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Cytochemical observation of the activities of diphosphopyridine nucleotide diaphorase (DPNH-D), triphosphopyridine nucleotide diaphorase (TPNH-D), succinic dehydrogenase (SDH) and a-glycerophosphate dehydrogenase (α-GDH) of Ehrlich ascites carcinoma cells were made and following results were obtained. The smeared cells showed moderate reactions and no marked difference in the intensity among the individual cells. The free floating cells were stained relatively faint but showed the differences in the staining intensity in individual cells. In the presence of benzalkonium, the reaction intensity proved to be intermediate between the smeared cells and free floating cells without benzalkonium and the differences in the staining intensity in individual cells were more marked. Observations revealed that the reaction intensity changes closely corelated with the stage of mitotic cycle of each cell. Namely, DPNH-D activity of the tumor cells, which generally hihger than that of leucocytes, increased remarkably in the end stage of interphase and decreases abruptly in mitotic stage reaching the lowest level in metaphase. After the metaphse the activity increased slightly and it is kept at almost the same level during the first half of interphase. This enzyme is localized mainly in the granules of the eytoplasm. The activity of TPNH-D showed the similar localizations as those of DPNH-D, though the reaction intensity is lower than that of DPNH-D. The activity of SDH of the tmnor cells is lower than that of leucocytes and its diformazan granules are localized in mitochondria. Its activity decreases in the mitotic stage the lowest level in metaphase and in the followed interphase it is kept in a almost constant low level. α-GDH activity of the tumor is lower than that of SDH but show the similar localizations as the latter.

Based on our original concept, a fibroblast-inhibiting agent, chloroquine, was used against various animal tumors. Among transplanted animal tumors, the drug was most effective on relatively connective tissue-rich Bashford and Brown-Pearce tumors, as reflected by prolongation of life span, inhibition of tumor growth, inhibition of lowering of liver catalase activity, improvement of iron metabolism, increase of tumor necrosis, inhibition of connective tissue formation, and decrease of acid mucopolysaccharide. On the other hand, it was of little advantage in Ehrlich, Yoshida and MH134 tumors which contain little connective tissue, except for a decrease of the amount of ascites and ascites tumor cells in the former two tumors. These results indicate that chloroquine suppress the growth of the tumors relatively rich in connective tissue. This effect of chloroquine appears to be due to the primary attack of the stromal connective tissue of tumors being followed by the degeneration of tumor cells, though its probable anti-tumor activity by the indirect effects through its anti-inflammatory and systemic humoral activities should be taken into consideration.

The effects of chloroquine on the growth and morphology of fibroblasts in tissue culture, and in vivo granulomas were investigated. As the result, the drug was shown to have a potent action to inhibit fibroblast growth, which has led to a possibility of its clinical use to patients with malignant tumors.

1. Attempts have been made to confirm how the formazan formation is affected in the presence of oxygen gas when the cells are incubated with neotetrazolium salt and the subsrtates for the enzymes to be tested. 2. In the cases of succinoxidase formazan formation is minimized under pure O2tension, it increases with decrease in O2 tension, and reaches its maximum value under N2gas. 3. This relationship between the oxygen tension and the diformazan formmation can likewise be observed even after pretreatment of the system with KCN. 4. In measuring enzyme activity of the DPN-diaphorase system with L-glutamate and DPN as substrate and NT as hydrogen acceptor, the same relationships between the oxygen tension and the NT-reduction can be seen as in succinoxidase system. 5. In the determination of enzyme activity of the cytochrome-c-cytochrome oxidase system with p-phenylene-diamine as substrate and NT as hydrogen acceptor, likewise the diformazan formation is markedly affected by oxygen tension and increased with the reduced oxygen tension but under pure Ns gas the value is reduced. When the systen is pretreated with KCN, however, the diformazan formation reveals its maximum value under pure nitrogen gas, the values of which correspond to those values of endogenous reaction without substrate. 6. The above results show that the neotetrazolium salt can compete with O2 as hydrogen acceptor, and less values of formazan formation may be obtained under higher oxygen tension and the higher values under lower oxygen tension independently from the true activity of the enzyme.

Since the classic work of Ranvier, it is well known that the mammalian striated muscle is composed of two types of muscle fibers, i. e., the red and white muscle fibers. In the previous paper1 it has been reported that the limb muscle fibers of mammals can be divided into three types from their activities of the histochemically demonstrable oxidative enzymes. Namely, the small red muscle fibers had a higher activity of oxidative enzymes, the large white muscle fibers a lower activity and the third type of muscle fibers being called "medium fiber" or "intermediate fiber" showed an intermediate activity between those of the red and white muscle fibers.

For the purpose to clarify the causes of X-ray disturbances a series of experiments have been conducted on biological and biochemical properties of compound lipids extracted from normal and X-ray irradiated rabbit organs with a special reference to the P³²-labeled compound lipids uptake, inhibitory action to L cell proliferation and uncoupling of oxidative phosphorylation, and the following results have been obtained. The compound lipids (lysophosphatide rich fraction) isolated from the X-ray irradiated rabbit organ have been found to possess a strong hemolytic action and also an action to inhibit the cell proliferation as well as to accelerate the respiration of the mitochondria in the rabbit liver and spleen. It has also been proven that they act as to induce a marked swelling of mitochondria, to impede the formation of high energy phosphate as well as to act as an uncoupler of oxidative phosphorylation in vivo. In the test to see the uptake of P³²-labeled compound lipids by various organs, a marked uptake has been observed in spleen, bone marrow, and liver of both irradiated and non-irradiated groups. Further, the uptake of P³²-labeled compound lipids in the rabbits given intravenous injections of compound lipid fraction for 30 consecutive days previously has been found to be greatest in pancreas followed by bone marrow, spleen, liver in the order mentioned in male group, whereas it is greatest in spleen, followed by liver and bone marrow in the female group. With these results the discussion was conducted concerning the relation between the lipid metabolism and X-ray disturbances.

As described in the foregoing, a certain degree of desirable effect can be recognized in every instance of the present trials. However, in order to apply these methods in clinics it is necessary to carry out further studies on the mechanism that operates in bringing about such an effect, but this paper is presented as a preliminary report.

A case of paroxysmal bundle branch block, two to one right bundle branch block followed by intermittent right bundle branch block, which is associated with chronic cor pulmonale secondary to active, far advanced pulmonary tuberculosis, is presented. The incidence and mechanism of the paroxysmal bundle branch block have been discussed.

A granuloma pouch was formed on the back of rats by the original method of SELYE. Seven days when granuloma tissue reached its maximum, 35S labeled ChS, 59Fe labeled ChS-Fe, labeled ferric ammoninum citrate and colloidal 198Au were injected into the pouch and their absorption and organ distribution examined and compared with the results in the case where 59Fe labeled ferric ammoninum citrate and colloidal 198Au were injected into the gluteal muscle. 1. When 35S labeled ChS was injected into the granuloma pouch, radioactivity of the organs per gram tissue was high in the kidney, liver, bone marrow and spleen, in descending order. The maximum activity was seen 12 to 24 hours after injection, which is slow compared to the results obtained by KISHIDA in intraperitoneal and oral administration. 2. The absorption of Ch S-Fe by pouch where the iron is enveloped by the large ChS molecule, is slower than that of ferric ammonium citrate, an inorganic compound. 3. The uptake of Fe from the blood by bone marrow is larger when the increase of blood Fe ion concentration is slow, rater than when the increase is rapid. 4. When conoidal 198Au is injected into the pouch and injected into the" gluteal muscle, the 198Au is phargocytozed by the reticuloendothelial system organs, the liver showing the largest uptake among all organs. 5. In the intramuscular injection of colloidal 198Au and 59Fe labeled ferric ammonium citrate, radioactivity of pouch fluid is lower than that of blood. However, the difference between the two is less in the case of colloidal 198Au. 6. In the granuloma ponch, radioactivity of the abdominal wall proves to be greater than that of the dorsal wall.

1. Rat liver mitochondria are swollen by inorganic phosphate in the medium of slightly hypotonic sucrose solution containing respiratory substrate and the mitochondrial swelling is inhibited or turned to shrink by ADP, respiratory inhibitor, anaerobiosis and uncoupler of oxidative phosphorylation. This mitochondrial swelling is not inhibited by the inhibitor of phosphorylating respiration such as oligomycin and tributyltin chloride. 2. Rat liver mitochondria are swollen by ATP in the presence of antimycin A, inorganic phosphate and 0.1 mM of CaCl2 and such a swelling is inhibited by oligomycin. 3. Accumulation of a small amont of P³² in acid soluble Pi fraction of rat liver mitochondria proceeds even in the medium containing neither ATP nor Ca++ but is inhibited by respiratory inhibitor, ATP, ADP and uncoupler of oxidative phosphorylation. The accumulation of P³² in mitochondria, however, is not inhibited by oligomycin. 4. The accumulation of P³² is induced by ATP in the presence of antimycin A and Ca++(O.l mM) and such an accumulation of P³² is inhibited by oligomycin. 5. It is suggested that the Pi-induced swelling of mitochondria is correlated to the accumulation of inorganic phosphate and both of them are tightly coupled to the initial step in the process of oxidative phosphoryaltion.

With the purpose to elucidate the cause and difference of blood fluidity in sudden death and natural one, we have observed the fibrinolysis of the blood in medico-legal and pathological autopsies by means of Fibrin Plate Method, a routine method devised in our laboratory. As the result it has been found that in the blood serum of sudden death and in some of natural deaths from tumors, leukemias, etc., the decrease in fibrinolytic activity is equivalent to the amount of proactivator that combined with the SK-like substance liberated into blood. On the other hand, in the blood of most of natural deaths, and in that bled from vessels and stored in body cavities, no natural fibrinolysis is observable and the same fibrinolytic activity with SK as normal one is demonstrated. Thus it is concluded that the cause of blood fluidity in sudden death is due to the fibrinolysis.

Some bile acids (dehydrocholic, cholic, chenodeoxycholic, ursodeoxycholic, and deoxycholic acids), and some hypocholesterolemic agents (22, 25 diazacholestanol, 20,25-diazacholesterol, triparanol, and SKF 525-A) are the inducers of isovalthinuria in guinea pig. Administration of methionine appears to increase the pool of sulfur compound which participates in the formation of isovalthine. Cholesterol appears to have no enhancing effect on the induction activity of isovalthinuria inducers. The mechanism of isovalthine formation and the role of sulfur amino acids in lowering blood cholesterol are discussed.

Using the method given by GOLDSTEIN (1961)9 for the determination of serum beta-glucuronidase activity, this value was determined in both normal and patients with epilepsy, neuroses, psychoses and multiple sclerosis. Of the patient groups examined, the group of those suffering from epilepsy is the only one showing any difference of statistical significance for all four methods of determination. The group of patients suffering from neuroses differs significantly from the normal group as regards the results got by the method of heat coagulation for removal of the proteins. The material is however too small to provide any explanation of the results, but it appears to show that a determination of serum glucuronidase activity may be of interest in groups of diseases other than malignant tumors.

A case of intramural pure pigment gallstones, which were fortuitously found in post-mortem examination, is presented. The incidence, mechanism of formation of the stones and roentgenological diagnosis of the intramural gallstones, porcelain gall bladder, are mentioned.

1. With strain L cells in culture the biological effects of oxygen rich environment have been observed with special reference to the cell growth, glycolysis, respiration, incorporation of P³² into Δ10 P and DNA synthesis. 2. Oxygen rich environment produces an increase in the vital activity of the cells at the initial stage of culture, i. e. increased activity of succinoxidase system, low glucose consumption and active cell growth, but in the later stage the activity of the cells is lowered likewise the activity of succinoxidase with the decreased oxidative phosphorylation, and an increase in the uptake of glucose and the enhanced lactic acid production. 3. The most adequate atmosphere for cell proliferation is found to be air though the reason for this remains unsolved. 4. Suppressed cell growth in the later stage of cell culture under the oxygen rich environment is accompanied by the increase of the cells containing smaller amount of DNA, but it is uncertain whether or not the decreased rate in DNA synthesis is responsible for the depressed cell growth.

Following Fibrin Plate Method of SZOLLOSY and RENGEI² , and ASTRUP and MULLERTZ³, the author conducted a series of experiments in an attempt to identify human blood by detecting the proactivator believed to be one of the enzyme proteins contained abundantly in human blood. As the results it has been found that with 0.1 mg. % SK-solution human blood alone responds to the reaction, showing almost absolute species-specificity within 4 hours but not with blood of monkey. In addition, the sensitivity is so high that it responds positively up to the dilution of 1: 8,000 to 1: 10,000 (human blood: physiological saline solution). By means of this method using 0.1 mg% SK-solution it has been clearly demonstated that the identification of human blood is possible in a variety of conditions and states as may be encountered in practical legal medicine such as with blood stains in cloth, wood, stone, leaves of tree even with a trace of blood stain, old human blood stain left standing for 20 to 30 years, old blood mixed with iron rust, blood stains soaked in various oils, and even the blood stained cloth washed thoroughly and left standing in room temperature for 6 months. Therefore, this Fibrin Plate Method seems to be the excellent one for the identification of human blood.

The synthesis of nuclieic acids in the liver and the lymphoid tissues of adult mice was studied during the restitution period after the 6-day starvation. The results obtained indicate that there occurs an unexpected rapid synthesis of DNA in the hepatic parenchymal cells during the restitution period without significant increase in the total amount of DNA in the liver. Most rapid DNA-synthesis in the liver appears to occur one day after refeeding. With respect to RNA in the liver as well as to both RNA and DNA in the lymphoid tissnes, on the other hand, there is a good parallelism between the rate of their synthesis and that of increase in their amounts, without apparent dissociation between both rates as seen in the liver DNA.

For the purpose to reveal the characteris6cs of the synovial fluid of the chronic rheumatoid arthritis the proteins of the synovial fluid and blood serum have been analysed by employing the methods of electrophoresis, gel filtration on Sephadex G-200 column and ultracentrifugation. Waaler-Rose test and latex fixation test have also been made on each protein fraction, and the following results were obtained. 1) The total protein level of synovial fluid, which is 3/5 of that of the serum, is slightly higher than that of control. 2) Fractionation of the synovial proteins by electrophoresis revealed nearly the same protein contents in each fraction in percentage as that of comparable fraction of the serum protein, with a slight increase in γ-globulin fraction. 3) The fractionation by Sephadex column G-200 give three peaks both in serum and synovial fluid, 19 S, 7Sand 4S. 4) 19S fraction of the synovial fluid, which is mainly of γ-globulin, showed a higher level than that of the synovial fluid from the controls. 5) Rheumatoid tests gave positive reaction in the 1st peak containing 19S γ-globulin from the synovial fluid and blood serum.

For the purpose of revealing the role of the reticuloendothelial system (RES) for the antibody formation, the rats which received the repeated intraperitoneal and subcutaneous injections of a vast amount of PVP were challenged by bovine serum albumin (BSA) introducing through 2 routes of intramuscular and intravenous, and then antibody formation was observed. Blood cell count and clearance rate of radiogold were observed for the purpose of obtaining the information of blockade grade of the RES by PVP. Phagocytic activity of macrophages ingesting PVP against iron colloid were also observed in vitro. 1. A severe anemia was induced by the administration of a vast amount of PVP, 15 ml of 3% solution daily or every other day for 63 days. Histological picture indicated the suppressed erythropoiesis probably by iron deficiency or the lowered iron transporting activity of the RES, as the anemia recovered after intraperitoneal iron injections. 2. With the generalized and marked swelling of the RES, the cells in germinal center of spleen and lymph nodes were extremely swollen and lymphocytes disappeared completely, suggesting that the macrophages in germinal center play an important role in reproduction and differentiation of lymphocytes. 3. The phagocytic activity of the RES as understood from the clearance rate of radiogold was suppressed only slightly even by a heavy deposition of PVP after the repeated injections. The state of blockade or the suppressed phagocytic activity persisted for 48 hours or more after the several PVP injections. However, complete blockade of the RES or inactivation of the phagocytic activity by PVP injection was not attained. 4. A prolonged treatment of animals with PVP caused delay in the appearance of circulating antibody but the final titration reached the same level as that of control. The data suggest that the blockade of the RES by PVP induces the delay in the transmittance of the information for the antibody formation from the macrophages to the immunologically competent cells but no delay in the ingesting antigen by the macrophages.