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1. Twenty.one patients with liver diseases were studied for their urinary mehylmalonic acid excretion after a valine load by means of an improved thin layer chromatography. 2. Methylmalonic acid positive cases were found in four out of the ten patients with cirrhosis of the liver, all four with cirrhosis and diabetes mellitus, and none with acute hepatitis of icteric phase. No apparent correlation was found between the methylmalonic acid excretion and the extent of hepatic damage. 3. A large amount of methylmalonic acid found in the case (S. I.) with cirrhosis of the liver and diabetes mellitus after the valine load was not corrected by cyanocobalamin but by DBCC, suggesting an impaired transformation from cyanocobalamin to DBCC. However, the nature of the impairment remains unknown.

The correlation of infectivity to the length of SV 40 DNA by dilute and undiluted passage was described. DNA was extracted from purified virions by Marmur's method, and propagation of virus was done using VERO cells. The infectivity of SV 40 (simian virus 40) ran parallel with the length of its DNA. Undiluted passage caused shortening of DNA and decrease in infectivity, but when these undiluted group was passaged dilutely, length of DNA approached the original length and the infectivity recovered. In undiluted passage group small circular DNAs under 1.0,11_, so far not reported in the SV 40 DNA, were found in low frequency. Replicating form and dimers were also noted from virions and the nuclei of SV 40 infected VERO cells.

1. The mixed lymphocyte culture test where live lymphocytes of female mice are cultured with supersonicated cell homogenate of isogeneic male mice does not reflect the difference in H-Y histocompatibility antigen alone. 2. When non-H-2 antigens are cumulated in various combinations, the rate of blastformation becomes greater than the combination of C3H ♀ themselves. 3. When non-H-2 antigens and H-Y antigens are added in various combinations, the rate of blastformation becomes significantly higher than in the combination of C3H ♀ themselves.

Antigenicity of clozapine was studied in rabbits, comparing with that of chlorpromazine as control. The results indicate that chlorpromazine produces antibody in rabbit as revealed by passive hemagglutination test, giving the titer of 1 : 2, 000 or higher in all the five cases observed, though specific precipitin lines has not been obtained and PCA test proves to be negative. Clozapine failed to produce anti.clozapine antibody giving negative passive hemagglutination test, passive cutaneous anaphylaxis and precipitin reaction, in all forms tested. Some remarks were made on the possible close relation between the antigenicity of the drug and its affinity to protein.

For the purpose to reveal the relation between cell death and nuclear stainability by supravital staining with basic dyes observations have been made on the cells of bone marrow, peripheral blood and lymph node from anemic and non-anemic rabbit, rat, mouse and chicken, and thymus from young mouse. The cells were stained supravitally in blood serum, isotonic saline, calcium chloride and sucrose solutions with the dyes; brilliant cresyl blue (B. C. B), Nile blue (N. B.), neutral red (N. R.), Janus green (J.G.) and eosin (E.). The following results were obtained: 1. In the presence of blood serum all the living cell nuclei observed were not stained supravitally, except some mature erythroblasts and nucleated red cells. 2. In isotonic saline, CaCl2 and MgCl2 solutions all the erythroid cell nuclei were stained deep by B. C. B., N. B., N. R., slightly by J. G. but not by E. In stainability the younger the cell is the deeper in its nuclear staining. The nuclei of other cell strains were not stained. 3. In isotonic sucrose the nuclei of mature granulocyte were also stained by B. C. B. and N. B. but not by other dyes. The nuclei of lymphoid cells and myeloid cells appeared pale without being stained by any dyes. The nuclei of erythroblasts in sucrose solution were stained deeper with B. C. B. and N. B. than those in isotonic saline. The differences between supravital stainability of the nuclei among the cells belonging to different strain and among those of the same strain but in different maturation stage and the nuclear staining after cell death have been discussed from the possible dissociation of DNA from histone.

1. Both EACA and AMCHA clearly showed an anti-inflammatory effect, by intravenous, intramuscular, or oral route, against inflammatory edema produced in rats by intracutaneous injection of rabbit's anti-rat serum, carrageenin, histamine, serotonin, or bradykinin, as tested by the punch method. 2. The two compounds also showed inhibitory action against the cotton pellet granuloma when used in a larger dose. 3. There was virtually no difference between the two compounds in their anti-inflammatory activity, in spite of the fact that antiplasmin activity of AMCHA is evidently greater than that of EACA. In addition, there was no increase in fibrinolysis at the site of antiserum inflammation in rats. Therefore, it would be difficult to presume that the anti-inflammatory action of these compounds is due to their antiplasmin activity. 4. Salicylates, pyrazolidine derivatives, and non-steroidal antiinflammatory agents like flufenamic acid inhibited degranulation of isolated rat mast cells induced by compound 48/80 and also inhibited ATP-32Pi exchange reaction in rat liver mitochondria, but such actions were not observed in EACA or AMCHA. 5. Anti-inflammatory effect of EACA and AMCHA did not decrease after adrenalectomy but did become weak in hypophysectomized rats. EACA did not increase blood sugar level in normal rats so that its antiinflammatory action is not due to hyperglycemia, and the effect of hypophysectomy may not be correlated to carbohydrate metabolism. 6. Anti- inflammatory effect of EACA and AMCHA appeared more rapidly after intramuscular or oral administration than by intravenous injection but the effect was not fortified after their in vitro incubation with tissues of stomach, intestine, or liver.

To obtain some information of the biological action of Kankohso 101dinicotinate and Kankohso 301-nicotinate, observations were made on the binding mode of these substances with protein, chondroitin sulfate and nucleic acids and the following results were obtained; 1. Kankohso 10 I-dinicotinate binds reversively with bovine serum albumin or serum r-globulin, resulting in metachromasia. By binding with proteins the absorption maximum of the dye shifts toward the long wave length side and the absorbance decreased distinctly. The data show that there are more than one kind of binding sites and the binding with bovine serum albumin is weak in acidic solution and strong in alkaline solution. 2. Kankohso 10 I-dinicotinate produces strong metachromasia with sodium chondroitin sulfate and the color of the solution changes from violet blue to reddish violet. The absorption maximum at 592 mp. decreases without shifting its wave length ,and the shoulder appears at 555 mp. be. comes distinct peak. The strongest metachromatical changes occurs at the concentration of the chondroitinsulfate whose sulfonate radicals is equal to the molecules of Kankohso 10 I-dinicotinate. 3. Kankohso IOI-dinicotinate produces metachromasia with nucleic acid, where absorption spectrum is shifted toward long wave length and absorbance is decreased at a certain concentration. 4. Kankohso 301.nicotinate binds weakly with bovine serum albumin, the binding of which is reversible and the maximum binding number is 1.1 per molecule of albumin. Metachromasia cannot be produced by binding. Kankohso 30I.nicotinate does not bind with bovine serum γ-globulin. This compund does not produce metachromasia with sodium chondroitin sulfate but produces weak metachromasia with nucleic acid, indicating some affinity to nucleic acid.

Transformation of Japanese encephalitis antibody from IgM to IgG in the sera of the experimental infected chicks with Japanese encephalitis virus and transmission of IgM or IgG from hen to chicks were examined by the gel filtration on Sephadex G-200 column. The following results were obtained. 1. Titer of hemoagglutination inhibiting antibody rose on seven days after inoculation of mouse brain homogenate infected with Japanese encephalitis, and that increased rapidly after the second inoculation of Japanese encephalitis. The maximum peak of antibody titer attained on 35 days after the first inoculation, on 7 days after the second inoculation and it maintained for a period of 2 months then decreased. Viremia was detected till 6 hours after the first inoculation. 2. IgM antibody by gel filtration appeared on 7 days after the first inoculation, kept on rising, reached the peak on 35 days after the first inoculation, then decreased, and disappeared on 120 days. IgG antibody appeared about 2 weeks after the IgM antibody appearance, and the titer of IgG antibody became higher than that of IgM antibody on 35 days after the first inoculation, then decreased gradually, and showed 1 : 16 of titer of peak on 150 days by gel filtration. 3. We could obtain the chicks by fertilization from experimentally infected hen, having IgM and IgG of hemoagglutination inhibiting antibody of Japanese encephalitis. And the localization of antibodies in the sera of its chicks was determined by Sephadex G-200 gel filtration. And IgG antibody was detected in chick serum, though IgM antibody was not detected by this method.

As to trial toward the elimination of Japanese encephalitis virus in natural surroundings, pigs received inoculation of inactivated Japanese encephalitis vaccine supplemented with complete Freund's adjuvant twice at one-week interval. Effect of adjuvant supplement on the magnitude of antibody and also prevention of viremia caused by natural infection by antibody induced with vaccine were investigated. The results of this study are summarized as follows. 1. In the group of pigs inoculated with vaccine containing adjuvant, titer of hemoagglutination inhibiting and neutralizing antibodies was higher than those inoculated with vaccine alone and their high titer persisted. 2. With respect to natural infection of pigs, on August 22 when the pigs were thought to have been infected, there was observed a rise in antibody titers. And on antibody formed in those pigs inoculated with vaccine with or without adjuvant proved to be all 2-ME resistant type, whereas the antibodies produced in control group were 2-ME sensitive antibody. 3. Viremia was detected in the blood of pigs naturally infected, but it was not demonstrated pigs inoculated with vaccine supplemented with adjuvant or without adjuvant. The virus of pig blood which was inoculated into suckling mouse brain and was separated after low suckling passage mouse was supposed to be JaGAr strain from optimum hydrogen ion concentration of its hemoagglutination reaction. 4. Effect of vaccination on antibody response of pigs having maternal antibody was not recognized.

The first successful electron-microscopic observation of a virus isolated from a patient with SMON was performed. The morphological and developmental characteristics of this virus suggests that this type of virus has not been isolated from humans. Hence, it is considered that the virus observed is of a new type and presumably the causative agent of SMON. The author wishes to express his profound thanks to Prof. TADASHI OFUJI for painstaking proof reading of the manuscript and also acknowledgement is due to Mr. NOBUO HAYASHI, Mr. NOBORU SAIHARA, Mr. TAKASHI NAKAMURA and Miss TOSHIYO OMIZU for their technical assistance of electron microscopy.

Epidemiological study on the prevalence of SMON III Ibara City, Yoshii Town and Yubara Town in Okayama Prefecture was conducted and the following results were obtained. "Epidemization precession" (Die Prazession der Durchseuchung), described by Rudder which shows that ages common to this disease is shifting to younger generation as incidence rate increases, in the endemic regions was recognized in Ibara City and Yubara Town. The mode of the distribution of the interval between the primary and secondary patients in a family suggests that it corresponds to the incubation period. Distribution of the patients to the size of family is not adaptable to binomial model, but adaptable to chain binomial model, and the intra-household transmitted rate is 5 per cent, and the true intra-household transmitted rate is about two times as high as extra-household transmitted rate by the modified chain binomial model. Intra-hospital incidence among SMON patients and non-SMON patients as well as medical workers were observed at the time when the number of new patients increased in a hospital. In SMON patients, the rate of appendectomized ones was higher than that of non-SMON patients, suggesting that gastrointestinal tract has some relationship to the development of SMON disease.

A non.specific esterase activity was demonstrated III the jejunum of rats by an azoindoxyl method. 1) Microvilli of the jejunal epithelial cells were remarkably stained in non-frozen specimens and feebly in frozen specimens. 2) The other cytoplasmic structures, i. e. mitochondria, endoplasmic reticulum, nuclear envelope, plasma membrane and multivesicular body showed a positive reaction product in frozen sections but not in non-frozen blocks.

The distribution and activities of five hydrolytic and eight oxidative enzymes were histochemically studied in 60 different tumors of the human intestines. Benign polyp showed similar activities of most enzymes as those in normal crypt cells of large intestine with exception of higher activity of succinic dehydrogenase in benign polyp than in crypt cells. Malignant polyp had higher activities of most oxidative enzymes. Reticulo-sarcoma had weak activities of all enzymes. Carcinoid had strong activities of glucose-6-phosphate dehydrogenase and isocitric dehydrogenase while very weak of succinic dehydrogenase. Carcinoma showed varying degrees of the activity of all enzymes. Alkaline phosphatase and aminopeptidase were almost negative in all cells but in the stromal elements their weak activities were sporadically observed. Most enzymes were decreased in the central area of the carcinoma cell nestle, while in the infiltrating area or in the margin of cell nestle they were not decreased and sometimes increased.

1. Isovaleric acid-1-C14, -4-C14, or C14-CaC03 with or without non-isotopic isovaleric acid was orally administered to rats and the incorporation of these isotopes into liver cholesterol, fatty acid, or urinary isovalthine was examined. 2. Isopropyl group of isovaleric acid was more efficiently utilized for cholesterol synthesis than carboxyl group, and also for cholesterol synthesis than for fatty acid. These results indicate that isovaleric acid is cleaved into two fragments before it is utilized for cholesterol synthesis. 3. Carbon dioxide was used for the synthesis of liver cholesterol and of liver fatty acid. Isovaleric acid seems to enhance the incorporation of carbon dioxide into cholesterol. 4. All the experimental rats received isotopic or non-isotopic isovaleric acid excreted isovalthine, but no radioactivity was found in it. Thus, isovaleric acid residue of urinary isovalthine molecule is not derived from isovaleric acid administered, and carbon dioxide is not the carbon source of urinary isovalthine. 5. Suspicious metabolism of isovaleric acid or of carbon dioxide was discussed. 6. Isotopic isovalthine which was synthesized from (± ) α-bromoisovaleric acid-4-C14 is administered to rat and it was found that the isotope did not incorporate into cholesterol or fatty acid of liver and of brain. About 15% of isotopic isovalthine was recovered in urine up to the next day after injection. The large part of isovalthine was missing.

1) Three different types of muscle fibers were clearly distinguished in the intercostal muscles and the diaphragm of human, cat and rat by histochemical demonstration of oxidative enzymes, phosphorylase and glycogen. 2) The intercostal muscles and diaphragm each presented dissimilar patterns of the muscle fibers. The diaphragm did not show any definite correlation between the diameter and the histochemical reaction of muscle fibers but its red fibers indicated a more intense uptake of Nitro-BT formazan deposits in the subsarcolemmal region. In this conection, the relationship between the motor innervation and histochemical evaluation of the diaphragm was described. 3) Phosphorylase and glycogen showed reciprocal reactions to the oxidative enzyme activity. They were generally high in large fibers but low in small fibers, and moderate in intermediate fibers.

With gastric carcinomas the activities of eight dehydrogenases; succmlC, lactic, malic, α glycerophosphate, glutamic, β-hydroxybutyric, glucose-6-phosphate and isocitric dehydrogenase were statistically estimated. Principal findings may be briefly summarized as follows. These enzymatic activities differed considerably even in the same classification of carcinomas and generally ranged from strong to weak in the following order: lactic, malic, glucose-6-phosphate, isocitric, succinic, α-glycerophosphate, glutamic and β-hydroxybutyric dehydrogenase. The activities of adenocarcinomas were stronger than those in simple ones, and these were not related appreciably to cell differentiation in adenocarcinomas except succinic, glutamic, glucose-6-phosphate and isocitric dehydrogenase. As for succinic dehydrogenase and NAD-linked dehydrogenases except for lactic dehydrogenase, the activities were strongest in intestinal metaplasia and early mucosal carcinomas, the next being in benign adenomatous polyps and weakest in the other carcinomas. As for NADP-linked dehydrogenases and lactic dehydronase, the activities were also strongest in intestinal metaplasia and early carcinomas, the second in the other carcinomas and the third in the benign polyps. Generally, these dehydrogenase activities were strongest in free carcinoma cells in blood and lymph vessels and in actively growing part of several carcinomas and weakest in the central area of tumors, especially almost negative in the central necrotic area.

Summing up the above problems we may group them as follows: 1) Szirmai's angio-myograph and myotonometer furnish us with means to evaluate the author's successful method or medical treatment in thrombophlebitic, postthromboembolic (ulcer, etc···) states on the basis of the blood circulation through the muscles, clearly registered by the angio-myograph. 2) Szirmai's medical preparation "HAH" serves as a quick and effective cure for thrombophlebitis. Results are very often reached within a few days. The patient's health is restored so as to make him able to work. 3) The above preparation assures full success in the cure of thrombotic esp. thromboembolic states of the lower limbs-cases of ulcus cruris includedwhich up to now could not be favourably influenced by any other method of treatment. Description of varieties of above problems and other types of cases of peripheral circulation (Endangitis, etc.) and their relationships with the subject will be given in additional papers. The author reports on registering and controlling thrombophlebitis, postthromboembolic states, including ulcus cruris, origin~ting either in above morbid conditions or in independent causes by means of the angio-myograph and myotonometer devised by the author. The reader is made familiar with the author's (Szirmai's) preparation "HAH" (Heacrin) and with the results achieved by applying it for the cure of acute thrombophlebitis and thrombotic states. Results are often showing up remarkably soon (2 to 6 days).

,The effect of infection of human embryonic skin-muscle cell cultures with adenovirus type 12 has been studied. When maintained in YLE containing 20 per cent bovine serum, human embryonic skin-muscle tissue culture cells developed little or no cytopathogenic effect for about 50 days after inoculation of adenovirus type 12, though a small amount of virus was always detected in the overlying medium. From day 50∼60, CPE started appearing and spread over 90 per cent of cells accompanied with the increase of virus in the overlying medium. The addition of human serum to the maintenance medium inhibited the virus release. After removal of human serum about 16∼37 days after its addition, virus-and, later, CPE also-again started appearing. The second virus release-and CPE also-was inhibited by addition of human serum to the medium. When maintained in the medium with human serum for about 200 days, the removal of human serum did not result in the appearance of virus or CPE. The virus isolated from the overlying medium of these cells during the whole process of the experiment was always highly oncogenic to newborn hamsters. Diluted adenovirus-12-immune rabbit serum also showed the effect similar to that of human serum. But, regardless of its much higher antibody titer, the effect of this diluted adenovirus-12-immune rabbit serum was weaker than that of human serum. In one of cell cultures, rapidly growing cells appeared 212 days after virus inoculation. But the available data suggest that these are the cells transformed rather spontaneously in tissue culture than by adenovirus type 12.

For the purpose to clarify whether minimal catalatic activity exists in Japanese acatalasemic cells or not and the manner how extrinsic hydrogen peroxide affects the acatalasemic cells, the author performed tissue cultures using the skin specimens from four acatalasemic persons affected with Takahara's disease and studied the nature of these cultured cells. The results are summarized as follows. 1. Between normal and acatalasemic cultured cells, no morphological differences could be seen and the growth rate of these cell-lines was similar to one another. 2. On the activity of succinoxidase and cytochrome oxidase there could be observed no difference between normal and acatalasemic cells. 3. In each acatalasemic cell line the minimal catalatic activity was observed and it seemed that this activity has an important role in decomposing hydrogen peroxide under normal metabolic pathway. 4. After treating with 10-4M hydrogen peroxide, respiratory enzyme activities and the growth rate in the acatalasemic cells were markedly disturbed, while in normal cells these remained almost intact. 5. There could be observed no differences between normal and acatalasemic cultured cells after X-ray irradiation (200 to 600 r) on the succinoxidase activity, catalatic activity and growth rate.

For the purpose to clarify the relationship between production of humoral antibodies and cellular reactions of the lymphoid system to allogeneic-transplanted cells in mice, a study on cross sensitization was carried out between inbred A(H-2a) and C3H(H-2k) strain mice. The median survival time of skin of C3H transplanted to A (C3H-to-A) was 14.1 ± 1. 4 days, and of A transplanted to C3H(A-to-C3H) was 11.8± 1. 6 days. Repeated A cell injections to C3H induced the formation of humoral antibodies, whereas the C3H cell injections into A did not. In A-to-C3H and C3H-to-A combinations, the immunization induced an increase in white blood cell number in circulating blood successively with the repetition of the antigen injection, and organ weights increased in thymus, spleen, and liver but not in kidney. Weight increases in the organs of A treated with C3H cell injection were less in extent, comparing to those of C3H treated with A cells. Histologic observations revealed that the cellular proliferation in the lymphoid system including plasmocytic responses were obviously predominant in the C3H treated with A cells comparing to those in the A treated with C3H cells. Hemocytoblasts also increased during the immunization in both cases showing no significant differences between the two series of experiments. These cellular reactions were observed not only in the draining lymph nodes but also in the generalized lymphoid tissues. The results of the present study suggest that the definitive factor for producing humoral antibodies is in the differences of the homologous antigenicity between the donor and the recipient but not in the degree of sensitization, and the Dk in H-2 loci is not so strong in antigenicity as to elicit sufficient plasmocytic responses for the formation of humoral antibodies in C3H strain mouse.