Experimental Biology and Medicine

Methods are described for the determination of chymotrypsin and rennin inhibitors in serum. The many physiological and pathological conditions which influence their concentration are discussed.

The close agreement obtained for the values of total body water by the use of the antipyrine method and desiccation, indicates the validity of this procedure for the purpose of measuring total body water.

1. By stenosis of the thoracic aorta in dogs, arterial hypertension can be produced above and below the stenosis.

2. This type of experimental hypertension has many of the characteristics observed in the hypertension occurring in coarctation of the aorta in man.

1. The blood volume of normal rats determined by the use of Fe⁵⁹ labeled red blood cells is essentially the same as with P³² labeled cells.

2. The use of P³² labelled red blood cells is hence satisfactory for the determination of blood volume.

3. Determinations with P³² indicate that the blood volume of the normal rat is 4.59 ± 0.57/cc 100 g body weight; the total red cell volume 2.16 ± 0.20 cc/100 g body weight; and the hematocrit 45.8 ± 2.6.

Observations on 18 cases of hyperthyroidism are presented. This condition is characterized by significantly increased levels of serum cholinesterase. These values returned toward the normal range with adequate antithyroid therapy. Of 3 patients with nontoxic adenoma of the thyroid, 2 were in the normal range and one showed a high value. No definite relationship was found between serum cholinesterase and basal metabolic rate, total serum protein, albumin, globulin or cephalin-cholesterol flocculation test. Due to the comparatively small series no conclusion can be drawn concerning sex differences in serum cholinesterase.

The average xanthine oxidase activities in CmmO₂/g dry weight/hour for adult rat tissues were: Liver 1862, small intestine 628, spleen 534, lung 479, kidney 135, stomach 42, skin 100-200, brain O, muscle O, testes O. Two-thirds of the total activity in the entire rat was in the liver.

Methylene blue added to the Warburg flask in the aerobic determination increased the xanthine oxidase activity in liver, kidney, stomach and small intestine, but not in lung or spleen. Added xanthine oxidase was recovered quantitatively in the presence of lung, spleen and kidney; greater than added amounts of activity were recovered from liver and decreased amounts from small intestine. As the liver xanthine oxidase was depleted by a purified low protein diet, the small intestine lost about 2/3 of its activity, and lung lost about 1/2. Losses from the spleen and kidney were small. The entire rat lost 3/4 or more of its xanthine oxidase activity.

1. Data are given showing the relationship between protein osmotic pressure and density in plasma from normal humans, cats, and dogs. Formulae are derived which allow the prediction of protein osmotic pressure from density in each of the three species (Equations 5 and 7). 2. For any given density human plasma has a higher protein osmotic pressure than does plasma from cats or dogs. Possible reasons for this species difference are discussed.

Mercuhydrin, salyrgan and theophyllin, in diuretic doses, do not interfere with the renal tubular transfer of glucose or p-aminohippurate in the dog.

Subcutaneous injection of manganese chloride into the guinea pig caused manganese to accumulate in all organs of the body tested, but the deposition was especially great in the thyroid gland where it began to appear within 30 minutes after administration. Absorption was particularly rapid during the first few hours, but the element continued to accumulate for at least 24 hours. Of the total amount of manganese deposited in the thyroid gland about 70%percnt; accumulated during the first 6 hours. Elimination of manganese from the thyroid, once it began, was rapid. Seventy per cent of the 24 hour accumulation disappeared within 48 hours after the injection; 85%percnt; of it vanished in 96 hours.

The theory that the acetyl-choline-cholinesterase system is concerned with membrane permeability is further substantiated by the finding that the L-isomer of isoamidone which has a greater inhibitory action on cholinesterase than has the D-isomer also changes permeability of dog erythrocytes to a different degree than does the D-isomer.

The conditions under which the L has a greater effect than the D-isomer on permeability are a medium containing a high proportion of K ions relative to Na ions.

1. The incidence and severity of renal injury in weanling rats fed diets low in choline and methionine were markedly decreased by supplementing the diet with a vitamin B₁₂ concentrate or crystalline vitamin B₁₂.

2. Under the conditions of these experiments, 30 μg of vitamin ₁₂ per kg of diet could replace about one-half of the supplementary choline or methionine required: for protection against kidney damage.

3. When sub-protective levels of choline were fed, the addition of vitamin ₁₂ caused a significant increase in weight gain. However, when an adequate protective level of choline was fed, no increase in weight gain was obtained from the addition of the vitamin.

4. The results established the existence of an interrelationship between vitamin ₁₂, and choline or methionine.

An inquiry has been made into conditions which may influence the viability of 1×1 cm areas of biopsied rabbit skin during refrigeration at 0° and 6-8°C. Since the availability of oxygen, as well as the nature of the storage medium, had an, important influence on the preservation of viability at these two temperatures, these relationships may be of interest to those wishing to store or ship tissues for surgical and other purposes.

Tissues separated from the circulation rapidly become anoxemic and necrotic at room or body temperatures. This condition may be prevented in uterine or intestinal strips by oxygenation or by chilling. 1 , 2 The survival of ligated limbs 3 and of the cells in whole embryos 4 , 5 or organs 5 , 6 is optimal (among the widely spaced temperatures which have been studied) at 0°. At this temperature respiration is minimal, while oxygen solubility in water is twice that at 30°.

Though Lambert 7 Carrel, 8 and Hetherington and Craig 5 found 0 to 7° favorable for preserving the small masses of crowded cells in embryonic tissue fragments, there is considerable evidence that the thin perimeter of migrating and dividing cells in established tissue cultures has maximal longevity around 30° 9 10 and are unable to re-establish growth after refrigeration for a few days. 9 , 11 , 13 Upon considering the fact that large tissues are killed more rapidly at higher temperatures and small groups of thinly spread cells at low temperatures, it seemed not unlikely that one of the common denominators might be a question of oxygen supply and demand.

Methods. After shaving and scrubbing, the chosen area of rabbit skin was covered for 2 minutes with a wet pack of tincture of iodine diluted 1:2 in water, and again scrubbed with alcohol. The strips of biopsied skin, either full- or half-skin depth were cut into 1×1 cm pieces.

It is frequently stated that in the strict bacteriological sense it is impossible to sterilize skin. This concept apparently arises from the use of methods which do not insure that an appropriate disinfectant is applied in active form for an adequate interval of time. Since the rich and; complex nutrients used for cell cultivation provide an excellent pabulum for a variety of microorganisms, long experience in sterilization of leprous and normal skin prior to biopsy for tissue culture induces us to present evidence that skin sterilization is not a difficult problem.

Early attempts to sterilize well scrubbed skin by painting with tincture of iodine were at times successful and at times failures. The inconstancy of the results suggested the desirability of controlling the action of iodine with respect to concentration, and time of action. By covering the area to be biopsied with a heavy patch of fabric, dripping iodine solution (tincture of iodine 1:2 in 70%percnt; alcohol by weight) on this patch until it was saturated, and then removing the patch and recleaning with alcohol after an interval of 5 minutes, the occurrence of scattered contamination was terminated. During the next several years it was noted that epithelial cells rarely if ever migrated from fragments ex-planted from the papillary or uppermost layer of the skin, while abundant epithelial cells appeared around fragments explanted from the deeper or reticular layer of the skin.

Upon the first occasion when a biopsy was taken after painting the skin with 2%percnt; Mercurochrome in Duponol as a wetting agent, good epithelial growth around papillary ex-plants appeared for the first time in our experience. Nevertheless, one after another of the culture tubes gave evidence of contamination by staphylococci until 40%percnt; of the cultures had been lost. The period of applying the iodine (1:2) patch was next shortened to 2 minutes, without trouble from contamination and with the result that the superficial layer of skin produced moderate to excellent epithelial migration.

1. Vit. B₁₂ was fed to Leghorn chicks receiving a choline-low basal diet supplemented with .6%percnt;, .2%percnt;, .1%percnt;, and .05%percnt; choline chloride. The 14-day increment in weight gain over the controls that did not receive vitamin B₁₂ was 26 g, 32 g, 63 g, and 21 g, respectively.

2. Crystalline vit. B₁₂ fed at a level of 15 μg per kg of diet and at a choline level of .1%percnt; was as effective as vit. B₁₂ concentrate in a 2-week feeding period.

3. The choline requirement of chicks under the conditions of these experiments was markedly reduced by the supplementation of the diet with vit. B₁₂.

4. It appears that dietary choline has a significant sparing action on vit. B₁₂.

1. An oxygenator consisting of 8 concentric revolving cylinders which utilizes a funnel to deliver the blood into a small cup is described.

2. Animal perfusions demonstrate that its efficiency as an oxygenator is high enough to make it possible to perfuse the entire animal body without using an impractically large amount of blood in extra-corporeal circulation.

Saline suspensions of stools obtained from both normal and poliomyelitis patients showed hemagglutination of group “O” human, chick, rabbit, guinea pig and sheep erythrocytes. The rabbit cells were agglutinated in highest titers. No significant difference in hemagglutination titer was noted with the normal and poliomyelitis stools. Normal human, rabbit and monkey serum and 20%percnt; bovine albumin inhibited hemagglutination. The hemagglutinin was thermolabile and absorbed by group “O” human, chick and rabbit erythrocytes. Stool suspension Seitz nitrates failed to show hemagglutination, but the titer could be partially restored by methanol precipitation. Various species of bacteria isolated from stool suspensions failed to agglutinate chick, group “O” human and guinea pig erythrocytes but showed some agglutination of rabbit cells in low titer. Mouse brain and cord suspensions infected with the Lansing strain of poliomyelitis virus also failed to agglutinate chick, human group “O” and rabbit erythrocytes.

Growth was markedly reduced in hypothyroid rats fed purified rations containing casein as the dietary protein and sucrose as the dietary carbohydrate. The retardation in growth was completely counteracted by the administration of a water-insoluble fraction of liver. Crystalline vitamin B₁₂ was ineffective. The protective factor in liver is distinct from any of the known nutrients including vitamin B₁₂.

Partial ligation of the portal vein resulted in the development of a small amount of ascites in 69%percnt; of a group of 110s rats. All rats showed an increase in portal pressure but the degree of increase did not correlate well with the amount of ascites.

The LD₅₀ for total body X-radiation in Wistar rats is 640 ± 5 r with a 30-day endpoint. The time elapsing before 50%percnt; mortality occurs at different doses is a more reliable measure of mortality than the dosage-mortality curve or the dosage-survival time curves for different X-ray doses. The dosage-mortality curve of rats has such a steep slope in the LD₅₀ range that variations in mortality of from O to nearly 100%percnt; may occur in this range because of variations in animals and in the X-ray machine output. Therefore in studies directed toward (therapy or protection of animals given total body ionizing radiation, if the mortality curves are extremely steep as in the rat, dosages in the LD₅₀ range should not be used. Other measures which might be employed are discussed.

1. A strain of swine influenza virus (Oti), isolated in 1939, has been re-investigated by serological means and found antigenically to resemble some members of the human A-prime influenza virus group.

2. Significant increases in the levels of anti-body against this swine strain of influenza virus were detected in the convalescent blood specimens of children who had had experience during the A-prime influenza epidemic of early 1947. High titers of antibody were also found against the current A-prime strains.

3. Adults involved in the same epidemic also showed significant antibody increases against the same swine strain and the current A-prime and Type A strains.

4. Results of cross hemagtglutination inhibition tests with specific ferret anti-sera supported the observations made from the serological tests with human sera.

5. The relationship of this swine strain to members of the A-prime group has been discussed.

Of the several compounds tested, only 1-arterenol has a hyperglycemic action of the same order as that of 1-epinephrine. Its activity can be placed at about 1/8 of that of epinephrine, while the d-isomer has about 1/20 of the activity of the l-isomer. Sahyun¹⁴ found that dl-arterenol, 1 mg/kg, produced an average rise iin blood sugar of 78 mg%percnt; 3 hrs after subcutaneous injection in 7 rabbits. This is considerably more activity than would be calculated from our observations on the separate isomers. However, if one of his rabbits (No. 8, Table II) is eliminated from consideration, the average rise becomes only 61 mg%percnt;, and the discrepancy is much less. Of the other compounds studied, Isuprel is clearly the least active. The approximate hyperglycemic activity of the whole series of compounds, taking l-epinephrine as 100, is as follows (all figures given have been calculated in terms in terms of the free base): l-arterenol, 12; Win 3046, 1.7; Butanefrine, 1.6; Win 515, 0.7; d-arterenol, 0.6; and Isuprel, 0.12.

D. granulomatis shows delayed fermentation of a few carbohydrates, chiefly monosaccharides, without gas formation. In this respect it resembles K. rhinoscleromatis. Its sensitivity is high against all the pure streptomycins and cases of granuloma inguinale should react as favorably to treatment with dihydrostreptomycin as to treatment with streptomycin itself.

The number of circulating eosinophiles of trichinous mice was decidedly reduced by the administration of cortical hormone. The same effect was induced by epinephrine in intact, but not in adrenalectomized, trichinous mice. Sex steroids in the doses used did not significantly reduce the percentage of circulating eosinophiles in 4 intact and 2 adrenalectomized trichinous animals. Eosinopoenia was induced by urethane in trichinous mice. Whether or not urethane-induced eosinopoenia is mediated by the adrenal cortex has not been determined.

Vitamin B₁₂ has been demonstrated to have a role in the growth of Rous sarcoma in chickens comparable to that previously shown for pteroylglutamic acid.

A method is described for the adsorption of gonadotrophic hormones from urine onto kaolin and their subsequent elution with ammonium hydroxide. The relatively small volume of the eluate reduces the amount of alcohol needed to precipitate the hormone from solution, the final extracts are less toxic and can be prepared in a shorter time. The hormone assays are comparable to those obtained with the usual alcohol recipitation method.

Evidence is presented that the original culture of S. griseus which was isolated from different soils, in 1915-1916, in the laboratories of the New Jersey Agricultural Experiment Station must have possessed the capacity for producing streptomycin. This capacity was lost upon continuous growth of the culture upon artificial media for more than 30 years. When irradiated, this culture yielded a mutant that was as potent a producer of streptomycin as the streptomycin-yielding strains of S. griseus isolated in 1943.

This assumption was fully confirmed by the characteristic antibiotic spectrum of the two cultures, their behavior to streptomycin-dependent and streptomycin-resistant mutants of E. coli and other bacteria, and sensitivity of the two cultures to the same actinophage.

These results further point to the accuracy of the identification of the streptomycin-producing cultures of S. griseus with that of the original 1915 isolate, based on literature description alone, in spite of the fact that the type culture available in two collection's showed distinct changes from the original description.

1. Tetraethylammonium bromide blocks the release of epinephrine from the adrenal medulla.

Following doses of radioactive iodine sufficient to cause destruction of most or all of the thyroid, tumorous enlargements were found in the pituitary glands and tracheas of mice. Hypophyseal growths as large as 240 milligrams (control, 2.1 mg) were found. Less than 10%percnt; of animals sacrificed more than 250 days after I¹³¹ administration had pituitaries smaller than 10 mg in weight.

Tracheal tumors of the fibrous tunica propria, obstructing the tracheal lumen, were of lower incidence (4%percnt;) but appeared earlier. Fibrous thickenings of the tunica propria, and metaplastic regeneration of tracheal epithelium were more common.

The shape of the preweaning growth curve of the rat is profoundly affected by the lack or availability of solid food. The divergence from conformity to Zucker's growth equation of rats raised on the Anderson and Smith diet may be due to the natural transition from a liquid (milk) to a solid diet and does not appear to be explained by a qualitative deficiency in this diet.

Precipitation occurs in solutions of egg-white or purified egg-white inhibitor of virus hemagglutination submitted to an electric field. The precipitate includes a great part of the inhibitor of the preparations in a state substantially purer than that of the starting material. This result suggests application of the phenomenon to the final purification of the inhibitor.

Studies are reported in which the Lederle stock strain of canine distemper virus was adapted and maintained through 74 serial passages on the chorio-allantoic membranes of developing chick embryos. The chick embryo adapted strain retained its capacity to produce lethal infection in ferrets through the 24th egg passage level but apparently lost its ability to do so between the 24th and 28th egg passages. Ferrets inoculated with chick embryo adapted virus of passage levels higher than the 28th transfer failed to show any signs of disease, but were found immune on rechallenge with fully virulent ferret spleen virus. Cross neutralization tests carried out in chick embryos and in ferrets indicate that the chick embryo adapted strain and the parent strain of ferret spleen virus are immunologically related.

Parenteral injections of chymo-trypsin had no effect on the course of 101 cases of neoplastic disease in man.

Treatment with crystalline chymotrypsin did not influence the course of neoplastic disease in experimental animals or in fifteen selected cancer patients.

Chloromycetin inhibits the growth of Salmonella organisms in vitro remarkably, but is not able even in large amounts to control Salmonella infections in mice. Neither the oral nor the subcutaneous form of treatment prevented the fatal outcome of the disease in most animals, although it prolonged the survival time. Its action in the animals was bacteriostatic and not bactericidal. Due to its absorption conditions, Chloromycetin exerts no influence on the intestinal flora, when given orally.

The lumbosacral ventral root outflow of the cat contains 20-25 p.c. small diameter nerve fibers. These cause no detectable shortening of muscles and no propagated muscle impulses. Stimulation of such fibers sets up discharges from muscle spindle receptors. The discharge is dependent upon the number and frequency of stimuli and on adjustment of muscle stretch.

Marked species variations in the staphylocoagulation phenomenon are encountered and can be explained in terms of several factors. Comparing with rabbit plasma as an empirical standard (shortest staphylocoagulation times encountered), the ability to shorten staphylocoagulation time by addition of co-factor (human serum albumin) is evidence for lesser amounts of cofactor in such plasmas as horse and bovine. Cofactor cannot be recovered completely or uniformly in plasma fractions obtained with the classical (NH₄)₂ SO₄ salting procedures. Nevertheless, the “albumin” is consistently rich in cofactor (for activation of prostaphylocoagulase) and devoid of thrombin (which interferes in tests on some euglobulins).

Excess of inhibitor (s), in part at least directed against active staphylocoagulase as shown by pre-incubation of prostaphylocoagulase with cofactor before adding to plasma (or plasma dilution + fibrinogen) is a feature of some plasmas, notably rat and chicken. Owing to this, the demonstration of cofactor requires plasma fractionation, in these instances.

Fibrinogenolytic and possibly other proteolytic effects may interfere with tests, especially at higher (37°C) temperatures, and are exemplified in guinea-pig and dog plasmas.

The quantitative evaluation of staphylocoagulation times must, therefore, take cognizance of variability not only in the two bacterial factors but, even more significantly, in the various plasma factors on which they operate in the systems devised for testing the staphylocoagulation and proteolytic 9 phenomena. There are marked species differences in these plasma factors.

Fibrinolytic tests are described which clearly demonstrate differences in the species reactivity of plasmas to the staphylokinase and streptokinase activation of the protease system. Whereas streptokinase acts only on human materials, staphylokinase works on plasma (or serum) of dog, guinea-pig, and rabbit also, but gives negative results with bovine and other species tested. Staphylokinase differs from streptokinase also in requiring an incubation period of many minutes before reaching optimal activation of the serum proenzyme.

A factor has been demonstrated in testis extract which inhibits the growth of a transplanted lymphosarcoma in AKm mice.

This factor is of unknown nature, but appears to be different from the Duran-Reynals spreading factor.

(1) Doses of urethane simulating those used in the topical treatment of infected wounds in man are not productive of toxic parenchymal changes in the organs of guinea pigs subjected to prolonged daily subjcutaneous injections. The animals remained healthy and gained weight.

(2) A marked mesenchymal cells reaction was consistently seen in the lungs and to a lesser extent in the liver and myocardiuim of urethane treated animals.

2,2-Diethyl-l,3-propanediol and certain other substituted 1,3-propanediols possess qualitatively similar pharmacological properties as myanesin. These compounds differ from myanesin in having a weaker paralysing action and a more powerful anticonvulsant action.

Golden hamsters (Cricetus auratus) from two different sources were found to present marked differences in their susceptibility to the Lansing strain of poliomyelitis virus. Both strains were found to be markedly inferior to the white mouse and to the cotton rat as experimental animals for poliomyelitis research.

The adjustment of the inoculum at a low pH did not increase the titer of the hamster passage virus consistently. The use of the autolyzed normal mouse brain technic did not increase the infectivity of the virus for the hamster.

The intraperitoneal inoculations of mouse-passage and cotton-rat passage virus resulted in higher titers of neutralizing antibody in the golden hamster than when hamster passage virus was used.

Mice exposed to 5O-20Or of x-rays can have one or several normal pregnancies and still be liable to the development of ovarian tumors in a stock in which this neoplasm is practically non-existent.

Irradiation at 1-3 days of life with 150r sterilized only 1/3 of the mice while ovarian neoplasms appeared in about 76%percnt;. Irradiation at 1-3 days did not hasten the onset of ovarian neoplasms as compared to irradiation at 4-10 weeks. In both groups the tumors developed in middle aged and old mice. The growth rate of the tumors was likewise slow, hardly interfering with the normal life span of these animals.

It is postulated that a specific delayed x-ray effect coupled with a hormonal imbalance provoked by x-rays leads to the development of ovarian tumors.

The steatorrhea of 3 bile fistula dogs given a fatty meal once daily was nearly completely corrected when the animals own bile was returned to the duodenum every hour. The steatorrhea persisted, however, when bile was returned every 4 or 8 hours. Since the total quantity of cholic acid returned during the 1- and 4-hour regimes was the same, the importance of continuous presence, apart from total daily quantity present, in the intestine is demonstrated.

The influence of pregnancy on caries activity has been studied in hamsters reared from weaning for 100 days on a sub-optimal diet, with their respective litter-mate controls. The results show that pregnancy does not influence caries activity in any direction.

A compound of steroid nature, having the probable formula of C₂₈H₄₆O, has been isolated from cane juice. This compound has therapeutic activity in alleviating an induced stiffness in guinea pigs. It has not been possible to further elucidate the structure of this compound, since not enough material was available at the time the project had: to be discontinued, due to circumstances beyond control of the two senior authors. The isolations, however, tend to confirm the findings by Oleson et at. and Petering et al., in that the antistiffness factor is steroid in nature.

1. Electron microscope studies of vesicular and spinal fluids from a patient with herpes zoster have demonstrated in both particles of similar size averaging 227 mμ in diameter.

2. These particles are compared to those in vesicular fluids from patients with herpes simplex and chicken pox. In herpes simplex particles averaging 213 mμ in diameter were found, and in chicken pox, particles of 245 mμ.

There is present in the plasma of tuberculous humans a heat-labile, non-dialyzable component which is responsible for the in vitro lysis of washed leucocytes by tuberculoprotein. Further properties of this lytic producing substance are its stability at 10°C for at least 7 days and its precipitation with the globulin fraction of the plasma proteins.

Five viruses (Eastern equine encephalomyelitis, MEF human poliomyelitis, mumps, influenza B, and Newcastle disease) were utilized in studying the precipitating effect on them of methanol. Optimal methanol concentrations and optimal pH for elution were determined for each virus and are tabulated (Table I). All of the viruses which were studied demonstrated a relationship between the pH of the elution fluid and solubility: as the pH was raised there was a corresponding increase in solubility of the virus. The methanol precipitation technic results in considerable loss of nitrogen from the virus suspension, but this loss does not appear to be at the expense of the virus.

Infected hamster brains from the 300th intracerebral passage and from the 4th intranasal passage titred lO^-4.77, whereas infected brains from the 7th intranasal passage titred 10^-3.66. Positive Newcastle chicken serum neutralized the virus from the 245th and 300th hamster passage, whereas normal chicken serum had no effect.

Nasal instillation trials were conducted from the 16th and 300th hamster passage but were not successful (except for the 1st passage) until after the 245th passage.

The incubation period of hamsters infected by intranasal instillation is longer than by intracerebral injection. Hamster-adapted Newcastle virus was carried through 9 passages intranasally, and brain material from this 9th nasal passage was neutralized by positive Newcastle chicken serum. Hamsters infected intranasally and intracerebrally showed symptoms of irritability followed by involuntary motor reactions and paralysis.

1. Testicular hyaluronidase in bulk assaying 150 TRU per mg is stable at room temperature for more than one year.

2. Hyaluronidase in concentrated aqueous solution is stable for several months at 10°.C Dilute solutions of the enzyme are less stable.

3. Hyaluronidase dried from the frozen state in vials can be stabilized by the addition of polypeptide. Its half-life at 60°C is thereby extended 10 to 20 times.

4. Certain bacteriostatic agents can be used throughout the production of hyaluronidase without decreasing the activity of the enzyme.

Doses of diethyl-aminoethanol sufficient to produce high blood concentrations resulted in a temporary inhibition of sympathetic vasoconstrictor reflexes in man. It is suggested that the effects of the drug on arterial pressure and ventricular tachycardias are secondary to its sympatholytic action.

WIN 1539, a new potent synthetic analgesic related chemically to Demerol, was found to have a stronger spasmolytic action than Demerol on the stimulated isolated intestinal strip. A decrease in tone of the intact small intestine of the dog was observed, WIN 1539 caused a greater delay than Demerol in the emptying time of the rat's stomach but morphine retarded emptying more than either of these drugs. Demerol caused a stimulation of the intact rat's stomach whereas WIN 1539 and morphine decreased the motility of this organ.

Folic acid whether commercial or aldehyde-free inhibits guanine metabolism and endogenous respiration in rat liver homogenates. Oxidation of folic acid itself in such systems is not clearly evident.

Oxidation of xanthine by a purified preparation of xanthine oxidase is strongly inhibited both by commercial and aldehyde-free folic acid. In a system containing only folic acid and the enzyme some oxidation of the folic acid appears to occur although this reaction is transient.

The antimalarial activity of chlorguanide against Plasmodium gallinaceum in the chick could be significantly but not completely inhibited by pteroylglutamic acid in vivo. Pteroylglutamic acid had no significant effect on the antimalarial activity of quinine, chloroquine or pamaquine, but completely inhibited the effect of sulfadiazine.

Degranulation of beta cells of the pancreatic islets and islet hyperplasia have been observed in, rats which were force-fed large amounts of high carbohydrate, otherwise balanced diets for periods of as long as 8 weeks. Glycosuria and, in rare instances, ketonuria occurred.

The adsorption of calcium phosphate complexes to plasma substrates, and the conditions required for their removal, indicate that the plasma gel is the site of an ionic exchange in which preferentially adsorbed ions may greatly alter the desired ionic balance. This process is one of the factors which causes plasma to become growth inhibitory during prolonged maintenance of cells in this type of substrate.

Eighteen extensively depancreatized male rats and 20 normal rats were force-fed a medium carbohydrate diet. Following periods of adaptation and control, all of the animals were tested for their resistance to twice daily injections of regular insulin. The initial dose was 5 units and was increased in increments of 5 units per dose every second day until the animal died. The average lethal dose for the depancreatized rats was 50.5 units per 24 hours as compared to an average of 68.5 units for the normal animals. This difference in averages was calculated to be statistically significant.

During December 1948, reports of widespread distribution of influenza began to appear in the Seattle area. The majority of illnesses were of relatively short duration and were accompanied in many cases by gastrointestinal disturbances. Proper specimens of throat washings and serum were not obtainable until the return of students to the University of Washington campus after the New Year holidays.

During the first 9 weeks of 1949, there were 116 admissions for upper respiratory infection to the University of Washington Health Center. It was apparent, through random questioning of the campus population, that many others were ill at home. Since approximately 60%percnt; of the 16,000 University students live at home in or close to the City of Se3attle, and there were a number of students known to be ill in living quarters on and off the campus, it seems obvious that the outbreak was much more widespread than the 116 admissions to the Health Center would indicate. A conservative estimate of 500 cases, of varying severity, during the period from the end of December to the first of March does not seem unreasonable. Observations of classroom attendance supported this estimate.

The majority of illnesses were mild, with the average period of hospitalization being between 3 and 4 days. The mildness of the disease further supports the idea that the cases seen in the campus infirmary represented only a relatively small proportion of the total number occurring in the Seattle area.

Materials. Blood specimens and throat washings were obtained during the acute stage from patients in the infirmary whenever possible. Serum was separated promptly and stored at −2°C until used. Throat washings were obtained by having the subjects gargle 15 ml of nutrient broth (Bacto) to which had been added pencillin G in an approximate concentration of 1000 units per ml.

Progesterone (2.5 mg daily) beginning on the 19th day of pregnancy in intact rats prevents labor, uterine bleeding and parturition. Estrone (25γ) started on the 19th or 20th day induces uterine bleeding, vigorous but ineffective labor and death of fetuses. Estrone (100γ) begun on the 20th day in 1 animal brought about delivery of young 1 day premature. Spontaneous delivery occurred in 4 rats given 35γ of estrone about 24 hours before time for delivery. Progesterone (2.5 mg) given simultaneously with estrone (25γ) beginning on the 20th day has the sams effect as progesterone alone, preventing the above effects of estrone alone. It is suggested that estrone plays a role in the prevention of parturition in the rat other than by way of its action on the corpora lutea.

The urinary excretion of phosphate is increased by the injection of sodium p-aminohippurate in man. The extent of the phosphaturia tends to vary directly with the plasma PAH level. At high PAH levels, the amount of phosphate excreted averages 26%percnt; of that filtered, a proportion comparable in magnitude to that reported after injection of parathyroid hormone.

Dietary cholesterol significantly decreased the oxygen consumption of thyroid-fed rats, but the reduction was not sufficient for the metabolic rate to approach normal levels. This effect of cholesterol cannot be attributed to a depression of the food intake.

1. Chick embryo juice, in concentrations suitable for chick cell cultures, induces calcification of the cell colonies and plasma in mammalian cell cultures.

2. Fresh embryo pulp and extract are saturated with Ca and P. Ca and particularly P increase with age of the embryos, their ratios being 1:2.4 at 11 days. Upon incubation the P increases an additional 30%percnt;.

3. Pasteurization of embryo extracts inactivates the phosphatases and prevents calcification in cell cultures receiving high concentrations of embryo juice.

4. Calcification of cultures in the presence of mammalian sera is due to combining the Ca of serum and salt solution with the high levels of P in embryo juice, to the further liberation of P during incubation, and to the limited Ca X P products held in solution by mammalian sera.

5. The calcifying propensity of chicken serum is much less than that of 3 mammalian sera, which may be arranged as follows: human < bovine < rabbit.