Casimir Funk

The Male Hormone.

The stumbling block in the successful demonstration of the testicular hormone has been the absence of a reliable animal test. Pézard and his collaborators found that caponized cockerels are good objects for demonstrating the presence of the hormone. They have shown that when ovarian grafts are introduced into such animals, the combs (and whiskers) diminish in size, become pale, and the feathers change in the direction of the female type. Testicular grafts, on the other hand, emphasize masculinity. Carindroit and Pézard 1 succeeded in demonstrating the presence of the male hormone in the blood of cocks. Busquet, 2 using the same test, concluded that the hormone exists in the blood of various young animals. He also claims to have obtained encouraging results by oral administration. In view of the apparent successful demonstration of the presence of the male hormone in the circulating blood, and of the estrus-producing hormone in the urine of pregnant women 3 it seemed of value to investigate the urine of young men, in the hope of showing the presence of the male hormone. Lowe and Voss 4 show the presence of “testiculin” in the urine of men. The method, however, is not described, and no details are given. Oslund 5 records the injection of fresh sperm from the vas deferens into 3 caponized cockerels, resulting in positive comb growth. We present our results which, though still in its initial stages, is already based on considerably more experiments than those recorded by Oslund. So far some 58 cocks, about 2 months in age, have been castrated. Of these, 13 died. Of the remaining 45, we believe the earlier 15 were badly castrated: but the later 30 seem to have been well castrated, judging by the results obtained.

The Male Hormone. II.

In a previous communication 1 we reported that the male urine contains appreciable quantities of the male hormone, as evidenced by the increased size of comb in castrated cocks. In the course of the work we noticed that the male hormone (testiculin) shows chemical properties similar to those of estrin. For example, testiculin, like estrin, can be extracted from urine by means of chloroform. The urine to be used is adjusted to a pH of 4.0-4.5 and continuously extracted with chloroform for 6 hours. The separated chloroform layer is evaporated and the residue dissolved in an appropriate amount of olive oil. Using urinary extracts, we find that the comb growth is proportional to the amount of hormone present (see chart). We have selected as our cock unit that amount of testiculin which, when injected daily into each of 6 castrated cocks, will give an average increase in comb of 10 mm. in 10 days. Such a cock unit is found in about 75 cc. of an active urine. The administration of chloroform extracts has permitted the use of more concentrated preparations, without the toxic effects of the urine. Changes in size of comb, in turgor and color are plainly noticeable after a few days of treatment. As a preliminary to the study of the functional test of gland activity, samples of mixed urines were extracted with chloroform, the residues dissolved in oil and each sample injected into six well-castrated cocks. Urine sample U.M.2, was obtained from men in the prime of life. The curves show how unifom is the effect of this extract on 3 different groups of animals. U.M.3 represents the urine from men of various ages, and U.M.4 from men between 50-70 years of age. The variation in hormone content is quite evident.

A STUDY OF HORMONAL FACTORS WHICH INFLUENCE THE PRODUCTION OF INSULIN

1. Insulinotropic effects were obtained with a protein fraction of the anterior pituitary, with estradiol and with stilbestrol. 2. Varying diabetogenic effects were obtained with highly purified prolactin preparations, with progesterone, with testosterone and with a fraction, probably also protein, from the anterior pituitary.

Role of the pituitary in growth of a transplanted rat tumor.

Summary Hypophysectomy in rats definitely, but incompletely, suppresses the growth of a transplanted tumor. It appears possible that a hormone, or hormones, of the pituitary are required for stimulation of tumor growth as evidenced by the effects of pituitary gland removal and the partial results of the administration of certain anterior pituitary extracts.

Enzyme levels in the growing and spontaneously regressing Flexner-Jobling carcinoma. II. ATPase and glycolysis☆

Abstract Glycolysis by homogenates from regressing tumors, in terms of lactic acid production, is reduced by an average of 54% as compared to homogenates from nonregressing tumors. This decrease is due to a sharp rise of the ATPase levels in the regressing tumor homogenates. Addition of fluoride, an ATPase inhibitor, will completely restore glycolysis as compared to homogenates from nonregressing tumors under identical conditions. In the presence of AMP, added either initially to the medium or produced enzymically from ATP or ADP, the lactic acid assay is higher than indicated by the CO 2 evolution from a bicarbonate-containing medium. The same is true of the phosphate assay in the incubation of ATP, MgCl 2 , and bicarbonate-phosphate with tumor homogenate. Additional evidence to that brought forth by other investigators suggests the enzymically catalyzed interaction between CO 2 and a split product of ATP.

A new pathological condition of probable dietetic origin in rats.

It has recently been observed in this laboratory, that when young or mature rats, of both sexes, which have been on the laboratory stock diet (milk, white bread and mixed grains) are transferred to individual cages with screen bottoms and are fed on an artificial food mixture, consisting of 18% casein (A and B free, British Drug Houses), 78% corn starch and 4% McCollums salt mixture, they develop, in the majority of cases, within 8 to 10 days, peculiar tail symptoms. These symptoms start with a slight segmentation of the distal end of the tail, followed by a more accentuated constriction, reddening of the segment, sometimes bleeding, necrosis and falling off of the tail tip. As the experiment proceeds the segmentation and the process extends. It has been thought first, that the symptoms present an analogy with the deficiency disease described by Burr and Burr. 1 However, the following reasons speak in favor of a new pathological entity. The disease observed by us develops much earlier and is neither prevented by a daily addition of 400 mg. lard, 2 drops of cod liver oil, nor by 2 drops of linoleic acid. Replacing the purified casein by ordinary commercial casein did not delay the symptoms.

Some Factors Influencing Production of Ulcers in the Shay Rat

Summary The paper presents data on the degree of ulceration developed in a large number of rats treated according to the Shay tech-nic, showing the incidence of esophageal changes, stomach acidity, and gastric liquid volume. The incidence of hematuria and pigmented urines is emphasized in their possible relationship to the etiology of Shay rat ulcers, as is the effect of preoperative diet in reducing hematuria and pigmented urines and increasing the incidence of ulceration.

THE ACTION OF CERTAIN HORMONES AS DIETARY CONSTITUENTS

The sex organs of the male are more resistant to dietary deficiencies, the animals showing only retrogression along with a significant loss in weight. Testosterone and gonadotropic hormone (of pregnant mare serum) produce a small but definite restorative effect. Oral administration of estrin to females depresses the food intake, while testosterone, and less so the gonadotropic hormone in males produces the opposite effect. This effect can be regarded as evidence of the dietary significance of these two hormones. The expenses of the present investigation were defrayed by the U. S. Vitamin Corporation, of New York City.

The chemical nature of insulin.

For the past two years we have suspected that the tendency to look upon insulin as a simple chemical compound might be erroneous, and that the diminution of the unit dose does not mean a purification in the strict sense, but rather a splitting off of some inactive amino acids. This conclusion was reached from the extremely small content of chlorine in the insulin hydrochloride. Some time ago we had a second proof that our contention is right. In studying the chemical composition of the insulin compound obtained with naphtol yellow S, or its sodium salt, it has been found that fairly pure preparations of insulin, containing about 10 clinical units to a milligram (impure insulin gave unsatisfactory results), give a precipitate with naphtol yellow S, the filtrates being absolutely inactive. The insulin compound is insoluble in water and glacial acetic acid, but soluble in dilute acetic acid and soda solution. It can be injected into rabbits with typical insulin action in the form of the sodium salt, which does not become inactive even on standing in alkaline solution. The activity is destroyed by trypsin. Judging from the content of naphtol yellow S in the compound we can imply that the molecular weight of insulin is about 700. On analysis, the ratio of nitrogen to sulfur was found to be 11:1. This means that in free insulin it would be N:S = 20:1. It seems, at present, that there is but one sulfur atom in the insulin molecule. The chemical nature of insulin is perhaps that of a complicated polypeptide. The data dealing with the recovery of free insulin from the compound, and full analytical results, will appear shortly. Some insulin preparations were supplied to us by the British Drug Houses, Ltd., London, through the kindness of Dr. H. H. Dale.

The nutritive value of the various layers of the wheat and corn kernel

Much work has been done on the relative vitamine content of the several layers in cereal grains. The peripheral layers and the germ are rich in vitamines. There is, however, not so much information available with regard to the relative content of the proteins in the various layers. The present contribution concerns itself with a chemical as well as a biological analysis of such proteins. The milling fractions of wheat and corn 1 were analyzed to determine their content of total protein, gliadin (the portion soluble in 70 per cent alcohol), globulin (the portion soluble in 5 per cent potassium sulfate solution) and glutenin (obtained by subtracting the protein representing globulin and gliadin from the total protein). The methods followed were those outlined in Methods of Analysis of the Assoc. of Official Agricultural Chemists, 1920. At the same time, the biological value of the wheat and corn fractions was determined according to Mitchells method, 2 using rats as the experimental animals. Chemical analysis showed that among the samples of wheat (bran, middlings, red dog, first clear, second clear and patent flour), the gliadin content increases as we go from bran to patent flour; the globulin content is much higher in the bran, standard middlings and the red dog than it is in the three grades of flour; and the glutenin content diminishes as we proceed from the outer layers to what represents the patent flour. The animal experiments, excepting for minor details, were carried out as suggested by Mitchell (see above). The diet consisted of the milling fraction (corresponding to about 1.3 per cent nitrogen), together with 2 per cent cod liver oil, 0.5 cc. aq. sol. of rice polishing (corresponding to 1 gm. of rice polishing), 7 per cent butter, 15 per cent lard, 4 per cent salt mixture (OSborne and Mendels), and enough starch to make a total of 100 per cent.