Experimental Biology and Medicine

It appears possible to correlate the urinary excretion of vitamin B₁ with the state of vitamin B₁ nutrition in man using a thiochrome method for determining the amount of vitamin B₁ in urine.

The usual difficulties of quantitative plethysmography are multiplied when the method is applied to the intestine by the occurrence of intestinal contractions which may compress gases trapped within the lumen of the gut, even though the ends of the loop are open, and which, if strong enough, may pull mesentery into the chamber. These difficulties are obviated by the substitution of a simple gravimetric method, which permits, as an added advantage, simultaneous recording of intestinal motility.

A loop 6 to 10 cm in length, whose mesentery is long, is separated by incisions which are converged to the root of the mesentery, care being taken to preserve the loop's blood supply. With the dog lying supine, the loop and its prepared pedicle are lifted over a horizontal rod held in a clamp as close as possible to the root of the pedicle, and the pedicle is sutured to the rod (Fig. 1, a). The loop is threaded upon a light supporting rod (b), and the ends securely fastened. The projecting ends of the supporting rod are attached by means of threads to the horizontal beam of a spring balance (c). The tension of the spring and the leverage are adjusted until the loop on its rod, and the entire pedicle are swung clear of the animal. The loop should lie approximately in the same horizontal plane as the rod which supports the pedicle at the root.

It has been found simpler to record the movements of the horizontal beam indirectly, through a second light magnifying lever (e) as this permits adjustment of leverages without disturbing simultaneous ordinates on the record. If long loops are taken, strong contractions may exert pull on the pedicle, and so interfere with the record.

1. Morphine sulfate added to blood in vitro produces no consistent change in choline-esterase activity. 2. Serum choline-esterase activity in the dog is consistently lowered following the injection of morphine sulfate. 3. The in vivo effect of morphine on serum choline-esterase furnishes another indication that morphine acts through a cholinergic mechanism.

Adult female starlings in the quiescent season are a favorable material for the study of the biological properties of natural and synthetic sex hormones. Their ovaries and oviducts are capable of increasing in weight 100 to 150 times and the wolffian ducts 10 to 20 times. The bill color and the wolffian ducts react only on stimulation by androgens while the oviducts react in an identical way on either male or female hormones. Stilbestrol produces purely gynogenic effects and is considerably more potent than equal doses of estrone. Progesterone even in high doses or in combination with estrone gives no effects; it does not seem to be one of the hormones that take a part, normally, in the endocrine physiology of passerine birds.

It has been shown that the acute toxicity of sodium sulfamethylthiazole in rats, as measured by intraperitoneal injection, is almost twice as high as that of sodium sulfathiazole, which in turn is slightly greater than the toxicity of sodium sulfapyridine.

The pH and buffer capacities of solutions of the sodium salts, and differences in solubility of the free compounds as factors influencing the toxicity, are discussed.

The depression of the Brunn Reaction in winter months reported by Boyd, Mack and Smith 1 was found accompanied by complete failure of pituitary (posterior lobe) extract to inhibit loss of normal body water in frogs. On the other hand, the loss of water injected into frogs was inhibited by pituitary (posterior lobe) extract in the winter though to an average extent less than in the summer.

Experiments conducted at the Hopkins Marine Station, Pacific Grove, California, during the summer of 1938 on the unfertilized and fertilized eggs of Dendraster excentricus have given certain interesting modifications in the early developmental patterns of this form.

Unfertilized eggs treated for 12 hours with a solution of NaCNS (20 cc .54 M NaCNS plus 100 cc Ca-free sea water) showed a retardation in cleavage rate when returned to normal sea water and fertilized. The developing larvae ranged from forms showing considerable inhibition (increased mesenchyme, shortened anal arms, undifferentiated oral lobe) to larvae (approximately 35%) which consisted entirely of ectoderm and ectodermal structures. These ectodermized forms were modifications in the same direction as the ‘animalized’ forms produced in the sea urchin by Lindahl 1 and Hörstadius 2 with NaCNS treatment and experimental isolation of animal blastomeres. Neither mesenchyme, pigment nor gut was present. With higher concentrations or exposure periods the number of ectodermized forms was decreased and the number of inhibited forms increased. When NaCNS was used in normal sea water only inhibited larvae resulted. Ca-free sea water alone had little effect.

Fertilized eggs treated with NaCNS (20-30 cc .54 M NaCNS plus 100 cc normal sea water) for 12-24 hours appeared as thick-walled radially symmetrical blastulae usually containing a surplus of mesenchyme and showing a loss of cells from the vegetal region. When these forms were returned to normal sea water as many as 95% developed into exogastrulae with large evaginated entoderms, often filled with mesenchyme, and with ectoderm reduced to a small pig-mented knob. Such forms were almost entirely entodermized and have been obtained by other workers (Herbst, 3 Child, 4 Horstadius, 2 etc.) with various methods in this and other forms.

Rats respond to short fasts (2 days) without change in arginase concentration in the liver tissue, and adaptations to changes in metabolism are reflected in decreases in the size of the liver. If fasted until near the moribund condition there is increased arginase concentration in the livers of both sexes, with loss of sex differentiation. Adjustment of the quantities of this enzyme by male rats when fasted 8 days is reflected by change in the size of the liver and by increase in its concentration of the enzyme. Adjustments to long fasts by female rats is accompanied by increase in enzyme concentration and after 4 days of fast the quantities of liver tissue are not further decreased. As shown in Fig. 1 fasts of intermediate duration give variable results in the concentration of the enzyme in the livers of male rats. Such variations may be correlated with the type of body materials, carbohydrates and fats (non-nitrogenous) or proteins (nitrogenous), that serve as primary energy sources at the time of sacrifice. Expressed as quantity of enzyme per unit of body weight, arginase activity changes during the adjustment to the expected utilization of proteins as energy sources.

Recent investigations of electrolytes in Thyone muscle by Steinbach 1 have led him to the conclusion that Ca is present in cells in lower concentration than that of the surrounding medium. Since these results are at variance with our own in a number of respects we have studied again the localization of Ca and Mg in mammalian and frog smooth muscle by the electron microscope as described by Scott and Packer. 2 Naturally, the results obtained cannot be compared directly with those of Steinbach as the same form was not used. Furthermore, our interest lay in determining the localization in smooth muscle cells which had been preserved as nearly as possible in their normal physiological state.

Thin strips of smooth muscle were taken from the duodenum and stomach wall of cats and frogs. These pieces were frozen in liquid air while still actively contracting from the mechanical stimulation of the manipulation. The frozen tissues were dehydrated in vacuo at -63°C and infiltrated in water-free paraffin without breaking the vacuum. This procedure precludes any salt shift during dehydration and the addition of water from outside sources at any time and thus maintains the spatial relations of the element in question. Thin sections (8μ) of muscle were prepared and examined by heating on a surfaced cathode in the electron microscope.

The technic of preparation of tissues for examination and the method of localizing Ca and Mg are believed to preserve the exact topical cellular relationships. Unfortunately, it is impossible by the method used to differentiate Ca and Mg, since both activate the cathode surface. Quantities of these elements in less than 1 × 10^-12 g are easily detected. In no instance was either Ca or Mg found outside the cell wall.

Sulfamethylthiazol apparently exerts a definite protective effect in mice which have been experimentally infected with a virulent strain of Salmonella enteritidis. This protective effect is manifested only by an increase in average survival time of medicated animals. Eventually all of the medicated animals succumb to the Salmonella enteritidis infection.

Astwood has demonstrated that 6 hours after the administration of estrogens to the 21-day-old rat. there is an increase in uterine weight. 1 This weight increase is largely due to increased hydration (edema) of the uterus. Pincus and Graubard have essentially confirmed these findings. 2 Zuckerman has stated that androgens also cause increased hydration of the uterus in the monkey. 3 Astwood in a brief report has stated that androgens will cause a uterine weight increase in rats similar to that caused by estrogens. 4

The Astwood technic as originally proposed for estrogen assays is a very rapid and convenient one. Reports have appeared in which it has been applied to urinary estrogen assays. 5 Inasmuch as androgens are known to be present in the urines of normal, pregnant or castrate women, it seems of interest to report our findings as to the uterine weight-increasing effects of various androgens.

Findings. All animals were given the androgen subcutaneously in 0.1 cc of peanut oil and were killed 6 hours after the treatment. Peanut oil has previously been tested in this laboratory and found to contain neither estrogen nor androgen. The cervico-vaginal junctions were cut, the uteri carefully removed and immediately weighed on a Roller-Smith balance sensitive to 0.02 mg. The tissues were preserved in Bouin's fluid and representative uteri were examined microscopically. Detailed weight-findings are presented in tabular form (Table I).

The animals of Groups 1 and 2 were 21 days old. Group 1 served as controls (A). As can be noted in the table, 0.5 mg testosterone (Group 2) caused a statistically significant increase in uterine weight, 22.8% over that of the controls.

It would appear that in these experiments the intra-peritoneal injection of crude peanut oil extracts of the brown fat of the woodchuck and the thirteen-lined ground squirrel produced a lowering of the heat production of the white rat either at once or after 1 or 2 days (as measured by the oxygen consumption). Such effects, with one exception, were not produced by the injection of peanut oil alone.

Typhus vaccine, consisting of killed rickettsiae from infected rat lungs prepared according to the method of Castaneda, protected guinea pigs against infective doses of virus injected intra-peritoneally. Immunization was accomplished by the subcutaneous injection of the vaccine in as small a dose as 0.2 cc. The virus used in the preparation of the vaccine and for the test for immunity was the “L” strain of Mexican typhus.

Heparin inhibits the action of trypsin and serum tryptase. The significance of this phenomenon in the blood clotting mechanism is briefly discussed.

(1) Contrary to Brakefield, and in agreement with Quick, we find that in humans an appreciable quantity of benzoic acid is excreted as the glucuronide. The ingestion of 5, 6, and 7 g of benzoic acid leads to an excretion of some 5% in the form of its glucuronide. The ingestion of 8 g gives a higher figure (7.7%). (2) The maximum excretion occurs in the first 3 hours. Detoxication by glucuronic acid is complete in 9 to 15 hours, depending on the amount of benzoic acid ingested.

Impairment of the normal movement of the legs frequently occurs in swine. This condition is variously termed lameness, stiffness, posterior paralysis or incoordination, depending on its severity and etiology. A characteristic stiffness occurs in rickets in pigs, a condition in which there is abnormal bone development. Incoordination occurs in pigs as the result of vitamin A deficiency, in which there are degenerative changes in the central nervous system. In addition, however, lameness of one type or another occurs in some instances in which calcium, phosphorus, and vitamins A and D do not appear to be the limiting factors.

In the course of growth studies of pigs conducted by the authors during the past few years, a type of stiffness or lameness has been encountered which appears to be etiologically different from any of the forms described in the literature. This condition occurred in pigs which were fed similar rations, all of which were comprised of yellow corn, tankage, soy bean oil meal, ground alfalfa hay and salt. The pigs were fed individually and indoors but had access to direct sunlight at all times in outdoor paved pens.

The growth of the pigs was good but lameness occurred at about the time they reached a body weight of 150 pounds. The condition was at first characterized by a slightly halting gait, but it slowly progressed in severity until it was only with difficulty that the pigs could rise to their feet. The behavior of the pigs indicated that the condition was painful. Gross manifestations, in addition to stiffness, were enlarged hock joints and crooked legs which occurred even with some pigs which never became stiff. It should be noted that only 30 out of a total of 60 pigs receiving the above treatment were affected with stiffness.

Ascorbic acid furthers the actinic transformation of tyrosine into dopa and that of synephrin into epinephrine but inhibits any further oxidation. It reduces melanin to a water-soluble yellow substance.

We have investigated the excretion of androgens and estrogens in 2 cases of male mammary carcinoma. The results obtained are recorded in Table I.

Twenty-four-hour urine specimens were extracted and fractionated according to the method described by Gallagher, Peterson, Dorf-man, Kenyon, and Koch. 1 Samples were assayed for their estro-genic potency by the vaginal smear technic, using mice. Androgenicf assays were done by a modification of the Zimmerman method as described by Friedgood and Whidden. 2 We endeavored to correct for the inherent color of the samples by subtracting the equivalent of the reading obtained when an aliquot (equal in amount to that used for the actual determination) was treated with all the reagents except m-dinitrobenzene. This method gives values higher than those which would be obtained by biologic assay, but we feel that it is probably acceptable for use in a comparative study.

It will be seen that the male patients with mammary carcinoma did not differ significantly from the controls in regard to the amount of androgens and estrogens excreted. The low excretion of both androgens and estrogens in the aged, found by several previous investigators, and the daily variability in the excretion of androgens reported by Gallagher, et al., 1 are confirmed.

We are indebted to Drs. M. B. Visscher, L. T. Samuels, and L. Earle Arnow for their suggestions and cooperation.

Various quinones with vitamin K activity have been assayed in order to establish the comparative potencies of these compounds in relation to a common reference standard. The reference standard is an hexane extract of dried alfalfa equivalent in potency to 1 g of dried alfalfa per cc. This reference standard at 2 or more levels has been employed in all of our assays for the past 2 years. A secondary reference standard consisting of a sample of dried alfalfa has also been preserved. Neither of these standards has shown any loss of total or relative activity. In periodic assays the potency of the second reference standard has been obtained as 1.09, 1.03, 1.07, and 1.06 cc of the first standard per gram.

Some assays have been conducted as we have already described, 1 that is, by placing the supplement in the diet of depleted chicks and determining the prothrombin clotting time after 7 or more days. More recently, 2 after a depletion period of 10 to 14 days, we have given the supplement orally for 4 days and have determined prothrombin clotting time on the fifth day. Certain supplements have been assayed by both procedures and have given identical results. In a few cases, where evidence of loss of the supplement in the diet was encountered, the oral feeding was relied upon entirely. In all asays, the reference standard was employed at 2 or more levels in order to establish values for interpolation and calculation of potencies in terms of the standard. This may be done by means of the linear relation between the reciprocal of the prothrombin clotting time and the logarithm of the vitamin K dosage. 1 No “master curves,” “response curves” or arbitrarily fixed values are required in our method.

The hibernating glands of woodchucks killed during the summer contained 1 International Unit of androgen, (100 μg androsterone) in 50 g of tissue, equal to the concentration in bull testis, the richest tissue source known. Human male and female axillary fat contained 1 International Unit in 1 kg and 3 kg respectively.

Fresh green plant material is frequently used to supplement standard stock diets in the raising of laboratory animals such as the rat. Recently, additional impetus has been given to the study of young green growing plants by observations on the previously unknown factor (factor pl of Wulzen and Bahrs, 1 grass juice factor of Kohler, et al. 2 , 3 , 4 ) necessary for the normal growth of guinea pigs 1-5 and on the nutritive value of cereal grasses. 6 Such studies would be greatly facilitated if it should be possible to grow young plants in the laboratory under reproducible conditions at any time and in the desired quantities.

A method has been developed in this laboratory for the purpose of adding the grass juice factor to relatively purified breeding diets for nutrition studies on mice which are handled with bacteriological precautions. The following qualities are essential: (a) a product which will provide a potent source of the grass juice factor, (b) uniform seed, (c) a nutrient solution composed of chemically pure ingredients, (d) a planting base which is as inert as possible, and (e) general cleanliness, with emphasis on freedom from insects, vermin and harmful bacteria.

After more than a year of study, the following method has been evolved and has proved satisfactory. It consists essentially of spreading soaked seeds on a layer of cotton placed on a screen which has been suspended over a pan containing nutrient solution. The roots grow down through the cotton. The grass begins to sprout within 24 hours after planting, and reaches a height of about 5 inches by the seventh day (Fig. 1).

The Rammstedt operation decreases the incidence of gastritis, hemorrhagic erosions and ulceration that follows cinchophen administration in dogs. This indicates that pylorospasm plays a significant but not an all inclusive role in the genesis of cinchophen ulcers.

The uptake of radioactive phosphorus was greater in spongy than in dense diaphyseal bone when salts of the element were administered orally to a dog. In dentine the amounts of radioactive phosphorus were constant for the various teeth and of the same order as that present in dense diaphyseal bone. No more than traces of P∗ were found in the enamel.

The administration of testosterone propionate in doses of 75 to 125 mg weekly to 8 hypogonadal adolescent boys ranging in age between 9 and 18 years was accompanied by an increase in average growth rate from 1.36 cm per 100 days during the control period to 3.6 cm per 100 days during the period of therapy. Following the cessation of treatment the average growth rate again fell to 1.56 cm per 100 days. This apparent stimulation of growth rate was unaccompanied by X-ray evidence of epiphyseal closure. The exact mechanism of this growth stimulation is not clear. Since all of these individuals exhibited evidence of hypogonadism, it must be pointed out that, on the basis of these observations, there is no justification for the use of testosterone propionate as a growth stimulant in normal adolescence since such therapy might result in permanent damage to the individual.

By paying special attention to a few fundamental characteristics, significant findings were noted which may have important practical implications. The concentration of the organisms in the cells of the yolk sac which can easily be separated by centrifugation would suggest a simple method for the removal of much undesirable inert materials contained in the yolk sac during the preparation of vaccine. The presence of these substances has been the main objection for employing this vaccine in human prophylaxis. Successful initiation of cultures from the blood of infected animals naturally leads to its possible application for diagnosis in patients, when test animals usually employed such as guinea pigs and white rats were not available. Preliminary studies in this connection have offered some indication as to its practical use. The fact that death is regularly produced in white mice by the yolk sac cultivated rickettsiae gives us an additional and convenient animal which may be employed for typhus studies. By its use, immunological and chemotherapeutic studies can be facilitated and extended.

Following subcutaneous and intramuscular infections of a Vi-phage and a non-specific antityphoid phage in a urinary typhoid carrier, only the Vi-phage appeared in the urine in amounts greater than those administered. The numbers of B. typhosus in the urine were transiently decreased as the result of phage-treatment but increased again considerably with the appearance of W-form and lysogenic V-forms in addition to the typical V-forms which had been persistently present. The presence of W- and lysogenic V-forms was attributed to the action of V-phage in the bladder-urine and the presence of the V-form organisms due to the intermittent discharge of organisms from a focus in which the action of the Vi-phage was probably inhibited. At the same time specific antibodies against both phages were developed in the blood. The type-specificity of the phage and bacteriophagic typing of the Vi-form organisms repeatedly isolated from the urine remained constant.

A phosphate buffer is described which has been found to maintain the skin-reactivity of dilute purified tuberculin for at least 4 months at 20 C and 6 months at 8°C.

Hamsters and white mice are suitable animals for testing virulence of Streptococcus hemolyticus when injected by the intracerebral route. It requires only 40-400 organisms to kill the animal when given intracerebrally, whereas it requires 400,000 when given interpleurally and 4,000.000-11,500,000 when given intraabdominally and intravenously.

In a case of glycogen disease, whose tolerance to creatine is known to be minimal, saturation with vitamin C caused a marked increase in the urinary excretion of creatine and creatinine. In 4 normal children and in a child with hypothyroidism, however, no significant increase was demonstrated. This is probably due to the fact that whatever increase in creatine formation due to the ingestion of ascorbic acid is covered by the normal capacity of storage. The mechanism of the action of vitamin C on creatine metabolism is not understood.

These observations on the rhythmic precipitation of pneumococcal antigen-antibody reactions suggest that the phenomenon is comparable to that occurring with crystalloids in a gel.

In earlier studies it has been demonstrated that deuterium may be retained on the betahydroxybutyrate molecule excreted after feeding deuterobutyric acid to rats. 1 When the α,β-dideutero acid was fed, only 4.2 atoms% were retained, while after feeding the β,γ-dideuterobutyric acid, an average of 22.5% of the excreted betahydroxybutyrate contained deuterium.

In order to determine which part of the caproic acid molecule is convertible to acetone bodies, a study of the deuterium content of betahydroxybutyric acid excreted in the urine after the feeding of the α,β- and β,γ-dideuterocaproate has been made. The deutero acids were prepared by saturation of the corresponding unsaturated acids.

The α,β unsaturated acid was formed by condensation of butyr-aldehyde and malonic acid under the influence of pyridine as a base while the formation of the β,γ hexenoic acid was a condensation of the same substances with triethanolamine as a base. They were identified by saponification numbers, boiling points, indices of refraction, and the rate of iodine uptake of the acids and their corresponding ethyl esters.

The acids were fed by stomach tube in the form of their sodium salts to female rats previously fasted 72 hours. The amounts administered were equivalent to 150 mg per 100 sq cm (calculated as acetone) per day in 2 divided doses. The urines of 8 animals were pooled, the betahydroxybutyric acid separated by acidification, extraction in butyl alcohol, and the subsequent extraction from this material as the sodium salt by means of sodium carbonate. The silver salt was then prepared. This was analyzed for betahydroxybutyric acid and its deuterium content. The results are shown in Table I.

1. The immediate effect of morphine on colon motility resulting from the subcutaneous injection of ¼ grain is essentially a marked increase in the tone, with a diminution, frequently a disappearance of quiet periods. 2. Alternating with the periods of high tone is an exaggeration of Type I components of Type II contractions. 3. As far as the colon motility is concerned the dog shows no increased tolerance to ¼ grain of morphine given daily. 4. No evidence of a cumulative effect on colon motility was seen to follow daily injections of ¼ grain of morphine for a period of 60 days. 5. Twenty hours after subcutaneous injection of ¼ grain morphine the colon motility is practically the same as that prior to the injection.

Dental plaque material was suspended in glucose solution and incubated with the quaternary ammonium wetting and detergent compound, alkyldimethylbenzyl ammonium chloride. One part of this compound in 3,000 (approximately M/1000) completely inhibited the production of acid by the plaque material.

In another type of experiment, a 1:500 solution of the detergent was applied to the plaque material in situ for 2 minutes. The plaque material was removed 10 minutes later, and did not produce appreciable quantities of acid when it was then incubated in glucose solution.

Prolongation of procaine HC1 anesthesia by addition of atropine sulfate in low concentration 10^-5 is reported; tests were made on rabbit's cornea after subconjunctival injection. The possibilities of an effect of low amounts of atropine sulfate on permeability are discussed.

All mice survived when fed, ad libidum, a complete diet containing 1 % of sulfathiazole for 2 or more days before and 7 days after injecting Shigella paradysenteriae Flexner. Seven of 18 mice survived when offered, ad libidum, 1% drug diets of sulfanilamide, sulfathiazole, and sulfapyridine immediately after injecting the organisms. Fifty-seven of 72 mice survived when given 4 feedings by stomach tube of these 3 drugs in doses varying between 2 mg and 1/10 mg. All of 72 mice survived when given 4 feedings of 2 mg of these 3 drugs by stomach tube, regardless of whether the first dose of drug was administered 2, 3, 4, or 5 hours after the injection of the organisms. Nine of 48 control mice survived.

From the accompanying graph it is seen that the fourth component is relatively more heat-stabile than the third component. The addition of 10% NaCl to the complement prior to heating markedly diminishes the thermostability of the fourth component, while this effect is not noted in the case of the third component, except that in the presence of 10% NaCl the third component is completely inactivated at 60° C while in the absence of the salt a temperature of 62° C is required. It is also observed that the fourth component is inactivated at 65° C while the third component is inactivated at 62° C. From these findings it is evident that the third component is more intimately associated with the serum proteins than is the fourth component. It is therefore suggested that in the routine inactivation of complement for various serological purposes, inactivation be accomplished at 54° C instead of the customary 56° C, provided the temperature control is adequate.

Discussion and Conclusions. The protective action of cooling during insulin shock has been reported previously. 2 These experiments demonstrate that when the body temperature is prevented from falling, the time necessary to produce damage is considerably shortened, and the degree of brain damage is greater. This is to be expected since the higher the temperature, the higher the metabolic rate and the greater the oxygen need of the brain. When the body temperature is kept at normal levels during insulin shock it would seem that a relatively greater degree of tissue anoxia results than when the body temperature is permitted to fall, due to the greater need of oxygen at higher temperatures. This may be a factor responsible for the prolonged comas and accidents frequently occurring during insulin shock therapy, since the drop in body temperature in humans is not as great as that which occurs in cats. Dogs also are found to be resistant to the hypothermic action of insulin. In this laboratory, it has been found that considerably shorter periods are necessary to produce brain damage in this animal than is needed for cats. In fact, brain damage has been produced in the dog without it showing the characteristic symptoms of the 4th phase. 7

Correlations between lengths of post-ejaculatory periods of inactivity in adult male rats were found to range between .65 and .83 for adjacent pauses. These correlations are remarkably high for complex psychobiological functions and are not exceeded by those for any other complex functions similarly studied. It seems probable that these post-ejaculatory pauses may be used as a basis for many fundamental studies of individual differences in male rats.

The coagulation time of normal human blood on a collodion surface is longer than the coagulation time on a paraffin surface even though the force of adhesion between blood and collodion is much greater than between blood and paraffin. There is no clot retraction in collodion lined tubes.

Numerous attempts have been made to “adapt” poliomyelitis virus to passage in the eastern cotton rat since Armstrong 1 reported that a strain of virus isolated from one of a number of cases occurring during an outbreak of poliomyelitis in Lansing, Michigan, could be propagated in the variety Sigmodon hispidus hispidus. These attempts, including those of Armstrong, to obtain takes with other strains of poliomyelitis virus have failed. Toomey 2 has recently reported unsuccessful attempts to propagate 9 other monkey adapted strains of this virus. He inoculated 9 groups of Sigmodon hispidus littoralis rats, each with one of 9 strains of virus and later when symptoms failed to develop, these animals were shown to be susceptible to the Lansing strain.

Duran-Reynals 3 , 4 has shown that testicular extract (“spreading factor”) added to the virus of vaccinia increased the extent of the lesions in the rabbit, and Hoffman and Duran-Reynals 5 showed that it had similar enhancing effects on the viruses of herpes, vesicular stomatitis and Borna disease. In this laboratory several experiments have been made recently to further investigate the effects of testicular extract on the pathogenesis of a number of filterable viruses. Our unreported and incomplete experiments, although of a slightly different character, tend to confirm those reported for the virus of herpes and also suggest that a somewhat similar effect is obtained when the spreading factor is combined with minimal doses of the virus of lymphocytic choriomeningitis given to mice intracerebrally.

It was decided to attempt to adapt a strain of poliomyelitis virus to the cotton rat by suspending the virus in testicular extract, in the hope that this factor might enable the virus to propagate in a tissue that presented conditions which possibly were partially though not entirely suitable for its growth.

In the course of plethysmographic studies on the peripheral vascular responses of hypertensive patients, it was noted that in about 50% of the cases the resting blood flow in the forearm was much greater than that in subjects with normal blood pressure. Such findings differ from those presented previously by Prinzmetal and Wilson 1 using a similar method, and from those obtained by Pickering, 2 who utilized Stewart's calorimetric procedure. 3 These investigators found that the average blood flow reading for the forearm in a series of hypertensive subjects was no greater than that in a normal group, and on the basis of these observations, together with some confirmatory studies, they concluded that the increased vascular resistance in hypertension is generalized throughout the systemic circulation rather than confined to the splanchnic area. Pickering 4 subsequently pointed out, as did also Stead and Kunkel, 5 that in the light of Grant and Pearson's work, 6 the data included in the two investigations represented not only arterial inflow to the forearm, but also venous return from the hand. It is well known that blood flow through the hand can be affected by a variety of stimuli, 7 and hence readings obtained under such conditions cannot be considered representative of peripheral blood flow generally. Further, as Prinzmetal and Wilson indicated, the opposite conclusion to the one presented by them, namely that the hypertonus is limited to the splanchnic region, could have been drawn if an increase in blood flow had been found in the forearm of hypertensive subjects of the magnitude of 1.5 times the normal figure, i.e., approximately 2.65 cc per min. per 100 cc of limb volume instead of 1.7 cc. 1

The fourth component of complement was not found to be in association with a lipid complex of serum.

In irradiated marrow cultures of nonleukemic cells the ratio of effect of 50 r of irradiation on progranulocytes to the effect on the more susceptible lymphocyte is the same as with a dose of 400 r of irradiation. An increase of 8 times in the amount of irradiation, that is from 50 r to 400 r, produced an increase of only about 1.7 times in the effect on either progranulocytes or lymphocytes. Leukemic progranulocytes showed a greater decrease from a dose of 300 r than nonleukemic progranulocytes from a dose of 400 r, but the character of the curves is similar which suggests that the mechanism of action of irradiation on leukemic cells is probably a prevention of the onset of cell division as was shown to be the case for nonleukemic cells. Leukemic progranulocytes which failed to mature in the patient also failed to mature in cultures in a medium containing normal human cord serum with or without irradiation. All of these observations are explained if the action of irradiation is to prevent the onset of mitotic or amitotic division and not by directly killing cells and if the leukemic process depends on a fundamental change in the cell affecting its rate of division and maturation and not on any alteration in the environment.

In endeavoring to determine the minimal prophylactic dose of alpha tocopherol which would insure the normality of the cross-striated musculature in suckling young rats, alpha tocopherol dissolved in ethyl laurate was administered at the 6 mg and 10 mg levels by stomach tube to a considerable series of mothers on the first day of lactation. The young were observed daily for evidences of the slightest impairment of gait, capacity to right themselves promptly when placed on their backs, et cetera. Thirty young were thus studied from mothers receiving 6 mg of alpha tocopherol and 41 from mothers receiving 10 mg. Sixty-four young from mothers receiving the solvent alone were also observed; in the last mentioned group all young showed paralyses or evident impairment of muscular function. Virtually all the young from mothers receiving either dose of alpha tocopherol appeared to be perfectly normal in their gait, righting reactions, et cetera.

Histological study of 16 muscles was made in the case of several animals from each of the 3 groups. To our surprise in the case of the 6-mg group, 12 of the 16 muscles examined showed the typical microscopic lesions of dystrophy, although these were not extensive; i.e. plenty of normal musculature remained, a fact in harmony with the complete lack of observable motor impairment of these animals.† In the 10-mg group microscopic lesions were entirely absent from all muscles studied. The muscles examined were diaphragm, gastrocnemius, adductor magnus, semimembranosus, extensor digitorum longus (communis), flexor digitorum profundus, biceps brachii, triceps brachii, psoas major, erector spinae group, sternomastoideus, serratus anterior, acromiotrapezius, pectoralis major, masseter and tongue. The masseter and tongue musculature proved themselves to be the least likely to be affected, the gastrocnemius and sternomastoideus the most likely to be affected.

The action of compounds of pharmacological importance on the oxidative systems of tissue cells has received widespread attention, particularly through the work of Jowett and Quastel 1 and of Krahl and Clowes. 2 The usual procedure is to determine the metabolism of excised tissue suspended in a balanced salt solution. However, since it might be expected that the respiration of tissue would proceed more normally if the tissue were suspended in a naturally occurring physiological fluid, our attention was first directed to the development of a simple, reproducible method of measuring tissue respiration in serum. A description of this method provides the content of the present paper. Rat liver was chosen as the standard tissue whose respiration was measured, and human blood serum as the nutrient fluid medium in which the tissue was suspended.

Experimental. Indirect methods for determining the respiration of tissue in serum are already available. 3 , 4 However, they have the disadvantage of being complicated and requiring more than the usual equipment for the determination of O₂ consumption by the direct Warburg technic.

The direct measurement of oxygen consumption of tissue in serum has been achieved by (a) alternately acidifying and evacuating serum until its CO₂ content has been reduced to about 0.5 mM per liter (2% of original value); (b) readjusting its pH to about 7.2-7.3; (c) using this CO₂-free serum as the fluid medium in which the tissue is immersed; (d) measuring the O₂ consumption of the tissue manometncally in the Erlenmeyer type Warburg chamber, with absorption of CO₂ by alkali. In this method, the pH of the fluid medium does not change outside the physiological limits, due to the buffering action exerted by the plasma proteins.

The effect on liver metabolism of adding methylene blue, chloral hydrate and sodium barbital to serum has been studied. Methylene blue has been found to increase the oxygen consumption of rat liver respiring in human serum. Chloral hydrate and sodium barbital both depress the metabolism. Following depression of the oxygen consumption by chloral hydrate, methylene blue increases the metabolism. Following sodium barbital, the dye further depresses the metabolism. This is interpreted as indicating a difference in the site of action of the drugs on the oxidative system of the tissue cells.

A method for the quantitative determination of cinchophen in bile is described. In 5 anesthetized dogs weighing about 10 kilos, it was found that 20% of 1 g of intravenously injected cinchophen is excreted in the bile in 5 hours. In 4 “chronic” bile fistula dogs an average of 55%, or from 28 to 78%, of orally administered cinchophen (100 mg per kilo body weight) was daily excreted in the bile. This shows that the liver is significantly concerned in the excretion of cinchophen and that an enterohepatic circulation of cinchophen may occur. By weight sodium cinchophen increases the volume output of bile more than sodium dehydrocholate, which is an excellent hydrocholeretic. Cinchophen in large doses orally or intravenously decreases cholic acid output. But, it cannot be concluded from our results on “chronic” bile fistula dogs that cinchophen specifically interferes with cholic acid synthesis.

Roentgen pigmentation in mammals and fish have to be considered as different phenomena. While the former is influenced directly by a photochemical process and therefore generally limited to the site of action of roentgen rays, the latter seems to depend on a disturbance of the chromatomotoric nervous system and, therefore, may extend over the total innervated area.

The development of micromethods for the determination of the individual phospholipids, lecithin, cephalin and sphingomyelin 1 has made possible investigations of the phospholipid distribution in blood. Characteristic patterns for the phospholipid partition among the 3 separate fractions appear to exist for normal plasma, erythro-cytes and tissues, 1 however, preliminary analyses presented in this paper demonstrate that certain anomalies occur in pernicious anemia and lipemia.

A marked reduction of the total phospholipid in plasma is a salient feature in pernicious anemia. 2 That some abnormality in the type of phospholipids likewise occurs has been indicated by lowered recoveries of the phospholipid with petroleum ether, following evaporation of the alcohol-ether extracts in air. 3 In view of data which showed that diminished recoveries accompanied lower fatty acid carbon: phosphorus ratios of the precipitated phospholipids (theoretical ratios for lecithin, cephalin and sphingomyelin being 13.9, 13.9 and 7.4, respectively) a higher proportion of sphingomyelin in pernicious anemia was indicated. Recent data from this laboratory have demonstrated that both lecithin and sphingomyelin may be refractory to petroleum-ether extraction under certain conditions, whereas cephalin is extracted completely. 1

The data given in the table were obtained on petroleum-ether extracts prepared under carefully controlled conditions of evaporation of the preliminary alcohol-ether extracts which effect a far more complete recovery of the phospholipids 4 and confirm the marked reduction of total plasma phospholipid in pernicious anemia. 2 This decrease is reflected in all three phospholipid components. With treatment and partial relief of the anemia the total phospholipid content of the plasma is raised, accompanied by corresponding increases in lecithin and sphingomyelin, but with a decrease in cephalin content. When complete remission is attained the total phospholipid is above normal; both cephalin and sphingomyelin are likewise elevated to levels far above normal, whereas the absolute amount and proportion of lecithin is conspicuously diminished.

That cerebrosides may exist in small amounts in blood has been shown by the development and application of a titrimetric microprocedure for this group of lipids. 1 , 2 , 3 This method, slightly modified, has been employed on lipid extracts of blood, stroma and brain. The data indicate that in erythrocytes, stroma, and brain, much of the lipid which formerly was designated as neutral fat∗ is cerebroside, whereas in plasma only negligible amounts of cerebroside are present. Furthermore, there are indications that the cerebrosides may comprise a significant fraction of the blood lipids in certain pathological conditions.

Method. Procedure for hydrolysis: The procedure of Kirk 2 was modified to facilitate hydrolysis of the cerebrosides and filtration of the hydrolysate. Aliquot samples of the lipid extract and blanks for initial and total reduction values 2 are measured into 10 cc glass-stoppered graduated cylinders. The solvents are evaporated carefully in a boiling water bath. To each of the samples for initial reduction are added 3 cc of water, and to each of those for total reduction 2 cc of 6 N hydrochloric acid. The cylinders are stoppered tightly and heated for 15 minutes in a boiling water bath to emulsify the lipids and hydrolyze the cerebrosides. After cooling, 1 drop of phenol red, 0.5 cc of 4% zinc sulfate solution (ZnSO₄.7H₂O), and 2 cc of 0.1 N sodium hydroxide are added to each sample for initial reduction, after which they are diluted to 10 cc and filtered through a medium weight filter paper. To each sample hydrolyzed with hydrochloric acid are added 1 drop of phenol red indicator and, in succession, the following: 40% sodium hydroxide (drop by drop) until the solution turns pink; 1 N hydrochloric acid until a yellow color is evident; and 0.1 N sodium hydroxide until a pink color develops.

Growth is stimulated in “plateaued” E-low female rats when their diet is supplemented by the daily administration of alpha tocopherol at the 0.25 and 0.50 mg levels. The growth corresponds approximately to that previously secured by the daily administration of 1 mg of alpha tocopherol.

Four generations of vitamin E-low rats were observed as to growth and reproductive behavior. In the female there was a delay in maturity in each successive generation. Growth was also affected.

The males in each generation were sterile but fertility was maintained in the fourth generation by the prophylactic administration of wheat germ oil.

Coarse, sparse, yellowish fur and dystrophy that appear in rats maintained on vitamin E-low diets appeared progressively earlier in each successive generation.

During the past few years at least two groups of investigators 1 , 2 have reported cases of upper-respiratory infection of unknown etiology, that resembled true “epidemic influenza” in clinical respects but which differed in that the convalescent serums failed to show any increase in capacity to neutralize standard strains of the influenza virus. Two similar cases of influenzalike infection occurred among the workers in this laboratory in February, 1940. The serums obtained from these cases 4 weeks after the infection and those obtained either at the time of or 3 weeks before the onset of illness were tested against the PR8 strain of the virus of epidemic influenza. Neither of the convalescent serums showed any detectable increase in capacity to neutralize the usual 1000 lethal doses of the virus; and neither of them fixed complement in tests against antigens prepared in the usual manner 3 from mouse lung suspensions. This apparent lack of development of antibodies reactive against this standard strain of influenza virus seemed to indicate that the influenza-like infection of neither of the cases had been due to the virus of epidemic influenza.

The throat washings from both patients when inoculated intranasally in ferrets under ether anesthesia evoked a fever of the type considered characteristic of influenza. The infectious agent from one of the cases was transmitted serially in ferrets, but even after 10 passages the lungs of infected animals showed no consolidation. The infection was transmitted by intranasal inoculation from ferrets to Swiss mice; in that species blue-gray areas of pulmonary consolidation were produced on the first passage. After 9 passages the virus increased sufficiently in virulence to kill some of the mice but although it has now been passed over 20 times it has not yet acquired the capacity of regularly killing all of the mice that are inoculated.

These experiments have revealed four main facts: (a) White mice are highly susceptible to intracerebrally-injected 48-hour broth cultures of rabbit-virulent strains of C. diphtheria; 99% of the mice so inoculated dying within 10 days, (b) Mice are like-wise susceptible, but in somewhat lesser degree, to similar injections of rahbit-avirulent, and hitherto supposedly harmless, strains of C. diphtheria; about 85% of the mice showing highly characteristic reactions within 10 days, (c) Mice receiving minute doses of toxin intracerebrally exhibit the same definitive and fatal results observed in the mice receiving the rabbit-virulent and rabbit-avirulent strains, (d) A number of cultures of diphtheroids (C. xerosis and C. pseudo-diphthericum) were harmless for mice when injected as indicated above. Several other organisms have failed to produce any reactions in any way resembling those produced by C. diphtheria and diphtheria toxin.

The effect of sulfathiazole, sulfamethylthiazole, sulfapyridine and sulfanilamide in concentrations of 10 mg per 100 cc on large and small inocula of E. typhosa, S. paratyphi, S. schottmuelleri, Shigella dysenteries, Shigella paradysenterice and Shigella sonnei in veal infusion peptone broth, urine and blood was studied. It was found that sulfanilamide had little bacteriostatic effect in broth cultures, whereas, if a small inoculum is used, sulfathiazole, sulfamethylthiazole and sulfapyridine showed a bacteriostatic action. In urine and blood, sulfanilamide usually caused some inhibition of growth. The action of sulfathiazole, sulfamethylthiazole and sulfapyridine in urine and blood was not only bacteriostatic but was often bactericidal. Sulfathiazole was only slightly superior to sulfamethylthiazole and sulfapyridine when urine and blood were used as media.

Of the 6 organisms studied, S. paradysenteriæ (Flexner) appears to be most susceptible to the action of the sulfonamide drugs. No further conclusions as to the comparative susceptibility of the strains used in this study seems justified.

Mizuta and Ikegami 1 have reported the presence in cattle bile of an ether-soluble substance which promotes glycogen formation in the liver. We decided to repeat their experiments because the data presented are not convincing and the concept may possess significance.

Methods. We have conformed as closely as possible to the general experimental plan of the Japanese observers. Young adult rabbits were fed our standard stock diet for 2 weeks. They were then placed in a stock in a wire cage to prevent coprophagia and fasted for 72 hours. The animals were then given 2 or 4 g of glucose per kilo body weight slowly intravenously (10 minutes) and 30 minutes later were given the bile extract similarly. 2.5 hours later, the animals were given pentobarbital intravenously, which induces anesthesia immediately. The liver was frozen in situ with CO₂ within a minute. The frozen liver was crushed to powder and a weighed portion placed in 30% KOH, digested and analyzed for glycogen by Somogyi's method. The results were calculated and expressed as the percent of glycogen in the powder.

The extracts were made from freshly frozen cattle bile. One extract, A, was made by rendering the thawed bile slightly alkaline (pH 8.0), evaporating to dryness in vacuo at a low temperature for 24 hours, and extracting the powder with ether and removing the ether in vacuo. A second extract, B, was made by slightly acidifying the thawed bile (pH 4.0) with phosphoric acid, steam distilling for 6 hours, the distillate being extracted with ether and the ether removed in vacuo. A third extract, C, was made as in the first extract, A, except the thawed bile was slightly acidified (pH 4.0) with phosphoric acid.

Inhalation of 100% oxygen has been shown to result in the elevation of the T wave in the electrocardiogram in 20 of 26 cardiac cases, and in 2 of a group of 8 normal and miscellaneous cases. When 97 or 96% oxygen with 3 or 4% carbon dioxide was tested, similar electrocardiographic changes were observed although the degree of change was frequently less marked; thus, in 7 of 20 cardiac cases the T wave was distinctly more elevated after inhalation of 100% oxygen than after the oxygen-carbon dioxide mixture. The significance of these observations is discussed. It seems likely that inhalation of 100% oxygen shortens the recovery period of cardiac muscle.

Using the uricase method for the determination of blood uric acid from 89 to 96% of the hyperuricemia of pre-eclampsia and eclampsia consists of true uric acid.

It has been shown by one of the authors that functional eyes can be successfully exchanged between the larvae of 2 closely allied species, Amblystoma punctatum and A. tigrinum. 1 Since we have also demonstrated that homoplastic eye grafts of adult A. puncta-turn 2 and Triturus viridescens 3 , 4 , 5 can be made with return of vision, it was of interest to see what success could be obtained in these 2 very different species if their eyes were exchanged. In the present experiments the right eyes were exchanged between 34 pairs of animals. The Amblystoma hosts were young adults around 10 to 11 cm in length. The Triturus animals were about 9 cm in length. The eyes were studied daily while living and the hosts were preserved from 14 to 141 days after operation for histological studies.

In Amblystoma eyes on Triturus hosts return of circulation was observed as early as the fifth day. In the majority of cases it was recorded during the first 2 weeks. In Triturus grafts it was first seen on the sixth day and recorded in one-half of the cases during the first 2 weeks. In some cases temporary opacity in the cornea in both groups delayed the observation of the first visible signs of circulation. In some Triturus eyes the opacity, when it appeared, lasted for many days. Corneal reflex was obtained in Amblystoma grafts after the first month. This appeared to be present in a few Triturus eyes also.

The experiments demonstrated that Amblystoma eyes were far better adapted to their new hosts than the Triturus grafts. Although there was sloughing and resorption of both types of grafts the Triturus eyes on Amblystoma if kept long enough beyond the sixth or seventh week ended in this way.

Harrison 1 demonstrated in Amblystoma punctatum that lens development was suppressed if the eye rudiment was excised just before or immediately after the closure of the neural folds, but that the same presumptive lens ectoderm would form lens tissue if grafted to other parts of the head. This indicated that the lens was established very early even though a prolonged association with the eye insured better development. He also showed that at this time body and head ectoderm was not transformed into lens if grafted over these eye centers. In stages after the closure of the neural folds this was corroborated by Beekwith 2 and Stone and Dinnean. 3

The present report involving 60 experiments demonstrates lens induction by the eye in this salamander and shows how the various lens-eye relationships in cyclopia can be explained on this basis.

The presumptive lens forming region in the open neural plate (Harrison stages 15 and 16) was completely excised on the right side and replaced by ectoderm which was taken from the ventral pole of Harrison stage 12 to 13, previously stained in Nile-blue sulphate. The blue ectoderm later covered the entire side of the head including the eye. The latter induced a lens in the grafted ectoderm. In his-tological sections the lens contained the blue dye, therefore leaving no doubt as to its origin.

In one group of experiments in this series the prechordal substrate under the neural plate (Harrison stage 15) was excised in the manner described by Mangold 4 and Adelmann, 5 while in another group, eggs (Harrison stages 8 and 9—when involution of blastopore begins) were placed in lithium chloride (0.4% solution) for 24 hours.

Blood of dogs with intact renal circulations reduces the blood pressure of those made hypertensive by partially occluding the renal arteries. Blood from hypertensive dogs gives no such effect. Blood of normal dogs contains an antipressor substance that is absent in hypertensive dogs. Normal blood pressures are not affected by it.

A method for perfusing the isolated right-atrium of the guinea pig heart is illustrated in the schematic diagram. The first step in making such a preparation is to insert a cannula connected to a reservoir of Locke's solution into the inferior vena cava. A second cannula is then inserted into the superior vena cava. A ligature is now tied around the right-atrium at its junction with the right ventricle. In performing this third step, I have found it convenient to pass forceps caudally and to the left between the superior vena cava and the large arteries, through the left atrial tissue near its caudal junction with the right atrium. A ligature is then drawn through and tied at the right atrio-ventricular junction. The greater part of the two ventricles and the left atrium is excised, and the preparation is dissected free from the supporting tissue.

At this stage the preparation resembles a tube with a cannula in each end. The 2 cannulae are now loosely tied together. The preparation is clamped in a vertical position to a ring stand, and connected to a light lever by means of a thread. An end of the ligature about the atrio-ventricular junction is a convenient place to attach the thread to the atrium. In the resulting preparation, oxygenated Locke's solution at 36 to 37°C flows through one cannula and out the other. The various pressure levels are approximately those shown in the diagram; the chief aims in adjusting these levels are to have the atrium moderately distended and to have sufficient Locke's solution flowing through the preparation. (I have used about 10 cc per minute).

The preparation will continue to beat with a rather constant rate and amplitude for several hours.

1. Feeding cinchophen to chicks produces gizzard lesions with the characteristics of peptic ulcers. 2. These gastric ulcers vary in severity with the amount of cinchophen fed. 3. The early lesions are similar to those produced by a dietary deficiency alone. 4. The lesions are less severe in chicks fed a dietary supplement containing the anti-gizzard erosion factor.

Serial observations on 18 patients reveal a rise of carbon dioxide-combining power of the plasma in 8 cases as their clinical status improves with treatment and operation. All values fall within the normal range. The excretion of total titratable acid of the urine in 9 patients on constant diets is reported. In the patients in whom decided improvement occurs shortly after operation, the preoperative amounts are from 2 to 4 times as large as those excreted postoperatively. Excretion of organic acids and ammonia nitrogen were also followed for nearly a month in one patient and in exacerbations of hyperthyroidism her excretion of these substances, as well as of total acid, is seen to rise; urinary pH values fall during exacerbations. In 2 patients who improved promptly following thyroidec-tomy, fall in total acid excretion is accompanied by a rise of alkali reserve; in a third patient who showed little early improvement and only a slight change of total acid excretion, there was no change in carbon dioxide-combining power of the plasma.

The susceptibility of the albino rat to chloroform poisoning is increased by the administration of crystalline thyroxin.

A mercury valve is described for obtaining samples of alveolar air over mercury at the end of expiration by a nasal catheter. The CO₂values are lower than the Haldane-Priestley values, even when respiration is voluntarily slowed.

1. The parotoid secretion of the spadefoot toad (Scaphiopus holbrookii holbrookii) has no digitalis-like effect, but it stimulates the isolated rabbit's intestines and the isolated guinea pig's uterus. 2. The spadefoot toad, like the nebulous toad, Bufo valli-ceps, is definitely more resistant to ouabain and cymarin, and only slightly more resistant to coumingine HC1, as compared with the Leopard frog, Rana pipiens. 3. The tree frog, Hyla cinerea cinerea, is less susceptible to cymarin than the frog, but almost equally susceptible to ouabain and coumingine HC1.

In a previous communication it was demonstrated that saline extracts of full term or immediately post-partum rabbit uteri contain a toxic substance lethal to other rabbits when injected intravenously in doses of 6 cc or less. 1 It was also reported that this toxic substance possesses some physiological properties similar to those of histamine. 2

Among the consistent characteristics of a full term or immediately post-partum rabbit uterus containing the toxic substance, there are: (1) increased color, of vascular origin; 3 (2) increased volume of the entire uterus; 4 and (3) increased amounts of intercellular fluid, or edema. 5 Similar uterine changes after administration of estrogens to non-pregnant animals have been adequately demonstrated by several investigators. Moreover, it has been demonstrated, at least in other species, that estrogens are present in both blood and urine in increasing amounts as pregnancy progresses. 6 The presence of high concentrations of estrogens in the final stages of pregnancy, and the similarity of uterine changes at term and after administration of estrogens, suggests the possibility that the primary female hormone may bear some relationship to the presence of a toxic substance in uterine tissues. For these reasons it was considered advisable to determine if the toxic factor can be recovered from uteri of non-pregnant rabbits previously injected with estrogens.

Ten adult non-pregnant rabbits were treated as follows: (1) 4 animals received theelin; (2) 3 received Progynon B; and (3) 3 received stilbesterol. These hormones were given subcutaneously in doses indicated in the table. Forty-eight hours after the last injection the animals were sacrificed, the uteri excised, and extracts prepared as described in an earlier communication, so that one cc of extract represented 0.5 g fresh uterine material. 2 Four non-pregnant untreated animals provided uteri serving as controls.

Gonadectomy exerted no influence on the degree of compensatory renal hypertrophy in the male albino rat. Testosterone propionate administration produced a marked increase in compensatory renal hypertrophy.

1. In all 3 types of experiment, skin tests, anaphylaxis, and precipitin reactions, the sulfanila-mide and sulfapyridine azoproteins produced heterologous antibodies for each other although the homologous reactions were always stronger. The animals injected with sulfapyridine azoprotein were more reactive to sulfanilamide azoprotein than vice versa. Therefore it might be predicted clinically that patients hypersensitive to sulfanilamide could tolerate sulfapyridine better than those patients sensitive to sulfapyridine could tolerate sulfanilamide. 2. Sodium sulfathiazole azoprotein did not form a precipitate with sulfanilamide or sulfapyridine azoprotein antiserums. This suggests the thiazole may be too far removed from the fundamental sulfanilamide structure to participate clinically in heterologous reactions. 3. The highest precipitin titer found was that of sulfanilic acid azoprotein antigen and sulfanilamide azoprotein antiserum. It seems difficult to account for this as a minor group phenomenon. Is it possible that the sulfanilamide in vivo is changed to a structure like sulfanilic acid which then acts as a precipitinogen? 4. There was no evidence that the simple chemicals uncombined with protein could serve either as immunizing or testing antigens. However, 2% sodium sulfapyridine inhibited sulfanilamide and sulfapyridine azoprotein precipitins without affecting the precipitins for beef serum.

Several workers have found large variations in the size of Trichomonas vaginalis in fresh preparations from vaginal discharges. Powell 1 reported lengths of 10 to 30μ and Wenrich 2 in 2 sets of preparations obtained lengths of 18 to 36 and 12.6 to 27μ. There has been some question concerning the cause of these variations, 1 and it was with the hope of clarifying this situation that this experiment was undertaken.

The materials and culture-methods used in this experiment are the same as those employed by Johnson. 3

T. vaginalis was grown in media at various pH values and the organisms were measured. Observations were made on the living organisms sealed with vaseline under a microslip supported by glass fragments. The organisms were thoroughly distributed throughout the media before samples were taken. Measurements of length and width were made on the first 35 motile organisms encountered in the field. No measurements were taken of the amoeboid forms which are considered degenerative. 2

Two-day cultures—The minimal mean size of the organisms (12.4±O.13μ in length and 9.4±0.11μ. in width) appeared at approximately pH 5.40. The maximal mean size (15 ±0.23ν in length and 11.6±:0.18μ in width) occurred at pH 7.60. Between these extremes there was a progressive although somewhat irregular increase in size with increase in pH. The extreme range was from 6.4 to 24μ in length and from 4.8 to 19.2μ in width.

Four-day cultures—After 4 days the media which exhibited a pH of 6.64 to 5.15 after 2 days had become more acid (pH 6.44 and 4.93), while the less acid media became slightly more alkaline (about 0.1 pH unit).

The minimal mean size of protozoa (11.4±0.12 by 9.0±0.10μ occurred at about pH 5.10 to 5.17.

1. Acute carbon monoxide asphyxia raises the blood sugar in direct proportion to the percentage of COHb induced. 2. Repeated exposure to carbon monoxide seems not to change the insulin tolerance significantly but to reduce the dextrose tolerance below the limits of normal variation. It reduces the fasting blood sugar 6-8 mg below that of untreated litter mates.

The spatial configuration of dextrorotatory canavanine has been determined by the method of Lutz and Jirgensons. It was found to be in agreement with that of the other naturally occurring amino acids and should therefore be designated as 1(+) canavanine. A simplified procedure for the purification of the cana-vanine flavianate prepared from Jack Bean has been described.

In a previous study of experimental hypertension in the rat, 1 it was found that subtotal nephrectomy was followed by a progressive increase in blood pressure over a period of 3 to 4 months. Daily vaginal smears from a few females that had developed hypertension as a result of this operation revealed marked abnormalities in the estrous cycles. In view of the latter observation, it seemed advisable to obtain additional and more conclusive data on the influence of subtotal nephrectomy upon the estrous cycle. Accordingly, the estrous cycles of 40 adult, healthy female albino rats were followed by means of daily vaginal smears for a control period of 50 days. These animals were then subjected to subtotal nephrectomy by a method described previously. 1 After sufficient time had elapsed for the development of hypertension, vaginal smears were made daily for a period of 35 days. Blood pressures were recorded during the control period and again 3 to 4 months after nephrectomy. Blood pressure was measured by the method of Diaz and Levy. 2 At the end of the experimental period the animals were sacrificed and the weights of the ovaries and pituitaries recorded.

Thirteen of the animals subjected to the above procedures survived for 3 to 4 months and remained in good physical condition throughout the experimental period. Observations on these 13 animals and similar observations on 8 rats which were subjected to the same experimental procedures but whose cycles were not observed prior to nephrectomy are presented in Table 1 and Chart 1. The changes in blood pressure (Table 1) were similar to those described previously. 1 Prior to nephrectomy, the estrous cycles were, with few exceptions, normal.

The average breaking-strength of femurs of estrogen-treated mice was 2499 g or 844 g greater than that of untreated controls.

1. Following surgical section of the olfactory tracts or treatment of the nasal mucosa with zinc sulphate, the monkey can still be infected by intravenous inoculation, whether or not a sterile inflammation of the central nervous system is produced. 2. An increase in susceptibility to intravenous inoculation in monkeys with a sterile inflammation of the central nervous system is indicated by the experiments; variation in susceptibility in individual animals is always, however, an important factor. 3. The failure of sterile inflammation of the central nervous system to alter the protective action of nasal treatment with zinc sulphate suggests that the action of zinc sulphate is localized in the olfactory mucosa. 4. The resistance of the monkey to poliomyelitis infection by the gastrointestinal route was not sufficiently altered by a sterile inflammation of the central nervous system to produce the disease.

Immune serum against the cells in a tissue culture inhibits their growth completely without necessarily killing them, whereas immune sera against the culture medium, plasma and embryonal extract have no influence on the growth of the cells (Lambert and Haines, 1 Hadda and Rosenthal, 2 Kimura, 3-4 Craciun and Sorescu 5 ). These findings open a new way for investigation into certain biological aspects of the cell that would be accessible by other means only with difficulty.

On immunization with such a complicated structure as a living cell we have to assume that several different antibodies are formed against the different chemical structures. The conditions become even more complicated when we are working not with a pure cellular material but with some crushed tissue containing not only various kinds of cells, but also connective-tissue elements, blood, supporting tissue of various kinds, etc. So the fact that an immune serum, for instance, against minced embryonal material inhibits the growth of homologous fibroblasts in vitro gives no exact information about the components taking an active part in the process.

Then the question is: What components of the cells are so vitally important that their blocking by antibodies is incompatible with the growth and proliferation of the cell? Or, in other words: What cellular antigens produce the growth-inhibiting antibodies?

Studies on the properties of the antibody, on the other hand, appear to be of no particular interest. A priori it is not reasonable to expect that its nature would differ essentially from that of other know antibodies.

In searching for the active antigens, two procedures are available. One is to treat a fairly large amount of cellular material (e. g., minced embryonal material, “brei”) in various ways and then use the purified product for immunization.

Rapid development of single colonies of a number of strains of C. diphtheriac on a “synthetic medium” agar occurs only in the presence of certain constituents of the blood serum of the horse and ox. Human and swine sera are ineffective. Activity is not impaired by removal of the protein by coagulation. The active material does not dialyze and evidently consists of at least two substances, one of which is soluble in acetone. Cow's milk is rich in these agents, which will be further characterized in subsequent reports.

One of the 2 (or more) factors concerned with colony development, (growth from small inocula) of C. diphtheriae appears to be oleic acid. The maximal effect is obtained with about 1 mg in 15 cc of basic medium. The other factor, present in the serum of certain species, in milk, and in commercial casein, is being further investigated.

Mammary carcinomas are readily induced by long-continued administrations of large doses of estrogens to mice of susceptible strains. In the numerous investigations 1 on the subject, the subcutaneous route of injection has been used almost exclusively. The induction of breast tumors following oral administration of estrogens has not been reported. The natural estrogens lose most of their estrogenic potency when given by mouth, 2 whereas the synthetic product, stilbestrol (4 :4'-dihydroxy-α :β-diethylstilbene) is effective orally, 2 and was used in this experiment.

Forty-one male mice of strain C₃H, raised in this laboratory, were used. Spontaneous mammary tumors develop in practically 100% of the virgin females of this line, but do not occur in the males. 3 The mice were maintained on a standard diet and an unlimited supply of water. They were about 2 months old and weighed from 21 to 23 g when the experiment was initiated.

Stilbestrol∗ was dissolved in sesame oil, and was fed to the mice by means of a curved metal tube passed into the esophagus. The animals were divided into 4 groups: (I) Eleven were given 0.125 mg of stilbestrol dissolved in 0.1 cc of sesame oil twice a week for 17 weeks, for a total dose of 4.25 mg of the compound. (II) Ten received 0.375 mg of stilbestrol in 0.1 cc of sesame oil twice a week for 17 weeks for a total of 12.75 mg. (III) Ten were fed 0.1 cc of sesame oil twice a week for 17 weeks. (IV) Ten were kept as untreated controls.

The first effect noted was the marked retardation in the gain in weight of the mice which were ingesting stilbestrol (Figure 1).

(1) Rabbit-anti-rat-placenta serum effectively interferes with the normal development of the placenta and foetus in the pregnant rat. (2) The same result is obtained with much smaller injections of rabbit anti-rat-whole-blood serum. (3) Rabbit anti-rat-serum serum fails to influence pregnancy unless injected in sufficient volume to induce anemia. (4) Anti-hormone serum and normal rabbit serum are without effect on pregnancy in the rat. (5) The immunological factor or factors responsible for the resorption of the foetuses following the injection of anti-placental serum and anti-whole-blood serum are as yet unknown.

The chief clinical value of the quantitative determination of direct serum bilirubin appears to lie in the detection of hepatic functional impairment in the presence of a normal serum bilirubin concentration. Values constituting more than 50% of the total with readings at 5 minutes or more than 70-75% with readings at 30 minutes are probably abnormal. There is no apparent significant relationship between the amount or proportion of direct bilirubin and the production of a positive direct van den Bergh reaction in sera with normal total bilirubin content.

Our experimental studies, 1 as well as those of MacLeod, 2 have shown that inadequate treatment of pneumococcal infections with sulfapyridine leads to development of organisms which are resistant or fast to that drug. Whether these sulfapyridine-fast pneumococci are equally fast to other sulfanilamide derivatives, such as sulfathiazole and sulfamethylthiazole, is a matter of theoretical and practical interest. Consequently, we have studied the reactions of 2 of our sulfapyridine-fast strains to these latter drugs.

Essentially identical experiments have been performed with each fast strain and with the parent strains from which these fast organisms were derived.∗ Mice in groups of 100 were infected intraperi-toneally, each mouse receiving 10^-6 cc of a 12 hour blood broth culture. Ten mice from each group were untreated. The remaining 90 were divided into 3 equal groups which were treated with sulfapyri-dine, sulfathiazole or sulfamethylthiazole†-20 mg doses of the respective drug being administered at 2, 8, 14 and 22 hours after infection and at 8 hour intervals thereafter for either 5 additional days (in the experiments with the parent strains) or as long as the mice survived (in the experiments with the fast strains). The drugs were administered as 10% suspensions in 10% acacia.

The results in the accompanying table show conclusively that sulfapyridine-fast pneumococci were also equally resistant to sulfathiazole and sulfamethylthiazole. Thus the mice infected with strain McGovern lived an average of 41 hours under sulfapyridine treatment and 40 and 38 hours under sulfathiazole and sulfamethylthiazole treatments. In the experiment with strain Wistuba, the average survival times were 39, 38 and 39 hours under therapy with the respective drugs.

The table shows also that these drugs were essentially equal in effectiveness against infections with the parent strains.

Lygranum antigen is superior to mouse brain antigen in sensitivity and specificity in the performance of the Frei test for lymphogranuloma venereum. Lygranum control is superior also to mouse brain control in producing non-specific reactions in only 2 of 35 persons as compared with 28 of 36 in the case of mouse brain control. In the group of 36 tested only one individual showed a reaction which could be regarded as resulting from hypersensitivity to chick material.

In experiments previously reported, it was found that application to the surface of the heart (cat, dog, rabbit) of squares of filter paper soaked in M/10 or M/5 KC1 solutions, produced changes in the electrocardiogram similar to those which follow various types of injury to the heart, except that the potassium effects quickly subsided following removal of the solution. 1 Generally accepted views of the action of potassium ions in the heart and elsewhere suggest that the electrocardiographic changes produced by the application of potassium to the surface of the heart result from the reversible extinction of the action current at the point of application. Potassium chloride has in fact been employed to abolish electrical activity at selected regions in muscle and thus to permit recording of monophasic action currents. 2

We have performed a series of experiments to determine whether potassium has a similar effect when applied to the surface of the heart, and found that we could obtain monophasic action currents from the surface of the dog's heart by use of potassium chloride, in M/10, M/5, or isotonic solutions. Electrograms were obtained by wick electrodes soaked in Ringer's solution placed on the anterior surface of the heart, one on the right and one on the left ventricle The wicks led to silver-silver chloride electrodes, and these were connected to a 100,000 ohm potentiometer from which one-fifth of the potential was recorded by an amplifier-type electrocardiograph (Cardiette). Sensitivity was approximately 2 mm per millivolt. After controls were taken, first one and then the other electrode was replaced by a wick soaked in the potassium solution. This procedure yielded monophasic records, and when the leads were arranged so that the spike was upright, the monophasic wave from the right ventricle was upright, while that from the left ventricle was inverted (Fig. 1).

During infusion of 1:75,000 epinephrine for as long as 45 minutes at the rate of 1 ml per minute the potassium content of the blood entering the liver, although actually below the resting concentration still exceeds that of blood leaving the liver. This strongly suggests that the cell source from which potassium is mobilized by epinephrine takes part, at least, in removing the excess from the circulation.

1. The thymus gland of adrenalectomized male rats maintained for 45 days on sodium chloride does not undergo the involution characterizing the gland in normal untreated animals or those treated with sodium chloride. Sodium chloride as employed appears to exert no consistent effect per se on the thymus or lymphoid tissues of the unoperated control. 2. The systemic lymph nodes of adrenalectomized animals maintained on sodium chloride were heavier than those of normal untreated animals or normal animals given sodium chloride. It is notable that the mesenteric lymph nodes do not undergo a comparable gravimetric increase.

With or without concomitant treatment with estrogen, the daily injection of 0.25 or 0.5 mg of progesterone into mice ovariectomized 3 weeks previously evoked mitosis in all the tunics of the uterus. The nuclei of the cells of the uterine tunica propria became larger and transformed from the fusiform, pyknotic type into vesicular nuclei. The latter condition was also seen in pregnant and lactating mice. Neither estrogen alone nor androgen provoked this morphology.

In the rat, desoxycorticosterone acetate (D.C.A.) causes marked diuresis, accompanied by increased chloride excretion, but not by a constant increase in the chloride concentration of the urine. The diuretic effect is much more pronounced in the hypo-physectomized than in the intact rat. Unlike in the intact animal, D.C.A. fails to decrease the blood chlorides in the hypophysecto-mized rat. The hyperchloruria and polyuria cannot be considered, therefore, as the consequence of the hypochloremia.

The kidney hypertrophy elicited by progesterone, D.C.A., and androgens is likewise not the cause of the polyuria since testosterone —which is most active among the steroids as far as kidney enlargement is concerned—does not influence diuresis. The kidney-enlarging action of all three above mentioned steroids is considerably decreased but not prevented by hypophysectomy.

The expenses of this investigation were defrayed by a grant received by one of us (H.S.) from the Committee on Endocrinology of the United States National Research Council. We are also greatly indebted to Doctors Gregory Stragnell and Erwin Schwenk of the Schering Corporation of Bloomfield, New Jersey, who donated the generous supply of steroid hormones which was necessary for the completion of these experiments.

1. Thyroxin and suprarenal cortical extract induce glycogenolysis of the glycogen normally present in the livers of 8-day embryos. 2. The injection of adrenalin in approximately physiological concentration appears to have no effect on liver glycogen. 3. Increased liver and skeletal muscle glycogen and a rise in the blood sugar level have been noted after the injection of crude anterior pituitary extract, suggesting a stimulation of gluconeogenesis. 4. Liver glycogen is increased in 8-day embryos by the injection of insulin and this increase is to be correlated with a rapid fall in the blood sugar level. 5. Injection of glucose fails to interrupt the normal disappearance of glycogen from the livers of 12-day embryos. 6. Both insulin and anterior pituitary extract cause an increase in the liver glycogen of 12-day embryos. The anterior pituitary extract also causes a rise in the blood sugar level at this time.

By means of a diet furnishing about 7 μg of Zn per rat daily, studies at the University of Wisconsin 1-4 have convincingly demonstrated the indispensability of Zn in the nutrition of rats, although the Zn-deficient diet permitted the experimental rats to gain about 8 g per rat weekly during the 10 week period on the diet. 4 The findings have invalidated the conclusion from our laboratory 5 several years ago that the element probably is not a dietary essential. We have recently prepared a diet furnishing not more than 2 to 4 μg of Zn per rat daily. Young rats restricted to it quickly developed extreme degrees of deficiency. Because the studies have been temporarily interrupted, owing to the transfer of one of us (H.G.D.) to another laboratory, we have decided to make a preliminary report on the work at this time.

Diet. The diet was : casein hydrolysate (tryptic)† 15.00, egg white (cooked) 3.00, sucrose 66.29, salts 5.71, butter fat 10.00, and Oleum Percomorphum 50%, 2 drops per 100 g diet (approximately 2800 vitamin A units and 400 vitamin D units).

Each rat was given a daily supplement containing the equivalent of 6.4 g liver, 40 μg of thiamin and 1.0 mg of choline. The liver concentrate,‡ dissolved in 6 volumes of water, was centrifuged. The supernatant solution was transferred to a Pyrex glass separatory funnel and the pH was adjusted to about 6. The solution was extracted repeatedly with dithizone (diphenyl thiocarbazone) dissolved in CC1₄. When no more Zn could be removed Zn-free dilute HC1 was added until the pH was about 4. The excess of dithizone was removed with redistilled CC1₄.

A change in the physiological activity of a nicotine solution accompanies a change in pH of the solution when the phenomenon of transport across membranes is involved. An increase in pH causes an increase in the proportion of free base present, and, as pointed out by Ellisor and Richardson, 1 an increase in the rate at which nicotine passes through membranes. This is believed to account for the fact that a solution containing equal parts of nicotine acetate and the free base is about three times more effective in inhibiting ciliary activity than a solution containing about 98% of the acetate.

Leukemia appeared at an earlier age than in controls when mice of the F strain were painted twice weekly with methylcholanthrene. Myelogenous leukemia, which does not occur in untreated F mice before 300 days of age, appeared as early as 97 days after birth in treated mice. Only 3 cases of leukemia occurred in 184 mice of non-Ieukemic strains treated in a similar manner. The effectiveness of methylcholanthrene in influencing the appearance of leukemia in young mice depended on the genetic susceptibility of mice to the disease.

Arginine, histidine, lysine, cystine, methionine, tyrosine, tryptophane, phenylalanine, glycine, alanine, and threonine can all be determined by available micro methods 1 , 2 in only 2 to 4 g of protein, but until recently methods were not available for the estimation of 3 of the essential amino acids : leucine, isoleucine, and valine in small quantities of protein. Last year Fromageot and Heitz 3 described a procedure for the determination of leucine and valine, involving deamination to the corresponding hydroxy acids with nitrous acid, oxidation to acetone with chromate, and measurement of the acetone colorimetrically after reaction with salicylaldehyde. Leucine and valine were estimated in the presence of each other by carrying out the oxidation with chromate on 2 aliquots under conditions such that the proportionate yield of acetone from the 2 amino acids differed considerably. We were unsuccessful in obtaining consistent results with this procedure. In the present study we have made extensive changes in and simplified the procedure of Fromageot and Heitz and in addition have found conditions which permit the determination of isoleucine by measuring the ethylmethylketone formed in the oxidation of the corresponding hydroxy acid. All 3 amino acids can be determined in as little as 100 mg of protein for no one of the other amino acids recognized as occurring in proteins yields acetone or ethylmethylketone under these conditions.

Hydrolysis and Deamination. The protein is hydrolyzed with 10 volumes of 8 N sulfuric acid and the excess acid is removed with barium hydroxide. The precipitate of barium sulfate is centrifuged and thoroughly washed with hot water. The filtrate and washings are reduced in vacua to 50 cc. Amino acid solutions prepared by other methods of hydrolysis are suitable provided the hydrolyzing agent does not yield compounds which would interfere in later steps.

Substances injected into the marrow cavity of the tibia of the rabbit and of the sternum of man are almost immediately absorbed into the general circulation. Blood and glucose solutions respectively, by intramedullary injection, corrected rapidly experimental anemia and hypoglycemia induced in rabbits.

The anti egg-white-injury factor (biotin) was found to be formed in the rumen of a cow receiving a ration which was low in this factor.

Typical rachitic changes in bones and teeth have been produced in young guinea pigs by a diet low in calcium and in vitamin D. In older animals the changes in the bones were obscured by arrested skeletal growth, but those in the teeth were constant.

To make contact with the scalp for electroencephalographic recording the practice has frequently been to cut the hair in small areas and fasten electrodes in place with collodion. The collodion and the electrodes were subsequently removed with ether. The attaching of a dozen or more such electrodes for an electroencephalographic exploration is laborious, the cosmetic effect is not too favorable, and the patient may not be enthusiastic.

A more expeditious procedure for placing of electrodes on hair covered areas is possible with an electrode which makes use of the hair itself to hold the electrode. Two simple electrodes are here described. The first, illustrated in Fig. 1, consists of a ring of metal I/2.” in diameter, 3/16” high, with a wire lead attached, and a slot 3/32” deep in the top into which fits a wire “hairpin” of No. 20 spring wire bent double like a cotter pin or a “bobby pin”. After massage of an area of scalp with electrode paste a wisp of hair from the area is passed through the metal ring and inserted in the slot of the wire hairpin. The wisp of hair is wound up, windlass fashion, on the hairpin until definite tension holds the pin against the top of the electrode ring and pushes the electrode ring against the scalp. The doubled wire pin is then permitted to slip edgewise into the slots in the top of the electrode ring. This prevents the pin from turning or the hair from unwinding, and the electrode ring is held snugly in place. The ring may be filled with electrode paste, insuring contact and providing assurance against drying of the electrode. Corrugations or comblike teeth may be milled on the lower edge of the electrode ring so that stray hairs beneath the ring may not prevent contact with the scalp.

It has long been a common practice to add charcoal to poultry feeds, under the impression that charcoal improves the health and reduces the mortality rate of chickens. Acceptable evidence for these alleged attributes has been lacking. Charcoal is claimed to act by adsorbing undesirable products from the digestive tract, for example, one manufacturer of poultry charcoal lists the following substances as removed: carbon dioxide, ammonia, bacteria, pigments, poisons and enzymes.

Charcoal is known to cause a bleaching of poultry flesh and of egg yolk by reducing the deposition of carotenoid pigments from the diet. This bleaching effect suggests the possibility that carotene and other vitamins in the diet might also be removed or inactivated by charcoal. For example, former experience in this laboratory showed that vitamin K is adsorbed strongly on charcoal and that it cannot later be removed without much destruction. The vitamin K-deficient chicken fed vitamin K adsorbed on charcoal seems unable to remove and utilize the vitamin. 1

A diet was formulated to provide optimum amounts of vitamins A, K, G (riboflavin) and the chick gizzard factor, without the considerable margins of safety that exist in most good commercial chick mashes. This diet was found capable of promoting normal growth and health of chicks.

Triphenyl phosphite has been used as a convulsive agent in experimental epilepsy. 1 Because it is relatively easy to prepare this compound synthetically 2 and to render it radioactive by synthesizing it from radioactive phosphorus,† it seemed feasible and desirable in our studies on epilepsy to combine the function of tracer substance with that of convulsant. Since it was eventually demonstrated that triphenyl phosphite hydrolyzes and the convulsant effects were due to the phenol fraction which is known to act at the cord level, 3 its use was abandoned. In the course of these studies, however, certain interesting observations were made in connection with the absorption, hydrolysis, toxicity and distribution of triphenyl phosphite. The results seem worthy of brief report, inasmuch as this compound belongs to the same series as triorthocresyl phosphite, which produces a combined system degeneration of the spinal cord 4 and is related to triorthocresyl phosphate, the contaminant responsible for the degenerative changes in cases of Jamaica ginger paralysis.⁵

The toxic doses of triphenyl phosphite as reported by Smith, et al., 4 were confirmed in rats and cats. In dogs 2 doses of as little as 0.5 cc per kilo of body weight, given one week apart, produced the typical ataxia and weakness in the hind legs within 2 weeks. The early convulsive reaction was found to depend upon the purity of the preparation. A commercial preparation proved most toxic and 2 synthetic preparations, in one of which a high degree of freedom from contamination with phenol was achieved, indicated that the convulsive effects could be closely correlated with the degree of phenol contamination of the preparation.

Smith and his coworkers 4 have adduced evidence purporting to show that the esters of phosphorous acid were absorbed very slowly from the site of injection.

Greying of the fur was first observed by Morgan, Cook and Davison 1 and Lunde and Kringstad 2 in black rats maintained on diets deficient in the vitamin B complex. Addition of preparations containing “filtrate factor”, 3 of liver extracts containing pantothenic acid 4 or of brewers' yeast 2 have been reported to prevent the development of greying or to restore the black pigmentation of the hair.

In our laboratory, greying of the hair was obtained in black and piebald rats on a diet consisting of casein, vitamin free, 18%; sucrose, 67%; butter fat, 9%; salt mixture, 4%; cod liver oil, 2%, and supplemented with thiamin, riboflavin, nicotinamide and vitamin B₆. On this diet greying of the fur developed in approximately 80% of the animals within 4 weeks. At this time, the animals reached stationary weights and showed, in addition to the grey symmetrical patterns of the fur, signs characteristic of pantothenic acid deficiency in rats, 5 namely, thinning of the fur, generalized scaly dermatitis, inflammation of the nasal mucosa, blood-caked whiskers, and hemorrhages in various organs, particularly in the adrenal cortex. The addition of graded doses of pantothenic acid (calcium pantothenate, Merck) to the diet demonstrated that a daily supplement of 80 or 100 μg of calcium pantothenate, e. g., doses representing the daily requirement for pantothenic acid in rats, 5 prevented, besides the above mentioned deficiency symptoms, the greying of the fur, whereas daily doses of 5, 10, or 20 μg were insufficient to prevent the greying, and 40 μg gave inconsistent results. Comparable results were obtained with various concentrates from rice bran and liver. Although the addition of these concentrates caused better growth responses than pantothenic acid alone, their efficacy in preventing the greying of the hair was found to parallel closely their content of pantothenic acid as measured by the bacteriological assay.

Pantothenic acid, first discovered as a growth factor for yeast, 1 has been established as a member of the vitamin B complex. 2 The vitamin was recently identified 3 , 4 as N-(α,γ-dihydroxy-β, β-dimethyl-butyryl)-β'-alanine and has been synthesized. 5

In continuation of our studies of the toxicology of the vitamins of the B complex, 6 , 7 the following investigation of the toxicity of pantothenic acid was carried out on animals maintained on completely adequate diets. Synthetic dextrorotatory calcium panto-thenate was used in all experiments. Calcium pantothenate is readily water-soluble and almost neutral in reaction, the pH of a 10% solution being approximately 8.

Local effects of calcium pantothenate were studied by instillation into the eye and by subcutaneous injection. Instillation of 0.5 cc of a 10% solution into the conjunctival sac of 3 rabbits did not produce any irritation. Likewise no irritation, inflammation or abscess formation was observed in 4 rabbits following the subcutaneous injection of 1.0 cc of a 10% solution. The infiltration of the subcutaneous tissues subsided about as rapidly as following a control injection of 10 cc of saline.

Acute toxicity was studied in mice, rats, dogs, and monkeys. In mice and rats the L.D. 50 was determined following oral, subcutaneous, intraperitoneal and intravenous injection, 10 animals being used for each dose level. Table I summarizes the results obtained from a total of 180 mice and 110 rats.

All 10 rats dosed with 10 g of calcium pantothenate per kg, when administered by mouth, survived without showing toxic symptoms. Lethal doses in mice and rats produced prostration and respiratory failure. Death occurred within one hour following intravenous or intraperitoneal injection and within 2 hours following oral or subcutaneous administration.

The choloresis produced in dogs by the administration of oxidized ketocholanic acids does not affect the pH, specific gravity, percentage of solids, or the cholesterol content of the gall bladder bile.

Pregnancy caused the thymi of young mice to undergo involution, both grossly and microscopically. With one exception out of 20, the degree of thymic involution varied between 45% and 80% as measured by gland weights. At parturition, the post-partum mice generally weighed more than their litter-mate controls.

Sulfathiazole is present in human red blood cells in lower concentrations than is sulfapyridine employed under identical conditions.

There is conflicting opinion concerning the motility of spermatozoa within the generative tract. It has been suggested that a high local tension of carbon dioxide prevents them from becoming motile in the epididymis and vas deferens. The carbon dioxide concentration of various tissues of the rat was studied and found even lower in the generative organs than in liver or skeletal muscle. Furthermore, the concentration of carbon dioxide found in the tissues of the generative tract was much lower than that which is required to immobilize spermatozoa in vitro.

The lethal effect of lanatoside C dissipates rapidly

Topping 1 has demonstrated that monkeys and guinea pigs fatally infected with Rocky Mountain spotted fever can be caused to recover by the administration of hyperimmune rabbit serum. This naturally suggests that such serum may have value in the treatment of human cases. The sera prepared by Topping were made by the injection of infectious tick-tissue as antigen. We have found that a serum of high neutralizing titer can also be obtained by injecting rabbits with yolk-sac material prepared according to the method of Cox. 2 We have determined that this can be refined by the same chemical procedures which are useful in purifying antipneumococcal rabbit serum and removing its toxic and reaction-producing elements. In such concentrates the neutralizing titer per gram of protein is increased about 20-fold as compared with the original serum. A similar antiserum and concentrate of high neutralizing value has also been made against the Breinl strain of European typhus. Sufficient quantities of these two sera are now being prepared and refined so that a thorough clinical study of their possible effectiveness in the treatment of Rocky Mountain spotted fever and epidemic typhus fever in man can be made.

The Western strain of equine encephalomyelitis has been successfully grown in whole minced chick embryo or chorioal-lantois in serum-ultrafiltrate and buffered salt solution (Simms') at room temperature and at 37° C.

The highest titers (10^-5 to 10^-7) have been obtained with the clear supernatant fluid of chick and serum-ultrafiltrate cultures maintained at room temperature. The titers of these cultures can be sustained with little decrease for as long as 10 and 15 days.

The Eastern strain of equine encephalomyelitis has been grown with a dilution activity of 10^-7 in whole minced chick embryo in serum-ultrafiltrate at room temperature and 37° C.

A vaccine, containing very small amounts of protein, prepared from the clear supernatant fluid of serum-ultrafiltrate cultures protected guinea pigs against 1,000 to 10,000 m.l.d. doses of virus (Western strain).

After 47 passages through the developing chick embryo, rabies fixed virus shows no enhanced virulence for the chick embryo, as judged by the complete development of the embryo, the relatively low virus titre in the embryo brain and the scanty pathological changes in the brain. The virulence of the chick embryo passage virus for mice and guinea pigs remained unchanged, while that for rabbits appeared reduced considerably, as demonstrated by the prolonged incubation period, and duration of the infection. The specific anti-genic character of the chick embryo virus remained unchanged during 47 passages over a period of 18 months.

Prolactin induces the water drive in normal, thyroidectomized or gonadectomized land phases of Triturus viridescens within 10 days, irrespective of their size or sex. It induces repeated molting in efts with normal thyroids but not in efts deprived of their thyroids. It causes also the rapid assumption of the adult coloration and finally the appearance of adult structural characteristics.

By a suitably fine dispersion in water benzocaine can be rendered efficacious as a topical anesthetic, to a higher degree than when powdered on the mucous membrane in pure form, or when dissolved in oil, or when rendered water soluble as a diethyl amino compound (=procaine).

Attempts at transmission of Theiler's virus to rhesus monkeys by means of intracerebral inoculation and, further, by consecutive passage in monkeys, failed; the virus, moreover, was not demonstrable in the CNS of monkeys 24 hours after inoculation. Thus, Theiler's virus has its own traits in monkeys, which fact conforms with Theiler's original conclusion 3 that the mouse virus is not identical with that of human poliomyelitis. Further proof of this finding can be deduced from results of immuno-logical studies. In the foregoing experiments two standard strains of poliomyelitic virus were used, the MV and Philadelphia, both monkey-passaged for several years; what would have been the outcome had strains freshly isolated from human cases of poliomyelitis been studied is, of course, unknown. It is apparent, however, that by the methods here used, such monkey-passaged strains are not only non-pathogenic in the type of mouse which is susceptible to Theiler's virus and which carries the latter in its intestines for a good part of its life, but also fail to produce serum-neutralizing antibodies in such mice against the homologous poliomyelitic viruses and against Theil-er's virus. Moreover, antipoliomyelitic sera of human and monkey origin that are capable of neutralizing homologous and other strains of poliomyelitic virus fail to inactivate Theiler's virus. Conversely, serum from mice harboring Theiler's virus does not neutralize the poliomyelitic viruses. On the other hand, serum from normal mice of certain adult age has antiviral bodies against Theiler's virus. In this latter relation, a similarity exists between the virus of Theiler's disease—a natural malady of mice—and that of poliomyelitis of man in that neutralizing antibodies against the latter virus develop with time in man in the absence of clinically apparent disease.

The production of immune isoagglutinins following repeated transfusions, although of common occurrence in some animal species, is very rare in man. The first instance of this sort in man was described by Landsteiner, Levine, and Janes. 1

Levine and Stetson 2 described a case in which an intra-group ag-glutinin was responsible for severe post-transfusion symptoms, with anuria after the first transfusion. This occurred in a woman who, after retaining a dead fetus for a period of about 2 months, finally delivered a macerated fetus. In view of the activity of the agglutinin at 37° C and because of its gradual disappearance, it was believed that the antibody developed as a result of immunization, and it was suggested that the products of the retained dead fetus served as the antigenic stimulus. This suggestion seemed plausible because the cells of the husband, of the same blood group, (who was the donor) were sensitive to the action of the agglutinin. Presumably, the fetus inherited from the father a dominant agglutinogen which was absent in the tissues of the mother, who could thus be immunized.

Our own recent experience and other cases from the literature 3 indicate that the isoimmunization-hypothesis may hold also in certain cases of pregnancy with complications other than the retention of the dead fetus. In each of the cases cited, atypical agglutinins could be demonstrated in the absence of a history of repeated blood transfusions.

In 5 cases 4 there was a history of varying degrees of toxic symptoms during the pregnancy (in 2 of which macerated fetuses were delivered). In one of these cases, the patient had some fever, and rupture of the membranes occurred a few days before delivery, which was normal but followed by a post-partum endometritis. In 3 patients 5 the agglutinins were observed following abortion, one of them of the septic variety. One patient, whose eighth pregnancy resulted in premature separation of the placenta, had stillbirths in the 4 preceding pregnancies. 6 In 2 of the remaining 3 cases there were histories of repeated miscarriages but the present pregnancies, apparently normal, had to be terminated by Caesarian section.† In one case the pregnancy was normal except for morning vomiting during the last six weeks; due to lack of progress in labor the delivery was by Caesarian section. 7

Zacho 1 reported an instance of premature separation of the placenta with a transfusion-accident, attributable to an atypical isoag-glutinin active on the donor's cells. This agglutinin had the unique property of greater activity at 37° C than at lower temperature. Although Zacho's case was the first and only published instance of this sort, it is likely that hitherto antibodies of this character may have been overlooked. In support of this, case reports are presented of 5 patients whose blood recently investigated were found to contain such agglutinins. A brief discussion of the properties and the origin of these agglutinins forms the basis of this communication.

One of these cases was observed in a patient† (B.M.) transfused 6 times because of bleeding from a duodenal ulcer. Apparently, this agglutinin was induced as a result of the antigenic stimulus of repeated transfusions. No such history could be elicited in the remaining 4 instances, nor in the case of Zacho, all of which were observed in women suffering from a variety of complications of pregnancy.

In one patient (G.B.) who gave a history of 2 miscarriages, the present pregnancy was complicated by death of the fetus during labor. A second patient (H.H.) had 3 consecutive miscarriages and in the present pregnancy delivery was by Caesarian section because of uterine inertia. A third patient (D.D.) who suffered from persistent vomiting from the fourth to the seventh months had fever and premature rupture of the membranes a few days before delivery which was followed by a parametritis. The fourth patient (A.H.) died of an induced septic abortion. (Zacho's patient gave a history of 4 consecutive stillbirths preceding the last pregnancy.)

The correlation of atypical agglutinins in general with certain complications of pregnancy has been mentioned elsewhere. 2

The action of type specific antiserum upon the pulmonary lesion of lobar pneumonia has been described in a previous paper. 1 Pneumonia was produced experimentally in white rats by intrabronchial inoculation of type I pneumococci suspended in mucin. The disease was uniformly fatal in untreated animals, and pneumococci were found to spread through the lung by way of edema fluid at the advancing margin of the lesion. Type specific antiserum penetrated the pneumonic lesion and apparently stopped its spread by agglutinating and immobilizing the invading organisms in the outer edema zone. The fixed pneumococci were then overtaken and were rapidly phagocytized by leucocytes.

Although sulfapyridine has proven beyond any doubt to be effective in the treatment of lobar pneumonia, the mechanism by which recovery is induced is not yet understood. Fully encapsulated living pneumococci are resistant to phagocytosis unless opsonized by specific antibody, 2 , 3 and phagocytosis is the only known method by which the host can destroy these organisms. 4 Since chemotherapy often causes a crisis long before antibodies appear in the blood, 5 and sulfapyridine in the usual dosage is mainly bacteriostatic rather than bactericidal, 6 it is not at all clear how the drug brings about the final destruction of pneumococci in the lung.

Thirty-eight albino rats were treated with sulfapyridine 6 hours after inoculation. The drug was suspended in 10% gum acacia and introduced into the stomach through a blunt cannula. Two hundred and fifty milligrams of sulfapyridine suspended in 4 cc of gum acacia mixture were given as an initial dose, and half of this amount in 2 cc of acacia was administered thereafter every 12 hours. In uninfected rats no toxic effects were noted after 1 week of treatment.

In 118 out of 122 cases where there were no indications of tumors, injection of the serum was followed by complete arrest of development of the chick embryo, the fertile egg substance undergoing degeneration, usually after the eleventh day of incubation. Confirmation, by waiting until the end of the hatching period, is advisable. The 4 negative results occurred in 2 male cases; one with mitral insufficiency plus migrating phlebitis, and the other with erythema multi forme and trigeminus neuralgia. The other 2 cases were women, one in menopause with atrophic arthritis, and tuberculous ulcers of both corneae; the other with a chronic paren-chymatous nephritis.

In marked contrast with these results, when serum from a case of malignant tumor or from a pregnant woman was used, the development of the embryo proceeded normally until a full term chick was hatched. In some cases, the growth was even accelerated, shortening the incubation period, the chicks being noticeably larger and more precocious. Positive results were obtained in 216 cases out of 220. In this group the sera which gave a negative result were 2 with squamous carcinoma of the larynx, one with spindle cell sarcoma of the scapula, and one with melanoma of the face.

The causes of the isolated failures cannot be predicated at the present time. It is to be noted that on the 11th or 12th day, chemical examination of the egg contents for carbohydrates shows marked quantitative differences between eggs injected with cancer serum and those injected with non-cancerous serum. Whatever the explanation, the fact has been established that non-cancerous sera inhibit the development of the chick, while cancerous sera sustain or even accelerate it.

The effect of acute anoxia on the economy of effort index of Wiggers and Katz was studied on medical students by allowing them to rebreath decreasing oxygen mixtures and holding these mixtures at 14, 12 and 10% oxygen, respectively, long enough to record subclavian pulses and determine brachial blood pressures leisurely.

In 20 subjects, the index increased during rebreathing of 14 and 12% oxygen; in 12, it increased further when the oxygen percent was reduced to 10; in 8 it began to show slight signs of decreasing. Three subjects showed no significant increase or a slight decrease. The possible use of the method in testing favorable and unfavorable responses of individuals to low barometric pressures is mentioned.

The dog's ventricles did not generally survive interruption of coronary circulation during fibrillation for more than about 1 minute, in the sense that useful coordinated beats sufficient for recovery are reestablished without massage. These results caution us not to entertain too high hopes of resuscitating human hearts by the countershock method. Countershock can stop fibrillation; but complete anoxia beyond one minute generally prevents a resumption of beats sufficiently vigorous to reestablish viable blood pressures.

One of the first bacterial growth factors to be studied in any detail was a substance necessary for the growth of the Johne's bacillus (Mycobacterium paratuberculosis) 1 . Twort and Ingranll showed that certain other acid-fast bacteria including the tubercle bacillus contained a necessary growth factor for the original cultivation of the Johne's bacillus. Crude concentrates of the active substance were prepared from Mycobacterium phlei. We have long been interested in the growth of the Johne's bacillus, and for some time have attempted to isolate the active growth factor. In this paper concentration of the active factor will be described and it will be shown that it can be replaced by the antihemorrhagic vitamins.

The strain of the Johne's bacillus grows as a dull, brittle, granular filni on the surface of liquid media. It always grows better in the presence of phlei cells or an extract of them, and we have not been able to subculture it more than 2 or 3 times on a purely synthetic medium although it has been artificially cultivated about 20 years. In our experience most strains of the Johne's bacillus become continuously cultivable on a synthetic medium, although the phlei factor is needed for priniary cultivation; but all strains always grow faster and give a better yield when the factor is present.

The basal medium was composed of the following substances per liter of solution :glycerol 70 cc, asparagine 5 g, sodium citrate 0.5 g, K₂HPO₄, 1 g, MgSO₄.7 H₂O 1 g, iron citrate 0.063 g. Twenty cc portions of this basal medium were sterilized by autoclaving in 50 cc Erlenmeyer flasks. Substances to be tested for growth factor activity were incorporated in the basal medium at suitable dilutions.

1. Our observations in 150 patients from a region of Alabama in which deficiency diseases are endemic show a relationship between these diseases and the resistance to, and presence of, infections with Staphylococcus aureus and Streptococcus hemolyticus. 2. The lesions at the corners of the mouth, characteristic of riboflavin deficiency, contained pure or nearly pure cultures of hemolytic strains of Staphylococcus aureus in 80% of the cases, and in the remaining 20%, Streptococcus hemolyticus predominated. Following the oral or intravenous administration of riboflavin or substances rich in it, the fissures healed rapidly and the organisms were no longer demonstrable. 3. When the bacterial flora of the conjunctival sacs were studied in cases of dietary deficiency disease and associated conjunctivitis, hemolytic strains of Staphylococcus aureus were found to predominate in 14 of the 30 cases. Smears and cultures demonstrated the presence of Corynebacterium xerosis in a pure state in all of the spots of Bitot which occurred in 5 cases. 4. In addition to masses of Vincent's organisms, 64% of the ulcerations of the tongue, gums, or buccal mucosa yielded Streptococcus hemolyticus, and the remaining 36% contained hemolytic strains of Staphylococcus aureus. Following specific therapy with anti-pellagric substances, the bacterial flora of these ulcerations, including the Vincent's organisms, promptly disappeared.

5. A low complement titre exists in acutely deficient patients, and in the subclinical and mild cases the titre is slightly subnormal or normal. Following clinical improvement the complement titre increases. 6. In the whole blood of acutely deficient patients there is a distinct depression in the bactericidal power for Staphylococcus aureus, whereas, only a slight diminution in staphylococcidal power was observed in the blood of subclinical and mild cases of vitamin deficiency.

Forty-five cotton rats were injected with 9 strains of poliomyelitis virus (Flexner M. V., Flexner W. E., Toomey, Philadelphia, Kramer, Harmon, Howitt, Trask, W. E., Australian). The doses injected were the same as those used by Armstrong 1 to produce poliomyelitis, i. e., 0.06 cc intranasally, 0.06 cc intracerebrally and 0.5 cc subcutaneously of a 10% cord brain suspension. After 6 weeks of observation, 11 animals had died—2 after the injection of Harmon's and one each after the injection of the other strains, except Toomey's and Trask's. The cords and brains of those animals that died were made up into 10% suspensions in saline and injected into other animals. None of the latter either died or developed paralysis even after 4 months of observation.

Flexner's M.V. virus strain, which was also our most potent monkey poliomyelitis virus strain and the one most recently passaged by us, was injected into 25 other cotton rats in the same manner as described before. The results were negative.

Previously, it was found that the addition of 10 day colon typhoid paratyphoid (list of organisms used, 2 ) toxic broth filtrate (called “enteric organism filtrate” for convenience) to poliomyelitis virus accelerated the production of the disease when the combination was injected intracerebrally into monkeys. 3 Perhaps such a virus filtrate mixture would produce poliomyelitis in Eastern cotton rats. Ten per cent virus suspensions in enteric organism filtrate were used throughout save where otherwise noted.

Ten animals were injected with the standard dose in enteric organism filtrate in the usual way. Three animals were living after 4 months. The 2 which died of shock on the first day and 2 which died on the sixty-ninth day were discarded.

Whatever vascular cells are affected, the induction of hemorrhagic lesions by the Rous virus indicates extreme susceptibility such as that usually found in the chick. 3 The present results show that adequate treatment of such hosts by serum from adult chickens—presumably through the antibody-like factor there present—causes either a shifting from hemorrhagic into neo-plastic lesions or suppression of all lesions. In other words the treated chicks react to the virus much as adult chickens do. This goes to show that in fowls the age differences in the response to the Rous virus are dependent upon factors in the blood, and confirms the supposition that the relative ineffectiveness of the virus upon the vascular system of the adult is due to the selective protection by the blood of the susceptible cells of the vascular wall whereas the susceptible extra-vascular cells are less protected. Presumably these cells in the chick are still less protected since after an injection of virus, large tumors always arise from them. 2 The present results show that treatment with adult's serum is also effective in protecting these extravascular cells as long as the virus has been injected in the tissues in very small amounts, as presumably happens in the procedure of the intravenous injection. or when the chicks have been prepared for several days before larger amounts of virus were injected intradermally.

The administration of normal human gastric juice to pregnant rats by intraperitoneal injections of concentrated juice and by stomach tube of unconcentrated juice was without effect on the mean red blood cell diameters of the newborn rats.

Since its use for the cultivation of a spirillum of fowls by Levaditi 1 and the fowl-pox virus by Woodruff and Goodpasture, 2 the fertile hen egg has been employed in the cultivation of many viruses. This medium was employed by Cox 3 for the cultivation of Rickettsiae, and has also been utilized in the study of bacteria 4 and protozoa. 5 Morrow, Syverton and their collaborators 6 employed the developing hen egg in the study of Leptospira icterohemorrhagiae, but Bessemans and De Meirsman 7 failed with Treponema pallida. Employing murine-passaged strains, both Oag 8 and Chabaud 9 were successful with Borrelia duttoni.

Owing to the difficulty of cultivating spirochetes of relapsing fever in laboratory media, we have attempted to grow these organisms in the developing hen egg. In this communication we are reporting successful cultivation of Texas relapsing fever spirochetes in this medium with blood from each of four patients. Specimens were received in the routine mail and after Giemsa preparations of the clots were examined, each specimen was divided in two portions. The first portions of the 4 inocula were introduced intraperitoneally in rats without delay, while an additional day of delay was incurred by sterility testing of the second portions prior to the inoculation of fertile eggs. Meanwhile, previously incubated white leghorn eggs were candled and scrubbed clean with warm soap and water.

For the first inoculum 10-day-old eggs were selected and the chorio-allantoic membranes were exposed by means of a modified dental drill, for the window method of study. 2 On each of 8 chorio-allantoic membranes was dropped 0.2 cc of the specimen and the membranes were observed through the window daily for possible changes or lesions.

These facts definitely prove in our minds that the bloody tears contain no erythrocytes, and the red pigment in chromo-dacryorrhesis is not hemoglobin. The minute pink corpuscular bodies are in reality colorless fatty globules, tinged red by the pigment that surrounds them. In other words, chromodacryorrhesis is not a dia-pedesis of erythrocytes, and the tears do not even contain hemoglobin. It is really a most surprising fact for those who are familiar with the phenomenon of chromodacryorrhea that such obviously bloody fluid coming out of mammalian tissue does not contain any blood.

On the other hand, these findings now seem to help explain many puzzling facts. One of us (with Stix) 1 found that dacryorrhetin causes bloody tear production only in rats (with occasional exceptions with mice), milky tears in guinea pigs, rabbits and mice, and clear tears in cats and dogs. Just why rats alone should shed bloody tears has been one of the perplexing problems. On the basis of the above findings, we can safely attribute the cause of chromodacryor-rhea to a peculiarity of rats in respect to Harder's glands.

Derrien and Turchini 6 describe a substance, a porphyrine, in Har-der's glands of rats and in lesser amount in those of mice. One wonders if this porphyrine is identical with the bloody pigments of the tears, or gives rise to it.

Preliminary experiments (by B) suggest that these two are probably identical as judged by ultraviolet fluorescence and the solubility characteristics of pigments as well as of their salts. Just exactly what the chemical nature of this pigment in the tears is and what the function of dacryorrhesis is, is another problem.

Strains of N. gonorrhoeae preserved by sterile mineral oil seals have been shown to be viable after transfer at 6 and 12 month intervals.

A method is presented for determination by the use of the photoelectric colorimeter of red cell volume of rat blood on a single drop of blood obtained from the tail vein. Hemoglobin content may be determined on the same sample.

Technical assistance of Dorrance Chase Deston is gratefully acknowledged.

By a modification of McCuIlagh's 2 method for extracting androgen from urine we have been able to improve the method of extracting specific substances from the urine of leucemia patients. These substances give specific cellular reactions in guinea pigs. We are now testing saponifiable and non-saponifiable fractions of the chloroform residue which have been prepared by Hause.

Circling to one or the other side has been observed in 13 animals with lesions in the medulla and pons. Lesions in Deiters' and in the descending vestibular nucleus produced circling toward the side of the lesion, while those in the medial vestibular nucleus, in the medial reticular formation, and in the medial longitudinal fasciculus resulted in circling toward the opposite side. Since the connections between the vestibular mechanism and the motor ocular nuclei and possibly, those between the former and some postural centers were interrupted, these observations are reported as confirmatory of the hypothesis put forward recently by Lurie and Dempsey to explain the circling observed in their experimental guinea pigs.

These findings indicate that exposure to high environmental temperature does not influence the digestion and absorption of carbohydrate in the dog, as far as the blood sugar content is concerned. Moreover, no relation is apparent between the blood sugar level and increased environmental temperature.

Fasting dogs normally excrete a urinary substance that appears to be iderctical with Vitamin C. This substance reduces 2:6 dichlorphenol indophenol and gives the color reaction used in the estimation of the vitamin. Moreover, it was established in these investigations that the reducing substance further resembles vitamin C in that it is not precipitated by mercuric acetate and is destroyed by incubation with the pressed juice of cauliflower. Dogs given a normal laboratory diet excreted increased amounts of substances that reduced the indophenol, but most of these could be removed from the urine by precipitation with mercuric acetate. After the dogs had been put on a fast, the excretion of reducing substance decreased over a period of about 4 days and thereafter the elimination remained constant throughout the period of fasting which usually was from 21 to 30 days.

The biological identity of the reducing substance excreted by the fasting dog has not been established, but Musulin, Tully, Longenecker and King 1 have shown that the reducing substance excreted in the urine by rats has antiscorbutic properties.

Experinzental. Female dogs were fasted for a preliminary control period of about 10 days and for the duration of the experiment but were allowed free access to drinking water. The animals were kept in metabolism cages permitting quantitative collection of all urine in dark colored bottles containing acetic acid. At the end of each 24-hour period the dogs were catheterized.

Hepatic injury was induced by 3 different methods. One method was the induction of deep chloroform anesthesia by inhalation for one hour, and another was the administration of carbon tetrachloride by stomach tube.

In a previous report 1 the excretion of vitamin C by the fasting dog was discussed, and it was shown that hepatic injury can increase the urinary output of the vitamin. In the following experiments, the effect of severe shivering, epinephrine, and iodoacetate on the excretion of various urinary substances was investigated.

Experimental. The technic was similar to that described previously. 1

Shivering was induced by anesthetizing the animals with intravenous nembutal and then placing them in a refrigerator at 12°C. In most instances the body temperature fell to around 31 °C. Although additional nembutal kept the animals in an unconscious state, severe and almost constant shivering occurred.

Observations. Data in typical experiments are shown in Table I.

Shivering produced considerable increase in the excretion of vitamin C and creatine, an increase in urinary nitrogen and urea and a decrease in creatinine.† Following periods of shivering lasting from 24 to 72 hours, the animals developed rigid paralysis of the extremities which persisted until food was given. One dog that was not fed had severe stiffness and paralysis of the extremities for 7 days when it died apparently with weakness of the muscles of respiration. In all animals muscular paralysis was accompanied by large output of urinary creatine and vitamin C.

The effect of nembutal in animals kept at warm room temperatures was determined. Complete anesthesia associated with slight shivering induced definite increase in urinary nitrogen but only a slight increase in vitamin C excretion and was without effect on the creatine.

Iodoacetate. Because of the similarity between the rigid paralysis induced by severe shivering and that seen in animals poisoned with iodoacetate, the effect of iodoacetate was studied. The data presented in Table I show that the administration of iodoacetate induced definite creatinuria but did not change the output of vitamin C.

The method of typing various strains of Bacterium typhosum, as reported by Craigie and Yen, 1 has received neither the recognition nor the widespread application that its useful possibilities seem to justify. The survey herein reported was undertaken in an effort to bring the advantages of this method to the attention of those most likely to benefit by its use.

The typing method is based on certain peculiarities of a strain of bacteriophage which is specific for the Vi form of B. typhosum. In addition this bacteriophage exhibits a selective affinity for the strain on which it is propagated and for epidemiologically related strains. By selective propagation of this bacteriophage on various strains of B. typhosum, Craigie and Yen 1 have shown that it is possible to divide freshly isolated cultures into a limited number of readily distinguished types, which have been designated by the letter system A to J inclusive, with the exception of type I. Later reports by Craigie 2 , 3 and by Yen 4 have added subtypes of B, D, E, and F and additional studies have brought to light types L and M, which are rarely encountered. 5 Among the typed cultures herein reported, 97.1 % fell into the 6 main groups, A, B, C, D, E, and F.

Cultures for this study were obtained through the cooperation of the directors and technical staffs of the 9 state laboratories concerned, and were typed by the method described in detail in the original report by Craigie and Yen. 1 Table I shows the distribution of the various types by states.

Table II is included to show the distribution of individual patients by states, since in many cases multiple cultures were received from single individuals.

Solid media for the rapid isolation of Bartonella bacilliformis are described. It is shown that this organism requires the × growth factor of blood. No growth was obtained with the v factor alone.

The intravenous galactose tolerance test offers valuable assistance in the differentiation of the parenchymatous or “medical” type from the obstructive or “surgical” type of jaundice.

1. The CD50 (Thayer-Doisy unit) by subcutaneous administration of tetra sodium-2-methyl-1,4-naphthohydroquinone diphosphoric acid ester (C₁₁H₈O₈P₂Na₄ · 6H₂O)⁴ is 1.34 γ. 2. The subcutaneous CD50 of 2-methyl-1,4-naphthoquinone (C₁₁H₈O₂) is 0.64 γ.

3. The ratio of these potencies is 0.48 but on a molecular basis allowing for 6 molecules of water of crystallization the phosphoric ester is 50% more potent than methyl naphthoquinone.

4. Five naphthoquinone derivatives not hitherto reported are shown to be inactive in doses from 200 to 1000 γ.

Although from the standpoint of treating prothrombin deficiencies lethal and sublethal doses of tetra-sodium-2-methyl-1,4-naphthohydroquinone diphosphoric acid ester are without practical significance, it is interesting to note their effects on the central nervous system. It seems likely that most of the peripheral effects observed are the result of central action but complete analysis of the manner of action has not yet been made. The anesthetic employed appears to possess a quantitative influence on the circulatory but not so much on the respiratory action. With ether anesthesia in cats there is less hypertension after substance N-123 than with Alurate anesthesia. The substance had very little influence on respiration in morphinized rabbits, but in cats under Alurate or ether anesthesia there was marked respiratory stimulation. As is true with most substances acting as biological catalysts (enzymes, vitamins, hormones) the safety margin of its antihemorrhagic effect is enormous. All doses are for the crystalline product containing a moisture equivalent of 6 molecules of water.

A decrease of venous pressure follows spinal anesthesia accompanied by intercostal paralyses irrespective of arterial pressure changes. A further decrease accompanies a fall of arterial pressure. No consistent changes accompanied low spinal anesthesia.

Conditioned reactions (CR) to angular acceleration and discrimination of the direction of the accelerated rotation were previously demonstrated (Spiegel and Oppenheimer 1 ). Experiments on laby-rinthectomized dogs indicated that discrimination of direction of rotation in the horizontal plane at low acceleration is an important criterion of the analyzing function of the labyrinth. Since the changes in the electro-corticogram produced by stimulation of the labyrinth (Spiegel. 2 Kornmueller, 3 Price and Spiegel, 4 Gerebetzoff 5 ) suggest that labyrinthine impulses reach the cerebral cortex, it seemed of interest to ascertain to what extent cortical centers participate in the development of these reactions, and the ability of dogs with cortical lesions (2 frontal, 2 parietal and 2 temporal lobectomies) to acquire these CR was studied. One frontal and both parietal lobectomies were performed in one stage, one frontal and both temporal lobectomies in two stages. 14-51 days after the last operation conditioning was started. The dogs were rotated as previously 1 described in clockwise or counterclockwise direction first at slow constant speed which later passed smoothly into accelerated rotation. During the counterclockwise accelerated rotation the unconditioned stimulus (electric shock) was applied, while it was omitted during clockwise rotation. The reactions of the leg and the respiration were recorded. The animals were usually tested every 2nd or 3rd day, but in some cases longer rest periods (up to several weeks) were intercalated.

In both dogs with bilateral ablation of the parietal lobes CR to angular acceleration were obtained in the first test series; discrimination of direction developed on the third day of experimentation in dog No. 506 in which conditioning was started 26 days after bilateral parietal lobectomy and somewhat later in dog No. 486 (inconstant discrimination on the fifth day, constant discrimination on the seventh day); in this dog conditioning was started 17 days after operation.

The role of the extrinsic nerves in the local intestinal response to filling and distention has been studied by subjecting innervated and denervated intestinal segments of unanesthetized dogs to identical distending forces. Distention acts as a stimulus to the smooth muscle of the denervated intestine so that an active pressure increase may be induced by the stretch and passive pressure resulting from a standard amount of filling. The stimulatory effect of filling is not equally evident in the innervated intestine. This result may be interpreted as indicating that the degree of motility observed at the site of a distention in the innervated intestine depends, in part, upon a balance between direct stimulatory effects and reflex inhibitory effects.

Of the numerous analytical methods employed for the determination of barbiturates, those making use of specific color reactions have been most extensively studied. Handorf 1 employed the mur-exide reaction for the detection of veronal, and Zwikker, 2 Bodendorf, 3 Herwick 4 and Koppanyi, et al., 5 , 6 have developed modifications of a color reaction with cobalt salts in conjunction with various bases.

Kozelka and Tatum 7 attributed this color formation to a specific reaction with substances containing one or 2 imide groups. In an investigation of amytal (isoamylethyl barbituric acid) excretion by dogs, using a modification 9 of the reaction designed by Koppanyi et al., 6 employing cobalt acetate and isopropylamine made up in absolute methanol solutions, it was observed in this laboratory, that of the more probable breakdown products of amytal 8 only isoamyl-ethyl acetic acid gave the characteristic color reaction. Since this compound contains no imide grouping, it was apparent in this case that color formation was not due to such a group.

This finding led to an examination of various classes of compounds a number of which have not been reported upon by other investigators. A partial listing is given in Table 1.

Since the acids tested gave a color apparently identical with that obtained with amytal, a number of these test solutions were submitted to spectrophotometric analysis.

The spectrophotometric curves given herewith were made with a Bausch and Lomb medium quartz spectrograph and sector photometer. The length of the absorption cells was 5 cm. To cobalt acetate and isopropylamine mixtures were added respectively, solutions of amytal, isoamyl ethyl acetic acid (RRC₂H₂O₂) and stearic acid in amounts sufficient to give an intensity of color suitable for measurement.

In 15 cases of induced hyperpyrexia, there were no changes in the potassium content of the blood which would indicate that an increase in the amount of potassium is a factor in shock. The blood potassium level tends to vary directly with the degree of hydration of the blood. There were no significant changes in the Na, Ca, or blood sugar levels, produced by this condition.

The reactions of the guinea pig ileum to the toxic substance indicate that the latter is not histamine. It is, however, inactivated by histaminase suggesting that histaminase is not specific for histamine but may attack other substances. 2 Zeller has shown that histaminase is capable of hydrolyzing several diamine compounds other than histamine. 5

The toxic substance is destroyed by mild treatment with acid or alkali. However, heating at temperatures sufficient to denature proteins present failed to effect complete destruction of the toxicity. Treatment of the extract with both boiled and unboiled solutions of pancreatin discloses no destruction of the toxic substance by enzyme action.

A method for the determination of diodrast and inorganic iodide iodine in blood and urine was presented previously. 1 The principles involved were acid permanganate digestion of the sample with oxidation of iodine to iodate, nitrite reduction of the permanganate, destruction of excess nitrite with urea, and titration of the iodate with thiosulphate in an excess of potassium iodide. This procedure could be carried out either by hand heating of the individual tubes over a micro burner or by heating a number of tubes in a boiling water bath. The digestion of individual samples by hand yields accurate results but requires the constant attention of the analyst. The water bath digestion, however, involves some error, particularly in the digestion of plasma filtrates at high iodine levels, and it is sometimes more difficult to remove all of the permanganate because some manganese dioxide adheres firmly to the walls of the tubes. It is shown here that with an alkaline permanganate digestion these difficulties with the water bath heating are eliminated.

With an alkaline digestion more permanganate must be used, and a longer heating period allowed than with the acid digestion. In order to keep down the volume of the digest at time of nitrite treatment and at subsequent titration, preliminary evaporation of filtrate samples is carried out.

Procedure. Reagents: 13% trichloracetic acid (A.R.), 72% sodium hydroxide (A.R.), approximately 0.4 M potassium permanganate, 4 N sulfuric acid, 1.0 M sodium nitrite, 5 M urea, granulated potassium iodide, 1 % starch, 0.0005 N sodium thiosulfate or 0.0025 N sodium thiosulfate.

Apparatus: circular test-tube rack, water bath, 18×150 mm test tubes, dropping bottles, burette calibrated to 0.01 cc or 0.05 cc.

In preparing the potassium permanganate make a 7% solution, boil for 5 to 10 minutes and let stand for at least 24 hours.

During the first 2-4 days of fasting the degree of ketosis in the human and the rat as measured by the blood acetone body concentration gradually increased to a level of 15-20 mg % (acetone) which then tended to be maintained during the rest of the 10-day fasting period.

The lungs of mice infected with the virus of epidemic influenza contain a soluble complement-fixing antigen which is separable from the virus. 1 , 2 The purpose of the work here reported was to investigate the conditions under which this antigen and its corresponding antibody are formed.

Mice were inoculated with the mouse passage strain PR8 of epidemic influenza virus in dilutions from 10^-2 to 10^-3 of lung. At different intervals of time from 1 to 10 days, 6 to 8 mice receiving each of the dilutions were killed by bleeding from the heart under chloroform anesthesia. The complement-fixing antigen was measured by titration of a saline suspension of the ground lung material against a constant dilution of 1:20 of a pool lof human convalescent serum according to the complement-fixation method previously described. 3

The results are recorded in the accompanying table. One day after inoculation none of the mice showed macroscopically visible lung lesions, but the complement-fixing antigen was already present in relatively high titer except in the group receiving a dilution of 10^-6. Between the second and fifth days the appearance of antigen at a maximum titer preceded by 2 to 3 days the maximum development of lung lesions. This suggests that the formation of the antigen is associated with a rapid multiplication of the virus before the appearance of the red pulmonary consolidation. With virus dilutions of 10^-3 or above, the maximum titer of antigen seemed to be related to the amount of virus inoculated. In the lungs of mice inoculated with dilutions 10^-5 and 10^-8 and killed after 4 days, the antigen titered 1 :40 and 1 :20 respectively and no macroscopic lesions were found.

Whole blood and plasma ascorbic acid determinations made on 70 patients in the nutrition clinic of the Hillman Hospital indicate that 55 of these persons had a whole blood concentration of ascorbic acid lower than in normal controls. In 8 of the 70 cases, the concentration was so low as to suggest that depletion may have been advanced. Most of these patients had clinical evidence of other deficiencies, such as pellagra, beriberi and riboflavin deficiency, but no significant correlation could be made between the blood ascorbic acid values and the symptoms of these deficiency states, nor indeed of scurvy itself. These observations seem pertinent in view of the fact that they were made in late June, at a period when leafy vegetables and berries had been available to the patients for some time, and probably indicate that ascorbic acid deficiency is still greater at other periods. These studies give strong support to the concept that natural-occurring deficiency diseases exist as complexities rather than as single entities.

Degenerative lesions of liver and kidneys following administration of sulfamethylthiazole are explained as a consequence of the damaging effect of high concentrations of the compound in the body. The accumulation of these toxic concentrations is believed to be due to severe impairment of the renal excretory function caused by precipitation of the acetyl sulfamethylthiazole in the collecting tubules of the kidneys.

The study of the nasogenital relationship has revealed the production of pseudopregnancy in the female rat following silver nitrate applications to the nasal mucosa 1 and following local anesthetization of the nasal mucosa. 2 The bilateral removal of the sphenopalatine ganglia in adult female rats is followed by pseudopregnancy. This condition is characterized by the persistence of leucocyte cell smears of the vaginal contents, by persistent corpora lutea in the ovary, and by decidual cell reactions to traumatizing of the uterine mucosa. The duration of this state ranges from 8 to 24 days, after which time the regularity of the 4-5 day cycle is resumed. 3

The present report embodies findings from the study of certain sexual functions in female rats which have been deprived of their sphenopalatine ganglia. Series I: The time of vaginal opening in immature rats from which the sphenopalatine ganglia had been removed, was studied in 30 animals. The ganglia were removed from rats ranging in age from 23 to 40 days. This is considerably before the normal time for establishment of the vaginal orifice. Fifty-six littermate controls were given the same care and handling as were the experimental animals, except for the surgical removal of the ganglia. The findings relative to the age and weight of the rats at the time of establishment of the vaginal opening reveal no significant difference in time of vaginal opening between normal and operated animals; mean age and body weight of experimental animals was 54.2±7.8 days and 99.7±14.8 g, and for the control animals the mean age and body weight was 51.8±6.1 days and 98.9±8.2 g.

Series II: The reproductive capacity of rats without sphenopalatine ganglia was tested in 17 cases by matings with virile males.

Bile acids facilitate the absorption of cholesterol, 1 certain of the fat soluble vitamins, 2-4 and certain drugs 5-7 from the intestinal tract. The similarity in chemical structure between male sex hormones and some of these substances suggests that the administration of bile acids by mouth may enhance the oral activity of the male sex hormone. Such a procedure has been used in the human 8-10 and in the monkey. 11 The studies in the human were such that evaluation of the response to therapy was based largely on subjective factors. The evidence in the monkey was based on the fact that one monkey failed to show postcastration bleeding when treated with testosterone propionate and bile acids by mouth. The purpose of the present study, therefore, was to investigate this question by using a well-established assay method.

Testosterone propionate was the hormone used in the previously quoted reports, and also in this study. This substance is not particularly active by mouth when compared with methyl testosterone, however, it does possess slight oral potency. 12 Desoxycholic acid was the bile acid chosen for this study. This substance has been shown to be the most effective in promoting intestinal absorption of the above mentioned compounds. 1 , 2 , 6

Male albino rats of 125-150 g in weight were castrated, and oral administration of the various substances was begun on the day after castration. The animals were divided into 4 groups: (1) those receiving 0.1 mg of testosterone propionate per day, (2) those receiving 0.1 mg of testosterone propionate plus 0.5 mg of desoxycholic acid per day, (3) those receiving 1.0 mg of testosterone propionate per day, and (4) those receiving 1.0 mg of testosterone propionate plus 0.5 mg of desoxycholic acid per day.

Under partial starvation some aerobic and facultative anerobic bacteria, yeasts, and fungi grow only in a fine, dense layer below the surface of the shake-cultures where the relation between nutrient-content and oxygen is an optimum. Under increasing starvation bacteria become increasingly sensitive against oxygen. Oxygen causes a bacteriostatic effect, the rate of which is in relation to the nutrient content of the medium. The less the medium contains of carbon compounds which may act as oxygen acceptor, the more inhibitory is the action of oxygen on the organisms. Variation in temperature below and above the optimal temperature and exposure to light increases the sensibility against oxygen.

The nutritional myopathy of ducklings which develops on vitamin E deficient diet is completely prevented through the experimental period by daily administration of 4 mg of synthetic α-tocopherol. A dosage of 1 mg daily does not afford protection.†

1. Fractionation of aqueous grass-pollen extracts with water-miscible organic liquids promotes purification of these extracts. 2. Precipitation of extracts purified by the above treatment with high concentrations of sulfates affords further purification.

Gramicidin, an alcohol-soluble, water-insoluble substance extracted from cultures of an aerobic sporulating bacillus, has been found to exert a marked bactericidal effect against Gram positive microorganisms, both in vitro and in vivo. 1 When injected by way of the cistern into infected quarters of cows suffering from mastitis caused by Streptococcus agalactiae (Lancefield serological group B), gramicidin apparently cured a number of the infections. 2

The method originally used for the administration of gramicidin in the treatment of bovine mastitis consisted in diluting 60-240 mg of the bactericidal substance in 1000 cc of aqueous media and injecting this material into the cistern of the infected quarter. As reported earlier, this method of administration caused an intense swelling of the treated quarter, accompanied by a marked elevation of temperature. Although this reaction lasted only a few hours, it was severe enough to prevent repeated treatments on successive days.

It has now been found that sterile mineral oil is a suitable, non-irritating vehicle for the administration of gramicidin. Of 42 quarters treated with gramicidin-oil mixtures, only 4, which had received 120-160 mg of bactericidal substance, exhibited a severe reaction. On the second and third day following treatment, the fore milk in 3 of these quarters was tinged with blood, whereas the secretion of milk was suppressed for 4 days in a fourth quarter. The reaction in the remaining 38 quarters was milder than in those injected with gramicidin in water.

On the basis of present experience, the following technique of treatment appears the most satisfactory. 2-3 cc of an alcoholic solution of gramicidin, containing 40 mg per cc, are emulsified in 15 cc of sterile double distilled water and the emulsion mixed thoroughly in 25 cc of heavy mineral oil.

The rapid production (within 10 minutes) of lactic acid in the oral cavity and in vitro is demonstrated as the result of the action of oral microorganisms on certain carbohydrates.

Previous work has disclosed that the arterio-venous oxygen difference of cerebral blood is diminished during insulin hypogly-cemia. 1 , 2 However, the arterio-venous oxygen difference is determined not only by the metabolic rate of the brain, but also by velocity of blood flow. Therefore, thd possibility exists that the smaller oxygen difference found may be due to a more rapid blood flow during hypoglycemia. Observations by Loman and Myerson 3 on human subjects and Leibel and Hall 4 on rabbits, have failed to demonstrate any marked changes in the rate of cerebral blood flow during insulin hypoglycemia uncomplicated by convulsions. However, for conclusive information regarding changes of brain metabolism, it is necessary to determine, simultaneously, the cerebral blood flow and the arterio-venous oxygen difference. The results of such a study on patients with schizophrenia are presented in this preliminary report.

Methods for the collection and analysis of the blood samples have been previously described. 2 The rate of blood flow in the internal jugular vein was estimated by a modification of the Gibbs thermostromuhr. This instrument proved to be at least as sensitive as the original and will be described elsewhere. Observations on the blood flow were made over long periods during which the position of the thermostromuhr in the vein was maintained constant. A special technique was devised which prevented error resulting from the formation of clots on the needle.

In 5 cases the blood flow and the arterio-venous differences were determined before injection of insulin as well as throughout the course of the coma and the subsequent administration of glucose. In 3 other instances the observations were begun during coma and continued until after arousal with intravenous glucose.

The magnitude of the blood pressure rises given by large doses of renin must be corrected for differences in starting pressure levels, while those given by smaller doses need not be. An assay based on repeated tests on 2 or 3 dogs is as reliable as one based on a larger series of animals. Nembutal anesthesia may affect, in some dogs, the magnitude of the rise after large renin doses, but usually has little effect on the rise after small doses. For most routine testing, the anesthetized dog is the preferable test animal.

In an earlier research upon neostriatal function' the present authors observed that “It is difficult to escape the suspicion that animals (cats) with neostriatal injuries suffer from some variety of vestibular disturbance”; the literature relating to this observation was reviewed and it was noticed that Bergouignan and Verger' found that the ipsilateral circus movement (dogs) produced by unilateral caudate lesion was accentuated by rotation to the same side while rotation to the opposite side gave no circus movement. Cocainization of the labyrinth of either side always produced an accentuation of the original circus movement, never one toward the opposite side. 2

In the further work of Bergouignan and Verger there is, so far as we are aware, no conclusive evidence to indicate whether these results are due to disregard of the labyrinth of the same side or over-emphasis of labyrinthine stimuli from the opposite side 3 In order to arrive at some information on this point a series of animals were prepared in which (1) both frontal regions exclusive of the caudate nuclei were removed; (2) both frontal regions inclusive of the heads Qf the caudate nucl'ei were ablated, and (3) complete, bilateral, labyrinthine destruction had been carried out.

Rotation of animals of the first or second group produced effects of the same type as those seen in normal animals though there was less obvious distress exhibited by either type of operated animal during rotation than was displayed by normal cats. Further, although no part of the usual post-rotation symptomatology was absent or perverted, this did not persist so long in the operated as unoperated animals. These differences cannot be explained upon the basis of postoperative weakness, for the operated cats were in good health and had attained a stable state.

(1) By means of intestinal intubation in man the absorption rate of hydrolyzed and unhydrolyzed protein from the jejunum has been estimated. (2) Most of the nitrogen of a 4 to 5% solution of casein and gelatin is absorbed 40 to 50 minutes after its introduction directly into the small intestine. (3) The nitrogen of an amino acid mixture (hydrolyzed casein) of similar concentration is fairly completely absorbed 15 to 25 minutes after its administration in the same manner.

We have reported here the failure to infect a great horned owl (Bubo virginianus virginianus) with the sporozoites of a strain of Plasmodium cathemerium of known infectivity and virulence.

Ascorbic acid, in the concentrations studied (up to 10 mg%), markedly increased the tone of the isolated guinea pig colon. This experimental observation is in keeping with the clinical impression that ascorbic acid administration may occasionally produce colicky pains of intestinal origin.

Rats receiving only our basal ration J36 showed an hemorrhagic cortical necrosis of the adrenals after 4-6 weeks. This condition was prevented by calcium pantothenate. Choline did not prevent but rather aggravated the condition. Acid autoclaved liver extract prevented the condition, but its action may have been due to its pantothenic acid content.