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To verify whether the administered isovaleric acid does directly incorporate into urinary isovalthine or not, a young male dog was given labeled isovaleric acid-1-C14 (0.2 mc.) and the urinary isovalthine was analyzed by means of paper electrophoresis and column chromatography. Observations proved that isovalthine appeared in urine but showed no radioactivity while glutamic acid gave a strong activity. The possible metabolic pathway of isovaleric acid has been discussed.

The ongm and characteristics in transmission and morphology of spontaneous lymphatic leukemia in a low-leukemic strain C3Hf have been described. The leukemia line is being currently subjected to a vigorous search for the presence of a filtrable leukemia agent.

For the elucidation of the relationship between the toluene concentration in the air and the amount of hippuric acid excreted in the urine as a metabolite after the work in the workshop where they use toluene, we have succeeded in measuring minute quantities of urinary hippuric acid by a modified method of Gaffney's paperchromatography and also by a simple mass screening examination method devised by us, and obtained the following results. 1. The average concentration of hippuric acid in the urine collected at 2 p. m. from healthy individuals proved to be 184 μg/ ml, and the amount of its excretion was 174 μg/ min. 2. After the three-hour inhalation of toluene whose concentration in the air ranged 20, 50, and 160 ppm, the average concentration of hippuric acid in the urine was found to be 190,863 and 1,930 μg/ml, and the avarage increment of hippuric acid, 33.2, 193.9, and 1060 μg/min respectively. From this results we have found that there is a linear relationship between the toluene concentration in the air on one hand and the concentration of urinary hippuric acid and the increasing amount of its excretion on the other. Thus it has become possible to calculate the toluene concentration in the air even in the workshop where the workers are exposed to toluene discontinuously from the curves of the linear relationships just mentioned. 3. For the control of health in the workshop where they use toluene (such as auto-tricycle factory) we have devised a simple mass screening examination method for the detection of urinary hippuric acid, in which urinary hippuric acid is extracted with ether alcohol, and the coloration of its spots on filter paper is conducted with aroylglycine reagent (DAB and acetic anhydride mixture) in order to find out those excreting 1,000 μg/ml (the maximum allowable concentration) or over.

The authors give the schema of the different zones of the body and their points of measuring. They report on some average values registered with Szirmai's myotonometer on persons before and after massage. These myotonometric schemas play a new and important part in systematic measuring of muscles in the field of "Myologia practica".

Glucuronide formations by mouse liver homogenate in several liver impairments were studied by using 4-methyl umbelliferone as a glucuronide receptor. The results were as follows : 1. Subcutaneous or intraperitoneal administration of carbon tetrachloride to the mouse produced a significant increase in the liver glucuronyl transferase activity 12 or 24 hours after the treatment regardless of histological and enzymatic evidences of liver-cell necrosis. This increase was not attributed to the increase in the 'activator' of glucuronide formation but to the increase in the enzyme activity itself. 2. In Ectromelia virus mouse hepatitis, the glucuronyl transferase activity of the liver tissue was markedly reduced in severe cases. In moderate or milder cases, a slight increase in the activity was observed in a few of them in the early stage of the disease, and the activity was significantly decreased on the recovery in all of the cases which survived. 3. In the early stage of carbon tetrachloride injury when the glucuronyl transferase activity of whole mouse liver was increased and the decomposition of uridine diphosphate glucuronic acid by the liver tissue was also enhanced, the glucuronide formation in vivo was rather increased. It was thus considered that the whole liver glucuronyl transferase activity rather than the uridine diphosphate glucuronic acid content was responsible for the glucuronide formation in vivo as a rate-limiting factor.

The effect of the boiled liver extract on the velocity of 4-methyl umbelliferone lucuronide formation by mouse liver homogenate was studied. The results were as follows: 1. Addition of mouse or rat liver boiled extract to the complete system for the glucuronide formation produced an increase in the velocity of the glucuronide formation. 2. The boiled liver extract produced the increase in the velocity of the glucuronide formation not as a substrate but as an activator. 3, The activator in the boiled liver extract was relatively heat stable, acid labile, and precipitated as a fairly ethanol-soluble barium salt. The solution of the activator partially purified by ethanol fractionation of the barium salt indicated its absorption maximum at 262 mμ. These results suggested that the most possible substance in the boiled liver extract responsible for the activation of 4methyl umbelliferone glucuronide formation might be a sugar nucleotide.

An attempt was made to apply the method devised by ARIAS for the determination of liver glucuronyl transferase activity using 4-methyl umbelliferone as a glucuronide receptor to the small amounts of liver tissue obtained by needle biopsy. This was accomplished by studying the kinetics of enzymatic 4-methyl umbelliferone glucuronide formation by mean(of mouse liver homogenates. The improved method was proved to be applicable to human liver and gave a satisfactory result. In addition, an assay method for the estimation of liver uridine diphosphate glucuronic acid content from the amount of 4-methyl umbelliferone glucuronide formed from the uridine diphosphate glucuronic acid contained in the liver homogenate used as a source of glucuronyl transferase was studied, and as a result it was proved to be also applicable to the small amounts of human liver tissue.

The method of identifying human blood is studied from the standpoint of the fibrinolytic enzyme system by means of the "fibrin plate method" and the following results are obtained: 1) "Fibrin plate method" is, in the point of its sensitivity and speciesspecificity, one of the most excellent methods for the identification of human blood. 2) A small amount of the blood stains left standing as long as for 5 to 30 years can serve in the determination of the human blood. 3) Putrefied fluid blood does not demonstrate fibrinolysis. 4) Blood stains absorbed in various objects can also he identified whether they are of the human origin or not. 5) The pieces of cloth stained with human blood give positive fibrinolysis even after four washings with soap or after heating at 100°C for one hour. On the other hand, positive results are obtained with the pieces of cloths after three washings by benzidine test and with the physiological saline-extracted solution obtained after two washings of pieces of cloth, by precipitation test. 6) Proactivator has been found to contain globulin fraction in human serum protein. 7) The blood type can be determined with the same materials previously examined by the "fibrin plate method".

By far the majority of studies and speculations on metabolism in man and animals have been concerned with the fate of proteins, fats, and carbohydrates. Even when interest was awakened in enzymes and vitamins, the transformation of the organic substances was still studied. Gradually, however, various methods of the analysis and measuring instruments used have been so perfected that other problems can now be included in the field of biological and medical researches, namely, the significance of inorganic substances for the living organism.

Histochemical studies of several oxidative enzymes of the muscle spindles in mouse limbs were carried out. Observations elucidated the existence of the three types of intrafusal fibers which can be distinguished from each other by the difference in the activity of oxidative enzymes as in the muscle fibers, i. e., the first one is the "red intrafusal fibers" and has the most intense activity of oxidative enzyme, the second one the "white intrafusal fibers" with a minimal activity, and the third, "medium intrafusal fibers" being distinguished by their oderate activity of oxidative enzymes from the others.

For the purpose to reveal the mechanism of the biological indirect action of X-rays the author has conducted the analysis of the compound lipids of the organs of the rabbits irradiated with X-rays. Silicic acid column chromatography was applied for the fractionation of the compound lipids. Component of each fraction was analyzed by silicic acid-impregnated paper chromatography and infra-red spectrophotometory. The result proved that the compound lipids showed some qualitative and 10+-+ quantitative changes. The changes occurred mainly in glycerophosphatides showing the formation of high level lyso-phosphatidylcholine and lysophosphatidylethanolamine. The fatty acid compositions of the compound lipids were determined by means of gas-liquid chromatography. Further, polyunsaturated fatty acids were analyzed as fatty acid bromide. Major differences found in fatty acid patterns included: in the irradiated group the amount of C18-monoenoic, C18-dienoic, C18-trienoic, C20-tetraenoic and C22-acids were decreased and of C18- and C18-monoenoic acids were increased. It has been elucidated that lysophosphatides not only posesses a strong hemolytic power but also it has an action to induce swelling of the rat liver mitochondria. And it has been suggested that the lysophosphatides and ethanolamine will be responsible, at least partially, for the cell damage induced by X-rays.

The effect of various fatty acids on the swelling-contraction and oxidative phosphorylation of mitochondria from rat liver and Ehrlich ascites tumor cell have been studied and the results are as follows: 1. The swelling of rat liver mitochondria is induced by fatty acid. The extent of this uncoupling action is in the descending order of myristate, laurate, parlmitate, stearate and behenate in saturated fatty acid and linoleate, linoleneate, richinoleate and oleate in the unsaturated fatty acid. This swelling action is stronger with unsaturated fatty acids than that of saturated ones and cis form is stronger than trans form. 2. The uncoupling oxidative phosphorylation of rat liver mitochondria is also observed with these fatty acids and the activities are proportional to the degree of the swelling action. 3. The degree of swelling of rat liver mitochondria is proportional to the concentration of oleate and is inhibited by anaerobiosis and respiratory inhibitor except amytal. 4. The mitochondria swollen by fatty acid can be recontracted reversibly by ATP, Mg++ and bovine serum albumin. 5. The swelling action of sodium oleate is the strongest on mitochondria from rat liver, followed by those from the liver of Ehrlich ascites tumor bearing mouse, Ehrlich ascites tumor cells and solid Ehrlich tumor cells. 6. Sodium oleate inhibits the incorporation of 32p into ATP, ADP, GTP and UDPG in mitochondria.

Our experience with Brachet test on twenty-four leukemic patients has shown a high degree of reliability of the test for differentiating acute leukemias. A standard method has been described which is simple enough to be carried out routinely. The test, however, is not without pitfalls and need be interpreted with some caution. From the fact that urine hydrolysis can be closely simulated by the enzymatic action of pure DNase solution, it is suggested that the urine factor responsible for the nuclear lysis is DNase excreted in the human urine. The possible mechanism and implication of the test have been discussed in relation to the results obtained.

The effects of high fatty acids such as oleic, richinoleic, linoleic, linolenic, palmitic and stearic acids, on the respiration, glycolysis, organic phosphate synthesis of Ehrlich ascites tumor cells, were studied. The unsaturated fatty acids added to the media enhanced the respiration of the tumor cells at the concentration lower than 0.2 mM, after a short incubation period and inhibited the respiration in a high concentration 0.4 mM. The saturated fatty acids did not show such effect. All the fatty acids, both of saturated and unsaturated, effected the increase in lactate formation in tumor cells, especially markedly at higher concentration being accompanied by the WQ increase and RQ around 1. The respiration lowered by the fatty acids was ameliorated by the addition of glucose. The lactate formation from glucose was greatly enhanced by the addition of fatty acids but hardly from pyruvate. The unsaturated high fatty acids proved to have a strong uncoupling action for oxidative phosphorylation. This effect could be recognized slightly in the saturated fatty acids. The addition of high fatty acid resulted in the striking decrease in ATP and ADP with the increase in AMP. With these results the discussion was conducted concerning the specificity of tumor cell related to the glucose and fatty acid metabolism.

1. Morphological observations were carried out on the strain HeLa cells cultured under various oxygen tension. 2. The growth of the strain HeLa cells in the present experiments was markedly inhibited when they were cultured under high oxygen tension or in nitrogen gas environment. It has been clarified that air offers the most optimal gas environment. 3. Effects of the changed gas environment on the fine structures of HeLa cells were studied by phase contrast microscope and electronmicroscope. As the results it has been found that in these cells cultured under a high oxygen tension there occurs a marked swelling of mitochondria. Under anaerobic condition, however, these cells undergo degeneration, as in oxygen environment revealing no swelling of mitochondria but rather contraction. 4. Endoplasmic reticulum (ER) of HeLa cells cultured under oxygen environment is transformed from its vesicular form to lamellar form. The mechanism of lamellar transformation of ER is obscure but no morphologic connection with nuclear, cytoplasmic and mitochondrial membranes has been detected. 5. A discussion was made on these findings concerning the changes in fine structures of the cells, with a special reference to the swelling of mitochondria and morphological changes of ER under oxygen environment.

The fibrinolytic activity of plasmin present in menstrual blood has been studied by means of the Fibrin-Plate Method, and utilizing the findings of such a study, the identification of menstrual blood stain in legal medicine has been conducted. As the result, it has become clear that the identification of a very small amount of menstrual blood is possible, for an example, only one thread from the menstrual blood stained cloth 1.0 cm2. in size, and with the blood stain left in a room temperature for as long as two years, or with the blood stain left in water for one month, it is possible to identify menstrual blood. Aside from menstrual blood, no other toco-gynecological blood responds to this Fibrin-Plate Method.

As the results of present study on plasmin in menstrual blood by means of the Fibrin-Plate Method, a large amount of plasmin as much as dilution of 1: 100 or 1:1,000 of the menstrual blood serum, has been found in a natural form and it is deduced that the incoagulability of menstrual blood is the result of the plasmin formation in the same blood serum. Further, it has been recognized that this plasmin is found in a rather large quantity in the blood of the second menstrual day and it is decreased by the fifth day. In addition, the plasmin of menstrual blood is contained in the globulin fraction of the same serum, especially markedly in β-globulin fraction. On the other hand, it has also been clarified that in the circulating blood during menstruation no plasmin is present in natural state and that a large quantity of inactive plasminogen still exists in menstrual blood.

The authors have succeeded in isolating a biologically-active leukemia virus from leukemic tissues of AKR mice with a fluorocarbon. From the chemical analysis of the biologically-active virus fraction it has been clarified that the AKR leukemia virus is of the RNA type.

<P>Daβ der anaphylaktische Antikorper mit dem Prazipitin identisch ist, ist zweifellos aus den zahlreichen Arbeiten vom hiesigen Institut ersichtlich. Der anaphylaktische Schock, welcher durch die Bindung des im Organismus vorhandenen Prazipitin mit dem eingefuhrten Prazipitinogen verursacht wird, ubt vielleicht auf verschiedene Organe eine Reaktion aus. Daβ die Blutdrucksenkung trotz der Leberausschaltung aus der Zirkulation auftritt, beruht wahrscheinlich auf der Veranderung der allgemeinen Zirkulation, die durch die Reaktion des anaphylaktischen Schocks auf Nervensystem, Blutgefassen, Blut usw. sekundar hervorgerufen wird. In kurzen Worten, man erkennt, daβ die Blutdrucksenkung beim anaphylaktischen Schock bei sensibilisierten Hunden trotz der Ent-fernung des Magendarmkanales und der Leberausschaltung aus der Zirkulation durch die Antigenreinjektion auftreten kann. Ich glaube, daβ die Blutdrucksenkung eine sekundare Erscheinung ist, bedingt durch die Antigen-Antikorperperbindung in vivo. </p>