Experimental Biology and Medicine

1. Mice having a transplanted interstitial cell testicular tumor developed sinusoidal dilatation in the liver, spleen and adrenal glands. 2. These changes were more marked in animals with intra-mesenteric transplants than in animals with subcutaneous transplants. 3. This testicular tumor did not have any hormonal activity in castrate animals.

In response to cytotoxic serum nephritis, bilaterally adrenalectomized rats develop hypertension as frequently as do animals with intact adrenals. In order for hypertension to appear among such adrenalectomized rats, it is necessary that a satisfactory state of nutrition be maintained over a period of weeks. In our laboratory, the difficulties encountered in fulfilling this provision have been considerable in spite of the employment of a high NaCl regimen. This experience has led us to wonder whether the conflicting results in the literature relating to the question of hypertension in adrenalectomized animals may not be due, in part, to similar difficulties.

From spectrophotometric examination of the tissue lipids of rats at 10, 16, 24 and 48 hours after being fed free conjugated linoleic acid or its glyceride, it is concluded that glycerides are absorbed more rapidly, and possibly more completely, than free fatty acids but that the pathways of absorption are the same.

The hemagglutinating factor of Hemophilus pertussis culture supernatants can apparently be eluted from human erythrocytes by suspending such erythrocytes in a distilled-water extract of frozen and thawed red cells. Following the removal of the H. pertussis factor from agglutinated cells, these cells can be re-agglutinated by addition of fresh H. pertussis culture supernatant. The factor cannot be eluted by incubation of agglutinated cells at various temperatures, or by suspending the cells in salt-free isotonic glucose solution.

Similar to carinamide, 3-hydroxy-2-phenylcinchoninic acid causes higher concentrations of penicillin in the plasma when injected before the administration of the penicillin. Both substances also decrease the rate of excretion of phenol red. The activities of the two drugs are roughly of the same order of magnitude. This increase in concentration of penicillin in the plasma and decrease in rate of excretion of phenol red is attributed to a blocking of the renal tubular secretory mechanism.

We wish to thank Sara Ann Verplanck and Irene T. Payne for technical assistance.

DCA pellets increase the concentration of ACTH in the adenohypophysis of the intact rat. The steroid inhibits the marked depletion of pituitary stores of ACTH which normally occurs 24 hours after adrenalectomy.

Cortisone failed to prevent the development of the polyarthritis of rats when its administration was begun on the day of infection or to improve it when given therapeutically. The animals receiving cortisone gained less weight than the unmedicated controls. Terramycin given in 0.3% in the diet prevented the development, and 0.3% in the drinking water favorably altered the course of the rat polyarthritis.

1. The mean blood and packed cell volumes of 22 adult male rats, as measured by transfused tagged cells, was found to be 4.95 and 2.32 ml per 100 g of body weight respectively.

2. The mean blood and packed cell volumes of six of the 22 rats, assayed by the hemoglobin extraction method, were found to be 4.92 and 2.32 ml per 100 g of body weight.

3. The efficiency of perfusion for the total rat was 95%. This corresponds to a possible error in blood volume, introduced by removal of tagged transfused cells from the recipients circulation, no greater than 5%.

4. Comparison of the blood volume of various tissues taken from maximally bled rats with the blood volumes of similar tissues taken from perfused rats indicates that perfusion is least effective for spleen and most effective for kidney and brain.

The authors wish to thank B. Baldwin, J. Gordon, and M. Kern for technical assistance.

Multiple transfusions of Pí₁ blood from one pig into two other pigs of anti-Pi₁ specificity resulted in the formation of high titers of “blocking” antibodies of anti-Pi₁ specificity. The implications in experimental hemolytic syndromes and in selective fetal death in the pig are discussed.

A microbiological method for the determination of “strepogenin” activity is presented that depends upon the essential nature of the material for a strain of Lactobacillus bulgaricus. The method is believed to possess advantages over existing methods that depend upon the stimulation of the growth of Lactobacillus casei.

1. Intramuscular injection of a dextrin protected radio-silver colloid results in a high concentration of the injected material at the site of injection. 2. Intravenous injection of a dextrin-silver colloid into rats and intraperitoneal injection of a gelatin-silver colloid into mice result in deposition of the silver in highest concentration in the reticuloendothelial system. 3. Considerable silver was eliminated through the intestines and feces. The possible causes of such elimination under the conditions of this experiment are discussed. 4. A method of preparing tissues of small bulk for determining radio-silver activity with the Geiger-Muller counter is described.

Experimental pulmonary edema occurs in the rat in a matter of seconds as a result of a blow to the calvaria causing fatal trauma to the brain. This edema is prevented or reduced in degree by agents which produce a deep narcosis and by adrenergic blocking compounds. The mechanism of its production is discussed in relation to other types of experimental pulmonary edema.

1. Chicken erythrocytes exposed to a dilute toxin obtained from Micrococcus aureus progressively become more fragile prior to hemolysis. 2. These cells hemolyze more rapidly in isosmotic solutions of ethylene glycol and glycerol. 3. Swelling and shrinking measurements show that the toxin has little or no effect on the permeability of these cells to glycerol. 4. The fact that more of the toxin-treated cells hemolyze following shrinking and swelling in 0.3 M glycerol in Ringer Locke than do controls suggests that the toxin may weaken the membrane in some way. 5. A comparison between erythrocytes exposed to the toxin for 10 minutes and then washed and erythrocytes continuously in the presence of the toxin indicates that the toxin effects an immediate and irreversible change. 6. Even though diffusible toxin is removed by washing, some toxin apparently remains and gradually the cells hemolyze. This process occurs more rapidly if the diffusible toxin is not removed.

Glutathione, when injected into normal animals which had been force-fed a high carbohydrate diet, induced glycosuria. The amount of glycosuria decreased with increasing adaptation to the high carbohydrate diet and after feeding the diet for 2 months, glutathione produced insignificant effects. By contrast the injection of glutathione into cortisone-diabetic rats caused a marked (in some cases up to 10 fold) increase in glycosuria which has been observed even after 3 months of the high carbohydrate diet. Cysteine and ascorbic acid were also studied. Glutathione potentiates the diabetogenic effect of cortisone. The mechanism of this potentiation has been discussed and its possible significance in the treatment of rheumatic disease has been suggested.

Both aureomycin and terramycin, started 48 to 72 hours before the expected onset of illness and continued for 9 days were found to be effective in preventing murine bartonellosis. Rats treated with these compounds developed no evidence of illness while the morbidity in the control groups was 100%, and the mortality 50 to 100%. A few bartonellae appeared in the blood of some of the treated rats, but anemia and hemoglobinuria were prevented. The chemoprophylactic effectiveness of these 2 compounds in murine bartonellosis is in contrast to negative results obtained with a wide variety of other antibiotics.

1. 42 compounds, of which 39 could be classed as triazines and 3 as ethylenimine ring compounds were screened for chemotherapeutic activity against Ak4 mouse leukemia. 2. 2,4,6-triethylenimino-s-triazine, 2,4-diethylenimino - 6 - amino - s - triazine, and hexamethylene diethylenurea significantly prolonged the survival time of mice injected with transplanted leukemia Ak4. 3. It would appear that the chemotherapeutic activity of the above compounds can be attributed to the ethylenimine structure. 4. Four other compounds containing either a triazine or an ethylenimine moiety showed a slight chemotherapeutic effect.

Hepatic tissue of normal mice inactivated 8.5 m.u. of estradiol per mg dry weight per hour. Hepatic tissue from castrated mice or from estrone-treated mice was slightly less effective.

Ethylenedinitrilotetraacetic acid is now available commercially from Bersworth Chemical Co., Framingham, Mass. It forms stable chelate complexes with a number of metal ions, including calcium(1). We find that neutral aqueous solutions of this reagent dissolve urinary calculi rapidly. Urinary calculi are composed principally of calcium oxalate, urate, phosphate, and carbonate, with other salts and organic matter(2). The solvent action of ethylenedinitrilotetraacetic acid is undoubtedly due to the formation of soluble chelated calcium complexes. One stone which was composed of cystine, with no calcium present, was unaffected by the acid. Because of its effect on urinary calculi, we propose the short name “calsol” for ethylenedinitrilotetraacetic acid in this application.

A 3% calsol solution was made up by dissolving 30 g of calsol and 10 g of sodium hydroxide in one liter of water and adjusting to pH 7.5 with additional sodium hydroxide. Approximately 2 g was required. For the purpose of comparing calsol with the currently used citrate solution of Suby, 3% sodium citrate was prepared according to his formula “G”(3). The solvent actions of these solutions were compared against 44 pairs of stones from 44 patients in the Squier Urological Clinic, Presbyterian Hospital, New York. The stones from any one patient were nearly identical in size and composition. They were classified into 3 types according to the principal anion present: oxalate, urate, and soft (phosphate-carbonate). Calcium was present in all stones, including the urates. The stones were individually immersed in the solutions for 24 hours and the percentage losses in weight determined. Care was taken to avoid breaking off small pieces of stones which had been softened by the solution. In each pair of stones the calsol-treated stone lost substantially more weight than the citrate-treated stone.

Osmotic diuresis produced by mannitol loading in hydropenic dogs does not significantly affect the glomerular filtration rate, renal plasma flow or filtration fraction.

The metaphase spindle of the maturation division in the crayfish testis seems to be composed largely of longitudinally oriented “chromosomal fibers” measuring about 1500–2500 Å in width. These fibers appear to be compound, i.e., made up of smaller fibers of about 300–600 Å in width. Both the “chromosomal fibers” and their smaller fibrous components may at times show banding, which is possibly the result of irregular contraction during fixation. The centrioles appear relatively dense, are irregular in outline, and are probably not homogeneous bodies. The “chromosomal fibers” and astral rays lose their usual orientation at the periphery of the centriole. “Interzonal fibers” are relatively small in the late anaphase stages of the crayfish testis. Though “continuous fibers” were not clearly observed, we do not wish to claim that such fibers may not exist. Astral rays are composed of fibers, oriented as described in the past. The coagulated cytoplasm presents the appearance of a sponge-like reticulum, which is probably mostly a protein “framework,” since much of the other materials had been removed by the method of preparation.

Cardiac damage was produced in rats by a severe chronic thiamine deficiency. Rats fed a natural polished rice diet and/or a purified diet with thiamine added to approximate that naturally present in rice had less severe physiologic and pathologic changes than rats fed a thiamine free purified diet. The results of this experiment indicate that the protection shown by the rice diet and a purified diet with thiamine approximately equal to that of the rice diet was due to the trace of thiamine and not to some unknown substance or property in rice.

1. Antisera satisfactory in the Quellung reaction were prepared for K. pneumoniae types A and B by injection of adult chickens with formalinized suspensions of the encapsulated bacilli. As few as 4 injections at weekly intervals were sufficient for this purpose.

2. Effective antiserum was similarly prepared for H. influenzae type B. Capsular swelling was readily demonstrable with sera collected from chickens after 3 or 4 weekly injections of living organisms from young cultures, while sera from rabbits undergoing the same immunization schedule were distinctly inferior at this stage.

3. Chickens are very useful for the production of Quellung antisera, with particular advantages where Gram-negative bacteria are concerned.

The assistance of Mrs. Frances Fuller during the early part of the work is appreciated.

Effects of intraperitoneal injection of the hamster with phosphorus³² are described. Animals treated with 10 μc/g of body weight died in 11 days, there were no apparent changes in the testis at this time. Toxic effects were observed in animals injected with as little as 6 μc/g. Damage to the testes of animals treated with 6 and 8 μc respectively, was extensive, consisting of marked decrease in size, absence of spermatogenic cells in tubules, and actual damage to the tubules themselves, suggesting permanent sterility. No changes in the testes of animals treated with 1 μc/g were apparent in 24 hours. However, some damage was observed 30 days after treatment with 1 μc. Spermatogenic cells were more sensitive than Sertoli cells. Some interstitial cells appeared to be affected.

It has been demonstrated that certain lymphoid cells of antibody forming lymph nodes agglutinate on their surface in vitro the bacteria with which the animals were immunized. The agglutinating cells which could be identified belonged to the plasma cell series, whereas typical small lymphocytes failed to show the phenomenon. These observations seem to show that it is the plasma cell rather than the lymphocyte which elaborates agglutinins.

The effect of 4-amino-N¹⁰-methyl-PGA in prolonging the survival time of mice with transplanted leukemia AK4 can be blocked almost completely by prior administration of 1/3 to 2/3 as much by dry weight of a preparation of citrovorum factor. Thus, citrovorum factor is at least 12 to 24 times as active as pteroylglutamic acid in preventing the antileukemic effect of 4-amino-N¹⁰-methyl-PGA.

1. During cardiac catheterization, a pressure gradient was found between a distal point in a pulmonary vein and either or both atrial chambers during the major portion of the cardiac cycle in 2 cases of atrial septal defect, in a third case wherein an anomalous pulmonary vein drained into the right atrium, and in a fourth in which an anomalous pulmonary vein emptied into the superior vena cava.

2. The magnitude of the gradient between pulmonary vein and right atrium varied from 3.0 to 5.8 mm of mercury with an average value of 4.2 mm in the 4 cases studied.

3. Evidence that this gradient could not be entirely due to an end pressure or Pitot effect was obtained from study of a model in which the end pressure effect was shown to be 0.5 to 0.8 mm of mercury at the same calculated flow velocities.

4. It is suggested that this pressure gradient is a result of the pressure transmitted by the right ventricle through the pulmonary circulation, and is an important factor in the left-to-right shunt observed at rest during quiet breathing in uncomplicated cases of atrial septal defect.

The degree of aortic atherosclerosis found in 14 male rabbits given 1 g of cholesterol in gelatin capsules 6 days a week for 9 weeks showed a highly significant positive linear correlation with the average blood total cholesterol level.

Five of these animals which received 1 g of choline chloride in gelatin capsules 6 days a week for the 9 weeks did not differ significantly from the 9 control animals in average blood cholesterol level, in body weight, or in the degree of atherosclerosis which developed.

The author wishes to thank Dr. C. H. Best, under whose direction this work was performed, for his interest and helpful advice.

1. The activity of cytochrome oxidase increased in the presence of convulsant agents used in concentrations of 1×10⁻³M and less. 2. The activity of cytochrome oxidase was not significantly modified by the anticonvulsant agents used in concentrations of 1×10⁻³M and less. 3. Convulsant agents, therefore, activate at least 2 processes, namely, a non-oxidative mechanism that initiates the seizures and the oxidative recovery processes occurring in each unit during the convulsive seizures.

The life span of the common fresh water cladoceran, Daphnia magna, is shortened in the presence of vitamin K in the medium, and lengthened with heparin. The possibility of relationship to protoplasmic coagulation phenomena is pointed out.

1. Inhibition of the Shwartzman phenomenon by HN₂ is confirmed. 2. Protection of the preparatory site from HN₂ fails to influence the inhibition. 3. Protection of the lower limbs from the action of HN₂ by aortic occlusion decreases or prevents the inhibition.

When yeast cells were exposed to 20,000 r of x-rays and cultured in liquid medium, they and their descendants attained a mean cell volume more than twice that of cells derived from nonirradiated controls. On the other hand, there were no corresponding differences in specific gravity and nitrogen content per unit of cell volume. It is accordingly concluded that enlargement of yeast cells after irradiation is largely a nonselective increase in cell constituents rather than a swelling due mostly to uptake of water.

The survival of Trichomonas vaginalis was determined by studying discharge from 50 untreated patients with vaginal trichomoniasis. Droplets of the discharge were placed on the enamel surface of wooden blocks and tested under natural conditions of drying at room temperature.

Tests at intervals ranging from 10 minutes to 7 hours were made by comparing the activity of the flagellates in direct microscopic examination with their ability to multiply in serial cultures. A close correlation was observed between the two procedures. The trichomonads survived in 100% of the tests at 45 minutes, in 96% at 1 hour, in 56% at 3 hours and in 4% at 6 hours. No survival was demonstrated at 7 hours.

These data, together with experimental and epidemiologic reports of other workers indicate that fomites contaminated with Trichomonas vaginalis may, under natural conditions, act as agents in the transmission of vaginal trichomoniasis.

1, Many, but not all, pituitary preparations produce an increase in liver fat content when administered to intact mice. This response is completely abolished by adrenalectomy.

2. Neither cortisone nor any other adrenocortical steroid tested to date is able, of itself, to cause a substantial increase in liver fat level of intact or of adrenalectomized mice. In the latter, however, the usual post-adren-alectomy loss of liver fat is prevented.

3. When pre-treated with adrenocortical hormone (cortisone), the ability of adrenalectomized mice to respond to pituitary hormone by mobilization of fat to the liver is equal to or greater than that of intact mice. Pituitary substances (growth hormone; weakly potent ACTH) inactive in either intact or adrenalectomized animals, became quite effective in mobilizing fat to the liver if the animals are pre-treated with adrenocortical hormone.

4. It is concluded that to mobilize fat from the depots to the liver the animal requires at least two factors, (1) adequate supplies of adrenocortical hormone (from either endogenous or exogenous sources) plus (2) a supply, either from its own anterior hypophysis or from exogenous sources, of the “triggering” pituitary factor.

5. The “triggering” action of the unidentified pituitary factor is not mediated via the adrenal cortex since it operates in the absence of the adrenal gland provided adequate amounts of exogenous cortisone are administered.

6. The possible identity of the “triggering” pituitary factor is discussed.

The authors are indebted to Miss Ilse Bauer for her kind assistance in performing the many adrenalectomies and necropsies.

The avidity of transplanted sarcoma in rats for labelled tyrosine (1) and rat hepatoma for labelled alanine and glycine(2) suggested the possibility that amino acids containing a toxic substituent might prove useful for the inhibition of tumor growth. Although n-ace-tylation of tryptophane does not prevent its utilization as a growth promoter(3). n-iodoacetyl derivatives of l-tryptophane, l-leucine and d, l-phenylalanine were considered suitable for an exploratory study because the iodoacetyl group, an inhibitor of certain respiratory enzymes, is a toxic residue. Accordingly, these derivatives were prepared from corresponding chloroacetyl derivatives by treatment with sodium iodide in acetone(4). For preparation of the radioactive analogues Nal¹³¹ was substituted for sodium iodide(4). For assessment of their distribution in tissues, their general toxicity and their specific toxicity for tumor tissue, control observations with iodoacetamide were necessary. Radioactive iodoacetamide was similarly prepared from chloroacetamide (2).

The distribution of radioactivity in tumor, liver and blood of Swiss mice bearing transplanted sarcoma 37 was determined at intervals after injection of the radio-active analogues of the three amino acid derivatives and of iodoacetamide. With the possible exception of radioactive iodoacetamide in tumor (Fig. 2, 3, 4) there were no consistently significant differences among the 4 compounds in the concentrations of radioactivity in liver, blood and tumor. Although the concentration of radioactivity in blood and tissues in the initial stages is not significantly different after administration of these 4 compounds than it is after the administration of Nal¹³¹, the radioactivity from the latter disappears within 4 hours(5) while from the former small but significant amounts of radioactivity persist for considerably longer periods. This persistence may reasonably be explained by the assumption that at least a small fraction of radioactive iodoacetamide and iodoacetylamino acid derivatives had reached tissue cells.

The variations in metabolic activity of the soil organism, Corynebacterium creatinovorans (Strain “NC”), grown in media containing different carbon sources is described.

The injection of heparin (50 mg intravenous) in the human causes a sudden fall in surface tension and a clearing of the lipemic plasma. The effect is believed to be due to the formation of a surface active heparin-phospholipid complex. The activity of the complex may be studied in vitro by mixing lipemic plasma with non-turbid plasma obtained after heparin injection. A gradual clearing is observed. The formation of a surface active heparin-egg lecithin complex has been demonstrated. Certain relations of these findings to pathological conditions are discussed.

‘Benemid', p-(di-n-propylsul-famyl)-benzoic acid, inhibits the conjugase responsible definitively for the over-all reaction involved in the conjugatior of glycine with p-aminobenzoic acid. It was demonstrated that the compound did not decrease the availability of energy derived from the phosphate cycle, which is a coupled component of the conjugative reaction, since ‘Benemid' did not inhibit glucose phosphorylation by phosphorylase plus preformed ATP.

The implication of such conjugative reactions was discussed and several pharmacological and clinical applications of the anti-catabolite hypothesis were indicated.

It is shown that glucose recovered from starch deposited in photosynthesis in presence of C¹⁴O₂ may be labelled primarily in 3 and 4 or 1 and 6 positions or evenly in all 6 positions, depending upon the periods of darkness and illumination to which the leaf is subjected.

Mature hens were given a single whole body X-irradiation of 300 roentgens. This treatment markedly reduced the bone marrow endogenous respiration, choline oxidase, and succinoxidase. The intraperitoneal injection of urethane also reduced marrow choline oxidase, and reduced succinoxidase but to a lesser degree. Both X-irradiation and urethane treatment rendered the chickens leucopenic.

Two compounds, 6-methoxy-8-(β - diisobutylaminomethyl) aminoquinoline dihydrochloride and 6-methoxy-8-(diisobutyl-aminoisopropyl) aminoquinoline dihydrochloride, which are chemotherapeutically effective in mice infected with schistosoma mansoni, have been shown to be potent inhibitors of hyaluronidase although they inactivate the serum inhibitor of the enzyme. Chemically related rosaniline and plasmochin, that have little chemotherapeutic effect, had no demonstrable direct effect on the hyaluronidase, although both potentiated the serum inhibitor to some degree.

1. It is confirmed in the dog, as has been shown for other species, that the relative weight of the hypophysis is greater in adult females than in adult males, and that this difference does not exist before maturity. 2. The hypophysis weight-body weight ratio of puppies is greater than that of adult dogs, and in adult dogs decreases significantly with increase in body size. 3. Relative hypophysis weight was smaller in senile than in adult animals. 4. No sex difference in hypophysis weight existed within any weight-group of adult animals. 5. No differences between groups could be detected with respect to water content of the hypophysis.

Insulin hypoglycemia causes in schizophrenics a significant eosinopenia and lymphocytopenia, which is considered to be indicative of adrenocortical stimulation. Insulin does not produce such changes if hypoglycemia is prevented.

1. The Brown-Pearce carcinoma of rabbits shows definite growth on the chorioallantois in 58% of embryos implanted on the 8th day of incubation, and in 68% of embryos surviving until the 17th day. The wet weight of the tumor increases 79 fold during the 9-day culture period. 2. The average percentage of successful grafts is increased by implantation two or more pieces of tumor within a single egg, and by incubating the excised tumor for several hours in rabbit serum before implantation onto the chorioallantois. 3. The tumor does not metastasize from the chorioallantois in spite of apparent direct contact between the embryonic blood and the tumor cells.

Antipyrine is distributed evenly in the various tissues of animals in close proportion to their water content. The values obtained for total body water by antipyrine and by desiccation agreed well and gave a correlation coefficient of 0.93. The rate of transformation and excretion of antipyrine as determined by plasma analysis was constant over a period of time sufficient for analysis.

Ease of analysis and lack of toxicity make antipyrine a suitable substance for use in measurement of total body water in animals.

1. From 100 mice, orally infected with Col.SK virus at various times during the period of gestation, only 18 viable litters were obtained. These litters were equally divided among 25 surviving animals and 75 animals which succumbed to the disease.

2. The rate of abortion for mice inoculated during the first half of pregnancy, regardless of their capacity to resist infection, was high, being 72% for surviving animals and 88% for those which died, since two-thirds of the litters found in utero at the time of maternal death gave evidence of having predeceased the mother.

3. Fetuses obtained at autopsy consistently contained demonstrable amounts of virus.

4. Living young, when delivered at the time of onset of maternal symptoms, had apparently escaped infection, while a majority of stillborn young yielded virus.

5. Surviving offspring of susceptible mothers, born during the maternal incubation period, were apparently normal and usually escaped infection.

6. Litters of immune mothers, with one exception, were fully protected against intranasal infection between weaning and the age of 44 days. When tested between 66 and 68 days of age only 2 of 7 young were resistant, while after 119 days there was no evidence of immunity.

7. By means of experiments in which the offspring of immune and normal mice were interchanged during the period of suckling, immunity in the young was shown to be related to the observed immunity in the nursing mother.

8. Milk collected from normal mice contained no antiviral substances while milk from immune mice was capable of neutralizing the virus in vitro.

9. The data indicate that the protection of young mice is passive in nature and probably brought about by the ingestion of antiviral substances (antibodies?) in the milk of immune mothers. There is no evidence in this work to suggest that the offspring of infected mothers may acquire a state of active immunity through inapparent infection.

The author wishes to express her grateful appreciation to Mr. Anton Samuel for technical assistance in collecting milk from lactating mice.

1. One hundred and sixty-three male Swiss mice weighing between 17 and 20 g were implanted sub-cutaneously with fragments of sarcoma 37 and then injected subcutaneously or intravenously beginning on the day of implantation or 7 days thereafter with hyaluronidase, heat inactivated hyaluronidase, or physiological salt solution.

2. The results on the effect of hyaluronidase administered subcutaneously or intravenously were somewhat variable, from which it is concluded that there was no effect on the growth or invasibility of implanted sarcoma 37. Neither was there metastasis to distant organs.

Marked retardation of growth of lymphosarcoma implants was observed in pyridoxine-deficient rats injected with 2 mg daily of cortisone or of methyl testosterone. Survival of pyridoxine-dencient, tumor-bearing rats was prolonged by testosterone and shortened by cortisone.

Thanks are due to Mr. T. C. Bielinski and Miss T. Visotsky for valuable assistance.

The subcutaneous injection of 20-methylcholanthrene into hamsters produced fibrosarcomas within 60 to 80 days. Examination of the injection sites, 24 hours following injection, showed fibroblasts which are morphologically different from normal. At first these cells show little active proliferation. The altered fibroblasts form a distinct strain of cells in a nodulus close to the area of injection and it is from these nodules of altered fibroblasts that invasive fibrosarcoma grows. In contrast to the oil solutions of carcinogen used, the saline suspensions produce cell necrosis and giant cell reaction.

A technic for the two dimensional chromatography of blood plasma and protein mixtures on filter paper is described. Separations of protein constituents are greatly facilitated by the addition of surface active substances such as the “Tweens” or “Spans,” etc. The technic at present adopted is to add “Tween 85” or “Tween 81” and, using hemin as protein marker, to employ M/10 sucrose solution in the first dimension and M/10 sodium potassium tartrate solution in the second dimension. Differences between the protein patterns of chromatograms of human, rat and guinea pig plasma are noted. With protein mixtures, associations of proteins may occur that may be detected by chromatography. The sera of hyperimmunized horses (as against diphtheria toxin) yield chromatograms which differ from, and exhibit fractions not present in, chromatograms of normal horse sera.

Grateful acknowledgment is made to the National Cancer Institute of Canada for a grant in aid of this work, to the Department of Microbiology and Hygiene of the University of Montreal for gifts of normal and immune sera, to Dr. J. Gross for the protein fractions obtained from the Cutter Laboratories, Berkeley, Calif., to the Atlas Powder Co. (per G. F. Sterne & Son, Brantford, Ont.), for gifts of “Spans,” “Tweens,” “NNO,” and “BRIJ-30,” to the Canadian Industries, Ltd., Montreal, for gifts of “Elvanols,” and to Mrs. Mary Hersey for her able assistance in this work.

1. Dromoran is less antidiuretic than morphine when anti-diuretic activity is related to analgesic potency. 2. In the range of doses employed in these studies, neither morphine nor Dromoran effected a chloruresis. 3. The lack of chloruresis in the presence of anti-diuretic action suggests that these drugs do not act by liberation of the anti-diuretic hormone.

Both poliomyelitis virus and Coxsackie virus (C virus) were isolated from the acute phase stools of several paralytic patients during an epidemic of poliomyelitis which occurred in Easton, Pa., in 1949. In 2 such cases, neutralizing antibody tests were carried out with acute and convalescent serum reacting with a suspension of virus obtained directly from acute phase stools. It was found that both patients responded to their illness by simultaneously developing antibodies both to poliomyelitis virus and to C virus.

1. The effects of compound E and ACTH on experimental hypersensitivity of the Arthus type and on antibody production in the rabbit are described. Both compound E and ACTH inhibited sensitization to repeated intracutaneous injections of crystalline egg albumin. Moreover the inhibition produced by compound E was much greater than that obtained with ACTH; with compound E sensitization was almost completely prevented in all of the 8 animals tested.

2. The data indicate that the inhibitory effect of compound E and ACTH on the development of the Arthus state results from the ability of these hormones to suppress antibody formation. As shown by antibody nitrogen determinations, compound E and ACTH suppressed antibody formation by an average of 100 and 50% respectively.

3. In contrast to their striking effect on the production of the active Arthus reaction, compound E and ACTH had no effect on the passive local Arthus reaction, when antibody is supplied to the animal. Thus, the capacity of the animal to react to antibody-antigen combination in the tissues is not altered by treatment.

The resistance of a strain of Klebsiella pneumoniae to chloramphenicol was enhanced 128-fold by serial subculture on the surface of agar plates containing graded concentrations of this antibiotic.

A chloramphenicol-dependent variant also emerged during the course of these subcultures. This variant grew best in the presence of a critical concentration of chloramphenicol; as the concentration of antibiotic was either increased progressively beyond this critical level or decreased below it there was a steady decline in growth until complete inhibition of growth occurred.

The chloramphenicol-dependent variant was first recognized soon after the resistance of the strain had been enhanced to a point where it was completely inhibited by this critical concentration.

Evidence was also obtained of possible “back-mutation” to variants which were as sensitive as the parent strain.

The observations indicate that under certain circumstances the treatment of rabbits with ACTH or Cortisone results in a type of local skin damage by meningococcus toxin which is not seen in untreated animals. The application of these findings to other varieties of tissue damage by bacteria and their products is a field which merits further study.

The technic for visualization of blood vessels by intravenous injection of the fluorescent dye, Thioflavine-S, has been used in a study of kidney circulation in normal rabbits and rabbits exposed to crush injury.

The results obtained show that the blood flow in the medulla is decreased. We have not been able to confirm Trueta's results indicating a by-pass through the juxta-medullary glomeruli.

The procedures described above utilize a previously undescribed property of plasminogen, solubility in dilute mineral acid, to extract it from placental and plasma fractions. Purification of over 250-fold has been obtained by successive extractions with 0.2N H₂SO₄ and precipitations at pH 5.2. The total yield has ranged from a minimum of about 10% to almost complete recovery of the enzyme present in the starting material, with an average recovery of about 30%.

The procedures described above utilize a previously undescribed property of plasminogen, solubility in dilute mineral acid, to extract it from placental and plasma fractions. Purification of over 250-fold has been obtained by successive extractions with 0.2N H₂SO₄ and precipitations at pH 5.2. The total yield has ranged from a minimum of about 10% to almost complete recovery of the enzyme present in the starting material, with an average recovery of about 30%.

Pain and temperature sensitivity of the structures in the sternal bone marrow cavity was tested on three normal subjects. Four injections of 1 ml of hot (40–45°C) and cold (16–22°C) saline were made through the anesthetized skin and periosteum of each subject. No temperature sensations were reported although a deep burning pain and an aching pain were reported at the beginning of each injection.

Two metabolites of pamaquine have been demonstrated in the plasma and urine of humans and dogs receiving pamaquine. One of these is an unstable compound which can, in vitro, convert hemoglobin to methemoglobin and lyse erythrocytes. The other is a stable compound which is highly fluorescent. Neither of these substances has been isolated in crystalline form.

It is possible that the substance with the oxidant and lytic properties is responsible for the toxic actions of the parent drug. The answer as to whether it also accounts for its therapeutic effect awaits definition of its antimalarial activity.

A homologue of pamaquine which differs only in its substitution on the side chain, yields metabolic products with properties similar to those obtained with pamaquine.

The incubation of slices of renal cortex with tetrazolium chloride (TTC) has disclosed patterns of formazan deposition in the proximal convoluted tubules with striking differences between normal and hypertensive kidneys obtained from dogs, rats and man. In the proximal convoluted tubules of kidneys from normotensive animals and man, the formazan is deposited as fine dust-like granules evenly distributed within the cytoplasm. In the hypertensive kidney the formazan is deposited in coarse clumps, plaques or needles, commonly at the cell margins and often extracellularly as well.

Despite the difference in formazan patterns the total amount of reduced TTC per mg of kidney tissue is approximately the same in normal and hypertensive kidneys, as is the oxygen consumption determined by the conventional micro-respiration technic. Concurrent studies of VEM metabolism show the uniform restriction of VEM production to anaerobiosis in normal kidneys, and the formation of VEM under both anaerobic and aerobic conditions by the hypertensive kidneys of animal and human subjects. When the VEM metabolism of a kidney from a normotensive rat, dog or human is transformed to the hypertensive type by a period of prior anaerobiosis in vitro, there is a parallel development of the characteristic hypertensive formazan pattern in the proximal convoluted tubules.

The reduction of TTC to the insoluble formazan results from an interaction between TTC and enzyme systems within the cell. The observations reported would indicate some alteration in the metabolic characteristics of the proximal convoluted tubules in experimental renal and essential hypertension. The association of these histochemical changes in the proximal convoluted tubules with alterations of the VEM mechanisms in experimental and human hypertension suggests, 1) that the proximal tubules may be the site of VEM formation, and 2) that the failure of the hypertensive kidney to limit VEM formation to anaerobiosis may be related to the enzymatic disorganization revealed by TTC.

Finally, the data presented provide specific evidence for a similarity between experimental renal hypertension and essential hypertension in man.

Vitamin B₁₂ has been found to enhance markedly the carcinogenic effect of p-dimethylaminoazobenzene in rats receiving a methionine-deficient diet. However, a control group of rats receiving this diet without p-dimethylaminoazobenzene showed no hepatic tumors.

The steroid Δ1 deoxycorticosterone acetate will maintain adrenalectomized immature rats for 20 days with dosages of 0.25 mg and 0.5 mg and will increase daily body weight 1 and 2 g respectively. Activity is approximately 1/8 to 1/10 that of deoxycorticosterone acetate. Liver water content was unchanged in maintained adrenalectomized rats and the relative liver protein content of adrenalectomized rats receiving Δ 1 desoxycorticosterone acetate was comparable to that of normal rats. This steroid did not alter organ weights in normal immature female rats over a 10-day period except for a tendency toward liver weight increase.

1. Differences in the vitamin requirements of susceptible strains of S. mitis, their artificially-induced resistant variants and a naturally-resistant S. fecalis, have been determined. 2. The greatest difference was in the PGA requirements, the mitis strains grew in its absence; the resistant mutants required PGA. 3. The induced resistance of the trained S. mitis strains appears to be the same as that of the natural resistance of S. fecalis, and may be due to the loss of ability to synthesize PGA, the synthesis of which is inhibited by sulfonamides.

The succinic dehydrogenase activity of beef muscle determined after different methods of homogenization of the tissue was investigated. The amount of beef muscle needed for a representative sample was too large to be conveniently used with the Potter-Elvehjem homogenizer; however, the samples could be prepared for assay by homogenizing in a Waring blendor, provided that the homogenization time is held to a minimum. This was evidenced from studies which showed that homogenizing the minced beef tissues in a Waring blendor for 1 minute or homogenizing Latapie minces for 1/2 minute resulted in a greater succinic dehydrogenase activity than that obtained when the tissues were homogenized for longer periods. Other experiments with rat tissues showed that the enzyme activity of rat muscle homogenized in the Waring blendor for 1 minute, in a Potter-Elvehjem homogenizer, or ground with sand was essentially the same.

1. The ratio of simultaneously determined renal clearances of hemoglobin and inulin, in successive clearance periods in a group of 5 control and 3 patients with the nephrotic syndrome, indicate that the overall glomerular porosity, for hemoglobin, of the patients with proteinuria is no greater than that of normals.

2. It is probable that glomerular permeability, in terms of “pores,” is not the factor determining the degree of proteinuria in the nephrotic syndrome.

The alkaline tide of the alligator has been studied. Although the plasma electrolyte changes were qualitatively similar to those reported for other animals the magnitude of these changes was much greater. Some plasma chloride levels as low as 15% of the fasting level were found and some plasma bicarbonates were elevated 5-fold. Although the average fasting blood pH is about 7.3, pH values of over 8 have been observed at the height of the “alkaline tide.” Plasma sodium concentrations were unaffected by feeding. No correlation was found between pH and plasma calcium content.

1. The effects of daily administration of 50 or 100 mg of dl-ethionine on the rat pancreas is described.

2. Degenerative changes in the acinar tissue occurred as early as 12 hours after a single injection of 50 mg dl-ethionine.

3. Daily injections of 50 or 100 mg in non-fasted rats, for 2 weeks, resulted in almost complete obliteration of the acinar tissue accompanied by fibrous proliferation.

4. No histological evidence of islet damage was observed with these dosages.

1. The intracutaneous injection of viable leucocytes obtained from the bloods of atopic subjects who respond with immediate whealing reactions to skin tests with common allergens may specifically and locally sensitize the skin of normal recipients.

2. Red blood cells obtained from the same donors lack this skin-sensitizing property.

This investigation indicates that cortisone produced well-characterized changes of serum lipids in man. The effects of ACTH on the serum lipids were less pronounced.

A positive inotropic effect of ouabain has been demonstrated on the rat ventricle strip. At a given ouabain concentration the magnitude of the response was quite uniform. The effects of variation in the concentration of ouabain, calcium, and phosphate, and in initial tension on the positive inotropic response have been investigated.

In cats under nembutal anesthesia, there is a selective depression of the cochlear response so that with threshold stimulations it functions as a band-pass filter for frequencies around 1700 cycles/sec. This depression does not appear to be due to the anesthetic, per se, nor to fall in body temperature, but occurs in association with depression of the respiratory rate.

Four patients with Addisonian pernicious anemia in relapse and one patient with nutritional macrocytic anemia were treated with aureomycin given orally, with definite although submaximal hematological improvement. A fifth patient with Addisonian pernicious anemia was treated with aureomycin given intravenously with no response.

We are indebted to Mrs. Helen Jakubowski for performing the blood examinations.