By 2,4-dinitrophenylhydrazine method the blood ascorbic acid level in the saturated state was found to be about 1.1mg/dl, which was a little less than that observed with indophenol method. The daily amount of ascorbic acid needed for saturation was about 100mg, and in a saturated state the amount of ascorbic acid consumed daily was estimated to be about 95mg. In later pregnancy, contrary to the view held at present, this amount decreased a little. During lactation it increased by the amount secreted in milk.

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In this paper, the decomposing mechanism of L-ascorbic acid by bacterium, which belong to Micrococcus species, isolated by this author, was studied. It was observed that when the cells were incubated in the synthetic medium which contained glucose and ascorbic acid as carbon sources, the decomposition of ascorbic acid began after the induction period succeeding to a complete decomposition of glucose. On the other hand, the cells grown in ascorbic acid containing medium could decompose ascorbic acid rapidly to CO_2 and H_2O, while the cells grown in glucose containing medium had no ascorbic acid decomposing activity. From the above mentioned results, it was supposed that the enzymes responsible for the decomposition of ascorbic acid were shown to be adaptive in nature, and this presumption was supported by the experiment that the adapted cells could decompose dehydroascorbic acid, while the unadapted ones could not. It was estimated, too, that the first step of ascorbic acid metabolism by adapted cells might be carried through the course of ascorbic acid→dehydroascorbic acid→ diketogulonic acid. Unadapted cells, also, were supposed to contain some enzymes which catalyzed the oxidation of ascorbic acid to dehydroascorbic acid under an aerobic condition and inhibition of which activity caused by 2,4-dinitrophenydrazine (10^<-3> or 10^<-4> M). As_2O_2 (10^<-3'>M), KCN (10^<-4> M), and NaN_3 was studied by Warburg's manometric method.

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Previously, the author isolated quinoid like yellow pigments as a thermostable antithiamine factor (SF) from horsetail. The thiamine decomposing activity of several oxyquinoid pigments was, therefore, studied in this experiment. The decomposing activity of p-henzoquinone resembled the one of flavonoid pigment in reactive conditions such as pH, temperature and time. The decomposing activity of 10μg of SF was found in 5.0μg p-benzoquinone, 4.7μg naphtoquinone, 4.9μg anthraquinone, 1.0〜1.7μg monohydroxyanthraquinone, 1.8〜2.5μg dihydroxyanthraquinone, and 2.1〜3.8μg trihydroxyanthraquinone. When the decompsoing products of thiamine by SF were studied by paper chromatography, the production of thiochrome was observed in the case of p-benzoquinone, but thiochrome was not produced by flavonoid. It was, therefore, supposed that each SF might have their own thiamine decomposing mechanism.

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Five hundred μg vitamin A was given daily for 2 weeks, and the amounts of the vitamin in feces, liver and blood were determined. It was found that the amount of the vitamin in feces and that in serum remained roughly the same without showing any tendency to increase. Therefore the saturation with the vitamin is not to be concluded from the finding of fecal vitamin. On the contrary, the amount of the vitamin in liver rose definitely. Twenty four hours after feeding 500μg vitamin A, the vitamin content in all organs was determined and a marked rise was found only in the liver. The vitamin found was only 22%, including the amount excreted in feces. The loss was 78% in 24 hours. After 100-400mg α-tocopherol was added to 1000μg vitamin A, the rise in the amount of the vitamin in liver was insignificant. The rise in vitamin A in serum and liver was determined at intervals after feeding a single dose of 1000μg vitamin A, and the highest value was obtained in 15 hours in liver and 3 hours in serum.

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Experiments on carotene metabolism was performed using male rats. Twenty four hours after an oral dose of 500μg β-carotene, the amounts of carotene and vitamin A in all tissues were determined. In all, 20 per cent was found as carotene or vitamin A and 80 per cent was consumed. Twenty four hours after feeding 100-400mg α-tocopherol in addition to 1mg β-carotene, the rise in amounts of carotene and vitamin A in each tissue was determined and the effect of tocopherol was found to be very slight. The highest values of carotene and vitamin A were found after feeding 1mg β-carotene, 12 hours in liver and 9 hours in blood. The increased amount of vitamin A in liver 15 hours after feeding varying amounts of β-carotene or vitamin A were compared and it was found that β-carotene was about 40 per cent as active as vitamin A on an I. U. basis.

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Correlation between menstrual cycle and blood level of carotene and vitamin A in the female was studied and it was revealed that in the ovulation and intermenstrual phase the levels of carotene and vitamin A were significantly higher than those in the other period.

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The P^<32>-labeled FAD was prepared from rabbit liver which was injected with a certain dose of P^<32>-phosphate and FMN. The extraction was made by Crammer's method, and the crude preparation was further purified by two phase chromatography (developers : 5% citrate・isoamylalcohol). P<32>-labeled FMN was obtained by hydrolysing FAD obtaining 5% trichloroacetic acid, and purified by the above solvent. In the present paper, the preparation method of P^<32>-labeled FAD and FMN, and the fate of these compounds in rats injected with them were deal with.

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Violet fluorescent substance was isolated by Crammer's method, column chromatography and paper partition chromatography. This substance is quite different from G and V-substances in Eremothecium ashbyii. From further examinations of various properties of the fluorescent substance it was presumed that it consists of pteridine body and saccharide.

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Observations were made in shaking culture of Escherichia coli communior in Anderson's medium added with the substance. Both riboflavin content and bacterial cell increased, but the ratio of two remained usually unchanged.

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The riboflavin content of the liver, kidney, muscle and heart of rabbits poisoned by arsenite was found to be reduced compared with normal animals. With regard to the distribution of the three types of riboflavin, it was recognized that FAD decreased and FMN and free riboflavin increased.

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On mice posioned by arsenite the riboflavin content was determined in the whole body and in various organs. Decrease was found in all cases compared with normal animals. Among the three types of riboflavin, FAD was reduced and FMN and free riboflavin were found to be increased.

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Two strains of pyridoxine requiring mutant obtained from E. coli Waksman's strain were available in this study. Pyridoxal and pyridoxol were highly active in this order in the medium of Davis-Mingioli or in that containing casein hydrolysate, while pyridoxamine was far less active. Casein hydrolysate, D-alanins and many of other amino acids spared pyridoxine and promoted the growth in media containing pyridoxine. The mutants, however, were u able to dispense with the vitamin even in the presence of casein hydrolysate and excess of D-alanine.

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In order to know the reducing substances for thiamine allyl disulfide in cerebrospinal fluid of a tuberculous meningitis patient, the effects of pH, temperature, reaction velocity, heat stability, dialysis, action of SH-inhibitor were investigated. It was considered that the reducing substances found in pathogenic cerebrospinal fluid consisted of SH compounds of lower molecular weight, such as glutathione and some SH-enzymes.

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Thiamine concentration in the cerebrospinal fluid was measured after intravenous and intramuscular administration of thiamine and thiamine propyl disulfide and the following results were obtained. (1) No difference of thiamine concentration in cerebrospinal fluid of normal children was found between the administration of thiamine and thiamine propyl disulfide. However, with meningitis patients, the concentration of thiamine in the cerebrospinal fluid was slightly higher when thiamine propyl disulfide was administered. (2) When thiamine propyl disulfide was administered intramusculary, the concentration of thiamine in cerebrospinal fluid was higher and maintained longer its level than when it was injected intravenously. (3) For patients in the acute stage of poliomyelitis an administration of thiamine propyl disulfide resulted in higher level of thiamine in the cerebrospinal fluid than an administration of thiamine hydrochloride. However, for patients in the convalescent stage there was no difference in thiamine concentration between thiamine and thiamine propyl disulfide administration.

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The correlation between the autonomic nervous and pituitary-adrenal system was investigated through the determination on the changes in ascorbic acid contents in the adrenal gland and liver of rabbits due to electrical and lead acetate stimulations to the Ganglion caeliacum. Total ascorbic acid concentrations in the adrenal gland and liver showed a marked decrease promptly 1-3 hours after the stimulations, thereafter there was a slight increase in 24 hours. When chlorpromazine (1 mg/100g body weight) was administered before the both stimulations, the adrenal and liver ascorbic acid had no significant diminution, as compared to the application of the above-mentioned stimulations alone. The stimulation by means of lead acetate on hypophysectomized rats did not bring about the lowered concentration of ascorbic acid in the adrenal gland. The injection of cortisone (5 mg/100g) has no appreciable influence on the adrenal glands. Only by the administration of ACTH (5 mg/100g) the adrenal ascorbic acid content showed a definite reduction.

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Method of detection of allicin homologues was investigated by means of paper partition chromatography. The results were as follows : 1) Iodine-azide, sodium nitroprusside and phosphomolybdenic acid reagents were suitable for the coloration of allicin homologues. Allyl radical of allicin homologues was colored by phloroglucin reagent. 2) Petroleum ether saturated with 95% methanol was best suited for the paper chromatography of [chemical formula] as the developing solvent, and the mixture of methanol and benzene (3 : 2) for that of [chemical formula]. 3) Cysteine was added to the extract of each spot on the paper chromatogram of [chemical formula] and these reactive liquids were submitted to paper chromatography with the upper layer of the mixture of butanol, ethanol, acetic acid and water (4 : 1 : 1 : 10) as the developing solvent. Alkyl radical of each spot on paper dhromatogram of [chemical formula] was confirmed by this method.

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Allicin homologues in the Allium plants were investigated by means of the paper chromatographic method stated in the first report, and the following results were obtained. 1) Allicin or its homologues are generally present in the Allium plants. 2) As the alkyl radicals of allicin homologues, CH_3-, C_2H_5- and C_3H_7-, but not C_2H_5-, are found in the natural products. 3) There are two types in allicin homologues. One has the two same alkyl radicals, and another has the two different alkyl radicals.

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Allithiamine, which is produced by the reaction of ingredient of garlic on thiamine, shows the more intense effects in the living body than ordinary thiamine. From this fact, considering whether the nutritional effects of garlic are due to the production of allithiamine in the living body, the author made experiments using the human blood and the rabbit. The results showed that when both of thiamine and garlic were taken together, similar effects to allithiamine were found, and the production of allithiamine in the living body was endorsed.

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The relationship of S-allylmercapto-L-cysteine and thiamine was investigated and the following results were obtained. S-allylmercapto-L-cysteine does not react with thiamine, but when hydrogen peroxide is added thiamine loses its characteristic thiochrome reaction and allithiamine is detected on paper chromatogram.

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Casting a doubt on the theory that when garlic is taken the synthesis of thiamine is facilitated by the intestinal bacteria and thiamine deficiency can be prevented, the author conducted supplementary experiments. The result of experiments carried out on animal and human body showed that when garlic was taken alone without thiamine, thiamine deficiency could not be prevented.

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Detection of alliin homologues in the several plants of Allium species was made by the paper chromatographic method, and it was found as the result, that allin homologues were present in all these plants. The amount of alliin homologues contained in these Allium plants was estimated by the measurement of the pyruvic acid and ammonia produced by the grinding of raw materials. The separation of alliin contained in garlic was attempted, and pure crystal of alliin was obtained.

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Although some studies on alliinase in garlic which belongs to the Allium species have so far been reported by Stoll et al., the author presumed that alliinase may be contained also in other Allium plants besides garlic. Accordingly researches on Allium plants have been carried out in an attempt to reveal the biochemical quality, content, and common feature of alliinase. The results thus obtained are summarized as follows : (1) Apparently a sufficient quantity of alliinase in proportion to alliin is contained in each Allium plant respectively. (2) A common feature is recognized in all Allium plants. (3) Alliinase considerably secures stability.

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In the course of studying on the distribution of alliinase and alliin in nature, the author has found several points of interest in the amino acids which Ipheion uniflorum Rof contains, Thus the author has conducted biochemical investigations designed to elucidate these points, and has obtained the following results. (1) Alliin homologues are contained in Ipheion uniflorum as its components. It has been demonstrated that they are S-methyl-L-cysteine sulfoxide ([chemical formula]), S-ethyl-L-cysteine sulfoxide ([chemical formula]) and S-propyl-L-cysteine sulfoxide ([chemical formula]). (2) No alliinase which decomposes alliin homologues is contained in Ipheion uniflorum. (3) Pyruvic acid, ammonia and allicin homologues are produced by addition of alliinase to the liquid extracted from Ipheion uniflorum just as to the liquid extracted from Allium plants. (4) By addition of thiamine to the liquid mentioned above, allithiamine homologues are produced.

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