Emil J. Baumann

Glycosuria. in rabbits following injections of saline extract of anterior pituitary.

A relation of the anterior pituitary to carbohydrate metabolism has often been suspected on the basis of the frequent occurrence of hyperglycemia and glycosuria in acromegaly, and of anatomical changes in the anterior pituitary in diabetes mellitus, but such a connection was not indicated experimentally until Houssay and Biasotti 1 showed that pancreatic diabetes could be prevented in toads by removal of the anterior pituitary. Subcutaneous implantation of anterior pituitary in such depancreatized toads re-established the diabetes. They were unable to show a similar relation in mammals either by gland implantation or by injections of anterior pituitary extracts, although depancreatized and hypophysectomized dogs survived longer and showed milder diabetes than did depancreatized controls 2 . Evans, Meyer, Simpson, and Reichert 3 reported observations in which they produced glycosuria and polyuria in 2 of 4 dogs by daily injections of the growth hormone fraction of anterior pituitary extracts for 8 or 9 months. We have made somewhat similar observations in rabbits. Hyperglycemia, polyuria, glycosuria, and lipemia developed after injecting a saline extract of ox pituitary daily for 6 or more days, in some instances of surprisingly great intensity. It occurred in each of the 4 rabbits used, though varying in degree. The extract was prepared twice weekly from fresh chilled glands, according to the method of Schockaert 4 except for the measures used in sterilization. It produced typical reactions among which were rapid hypertrophy of the mammary glands with lactation, luteinization of the ovaries, hypertrophy of the thyroid and suprarenal cortex. The rabbits were kept on our stock diet of alfalfa hay and oats. Two were injected with 2 cc. and 2 with 4 cc. daily, equivalent to one-half and one anterior pituitary respectively.

Effect of ascorbic acid on thyroid and suprarenals of guinea pigs.

In previous publications from this laboratory1-5 it has been shown that fresh plant and fruit juices contain an antigoitrous agent when tested on immature rabbits maintained on a goitrogenic diet of alfalfa hay and oats. (The goitrogenic effect of this diet is greatly enhanced by cyanide.) It was shown that this antigoitrous effect was in general inversely proportional to the iodine reducing capacity and that treating the fresh juice with copper salts destroyed or lessened this antigoitrous effect. Our data further indicated that the antigoitrous agent was probably identical with ascorbic acid but because of the lack of crystalline ascorbic acid on the one hand and the presence of iodine (also an antigoitrous agent), on the other, in all these extracts we were unable to prove it. About 2 years ago one of us 6 simplified Szent-Györgyis and Waugh and Kings method of extracting ascorbic acid and by utilizing iris leaves, which are extremely rich in ascorbic acid, has prepared sufficient quantities of this hormone in crystalline form for these experiments, which were carried out as follows: Young guinea pigs weighing between 150 and 250 gm. were placed on a diet of alfalfa hay and rolled oats. (This diet produces scurvy in from 15 to 20 days. Pigs maintained on this diet when injected with the thyrotropic factor develop more marked exophthalmos and thyroid hyperplasia and more quickly than when given greens daily in addition.) They were injected with 1/2 to 2 cc. of ox anterior pituitary extract prepared after Loeb and Bassetts method and given 25 to 100 mg. of crystalline ascorbic acid by mouth daily. The pigs were sacrificed at various intervals and the principal data of 22 are given in Table I.

Behavior of the Thyroid toward Elements of the Seventh Periodic Group. I. Halogens and Thiocyanates.

Summary 1. Thyroid filters preferentially and accumulates thiocyanates as well as all the elements of the seventh periodic group that have been studied, viz., Cl, Br, I, Mn, At. 2. All halides and thiocyanates when administered in relatively large amounts compared with iodine wash inorganic iodide from the body and prevent new supplies of iodine from being received by the thyroid, thus producing thyroid hyperplasia and goiter. It is suggested that these ions exert, at least in part, a mass action-like effect. 3. It is further suggested that similarity in arrangement of electrons in the outer shell of the elements of the seventh periodic group enables the thyroid to differentiate between them and other elements.

Behavior of Thyroid Toward Elements of the Seventh Periodic Group. II. Rhenium.

Summary Rhenium is preferentially and actively filtered from the blood by the thyroid. After injecting a single dose, the Re∗ concentration in that organ is 25 to 100 or more times greater than that found in any other tissue. These findings therefore add support to our hypothesis that the thyroid will selectively filter from the blood all the elements of the seventh periodic group. The greatest concentration of Re∗ in tissues occurs within one hour. For several hours it decreases slowly and then more rapidly so that more than 90% is recovered from the urine in 24 hours.

The Active Fraction of Rous Chicken Sarcoma

Recent attempts to isolate the active agent of the Rous chicken sarcoma have resulted in conflicting views as to the nature of the causative agent. Jobling and Sproul 1 - 2 have reported that repeated injections of a lipid fraction obtained from fresh or dried tumor tissue, together with nonspecific protein, produce sarcomas which appear in the more responsive birds within 3 to 4 months after the third injection. Fraenkel and Mawson, 3 on the other hand, were unable to obtain tumors by injecting acetone extracts of fresh or dried tumor tissue but were successful with the residue. In 1936 we attempted to isolate lipid fractions from a very active preparation of Rous chicken tumor No. 1 with pentane and chloroform as solvents. With neither were we successful in producing tumors in young birds. The residue of the pentane extract, however, invariably produced tumors after a single injection in spite of 4 extractions of the powder with 100 volumes of solvent (see Table I). The residue from the chloroform extract was inactive, possibly due to denaturing the protein. The whole powder, used as a control, was very active; birds had to be sacrificed 12 days after injection. We then believed that the active fraction was not to be found among the lipids; that it was water soluble and very likely protein in nature. In view of the recent report of Jobling and Sproul, 2 we repeated our experiments with a portion of the powder previously used. It was kept at about 5°C. for 11 months in a sealed but not evacuated tube. Pentane was used for extraction in one series of experiments; a mixture of equal volumes of chloroform and pentane in another. In each case 0.5 gm. powder was extracted 4 times with 100 volumes of solvent, taking measures that would tend to avoid denaturing the protein.

Ascorbic Acid from Iris and Other Plants by a Simplified Method.

We have had great difficulty in obtaining more than a few milligrams of the antiscorbutic vitamin, ascorbic acid, from either plant sources or suprarenal glands by the published methods of Szent-Györgyi 1 or of Waugh and King 2 and Svirbely and King. 3 Large and uncontrolled losses occur at many places in both processes as published. Svirbely and Szent-Györgyi 4 have just reported a somewhat modified procedure, using the Hungarian sweet pepper from which paprika is made, which contains considerably more vitamin C than we have obtained from the American sweet pepper—another variety of Capsicum annuum. We have studied the causes of these losses with a view to developing a more certain method of preparation. A survey of common plants was also made to find richer sources of the vitamin. Some of these results have been published by Marine, Baumann and Webster. 5 Leaves of German iris (Iris germanica) are by far the most satisfactory material from which to extract vitamin C, both because the vitamin is present in high concentration and because there are fewer substances in this plant that interfere with its separation. Most plants have a relatively high concentration of ascorbic acid in the young actively growing portions but on maturation, the vitamin rapidly disappears. In German iris leaves, however, the concentration of the vitamin, which is 600 mg. per 100 gm. of fresh leaf in early spring, remains as high as 250 or 300 mg. per 100 gm. in late summer, and this plant offers a very rich source during the entire growing season. Skunk cabbage (Symplocarpus foetidus) gathered during April and May and sweet green peppers as purchased from January to May both contained between 100 and 200 mg. per 100 gm.