Wm. R. Lyons

The Direct Mammotrophic Action of Lactogenic Hormone

Summary Thirty oöphorectornized, virgin rabbits, injected subcutaneously 5 days weekly for 4 weeks with 200 I.U. of estrone and 1 I.U. of progesterone showed prolactational mammary growth approximating that of a 3-weeks′ pregnant rabbit. Three days after the last of these injections, 5 different levels of lactogenic hormone were tested on groups of 6 rabbits. The hormone was given intraductally in such a way that an individual sector in each animal received a constant volume of fluid (1.0 cc). Of the different doses of hormone tested, 6 and 3 I.U. caused localized sector lactation in all 6 animals; 1.5 I.U. in 5 of 6; and 0.75 I.U. in 3 of 6 animals. None of the 6 animals injected with 0.37 I.U. lactated; nor did the uninjected or control-injected sectors in any animal lactate. This experiment demonstrates for the first time that the hypophysial lactogenic hormone has a direct mammotrophic effect just as it was previously shown to have a direct crop sac stimulating action. With all other factors essentially equal in any 2 adjoining sectors in a mammary gland, the mere bathing of the parenchyma of one with 25 μg of this purified protein brought about the complex cell-growth and cell-renewal processes of lactation, whereas the introduction of a control solution or no treatment whatsoever, caused no detectable change in the other. The estroiie and progcutcronc used in this study wcre generously supplied by Parke, Davis and Company, Detroit, Mich.

Pregnancy Maintenance in Hypophysectomized-Oöphoreetomized Rats Injected with Estrone and Progesterone.

Summary Levels of estrone and progesterone have been determined which, when given in combination to rats oöphorectomized and hypophysectomized on day 7 or 8 of pregnancy, will permit most of them to carry a slightly less than average number of apparently normal young through the usual gestational period. Of 7 rats given 1 fxg of estrone and 4 mg of progesterone daily, one died on day 22 following an attempt to deliver 6 large and 2 resorbing fetuses, and the other 6 when sacrificed on days 22 or 23 were found to have a total of 39 (3 to 9) living young and 16 (1 to 6) fetuses in various stages of resorption. Of the 10 rats given 1 μg of estrone and 3 mg of progesterone, one died on day 21 without being able to deliver 9 term-size fetuses, 6 when sacrificed on days 21 or 22 were found to have a total of 36 (4 to 8) living young and 8 either recently dead or in various stages of resorption, and the remaining 3 when sacrificed on days 22 or 23 were found to have resorbing implants or no visible-implantation sites. It may be concluded, therefore, that 1 μg of estrone and 4 mg of progesterone substituted for the ovaries and pituitary during the period of pregnancy between days 7 and 11, and for the ovaries alone from day 11 until term. The pituitary is only necessary during the first half of pregnancy in the rat, and then probably because of its gonadotrophs triad, FSH, ICSH and lactogenic hormone.

Effects of Estrone and Progesterone on Male Rabbit Mammary Glands. I. Varying Doses of Progesterone

Summary Eighteen doses of 0.25, 1.0, 4.0 and 8.0 I.U. of crystalline progesterone were given, simultaneously with 120 I.U. of estrone, to immature male rabbits during a period of 4 weeks. Of these 4 levels of progesterone, the 1.0 I.U. dose synergized best although the prolactational proliferation induced was not maximal. The 4.0 and 8.0 I.U. doses of progesterone were apparently inhibiting as shown by the relatively poor mammary growth obtained.

Direct action of estrone on the mammary gland.

Summary The proper dose of estrone in oil rubbed into the skin over the rudimentary mammary glands of young male rabbits caused growth only of those glands, and not of the control glands treated with oil. Such evidence supports the view that estrogenic substances are directly mammary-stimulating.

Induced Postpartum Lactation in Hypophysectomized Rats.

Summary It was possible to induce postpartum lactation in hypophysectomized rats, and to maintain a functional development of their mammary glands by injections of combined mammotropic and adrenocorticotropic hormones. That this treament did not constitute complete substitution therapy was shown, however, by the fact that sufficient milk was not available for sustenance of the young.

The Blood of the Normal Guinea Pig.

It is suggested that inconsistencies in the work of previous investigators on the blood of the normal guinea pig have been due, for the most part, to the use of the usual fixed staining technic. The supravital method of blood study, first used by Pappenheim, 1 and reintroduced by Simpson, 2 overcomes the common difficulties of cell distribution, and identification, as met with in the smear method. The peripheral blood of the normal guinea pig has been studied by the Neutral Red-Janus Green supravital method. Sixty-four guinea pigs from four different sources, of both sexes, and weighing from 500 to 1000 grams, comprise the series. From 200 to 400 white cells per preparation were counted on each. To compensate for normal fluctuations, counts were made at all hours of the day and night. The normal guinea pig has been found to have a leucocytic formula peculiarly its own, especially in that an atypical mononuclear is normally present in the circulation. In our counts this will be termed the Kurlow cell, because of its inclusion, the so-called Kurlow body. Considerable evidence has been obtained upholding the belief that the Kurlow inclusion is the result of a symbiotic protozoan. An average total leucocyte count of 9,600, and a total erythrocyte count of 5,565,000, were established. The differential count showed the following: Neutrophile, 34.9%; Lymphocyte, 49%; Monocyte, 7%; Eosinophile, 3.1%; Basophile, 0.8%; Kurlow cell, 5.0%; Unclassified cells, 0.2%. It is believed that the above data establishes a fairly accurate means of ascertaining the normalcy of a fully matured guinea pig.

Tutocain as a Shock Preventative in Tuberculous Guinea Pigs Injected with Tuberculin.

The symptoms following the rapid absorption of tuberculin in the tuberculous guinea pig, in which the subcutaneous inoculation of small amounts of tuberculin invariably prove fatal, are well known. Fischel 1 has suggested that the severe tuberculin reaction can be mitigated by introducing tuberculin into the skin with a view to retarding absorption and slowly removing the toxic factors elaborated upon the contact of tuberculin with tuberculous tissues. It had occurred to us that if conditions could be set up subcutaneously which would retard absorption of this toxic factor, and at the same time permit the use of larger amounts of tuberculin, some therapeutic results might be developed, without the danger of fatal issue. To produce these conditions, a local anesthetic, hydrochloride of p-amino-benzoyl-di-methyl-amino-methyl-butanol or by its trade name, the “Tutocain” brand of Butamin, which induces an edema and at the same time renders it painless, was selected. As a rule 4 cc. of a 1% solution was injected subcutaneously. The guinea pigs had been rendered tuberculous by the subcutaneous inoculation of 1 cc. of a suspension of tubercle bacilli (virulent strain No. 427) into the inguinal region 4 weeks previous to the tests. Animals were subjected to an intracutaneous tuberculin test before they were employed in the experiments, only animals showing a strongly positive reaction being used. One half hour after the application of the drug 0.4 cc. of old tuberculin was injected subcutaneously into the infiltrated area. Previous tests had shown that 0.2 cc. of the same tuberculin was sufficient to cause death of tuberculous guinea pigs within 24 hours. Guinea pigs, from numbers 1 to 10, were autopsied one week after the test inoculation and the extent of the infection noted. All guinea pigs showed extensive and typical tuberculous lesions in the liver, spleen, lymph nodes, and lungs. The remainder of the guinea pigs were kept under observation for subsequent tests.