protein depleted rats which received only sesame oil
injections served as controls.
The results Indicated, in confirmation of earlier
experiments (18), that the loss of body weight on
protein-free diets was not affected by testosterone
propionate injections. In contrast, the weight of the
levator ani musele, the seminal vesicles, and the kidneys
increased considerably, (p < 0.01) .
The same effect could also be demonstrated in
protein depleted rats which had been castrated a few days
before the start of depletion. The determination of the
water and of the nitrogen content of the levator ani
muscle, showed further (Table I, Group II) that an
38
increase of the organ weight after testosterone
treatment was not due to water accumulation but repre
sented a real increase of organ protein*
In order to ascertain how testosterone propionate
affects the growth of another muscle, we also determined
t
' (Group II) the weight of the diaphragm muscle* This
i I
muscle was selected because it could be easily dissected*
The results indicated that, in contrast to the increase
of the leva tor ani* the weight of the diaphragm muscle
was not affected by testosterone injections*
We then sought to determine why the weight loss of
!
the levator ani muscle on protein-free diets was
relatively much larger than the loss of total body weight,
(Table II). One possible explanation of this phenomenon is
that the secretion of gonadotrophic hormones decreases in
rats on protein deficient diets (53), and as a consequence
i
the stimulus for androgen production is diminished* In j
order to investigate this possibility, normal and
castrated rats were protein depleted, and for 11 days on
4
a protein free diet they were injected subcutaneously with
2 to I 4. rat units daily of pituitary gonadotrophin (Squibb).
It was found (see group II and III, Table II) that the
weight of the levator ani muscle, of the penis, and of the
seminal vesicles of the intact gonadotrophin-treated
increased considerably over the weights of these tissues
EFFECT OF TESTOSTERONE PROPIOMTS OK ORGAN ’ AMD MUSCLE WEIGHT OF HORMAL A® CAS1A®
. . . . MALE RATS ON M E M E E DIETS.... , .
GROUP RATS CONDITION PERIOD DIET ,MENT m
— — — -- -- . - - - - - - - - „ later con-'; Nitrogen
Levator Seminal " tent of ; content of
Ani Vesicles Kidney Liver Testes Diaphragm L,Ani dry L, Ani
I .
4
Normal Stock -- 119 18! • 70.4 143.1 1756 9505 2430 78.5 1
(*13)
(*29)
(*-30.0) (59)
(t202) (*-1310) (1212)
(t.91)
/
k
1 1 12 days Prot. - '118 83 13.2 28,7 833 4050 1685 74.5
Free
(5)
(12.7)
(*-2.9) (i6.5) (153) (448) (1272)
(11.4)
II 5
Normal 15 days Prot, 15 mg 170 137 94.5 1103.0 1307 5340 2590 354.2 78.5 15.5
Free
M ) (*U) (122.9) (1356) (*94) (1605) (112?) (H6.2) (i.7) (*1.2)
5
t r i t i t 1 1 —
170 131 33.1 102.2 1137.5 5590 2355 387.0 76.5 14.8
* * (6)
( 18)
(410.9) (122,8) (65.3) (515) (*179). P52.3)
(1.2) (1,9)
III 5
Castrated 12 days Fret. i; mg
99 71 51.9 501.8 948.5 2781 78.6
tee T.P.* ('.?)»
( 18) (i6.7) (*43.5) (176.51 (420) P.42)
5
1 1 i t i t I I —
98 ■
72 7.6 17.8 783.7 2891
(19)
(13.8) (12.5) (15.2) (*44.51 (1274), 77.8
( * 2. 1)
# 2.5 og every second day * Vaises in prentesis. standard deviation
in the control animals despite the protein depletion#
In protein depleted castrated rats (©roup IV), the
injection of gonadotrophic hormone proved to be ineffective
thus suggesting that this hormone elicited its effect
through the testes# In order to eliminate the possibility
that traces of growth hormone present in the gonadotrophin
might be responsible for the described effect, another
group of animals (©roup V) were treated with 10 rat units
daily of Antuitrin ©rowth (Parke Davis)# Neither the total
body weight nor the investigated organ weights were
influenced by the injection of soma to trophic hormone on a
protein-free diet. It should be mentioned here that an
increase of the levator ani muscle by injection of growth
hormone had been found earlier in animals on normal diets*
(2$).
Table III contains the results of experiments in
which the effect of Norethandrolone on protein-depleted
castrated rats was Investigated in comparison with
testosterone propionate# Norethandrolone has been
recently introduced under the trade name of Nilevar
(Searle) and, according to the manufacturer, has a high
anabolic but low androgenic activity.
For these experiments the rats were castrated at the
age of 31 days and kept on stock diet for an additional
25 days. They were then protein depleted for 21 \ . days, at
kl
the end of which time they were sacrificed* Beginning on
the eleventh day of the protein depletion diet, the
hormones were injected daily at two different dose levels*
The results show that norethandrolone increased the organ
weight of the protein depleted castrated rats in the same
| way as did testosterone propionate* The body weight was
not affected by the injection of these hormones*
TABLE II
E F F E C T O F G R O W T H H O R M O N E A N D PITUITARY G O N A D O T R O P I N O N N O R M A L A N D C A S T R A T E D M A L E
R A T S O N A PROTEIN-FREE DIET F O R E L E V E N ! D A Y S
GROUP No.OF CONDITION TREATMENT INITIAL FINAL WEIGHT OF TISSUES
RATS BODY BODY
HEIGHT WEIGHT’
LEV.ANI SEM.VES. PENIS KIDNEY LIVEK DIAPHRAGM
(g®) (g®) (®g) (mg) (®g) (mg) (mg)
(mg)
I 5 ' Castrated 137 107 16.5 37 94 1002 4580 350
(±13) (*13) (±5.7) (±12) (±9) (±193) (±1130)
II 4
Normal 20 rat u. 133
112 52.5
680 166 1168
4195
gonadotrop. (±6)
(±1)
(±8.1) (±88)
(±14)
(±106) (±173)
III
4
Normal 40 rat u. 134
101 57.5
750 179 1079
4006 322
gonadotrop© (±3) (±4) (*8.4) (±229) (±10) (±59) (±630) (±17)
IV
4
Castrated 20 rat u. 131 107 15.7 30 95
1030 4000
gonadotrop© (-3) (±2) (±1.6) (±5) (±5) (±220) (±550)
V
4
Castrated 100 rat u. 140 108
14.1 54 99
1168
5505 372
growth h© (±4) (±5) (±1.5) (±13) (±12) (±83)
(±387)
(±15)
VI
4
Normal — 140 115
21.6 65 110
995 5145 348
(±4) (±4) (±1.5) (±15)
(±8) (±65) (±450) (±25)
fc
TABLE .III
EFFECT OF TESTOSTERONE PROPIONATE AND NORETHANDROLONE*
ON ORGAN AND MUSCLE WEIGHTS OF CASTRATED PROTEIN DEPLETED RATS**
GROUP TREATMENT
WEIGHT OF TISSUE
Levator Ani
(mg)
Seminal Ves.
(mg)
Kidney
(mg)
Liver
(mg)
Penis;
(mg)
I
I I
I I I
IV
V
T.P. 12 m g
Nilevar 1 1
T.P, 2.4 m g
Nilevar w
46.2
37.1
44.3
33.1
4.2
488
408
270
126
18
1100
1045
1083
1023
988
4435
5365
3873
4313
4820
173
150
153
138
50
* Nilevar (Searle) which was supplied by the courtesy of Messrs. Searle & Co,
** Average body weight after depletion, 111 (±9) gm.
&** 2 rats: per group
hk
DISCUSSION
It seems evident from the nitrogen equilibrium 7
experiments that the subcutaneous administration of
testosterone propionate was able to indue© a nitrogen j
t
retention. In all instances animals were fed diets that
because of the inadequacy of their amino acid composition
could not support growth, as is apparent from the observa
tion of the body weight loses*
There are several possibilities that would explain
the fact that nitrogen was retained under these circumstan
ces.
1. The nitrogen retained is not necessarily
associated with the process of growth, and is stored in
a form different from that of tissue protein. We have no
information bearing ©n this theory• No changes have been
observed in the plasma non-protein nitrogen and amino acid
nitrogen (19) after administration of testosterone,
although the possibility of non protein nitrogen storage
in tissues other than the blood has not been excluded*
2* The injection of testosterone promotes the
reutilization of amino acids coming from the breakdown
of tissue proteins as a consequence of the "wear and tear”
metabolic processes. Further investigation would be
required to support this possibility, althoug it is evident
/
from th© observation of the nitrogen balance data, that
the greatest degree of nitrogen retention was promoted on
animals fed the gelatin diet, from which the only
missing amino acid is tryptophane • When animals were fed
zein, which lacks tryptophane and lysine, and from which
valine and threonine are physiologically not aveilable,
(5b) they showed a lesser degree of nitrogen retention
after receiving testosterone propionate; while the
animals fed a diet containing a mixture of glycine, alanine
and glutamate did not show any significant decrease of
nitrogen excretion*
3# Another explanatory mechanism would be that of
hormone antagonism which would lead to a protein sparing
effect* Androgens are assumed to antagonize the excess of
circulating adrenal hormones and suppress their formation
by inhibiting ACTH secretion* Several animal experiments
by numerous investigators have shown that the administra
tion of testosterone has a definite effect on the anterior
pituitary, as well as other glands of internal secretion,
such as the adrenals, thyroid, testes and ovary (55)• In
rats testosterone propionate was shown to cause atrophy of
the pituitary and involution of the adrenals. A decrease
of glucocorticoids after testosterone propionate was also
shown*
It is possible then to consider testosterone as an
1 * 6 !
anticatabolie hormone, which by inhibiting the secretion
of adrenal cortical steroids (of known eatabolic activity)
may decrease the breakdown of body tissue. This would
explain the nitrogen retention which took place on feeding
a diet which cannot promote growth. Under these conditions
direct anabolic effects were only manifest for the
accessory sexual organs.
The increase of protein synthesis (or decrease of
catabolism) postulated under 1 and 2, should of affected
the loss of body weight. This was never observed in our
experiments, although an increased fat utilization may j
j
be the cause of this apparent contradiction.
The preceding results of the effects of androgenic
steroids on the organs and muscles of the rat show that
testosterone propionate and norethandrclone induced
growth of the levator anl muscle and of some other
accessory sex organs in normal and in castrated male rats,
even in those animals on protein-free diets. Such
deficient diets cannot support somatic growth; therefore,
the animals lost weight. The fact that the growth of
certain target.organs can be stimulated by hormones even
on such an unfavourable dietary regime must be attributed
to the androgenic effect of testosterone. These results
also suggest that on a depletion diet the proteins
necessary for the hormone stimulated growth of the levator
1 4 - 7
anl muscle and other sex organs must be obtained from
other tissues, possibly from the liver or the skeletal
muscle. These findings indicate that the hormone-induced
anabolism In the levator ani muscle is associated with
protein catabolism in other organs. Further work involving
a greater number of animals would be required to identify
the tissues from which the protein was shifted to the.
sex organs.
The fact that gonadotrophic hormone and growth
hormone did not affect the weight of the accessory sex
organs or of the levator ani muscle of castrated male
rats during protein depletion further indicates the
specificity of the hormone effect on the growth of this
muscle.
Our conclusions that the size and weight of the
levator ani muscle is a sex-linked function is supported
by earlier observations. For example, the weight of this
muscle in females is only a small fraction of that
observed in adult males. After castration in males the s
levator anl loses about 60 to 70 % of its original weight
within 10 to l l j . days• The loss of the skeletal muscle
during this time is^ however, usually insignificant.
Our experiments do not explain why the relative
activity of tie various androgenic steroids differs
according to the organ selected for comparison,
.i.e. why some of them have a greater effect on the weight
of the seminal vesicles, while others have a greater
effect on the levator ani muscle*
Such differences have been observed in the past,
£*g* testosterone is ten times more active than
androsterone when tested on the seminal vesicles but only
two to five times as active when compared on basis of the
increase of prostate weight*
On the basis of such observations and of our results,
we must assume that the threshold levels, the speed of
absorption and destruction and similar factors, rather
than differences in androgenic and anabolic activities of
the compounds, must be responsible for the differences in
the effect on various organs.
To account for this specificity of action the
following theory is proposed.
The difference of relative stimulation of androgenic
steroids on the accessory sexual organs is well established
in the literature. Slight changes in the structural
configuration of the basic androgenic substance testoste
rone, seems to produce compounds which would apparently
have a preferential effect in stimulating a specific
accessory sexual organ. The comparison of androsterone and
testosterone has been discussed above.
Now that we have supplied evidence to consider the
levator anl muscle as an accessory sexual muscle, we can
assume that we are dealing with a similar mechanism when
we talk about preferential growth stimulation. To my
knowledge no theory has ever been proposed to explain the
reason for this difference of relative potency.
It is evident that on a weight basis testosterone
esters are the most active compounds on all the accessory
sexual organs. The fact that changes in the structure of
testosterone can render available compounds that stimulate
the accessory sex organs at a different ratio can probably
be explained by the following considerations.
Assuming that the required structure for androgenic
stimulation is that of testosterone as such, then in order
to get a response from the administration of any other
closely related steroid, it would have to be transformed
at the cellular level of the accessory sex organ, by an
enzymatic process, into the active substance testosterone.
The greater or lesser efficiency of the particular organ
in producing this conversion could be the limiting factor
for the organ* s growth.
j
SUMMARY
1* Subcutaneous administration of testosterone
propionate induced a nitrogen retention in castrated male
I rats fed amino acid deficient diets* These diets cannot
j support growth* Several theories to explain the mechanism
R
of retention under these circumstances have been proposed*
2* During the course of all these experiments
j animals depleted their body weight at a constant rate,
! '
I
| due to inadequacy of nitrogen source, although caloric
' intake was satisfactory* This loss of body weight did
1 not seem to be affected by temporary or total periods of
positive nitrogen balance induced by the injection of
testosterone propionate*
3* Androgenic steroids were shown to be able to
increase the weight of the accessory sexual organs,
levator ani muscle and kidneys of rats depleted on a
protein free diet*
I } . * Evidence for the non-suitability of the levator
ani bioassay technique for screening the anabolic
effects of androgenic steroids is furnished*
BIBLIOGRAPHY
1* Koehakian, C. D., and Mur1in, J., J* Nutrition, 10, I
- 1+36, (1931) “ I
I
2• Koehakian, G. D., and Mur1in, J*, Am. J. Physiol., I
117, 61+2, (1936) |
i
! 3. Koehakian, G. D., Endocrinology, 21, 750, (1937) i
i \ i
| I j . . Thorn, G. W. and Engel, L. L., J. Exp. Med., 68, 299, !
j (1938) |
j
i 5* Gaebler, 0. H. and Tarnowski, S. M., Endocrinology,
| 13, 317, (191+3)
| 6. Koehakian, C. D., Conference on Metabolic Aspects of j
| Convalescence, Transactions of the 12th Meeting, |
j E. G. Reifenstein, N. Y., 12, lOij., (19lj.6), J. Macy jr.j
7* Braasch, J. W., Wakerlin, G. E., Bell, J. H. and j
Levenson, S. M., Proc• Soe. Exp. Biol. Med., 75.
183, (1950) S
! !
i . !
I 8. Koehakian, G. D., and Beall, B., Am. J. Physiol.,
j 160, 62, (1950)
9* Samuels, L. T., Henschel, A. P., and Keys, A.,
J. Glin. Endocrinol., 2, 6^9, (19^)
| 10. Bassett, S. H., Kentmann, E. H., and Koehakian, G. D., !
Conference on Metabolic Aspects of Convalescence, r j
Transactions of the 12th Meeting, E. C • Reifenstein,
N. Y., 10, 26l, (191^5) , J* Macy jr. Foundation.
11. Borum, E. R., and Geiger, E., J. Glin. Invest.,
1109, (1956)
12. McCullagh, P., and Rossmiller, H. R., J. Endocrinol.,
1, 507, (19) 413
13* Wilkins, L., and Pleischmann, W., Endocrinology, 6,
383, (191*6)
ll*. Kenyon, A. T., Knowlton, K, Sandiford, I, Koch, P. C.,
and Lotwin, G., Endocrinology, 26, 26, (I9I 4 .O) |
j
15. Brown, H., and Samuels, L. T., J• Clin. Endocrinol. j
and Metabolism, 16, 775, (1956) j
52
16. Koehakian, C• D., Tribalsky, E., and Bartlett, M.,
Conference on Metabolic Aspects of Convalescence,
Transactions of the 13th. Meeting, E. C. Reifenstein,
N. Y., 11, 65, (1914.6) , J. Macy jr. Foundation.
17♦ Koehakian, G. D., Cohn, L., Quigley, E., and
Tribalsky, E., Am. J. Physiol., lgg, 272, (19^8)
18. Geiger, E., and El Bawl, I., Metabolism Clin, and Exp.,
1, llt-5, (1952)
19* Koehakian, C., Robertson, E., and Bartlett, M. N.,
Am. J. Physiol., 163. 332, (1950)
20. Rubinstein, H. S.. and Solomon, M. L., Proc. Soc.
Exp. Biol. Med., glu Uh2, (191+0)
21. Korenchevsky, V., Denison, M., and Brovsin, I.,
Bioehem. J., JO, 558, (1936)
22. Koehakian, C. D., Am. J. Physiol., lij.2, 315# (19144-)
23* Papanicolaou, G. N., and Falk, E. A., Science, 87,
238, (1938)
2I 4 .. Wainman, P., and Shipounoff, G. C., Endocrinology,
29, 975, (1914.x)
25. Eisenberg, E., and Gordan, G. S., J. Pharmacol. Exp.
Therap., 21, 38, (1950)
26. Preston, P. L., and Cassidy, J. W., Pro©. Sec. Exp.
Biol. Med., 8£, 6 7 l j . , (1953)
27. Wijnans, M., and De Groot, G. A., Acta Physiol, et
Pharmacol. Neerl., J, 85, (1953)
28. Koehakian, C. D., and Van der Mark, W., Proc. Soc.
Exp. Biol. Med., J2, 7k, (1952)
29. Grayhack, J. T., and Scott, W., Endocrinology, 50,
1 4 . 06, (1952)
30. Leathern, J. H., Am. J. Physiol., 165, 73, (1951)
31. Forsyth, B. T., J. Lab. Clin. Med., IjJ, 732, (195^)
32. Aschkenazy, A., and Dray, F., Compt• Rend. Soc. Biol.
1 1 4 . 7 . 1772, (1953)
33• Koehakian, C. D., Garber, E. E., and Bartlett, M. N.,
Am* J. Physiol., 155* 265, (19k8)
3k« Leathern, J* H*, Am. J. Physiol., 15k. k59* (19k8)
35* Schilling, W., and Laqueur, G. L., Endocrinology,
^0, 753, (19k2)
36. Selye, H., Can. Med. Assoc. J., k2. 113, (19k0)
37* Talesnik, S., Rev. Soc. Argentina Biol., 2£, 87,
(1953)
38. Koehakian, C. D. and Stettner, G. E., Am. J. Physiol.,
155. 255, (19k8h
39* Koehakian, C. D., Am. J. Physiol., 160, 66, (1950)
I 4 .O. Kenyon, A. T., Knowlton, K., and Sandiford, I.,
Ann. Internal Med., 20, 632, (I9kk)
kl. Koehakian, G. D., Am. J. Physiol., 160, 53, (1950)
k2. Rupp, J. J., and Paschkis, K. E., Metabolism Glin.
and Exp., 2, 268, (1953)
k3. Koehakian, G. D., Moe, J. G., and Dolphin, J.
Am. J. Physiol., 162, 581, (1950)
kk* Parkes, A. S., Brit. Med. Bull., 11, 105, (1955)
k5. Mieseher, K., Wettstein, A., and Tschopp, 1.,
Biochem. J., ^0, 1977, (1936)
k6. Burn, J. H. Biological Standardization, p, 152,
Oxford University Press, (1937)
k7. Deanesly, R., and Parkes, A. S., Lancet, 230* 837,
(1936) ,
k8. Stafford, R* 0., Bowman, B. Jand Olson. K. J.,
Proc. Soc. Exp. Biol. Med., 86, 322, (195k)
k9. Barnes, L. E., Stafford, R.O., Guild, M. E., Ihole,
L. C., and Olson, K. J., Endocrinology, 55* 77, (195k)
50. Engle, E. T., and Pincus, G*, Hormones and the Aging
Process, Academic Press Inc. N* Y., (1956)
5k
5l« Koch, F. C. and McMeekin, T. L., J. Am. Chem. Soc*,
1 ±6, 2066, (19%k)
52* Hershberger, L. G*, Shipley, E. G., and Mayer, R* K.,
Proc* Soc* Exp. Biol. Med., 8£, 175, (1953)
53* Samuels, L. T., Progress in Clinical Endocrinology,
P* 5^7, Grune and Stratton, N. Y., (1950)
51+. Geiger, E., Courtney, G. W., arid Geiger, L. E.,
Arch. Biochem. and Biophys., 1+1, 7l+, (1952)
55• Venning, E. H., and Browne, J. S., J. Clin.
Endocrinol. 7, 729, (191+7)
0nlvers3,ty of Soutfiern Caiiforai®
UMI Number: EP41334
All rights reserved
INFORMATION TO ALL USERS
The quality of this reproduction is dependent upon the quality of the copy submitted.
In the unlikely event that the author did not send a complete manuscript
and there are missing pages, these will be noted. Also, if material had to be removed,
a note will indicate the deletion.
Dissertation Publishing
UMI EP41334
Published by ProQuest LLC (2014). Copyright in the Dissertation held by the Author.
Microform Edition © ProQuest LLC.
All rights reserved. This work is protected against
unauthorized copying under Title 17, United States Code
ProQuest LLC.
789 East Eisenhower Parkway
P.O. Box 1346
Ann Arbor, Ml 48106-1346

Linked assets

University of Southern California Dissertations and Theses

Conceptually similar

$Studies on the fractionation of rat plasma proteins: The separation of albumin$

PDF

Studies on the fractionation of rat plasma proteins: The separation of albumin

PDF

The comparison of the metabolism of C¹⁴ carboxyl and methylene labeled glycine in the intact rat

PDF

A study of methods of cholesterol analysis

PDF

Properties of the myosin-like proteins of sea urchin eggs

PDF

Some Aspects Of The Intermediary Metabolism Of Mycobacteria

PDF

The Biosynthesis Of Ergothioneine

PDF

A comparative study of halodeoxyuridines

PDF

Studies On The Metabolism Of Iron

PDF

Altered interaction of human endothelial cells to the glycosylated laminin

PDF

The effects of the non-dialyzable products of pepsin digested serum proteins on the growth of tissue cells in vitro

PDF

Studies On The Mechanism Of Action Of 5-Hydroxyuridine

PDF

Studies of trypsin-binding ɑ₂ macroglobulin of human plasma

PDF

A comparative study of the utilization of jojoba and cottonseed oil in the rat

PDF

A study of the ketone body metabolism of excised rat tissues with respect to labile phosphate content

PDF

A comparison of glucose tolerance in normal rats and those with fatty livers

PDF

Microheterogeneity of fetuin

PDF

The isolation and characterization of the porcine submaxillary glycoprotein

PDF

Isolation And Characterization Of Hageman Factor And Plasma Thromboplastin Antecedent

PDF

Sociological aspects of the novels of Edith Wharton

PDF

Galactose metabolism in human blood cells

Asset Metadata

Creator Nimni, Marcelo Efraim (author)

Core Title Some metabolic effects of androgenic steroids on rats fed different inadequate diets

School Department of Biochemistry and Nutrition

Degree Master of Science

Degree Program Biochemistry and Nutrition

Degree Conferral Date 1957-06

Publisher University of Southern California (original), University of Southern California. Libraries (digital)

Tag health sciences, nutrition,OAI-PMH Harvest

Language English

Contributor Digitized by ProQuest (provenance)

Advisor [Seyer] (committee chair), Mehl, John W. (committee member), Stowell, Ellery Cory (committee member)

Permanent Link (DOI) https://doi.org/10.25549/usctheses-c20-296922

Unique identifier UC11259983

Identifier EP41334.pdf (filename),usctheses-c20-296922 (legacy record id)

Legacy Identifier EP41334.pdf

Dmrecord 296922

Document Type Thesis

Rights Nimni, Marcelo E.

Type texts

Source University of Southern California (contributing entity), University of Southern California Dissertations and Theses (collection)

Access Conditions The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the au...

Repository Name University of Southern California Digital Library

Repository Location USC Digital Library, University of Southern California, University Park Campus, Los Angeles, California 90089, USA