Pregnancy Urine Extract Research Articles

Lack of Maturity Hormone in the Hypophysis of the Infantile Rat

RECENT work on endocrines has brought out two facts interesting in their implications. One is that pregnancy urine, blood serum and placental extracts produce lutein cells in the hypophysectomised animal but do not induce follicular growth1,2,3; and the other is the discovery4 of an activation factor in adult beef hypophysis which is not identical with either the growth or maturity hormones. Since it is well known that these pregnancy extracts will induce precocious sexual maturity in normal immature animals (in other words, follicular growth), it seems certain that they contain an activation factor which stimulates the hypophysis of the host, thus initiating growth in the ovary. There is as yet no basis for comparing these two factors, but, since the hypophysis of the pregnant rat shows but little increase in potency over that of the non-pregnant one5, this organ would not seem to be responsible for the activator in pregnancy extracts, this probably coming from the placenta.

Mating Reaction of Hypophysectomized Male Rats Treated with Pregnancy Urine Extracts

Adult male rats are known to lose all libido immediately after hypophysectomy (Smith1), though matings for some time after castration have been reported. Also motile sperm have been demonstrated in the epididymis of hypophysectomized male rats no longer than 21 days after the operation (White2). Complete fertility is restored to hypophysectomized males by giving rat pituitary implants (Smith1).Since it was observed that pregnancy urine extract (P.U.) slowed the degenerative changes in the testes of hypophysectomized rats (see previous article), matings were attempted at various times with oestrus females where treatment was begun immediately fob lowing the operation. Of 7 hypophysectomized males, 6 mated one or more times. In a total of 25 attempts at mating, 13 were positive and, furthermore, 11 of these were fertile, producing normal litters. It was most interesting that of these fertile matings some took place 18, 20, 32, 34, and 47 days following hypophysectomy. It was not surprising because of the en...

A Quantitative Procedure for Determining Normal Excretion of Prolan

Several methods have been developed for the preparation of active extracts of the gonad-stimulating substance of pregnancy urine. Although some of these procedures give good recoveries from pregnancy urine no one has reported a satisfactory procedure for the quantitative estimation of the small quantities of prolan that occur in normal urines. The excretion of prolan in conditions unassociated with pregnancy has been determined exclusively with untreated urine or by means of Zondek's alcohol precipitation method which gives only a 5-fold concentration. Since alcohol causes an appreciable loss of activity (Wiesner and Marshall, Katzman and Doisy,) this procedure is not suitable for use in normal excretion of the anterior pituitary-like substance. In attempting to purify pregnancy urine extracts by means of various protein precipitants we observed that precipitation with tungstic acid invariably gave remarkably complete recovery. Wiesner and Marshall and Zondek, Scheibler and Krabbe found that phosphotungstic acid quantitatively precipitates the active material of pregnancy urine but they have not reported studies on the recovery of the small amounts of prolan which occur in normal urine. We have not used phosphotungstic acid but it is evident from Table I that our tungstic acid method gives a surprisingly good recovery of minute quantities. The procedure is as follows: A 24-hour urine sample to which has been added 10 cc. of 10% sodium tungstate and 10 cc. of 0.5% casein is made faintly acid to congo red with dilute H2SO4. The precipitate is collected by centrifugation, washed with acetone, freed from acetone by reduced pressure, and dissolved by adding dilute NaOH until the mixture is faintly alkaline to phenolphthalein. BaCl2 and Ba(OH)2 are then added in the proportion necessary to prevent the solution from becoming acid, until precipitation is complete.

Effect of Injecting Pregnancy-Urine Extracts in Hypophysectomized Rats. I. The Male.

The injection of pregnancy urine (P.U.) in normal young rats causes an increase of the interstitial cells of the testes, an enlargement of the accessories and occasional injury to the tubules (Engle1). Collip et al.2 reported that injections of placental extracts (which he considers physiologically similar to P.U.) in hypophysec-tomized male rats caused a hypertrophy of the interstitial tissue of the testes, prevented atrophy of the accessories, but did not prevent a loss in weight of the testes or degeneration of the tubules. Freud3 reported that P.U. increased the size of the testes in hypophysectomized rats but gave no structural details.We have injected several series of hypophysectomized male rats with Antuitrin S (P.U. extract) starting either immediately after hypophysectomy or after a 20-75 day post-operative period. For controls, either one testis was removed before injections were started or littermates were autopsied at that time. Untreated operated littermates were also autopsied at the end of...

THE PHYSIOLOGY OF PREGNANCY IN THE RAT.—I. THE PROLONGATION AND INTERRUPTION OF PREGNANCY

1. The gestation period in the rat can be prolonged by from 4 to 10 days by (a) administration of anterior pituitary tissue; (b) extract of human pregnancy urine; (c) alkaline extract of pituitary; (d) extract containing “purified” growth hormone.2. Pregnancy is apt to be interrupted when injection of the urine extract is made in the early stages.3. The evidence adduced suggests that the pregnant condition is not maintained solely by the corpus luteum, and that parturition is not due merely to degeneration of that body.

THE ACTION OF EXTRACTS OF ANTERIOR PITUITARY AND OF PREGNANCY URINE ON THE TESTES OF IMMATURE RATS AND MONKEYS*

Several score of papers dealing with the action of extracts of pregnancy urine and of the anterior pituitary on the male genital system bear witness to the attention which has been focused on this point since 1927. In our earlier work on the effect of implants (Smith and Engle), we noted that the distinct growth of the seminal vesicles of implanted animals demonstrated an activity of the interstitial cells, though morphological changes, were not directly observable on these cell bodies. These observations have been greatly extended by many investigators,1 who note varying degrees of hypertrophy and hyperplasia in the interstitial cells after treatment with extracts of anterior pituitary or pregnancy urine. With the exception of the report of Balawenetz (1930), no one has reported success in hastening the formation of spermatids or spermatozoa, in an immature male mammal, and it is generally agreed that, in the male at least, none of these gonad-stimulating factors are directly concerned with gametogenesis. These accumulated observations on the prepubertal male emphasize the position taken a year ago in regard to the action of anterior pituitary principles on the female (Engle, 2). We are dealing with a genital acceleration mainly of the hormone producing structures but have not produced true precocious puberty. That phenomenon is more complex than may be explained by the action of a single hormonal factor.

Prolongation of Pregnancy

RECENT experiments upon rats demonstrate that pregnancy can be prolonged by 4–10 days by any of the following treatments: implantation of anterior pituitary tissue (cattle), the injection of an alkaline extract of that gland, and of extracts of human pregnancy urine prepared by precipitation with barium-alcohol and/or phosphotungstic acid. Judged by the maternal weight curve, full development of the fœtus was reached at normal term, but the birth mechanism failed. Where pregnancy was prolonged for more than 3½ days, the fœtuses were invariably still-born; in several instances parturition was protracted for 12–70 hours. Since the ovaries of such animals were found, on biopsy, to be highly luteinised, prolongation was thought to be due to the persistence of the corpora lutea formed as the result of the treatment administered. Such was the view of Teel1 and of Levin, Katzman, and Doisy.2 There are, however, certain indications that another factor besides the corpus luteum is concerned in maintaining the conditions of pregnancy.

Uterine Bleeding of the Interval Type in Macacus Monkey During Injections of Extracts of Pregnancy Urine.

Uterine bleeding in the spayed or immature macacus monkey is known to occur from an interval type endometrium about 6 days after cessation of an adequate treatment with oestrin (Theelin, Allen1, Maddux2;) (Amniotin, Morrell et al.3). These observations have been repeatedly confirmed in this laboratory. Uterine bleeding in the adult also occurs after bilateral ovariectomy (Allen; Van Wagenen and Aberle4) or occasionally after unilateral ovariectomy (VanWagenen and Aberle). (Uterine bleeding indicates a hemorrhage of uterine origin macroscopically visible on the external genitalia of the monkey, but which may not be homologous with menstruation). Saiki5 has shown that uterine bleeding may be induced in immature macaques 4 to 9 days after the cessation of 8-17 days treatment with an extract of anterior lobe (P. D. & Co.). Our work confirms these results. However, if water soluble extract of pregnancy urine or the “luteinizing hormone” (P. D. & Co.) be administered in adequate doses for from 19 to 22 days, in...

PREGNANCY CHANGES IN THE RABBIT'S UTERUS AND THEIR RELATION TO ENDOCRINE ACTIVITY.—II. THE ACTION OF GONADOTROPIC PREPARATIONS OF THE PITUITARY AND OF PREGNANCY URINE

Minced bovine anterior pituitary lobe substance, alkaline extracts of the same, and gonadotropic extracts of the urine of pregnant women have been injected into rabbits and the duration of activity of the luteal tissue formed has been determined; progestational proliferation of the endometrium and inhibition of the in vitro reaction to a posterior pituitary lobe extract of the uterine musculature were used as criteria of luteal secretion. A single injection (intramuscular or intravenous) of the above preparations was, as a rule, followed by a period of luteal activity more or less equivalent to the duration of normal pseudopregnancy. Certain exceptions to this type of effect are described.Repeated injections have prolonged the period of luteal activity; towards the end of this period it has been possible, in a limited number of experiments, to obtain inhibition of the reaction of the uterus to the posterior pituitary lobe extract in the absence of progestational proliferation of the endometrium. The significance of these phenomena is discussed, and it is suggested that they may be explained on the assumption that the corpus luteum secretes two hormones responsible for the progestational and the inhibitory effects respectively.I wish to express my acknowledgments to Dr B. P. Wiesner for help and advice; and to thank him and Mr R. E. Illingworth for the pituitary and pregnancy urine extracts used.

Difference Between Gonad-Stimulation by Extracts of Pregnancy-Urine and of Pituitary Body.

The source of the gonad-stimulating substance of pregnancy-urine is still undecided; the weight of evidence, however, indicates that it is extra-hypophyseal in origin. In studies of the comparative effects of extracts of sheep or pork pituitary bodies and of pregnancy-urine or oestrin-free extracts of pregnancy-urine or chorionepithelioma urine, it was found that the minimal effective doses of the urinary preparations were practically the same for both the male and the female. In the cases of crude or refined extracts of pituitary bodies, however, the minimal effective dose for the female was but a fraction (less than 10%) of that required for the male.Assay was performed on rats exactly 21 days old when the first injection was made; one injection was made daily for 4 days; necropsy was performed at the age of 26 days or 120 hours after the first injection. Assay-results were appraised by the weights of both ovaries or of both seminal vesicles and coagulatory glands. The extracts of sheep and pork pituita...

The Ovary Stimulating Hormone of the Placenta

THE production of prematurity in young female rats and mice by Smith and Engle in America, and by Zondek and Aschheim in Germany, through the use of the anterior pituitary gland transplant or emulsion has afforded a great stimulus to further investigation of the hormonal control of gonadal activity. The latter investigators have demonstrated that effects somewhat similar to those following pituitary gland implantations can be elicited in immature rats and mice by extract of pregnancy urine which has been rendered œstrin-free. They have likewise shown decidua, placenta, the corpus luteum of pregnancy, as well as pregnancy blood, to contain an ovary stimulatory hormone analogous to that of the anterior pituitary gland.