Abstract: SummaryIn minced and washed frog muscle incubated in phosphate buffer, added adenylic acid transfers inorganic phosphate to carbohydrate resulting in the formation of hexosemonophosphate. The first phosphorylation product proved to be a new ester which was isolated as the crystalline brucin salt and had the properties of glucose-1-phosphoric acid; when added to frog muscle extract it was converted in a few minutes to the Embden ester.

Abstract: We previously presented data as to the quantity of reduced ascorbic acid in blood plasma and showed its dependence upon the dietary supply of vitamin C, or of ascorbic acid administered as a medicament.1 The data were obtained by deproteinizing plasma by means of tungstic acid and titrating immediately the reduced ascorbic acid present with 2:6 dichlorobenzenoneindophenol.In working with infants, and when making repeated observations on the same individual, it is desirable to have a method which requires a minimum of blood. We have, therefore, developed a micro-method requiring but 0.3 ml. blood, which may readily be secured from the finger of an adult, or the heel of an infant. In the guinea pig, blood is obtained by cardiac puncture, or sometimes by incision of the marginal vein of the ear. In the latter case, a special blood pipette is required.Collection of Blood. Collect approximately 0.3 ml. blood from a lancet wound, into a small phial (Fig. 1, B) containing sufficient powdered potassium oxalate to...

Abstract: A physiologically inactive solid alcohol was prepared from human pregnancy urine by Marrian1 in 1929. This substance was identified by Butenandt2 in 1930 and was called by him pregnandiol. It was insoluble in water and showed a melting point of 233-235°C. (uncorrected). Recently O'Dell and Marrian3 have obtained evidence for the existence of an acid-hydrolyzable form of pregnandiol.We have been able to isolate a water-soluble complex of pregnandiol from pregnancy urine. The method of preparation was as follows: Pregnancy urine (9th month) was extracted with butyl alcohol. The extract was evaporated to dryness under reduced pressure. The residue was taken up in N/2 NaOH and re-extracted with butyl alcohol, the butyl alcohol fraction washed twice with water and evaporated to dryness. The residue was dissolved in a minimal amount of water and the substance was precipitated with acetone. This was collected by centrifuging and purified by crystallization from hot water and several times from ethyl alcohol.The ...

Abstract: Two cultures of Clostridium botulinum were sent to this laboratory in March, 1936, by Dr. L. Bier of the Bacteriologic Institute at Dniepropetrowsk, Ukraina, U.S.S.R. Toxin-neutralization tests had suggested a new type. The original cultures were toxic for mice in doses of 0.001 cc. and antitoxin of types A, B, C and D failed to protect against the the toxin. The organism is a Gram positive, motile, granular, pleomorphic rod and forms oval, subterminal spores which swell the rods very slightly. In liver-agar shake-cultures small, disc-like colonies with or without small polar fluffs are formed. Colonies on the surface of glucose blood plates are non-hemolytic, greyish, translucent, smooth and flat with a marked tendency toward confluence. These cultures are not ovolytic and do not liquefy coagulated egg-white even after a month's growth. In beef heart medium there is slight reddening of the meat and a large volume of gas is evolved. Brain is not blackened or digested. In milk, slight acid is produced but the casein is not attacked. Slight liquefaction of gelatin occurred after 23 days. The peptolytic properties are extremely low and the Sörenson figure was 2.00 after 21 days' growth. Dextrose, levulose, maltose, sucrose, arabinose, xylose and adonite are fermented with little or no gas production in a medium composed of 0.3% Liebig's extract, 0.5% Difco peptone, and 1% carbohydrate. Lactose, rhamnose, galactose, dextrin, raffinose, glycerin, salicin, mannite, inulin and dulcite are not fermented. The thermal resistance of the spores is extremely low. Spores were destroyed in a suspension in buffer solution (pH 7.4) containing 5 million spores per cc. after heating at 100°C. for 2 minutes or 80° C. for 6 minutes. A suspension containing 50 million spores per cc. failed to show growth after 5 minutes at 100°C. or after 40 minutes at 80°C.

Abstract: The study of mutual interactions between developing fish and amphibian tissue was tested by implanting parts of fish blastulae into the blastocœle of developing amphibians. This method applies also to the study of the capacity for differentiation of isolated embryonic cells of the teleostean. Halves of blastulae of the eggs of Danio rerio (the Zebra fish), isolated from their yolk an hour before the commencement of the visible processes of gastrulation, were implanted into the blastocœle of Triturus torosus eggs. The hosts were fixed after 10 days of development.The developing fish and amphibian cells are mutually compatible during gastrulation and the stages immediately following. Thus developing tissue of young embryos from different classes of vertebrates at first possess considerable tolerance, even though a marked tissue specificity characterizes the adults. In the present series of experiments, the implanted fish material developed wherever located: in the center of the yolk, between yolk and epidermi...

Abstract: Experimental production of exophthalmos has been accomplished repeatedly by stimulating the sympathetic innervation of the eye. MacCallum and Cornell,1 Code2 produced, by electrical stimulation of the cervical sympathetic ganglion of dogs and cats, a contraction of the smooth muscle elements in the orbit which caused exophthalmos. Schockaert,3 Loeb,4 Friedgood,5 and Marine6,7 have reported exophthalmos resulting from the injection of thyrotropic anterior pituitary extracts into ducks, normal and thyroidectomized guinea pigs, and by the injection of methylcyanide into normal and thyroidectomized rabbits. Loeb and Schockaert noted that the exophthalmos produced was functional, i. e., did not persist during anesthesia or after death. Marine showed that the protrusion resulting from injection of either pituitary or cyanide was accomplished through a nervous mechanism, presumably upon the unstriated orbital muscle of Muller, and that it could be prevented by removing the cervical sympathetic ganglion.In the fo...

Abstract: The importance of light as a regulating factor of the sexual cycle in birds has been demonstrated by Rowan1 (junco), Bissonnette2 (European starling), Miyazaki3 (Zosterops palpebrosa) and Kirschbaum4 (English sparrow). These investigators have observed a precocious development of the testes following daily increases in the period of illumination. Such experiments were usually carried out during the early winter. The present author has undertaken to determine whether lengthened daily periods are equally effective at a time further removed from the normal breeding season. In this paper, a group of male sparrows, exposed to additional light beginning November 18th, is compared with another group first exposed on September 30th.The November group of sparrows had been in the laboratory 16 days previous to the artificial lengthening of the days and all possessed the horn-brown bill of the sexually quiescent period. The birds received 4 1/2 additional hours of light, daily, from a 100-watt incandescent bulb thro...

Abstract: The current opinion of many investigators is that there is no unequivocal evidence that the virus of poliomyelitis has as yet been successfully cultivated outside the body. The recent demonstration that the virus is of very minute size (8 to 12 mμ1; 12 to 17 mμ2) emphasized the improbability that certain minute, visible microorganisms, which have been cultivated from poliomyelitic tissue, are etiologically related to the disease, while a critical analysis of presumably successful multiplication in chick embryo media3, 4 indicates that insufficient care was taken to rule out contamination with other viruses. Investigations by workers at this Institute and elsewhere are in accord that no propagation of the virus has been demonstrated by methods which have proved successful for the cultivation of most of the other viruses.5The problem of cultivating the virus of poliomyelitis is being pursued not only for the further elucidation of the nature of the virus but also in the hope that successful in vitro propaga...

Abstract: In continuation of our experiments on sulfur metabolism, the behavior of carboxy-methyl-S-cysteine1 (I) was investigated. (We are indebted to Dr. L. Michaelis for suggesting this compound for study.) Carboxyl-methyl-S-cysteine, like methyl-S-cysteine2 does not support the growth of rats on a sulfur deficient diet, indicating that (I) does not yield cysteine (IV) in the course of its metabolism.Three and six-tenths grams of (I) were administered to a normal human being, while 7.2 and 14.4 gm. respectively were given to 2 cystinurics. The substance was not toxic and yielded in the urine neither cystine nor -SH compounds. In the normal, the sulfur of (I) was only partially oxidized (40%), the larger portion (60%) being excreted as undetermined neutral S. Part of this neutral S was apparently a disulfide, since there appeared in the urine a strong cyanide-nitroprusside reaction, while the Sullivan test remained negative. The urine was discarded before the probable nature of this disulfide was realized.Followi...

Abstract: It has been shown by the author1 that sea-water inhibits the multiplication of freshwater bacteria while marine bacteria require sea-water media for their initial isolation, although these differences rapidly disappear following the laboratory cultivation of the bacteria. Similar observations on the selective bacteriostatic action of sea-water have been reported by Berkeley,2 Lipman,3 and Korinek.4It is generally known that intestinal and other bacteria of public health interest do not survive very long when emptied directly into the sea unless there is appreciable organic matter present or considerable freshwater dilution. Colon bacilli are rarely recovered from the open sea and they occur far less frequently in the vicinity of sewage effluents than can be accounted for by dilution or oceanic circulation. The failure of freshwater bacteria to survive in the sea has been attributed by various investigators to predacious protozoa, lack of nutrients, bacteriophage,5 or to the lethal effect of sunlight. The ...

Abstract: SummaryDaily administration of 4 mg. of synthetic progesterone to iiormally cyclic iemale rats causes immediate cessation of cycles, without vaginal mucification. There was ovarian atrophy, hypertrophy of the pituitary, and slight atrophy of the thymus. It has no effect n-hatsoever upon the condition of the mammary gland. Since cestrin is also without any direct effect upon the mammary gland (as shown in hypophysectomized animals) the effect of ovarian hormones on the mamma must be largely dependent upon the presence of pituitary secretions.

Abstract: ConclusionsIt has been found that single fowl pituitaries implanted over the crop glands of 8- to 10-weeks-old pigeons (age computed from time of conception) will, in many instances, cause a prolactin-like reaction. It is assumed that this reaction is caused by prolactin in the pituitaries of the donating fowls.Pituitaries from broody hens cause a greater reaction of the pigeon crop gland than the pituitaries of laying hens. Pituitaries from males cause a reaction about equal to, or slightly less, than the pituitaries from laying hens.It was noted in a few cases that the pituitaries of hens just becoming broody gave a greater reaction than those of hens nearly over their broody period. More data are being gathered on this point.

Abstract: Numerous investigations on mammary gland development in recent years have indicated that the ovarian hormones, estrone and progestin, are responsible for the growth of the glands. However, since increasing knowledge of the physiology of the pituitary gland seems to give it a central position in the endocrine system, investigators have become interested in determining whether or not the mammary glands could be experimentally developed in hypophysectomized animals.Pencharz and Long1 who hypophysectomized rats in late pregnancy (20th and 21st days) concluded that mammary development continued in the absence of the pituitary. However, at this stage of pregnancy growth is complete and lactation is being initiated. In fact, Selye, Collip and Thomson2 observed that milk secretion always set in normally after parturition, but stops after a few hours, in rats hypophysectomized during the latter half of pregnancy. The mammary glands of rats hypophysectomized during the second half of pregnancy have been studied cyt...

Abstract: The experiments of DuShane1, 2 have proved conclusively that the dermal melanophores of the amphibia originate in the neural crest, as suggested by Harrison,3 and later by Weidenreich4 and Holtfreter.5 The differentiation of large numbers of typical pigment cells in tissue cultures of neural crest taken from embryos of pigmented fowls indicates the probability of a similar source of origin in the chick.Embryos of various breeds have been used, ranging in stage between 6 and 10 somites. Thin strips of the rising folds anterior to the first somites were removed with fine scissors and explanted in a medium consisting of equal parts of plasma and embryonic extract. For controls, whole embryos minus the neural crest were cut into small pieces and explanted on a single slide; or specific areas, such as the eye, the brain floor with underlying endoderm, and the ectoderm and mesoderm lateral to the crest region, were explanted separately. Control cultures never contained more than one or 2 isolated pigment cells,...

Abstract: The fact that derivatives of the male hormone may exert an action on the female sex organs is known from the work of Butenandt and Hanisch,1 who showed that androstenedione which is inactive in castrate females produces vaginal oestrus in the presence of an ovary, and from the work of Parkes,2 who found that the same is true in the case of androstanediol. The action of these compounds might possibly be explained by a transformation of these substances into oestrin by the ovary. This would seem all the more likely since Collip, Browne and Thomson3, 4, 5 have shown that the ether-insoluble fraction of oestrin extracted from pregnancy urine (Emmenin) is relatively inactive in the spayed female, while it is much more active in the presence of an ovary. Several compounds of the androsterone-testosterone series, however, do also cause vaginal cornification in ovariectomized female rats, as shown recently by Deanesly and Parkes6 and Korenchevsky and coworkers,7 and it is still to be seen whether this is due to a...

Abstract: Recent experiments by Hohlweg and Junkmann,1 Haterius, Schweizer and Charipper,2 and Hill and Gardner3 have indicated quite conclusively that pituitary tissue will function as a graft.It is the purpose of this report to present summarily a series of experiments in which the empty sellae of hypophysectomized 28-day-old male and female rats were utilized immediately after the operation as sites for grafting pituitary tissue. The types of grafts included: 1. male hosts bearing (a) their own pituitaries as grafts and (b) the pituitaries of 28-day-old female rats; 2. female hosts bearing (a) their own pituitaries as grafts (b) the pituitaries of other 28-day-old female rats (c) the pituitaries of 1 to 5 newborn male rats; (d) the pituitaries of male rats 20 to 40 days old. Hosts and donors were taken from an intensively inbred strain of rats. In a group of 37 such hosts (5 males and 32 females) which were observed for 14 weeks, 73% takes occurred as judged by marked body growth and attainment of a functional s...

Abstract: By the combination of 3 different methods the analysis of bile for taurocholic acid, glycocholic acid, total conjugated bile acids, cholic acid, deoxycholic acid, total bile acids, and free bile acids has been achieved. It has also been possible to analyze duodenal drainage material for cholic acid, deoxycholic acid and total bile acids. Methods. 1. The Cuny 1 modification of the Schmidt-Dart 2 method is used to determine the taurine and glycine nitrogen percentage. The first is multiplied by 33.87 and the second by 28.29 to give the bile acids conjugated with taurine and with glycine. The sum of these 2 figures gives the total bile acids that are conjugated. 2. Cholic acid is determined by a method outlined in the previous paper. 3 3. The total bile acids are estimated by an iron precipitation method based partly on principles devised by Harwood. 4 After the bile acids have been extracted and made up to 100 cc. in a volumetric flask and the cholic acid determined, the amount of solution in the volumetric flask, estimated to contain between 40-60 mg. of bile acids, is measured out into a 150 cc. beaker. This calculation is derived from the known amount of cholic acid present in the volumetric flask. The bile acids of dog bile may be considered to be approximately 100% cholic acid, while those of human bile can be assumed to be 50% cholic acid. Thus if the volumetric flask is found to contain 100 mg. cholic acid derived from dog bile, 50 cc. of the solution is measured in the beaker. If the extract is derived from human bile, only 25 cc. is taken.

Abstract: Summary Oral papillomatosis is common in domestic rabbits bred in the New York area. The disease is due to an hitherto undescribed virus which is evidently pathogenic for the oral mucous membrane only.

Abstract: It has been shown by Wigglesworth1 that in Rhodnius prolixus the corpora allata secrete a hormone which is necessary for the production of eggs. In order to determine whether these glands have a similar function in the grasshopper, females were operated as shown in Table I. For comparison, unoperated adults were killed at intervals from one to 42 days after moulting. Camera lucida drawings were made of the largest eggs in each of the above animals, egg length was measured, and the oviducts were examined to determine whether the secretion produced by the anterior portions of the oviducts was present. This secretion forms the ootheca.There was great variation in the development of the ovary in unoperated adults autopsied from 28 to 42 days after moulting. However, of the 43 specimens examined, 35 contained eggs which were more than 1 mm. in length. Of these, the eggs of 8 were fully developed and had passed into the oviducts. The operated controls at autopsy also contained eggs exceeding 1 mm. in length and...

Abstract: A number of observers have found that extracts from the kidneys of various animals may have pressor effects.1–7 The object of the present investigation was to determine whether extracts of kidneys from human beings and dogs with hypertension had greater pressor effects than those of control kidneys.Kidneys were obtained at autopsy from 15 patients with hypertension and from 17 control subjects. The hypertensive group consisted of 9 patients with benign hypertension, 2 with malignant hypertension, 2 with chronic glomerular nephritis, and 2 with pyelonephritis and secondary contracted kidneys. The material was put through a meat grinder, crushed in a mortar, shaken for 30 minutes with saline (one cc. per gm.), filtered and kept on ice. The tests were most satisfactorily performed on an unanesthetized dog with a Van Leersum (carotid) loop.8 In several instances records were obtained by direct cannulation of the carotid artery of anesthetized dogs and cats. Injections were made intravenously, using the equivl...

Abstract: The determination of the threshold of vibratory acuity is a valuable part of the neurological examination, particularly in the diagnosis of lesions affecting the posterior columns of the spinal cord. There have been numerous studies in which an attempt has been made to obtain a quantitative measure of acuity. The tests have all been made with a tuning fork of some description, usually with some device designed to measure the intensity of the vibration, an end which, in the opinion of the workers themselves, was not satisfactorily attained. We have, therefore, constructed an instrument which produces a vibration of constant frequency, the intensity of which can be varied over a considerable range, and measured accurately. It consists of an iron pole-piece which constitutes the vibrating member, and to which is attached a round, metal button 12.5 mm. in diameter which is placed over the bony prominence to be tested. This pole-piece is actuated by an electromagnet energized by an alternating current of 60 cycles per second. By means of a potentiometer this current may be varied, and its strength measured by means of an alternating current milliameter in series with the magnet winding. Thus the amplitude of vibration, roughly proportional to the current flowing through the electromagnet, is subject to control and measurement. Since the intensity of vibration bears no absolute relation to the amperage, depending on the design of the coil in the particular electromagnet, this intensity is expressed in arbitrary units from 0 to 10. Each individual instrument should, therefore, be calibrated to correspond to the original one used as a standard. An even pressure is maintained by allowing the instrument to rest by its own weight over the bony prominence, the operator merely supporting the end of the handle.

Abstract: ConclusionsIn the regulation of the ovarian activity of rats prolactin is shown to be capable of playing a part, directly or indirectly. Prolactin can not be summarily omitted from lists of pituitary principles having action on the gonads of mammals. The suggestion of Dresel that lactation anestrus in mice is induced by prolactin is supported by these tests on mature non-parous rats. But under effective prolactin dosage the corpora of rats remain large, and at the age of 10-12 days their large cells show no clear evidence of degeneration. The failure of daily injections of low or moderate doses of progesterone to affect the estrous cycles of rats suggests that in this rodent, though the corpus luteum hormone plays its part, it is perhaps not the hormone primarily responsible for the anestrous of either gestation or lactation.

Abstract: A rise in blood pressure may be produced in dogs either by partial obstruction of the renal arteries1 or by ligation of the ureters.2 Since the latter procedure often causes a marked decline in the renal blood flow,3 it seems likely that the hypertension is in both instances related in some way to ischemia of the kidneys. It was shown by Tigerstedt and Bergman4 that saline extracts of the kidney of rabbits produced a sustained rise in blood pressure when injected into other rabbits. The object of our experiments was to determine whether extracts prepared from ischemic kidneys, removed from dogs with hypertension, had a greater pressor effect than similar extracts of normal kidneys.The freshly removed kidneys were chopped up with scissors and ground with carborundum and 0.9% salt solution. The suspension so obtained was centrifuged and the supernatant fluid was kept in the ice box until used. Amounts corresponding to 5 gm. of kidney tissue were administered intravenously to normal unanesthetized dogs. Chan...

Abstract: For a number of years 1 it has been known that tendon gives excellent crystalline reflections. Recently 2 large spacings have been observed which provide strong evidence for the view that collagenous structures may be fibrous arrays of true crystals composed of very large molecules. Most forms of tendon from adult animals cannot be put into solution without obvious chemical disintegration. The work of Nageotte and of others 3 has, however, demonstrated that optically active fibrous precipitates can be obtained from weakly acid solutions made from the tail tendon of adult rats and from many connective tissues of foetal animals. The optical appearance of these precipitates suggests that they are at least similar to the tissue from which they were dissolved. We have carried out a series of experiments to determine: (a) whether reprecipitated tendon is really crystalline and (b) whether its X-ray diffraction pattern is like that of the original tissue. Reprecipitated connective tissue was obtained according to the procedures outlined by Nageotte. Pieces of freshly excised tail tendon from adult rats were placed in dilute (1 to 25,000) acetic acid. In such acid the tendon swells and a portion of it dissolves to form a clear but highly viscous solution. The reprecipitated tissue is obtained from this solution by salting out with 5% NaCl. Series of X-ray diffraction studies have been made of a number of these preparations. Some of these diffraction photographs were made with the usual short crystal-to-film distances; others were made with very small slit systems and with the large camera radii needed to record spacings of the order of 100 A.

Abstract: SummaryThe blood plasma ascorbic acid (reduced form) in active cases of rheumatoid arthritis is regularly low if the individuals have not been maintained on a high vitamin C supplement. Unadvised cases were found to show uniformly lowered levels. The reduction is striking. Such levels rise in response to extra supplements of vitamin C. In many this rise is refractory. Our studies indicate that in some cases the intake required to maintain adequate vitamin C levels in the plasma are much above the average requirement for normal individuals. The mechanism involved is unexplained.

Abstract: The active fraction obtained from B. coli filtrate by the method described in the preceding paper was dissolved in water and evaporated to dryness in vacuo. A white residue was obtained, largely crystalline in nature. Strong heating produced some charring, but much of the crystalline residue was unaffected indicating that a considerable portion was inorganic in nature.On repeated treatment of the active fraction with methyl alcohol, most of it (82%) went into solution. The methyl alcohol solution was evaporated to dryness and taken up in water, but most of it now failed to dissolve. However, addition of sodium hydroxide effected solution. Acidification with hydrochloric acid produced a heavy, flocculent precipitate which dissolved again on further addition of alkali. This solution was neutralized and tested on tumor-bearing mice; it was completely inactive as far as production of hemorrhage was concerned.The methyl alcohol-insoluble portion (18%) dissolved readily in a small volume of water, with formatio...

Abstract: Avery and Cullen1 obtained from both rough and smooth pneumococci a ferment capable of lysing heat-killed pneumococci. In unpublished experiments (Dubos) it was found that the same ferment preparation renders ineffective in rabbits the capsular type specific antigen of killed cells of virulent pneumococci.A hydrolytic action of the ferment preparation of pneumococcus on two tissue polysaccharides is described here. This action seems to be identical with that of the same ferment on the pneumococci.From bovine vitreous humor and from human umbilical cord two polysaccharides have been obtained in pure form, seemingly identical in composition, rotation, and general physical behavior.2 They are composed of acetylglucosamine and glucuronic acid, the components having been shown to be present in equimolar ratio by isolation or quantitative analysis.The 2 polysaccharide acids yielded reducing sugar following hydrolysis by the ferment obtained from pneumococcus autolysates. The hydrolysis is optimal between pH 5 a...

Abstract: Three cases of yellow atrophy of the liver have occurred in each of 3 widely separated plants and under different management within a year or two. In each instance where the illness appeared, the men were exposed to a chlorinated naphthalene heated above the melting point and giving off fumes. Yellow atrophy of the liver is a rare disease and there have been no cases reported where chlorinated naphthalene was the proven etiological agent.The following experiments were conducted to determine if some of the chlorinated naphthalenes might produce such lesions. Three naphthalenes obtained from one of the above factories were used in this work. They are designated then as A, B, and C.Compound A is a mixture of tri- and tetra-chlor-naphthalene. Compound B is a mixture of tetra- and penta-chlor-naphthalene and may contain some tri-chlor-naphthalene. Compound C is a mixture of penta- and hexa-chlor-naphthalene which is plasticized with a relatively small percentage of asphalt.Groups of 5 rabbits weighing approxim...

Abstract: The mechanism by which choline chloride influences the deposition of fat in the liver is by no means clear. Best, Channon and others have shown that this nitrogenous component of lecithin is capable of preventing or curing the fatty infiltration of the liver which is produced in the rat on a high fat diet or in the dog on a lean meat-sucrose diet. The finding that diets essentially free of choline cause an infiltration of fat into the livers of rats, a condition curable by the addition of choline chloride to the diets, led Best, et al., to suggest that choline may be an essential dietary factor.The most obvious hypothesis as regards the manner in which choline produces its “lipotropic” effect would involve perhaps the formation of lecithins and other choline-containing phospholipids from fat, phosphate and ingested choline, thus favoring the transport of lipid materials. There are obstacles to the immediate acceptance of such a view, of possible significance among which is the lipotropic inactivity of ami...