Excretion of thiamine and its degradation product, pyramin, in human urine has been investigated by several authors. Although the excretion of pyramin seems to be proportional to thiamine intake, it has so far been measured only by the fermentation method, and pyramin has neither been isolated in its pure form, nor its chemical constitution proved. Three persons took 50-100 mg each of dibenzoylthiamine daily and urine samples for 8 or 24 hours were collected. In total 56 l urine after the dosage of ca. 4 g dibenzoylthiamine was obtained. It was diluted, adjusted to pH4-4.5 and shaked with an adequate amount of acid clay. The clay was eluted by baryta solution, the acidified eluate was evaporated nearly to dryness and treated with alcohol. The extract was dissolved in water, shaked with hot n-butanol and the butanol solution was poured into a cellulose column. The eluate was divided into 3 fractions by coloring with Dragendorff's reagent or thiochrome reaction and the first fraction was proved to contain 2-methyl-4-amino-5-hydroxymethyl-pyrimidine (OMP) by paper chromatography. Its further purification and concentration were carried out by partition chromatography and finally the presence of OMP was proved by paper chromatography and absorption spectrography, comparing to the authentic specimen of OMP. Also 2-methyl-4-amino-5 (2', 4'-dinitrophenyl)-hydroxymethylpyrimidine was isolated after the incorporation of 2,4-Dinitrofluorobenzene on the alcoholic extract.

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The photooxidation of reduced diphosphopyridine nucleotide to diphosphopyridine nucleotide in the presence of various flavin compounds and methylene blue was demonstrated. In this photooxidative reaction, the potocatalytic activity was most intense in riboflavin, somewhat less intense in flavinmononucleotide and still less in flavinadeninedinucleotide. A strong activity was also found in methylene blue.

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A new S-derivative of riboflavin was prepared by the reaction of riboflavintetraacetate with P_2S_4 in benzene or pyridine solution. This compound, mp. 234〜235℃, C_<25>H_<23>O_3N_4S, reacts readily with 2NNaOH in methanol solution to form free thioriboflavin, mp. 232〜234℃, C_<17>H_<20>O_3N_4S.

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A synthetic diet supplemented with 100μg of vitamin B_<12> per 100 g ration showed a growth promoting effect on young mice after a continuous intake for 30 days.

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The relationship between the reduction rate of TAD or TPD by the cerebrospinal fluid in the patients and their clinical patterns has been studied. (1) No reduction of TAD and TPD occurred by the cerebrospinal fluid in the patients such as poliomyelitis, cerebral palsy or hydrocephalus. (2) However, it was demonstrated that TAD and TPD were reduced significantly by the cerebrospinal fluid in tuberculous meningitis and purulent meningitis, which revealed pleocytosis and high protein level of cerebrospinal fluid. (3) Reduction rate of TAD and TPD decreased as the clinical condition was improved.

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Comparative studies were made on the ingestion of thiamine and its derivatives by human erythrocytes, erythrocyte stroma, and yeast cells. The experiments were carried out in vitro, and S, S-alkyl, O, S-diacyl, O-acyl, S-alkyl, and various other derivatives were tested. 1) First, the intensity of the thiochrome reaction of thiamine analogues was compared with that of thiamine using the cyanogen bromide method. 2) Next, using thiamine propyldisulfide the conditions suitable for comparing the ingestion of various derivatives by erythrocyte were determind. 3) Then the ingestion of various derivatives by erythrocyte was compared. The order of the incorporation degree was as follows : S, S-alkyl derivatives>S, S-alkyl-O-acyl>O, S-diacyl>thiamine>O-acyl. 4) The incorporation of the derivatives into erythrocyte stroma was also examined. In this case, remarkable differences such as in the case of erythrocyte were not observed. 5) The ingestion of the derivatives by yeast cells was examined. The appearence of the incorporation and the order of the incorporation degree of the compounds were considerably different from those in the case of erythrocyte. 6) The mechanism of the ingestion of thiamine derivatives through cell membrane was discussed.

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In previous reports, metabolic disturbances of pyruvic acid and α-ketoglutaric acid have been shown to exist in liver diseases. Present study deals with the therapeutic effect of thiamine, thiaminepropyldisulfide and cocarboxylase on these metabolic disturbances in patients with chronic hepatitis. The influence of these vitamins in rats with experimental chronic liver damage caused by CCl_4 was also studied. The metabolic disturbances of α-keto acid were often normalized by the administration of thiamine, thiamine propyldisulfide and cocarboxylase, the latter being most effective. Patients with abnormalities in α-keto acid metabolism, showing various symptoms including fatiguability, nervousness, abdominal discomfort, in spite of the ordinary treatment, were given daily injection of cocarboxylase, flavinadeninedinucleotide and flavinmononucleotide for seven days. By this type of treatment, remarkable improvement in clinical symptoms and the metabolism of α-keto acids were observed in most cases. The significance of the disturbance of α-keto acid metabolism in chronic hepatitis was discussed.

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Experiments were made on about 100 each of normal subjects and tuberculosis patients to see if there are any relations between the metabolism of thiamine, the metabolism of pyruvic acid and the liver function. 1) There was correlation between thiamine deficiency and disturbances in liver function and a more closer interrelation between thiamine deficiency and disturbances in metabolic function. 2) Sawada's urinary test was found to be interdependent upon disturbances in liver function and vagotonia. 3) In the group of subjects with liver function disturbances or vagotonia, there was a marked correlation between the results of Sawada's urinary test and those of thiamine administration test. 4) The cases showing an increased urinary excretion of pyruvic acid were all positive for Sawada's urinary test. 5) Changes in total protein and gamma globulin levels were most closely related to thiamine deficiency. 6) The group with positive Takada's test showed a rise in gamma globulin level, while the group with thiamine deficiency showed a fall in gamma globulin level.

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When an excessive amount of thiamine was loaded with perfect meals to men, the urinary excretion of the vitamin varied by the way of loading. Administration of three times a day (A) showed a larger excretion than that of once a day (B). Changing the diet to thiamine deficient one, the urinary excretion of the vitamin reached the minimum level after a few days in both conditions, but it was always higher in A than in B. This difference seems to be resulted not to difference in the leakage of the vitamin but from a lower thiamine requirement in the A condition. Loading the vitamin with meal is supposed to lower the thiamine requirement by about 20 per cent compaired with that made between meals.

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The effect of 2-methyl-4-amino-5-hydroxymethylpyrimidine (OMP) and hydroxylamine on the enzymic system related to glutamic acid metabolism in the brain and involving vitamin B_6,that is, glutamic decarboxylase and glutamic-aspartic transaminase was studied. It was found that when a running fit is induced by OMP, the OMP is phosphorylated in the body and inhibits glutamic-aspartic transaminase but this enzyme system is not affected by hydroxylamine. Glutamic decarboxylase, on the other hand, is inhibited by both OMP and hydroxylamine and this inhibition takes place prior to the onset of the running fit. It is suggested that the inhibition of glutamic decarboxylase plays an important role in the inducement of the running fit.

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Using Crammer's concentration method and paper partition chromatography, several fluorescent substances were obtained from the acetone powder of Mycobacterial cells. These fluorescent substances were purified into single components, and their chemical structures and biological significances were attempted to determine. The results obtained are as follows : 1) Six kinds of fluorescent substances were detected with certaincy from Mycobacterium avium. The red fluorescent substance is coproporphyrin ; the two yellow fluorescent substances are FAD and FMN ; the two bluish green fluorescent substances are both folic acid derivatives, the one has growth promoting activity of Leuconostoc citrovorum but has little activity of Coenzyme F, and the other is the reverse ; the bluish violet fluoresnt substance is an unknown compound. 2) Any difference of distribution of these fluorescent substances were not observed between the resistant strains of both streptomycin and p-aminosalicylic acid and the sensitive strains of them. The red fluorescent substance was not detected specifically from the resistant strain of INAH. This phenomenon suggested us the nature of the resistance and the antibacterial mechanism of INAH. 3) Making clear the distribution of the fluorescent substances, it may be able to classify Mycobacteria.

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It was observed by human experiment that the folic acid excreted in feces as well as the vitamin synthesized markedly increased with the change of an ordinary diet to a vegetable one. On the contrary, both values of the vitamin decreased after the diet was switched from an ordinary diet to a meat one. The addition of cellulose to each diet caused a definite rise in both amounts of the vitamin, suggesting that the main promoting effect of the vegetable diet is due to cellulose contained therein. The rise in the vitamin is roughly proportional to the increase in the amount of feces. The vitamin in urine was very low in amount as compared with that in feces.

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