The effect of vitamin E on the experimental arteriosclerosis of rabbits caused by the administration of cholesterol was observed. In the groups treated with vitamin E, the change of aortic sclerosis was not so much remarkable as the control. It is supposed that the increased function of thyroid gland by vitamin E plays an important role in this effect.

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Following researches were made on serial and individual fluctuations of the nicotinic acid metabolism in healthy body by measuring the excretion of N'-methyl-3-carboxylamide-6-pyridone in urine. The data reported in the previous papers should be corrected as follows : 1) The average excretion of N'-methyl-3-carboxylamide-6-pyridone in urine per day (12 men and 6 women) was 11.8±2.30 mg. The individual and serial fluctuations were remarkable. 2) Proportion of excretion of the three products from nicotinic acid metabolism in urine was estimated as 100 : 192 : 492 for nicotinic acid : N'-methylnicotinamide : N'-methyl-3-carboxylamide-6-pyridone. 3) The average total excretion of nicotinic acid metabolism products in urine per day (12 men and 6 women) was 15.3±2.56mg.

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The author observed that by the administration of 200mg CaCO_3 to subjects for five days the urinary excretion of riboflavin increased ca. 2.6 times that of standard diet. In order to ascertain the cause of this findings, the following studies were made. Using homosulfamine as an inhibitor of riboflavin synthesis in intestines, the factor was assayed for 6 subjects for 22 days, dividing 4 periods of time, the 1st, standard diet ; the 2nd, CaCO_3 administration, the 3rd, CaCO_3+homosulfamine added ; and the last, standard diet. (1) Six experimental subjects showed maximum excretion of riboflavin in the urine at the CaCO_3 administration period. Only one subject showed some reduction in the urine output at the CaCO_3+homosulfamine period, but not so low as homosulfamine alone. (2) The concentration of riboflavin in blood increased about all subjects in next order. 2nd period (CaCO_3 administrated)>1st period>3rd period. (3) The fecal excretion of riboflavin could not show any evident results. (4) An attempt for rabbits to inhibit the biosyntnesis of riboflavin in the intestine by the administration of homosulfamine for a period of five days met with success as well as in man.

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The mouse injected with TXP-acetylpyridine died with a quite different manner from the case of individual injection of either TXP or 3-acetylpyridine. Expecting for the competition with TXP-acetylpyridine, we examined the action of pyridoxamyl niacinamide synthesized from pyridoxamine and niacinamide, which antagonized to TXP and 3-acetylpyridine respectively. But no evident antagonism was seen between these two substances.

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The present experiment was designed to ascertain the availability of dehydroascorbic acid as a source of vitamin C for the human subject. Comparison of results with oral administration of dehydroascorbic acid and of ascorbic acid showed interesting differences. Solutions of dehydroascorbic acid, freshly prepared by oxidation of ascorbic acid with bromine were used. After giving equal doses of dehydroascorbic acid and ascorbic acid, equal amounts of ascorbic acid were excreted in urine, but after oral administration of dehydroascorbic acid the blood and urinary vitamin C increased more rapidly than in the case of ascorbic acid. The fact that the same dose of ascorbic acid and dehydroascorbic acid given orally leads to the excretion of similar amounts of ascorbic acid in the urine suggests that the latter is almost quantitatively reduced to ascorbic acid and is satisfactorily utilized in the body. Dehydroascorbic acid readily penetrates erythrocytes and is reduced to ascorbic acid. After adding 2.26 mg of dehydroascorbic acid to 10 ml of leucocyte-free blood, ascorbic acid content per 100 ml in plasma was 4.4 mg, and in erythrocytes 14.8 mg after 15 minutes at 37℃.

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Distribution of thiamine-degrading bacteria was tested on 291 rat feces collected in Osaka Prefecture. Thirteen strains were found to belong to Clostridium thiaminolyticum, three to Bacillus thiaminolyticus and one to Bacillus aneurinolyticus.

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A microbiological assay method of vitamins in samples containing colistin and bacitracin, cyclic peptide antibiotics, was described. Antibacterial activities of these antibioties to St.faecalis, Leuc.mesenteroides P-60,L.arabinosus 17-5,L.casei and L.fermenti 36 were tested, in order to screen a species of which growth was not inhibited. St.faecalis R and L.casei were naturally resistant to colistin and not inhibited even up to 1000μg/ml of colistin. On the other hand, the growth of all these lactobacilli was inhibited in the presence of relatively low concentration of bacitracin. The bacitracin resistant strains of St.faecalis R and L.arabinosus 17-5 were isolated by successive transfer in the medium containing graded amount of bacitracin. Assay range of pantothenic and folic acid with the former and that of pantothenic acid with the latter were different from that with parent strains. In the case of resistant strains, there was observed a little shift towards high concentration of vitamins. It was shown that when the colistin was contained in the sample, pantothenic, folic and nicotinic acid were able to assay with St.faecalis R, and when the bacitracin was contained, these vitamins were determined by bacitracin resistant strains.

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The growth of St.faecalis R was inhibited by low concentration of erythromycin. The inhibitory concentration was approximately 20〜200 mμg/ml. The extent of inhibition was decreased competitively as the Ca-pantothenate was increased. The growth of L.arabinosus 17-5 was inhibited by 100mμg/ml of erythromycin. But this inhibitory effect was not reversed by Ca-pantothenate. Inhibitory effect of erythromycin to erythromycin resistant strain of St.faecalis R, was also observed. It was suggested that erythromycin inhibits the utilization of Ca-pantothenate, but the mechanism is not single one as an antimetabolite.

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Dihydrostreptomycin resistant strain of L.casei is isolated by successive transfers in the bouillon medium containing glucose, yeast extract and graded amount of dihydrostreptomycin. This strain grows well in the bouillon medium, whether dihydrostreptomycin is present or not, but can not grow well in the semisynthetic medium that is used for vitamin assay, if it does not contain dihydrostreptomycin. L-Asparagine and L-glutamine are able specifically to substitute dihydrostreptomycin as a growth factor in the dihydrosteptomycin resistant L. casei. It is also observed that the growth inhibitory effect of dihydrostreptomycin to L. casei is reversed by the addition of L-asparagine or L-glutamine. It is suggested that asparagine and glutamine have important role for this strain to acquire resistance to dihydrosteptomycin.

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Kamikubo and Tanaka reported that vitamin B_<12> activity is eluted from carbon cake, on which vitamin B_<12> in activated sludge was adsorbed previously, by using of 60% acetone after washing of the cake with 15% acetone with good yield. In order to economize the solvent and the cost of equipment, experiments were made by using a low concentration of acetone. When 60% acetone is mixed with 10% butanol, carbol or ethyl acetate and the mixture is heated with the cake, vitamin B_<12> is eluted in a higher yield than when 60% acetone is used. When 10% acetic acid, dimethyl amine or 0.1 N sodium bicarbonate is added to the above mixture, the yield of vitamin B_<12> is higher than by using any of the above mixed solutions alone.

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It was observed that in the induced rachitic rats the thiamine contents in liver and blood were considerably lower than those of the normal rats. When the rachitic rats were given intramuscularly thiamine and vitamin D, two days later the thiamine contents of the rat tissues, especially liver, developed equally those of the normal cases. But when they were given thiamine only, the increase of thiamine concentration was not obvious. This result suggests that the administration of vitamin D has caused the accumulation of thiamine in liver tissue.

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The hourly change of thiamine content of the induced rachitic rats was investigated by dividing into two groups. In one group, thiamine contents in liver and blood were measured after administration of thiamine and vitamin D, in the other group thiamine alone. Twelve and 24 hours later after loading, there was no difference between them, but 48 and 72 hours later the thiamine content of the first group was considerably higher as compared with that of the second group. A similar experiment was made in human infants, and the thiamine content in blood of vitamin D administered group was higher than that in the control group.

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In order to exclude the action of phosphatases, thiamine extraction from animal tissues was carried out in the ice-cold. The tissues were kept in the cold and homogenized, then heated. The estimation of thiamine by thiochrome method resulted in showing a negligible amount of free thiamine in kidney and intestine, while total thiamine was almost same as that of ordinary extraction procedure. In liver free thiamine was not found. The fair exsistence of free form of thiamine previously reported in these organs might be due to liberation during the process of extraction.

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During the last few years we have been studying the pharmacological and therapeutic effects of pyridoxal phosphate. In this paper the author examined its effect in the treatment of patients with hyperthyroidism. Moreover, the function of pyridoxal phosphate upon hyperthyroidism was studied. Twenty-nine cases of hyperthyroidism were treated with pyridoxal phosphate. In 15 cases the toxic manifestation was improved satisfactorily and the basal metabolic rate returned to a normal range. Blood-iodine, examined on 5 patients, showed in 4 cases a fall of the protein-bound iodine to a normal level. In experimental studies, pyridoxal phosphate inhibited decarboxylase and tryptophanase of Escherichia cold and oxygen consumption of the tissues of rats. The synthesis of organic iodine compounds by surviving slice of thyroid tissue was inhibited by pyridoxal phosphate.

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Investigations were made on the absorbing mechanism of vitamin B_6. Vitamin B_6 was rapidly absorbed from small intestines in the order of pyridoxal, pyridoxine and pyridoxamine. Kallikrein and pituitrin acclerated the absorption, epinephrine inhibited it and insulin, thyroid hormone, cardiac hormone, sexual hormone and parotic hormone had no influence on the absorption. The effcts of bile and cholic acid on the absorption were uncertain. From these observations, the author supposed that vitamin B_6 was absorbed mainly by the physico-chemical processes, such as diffusion, osmosis etc., and a smaller part by phosphorylation.

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Injection of horse serum caused an anaphylactic shock on guinea pigs. Changes of the blood riboflavin level, taken out by cardiocentesis were observed. Within 10-20 minutes after the injection, the total riboflavin level in the blood decreased by 1.2-3.5 μg/dl, while the level of ester form decreased by 2.0-4.3 μg/dl. Within the next 120 minutes, each was nearly restored to the value before the shock. Namely in this case, riboflavin level rather increased slightly. Free riboflavin was administered 15 or 30 minutes before, and shortly after the injection of horse serum. In such cases, the total and esterified riboflavin level decreased by 5.0-7.4 μg/dl and 7 μg/dl, respectively. In mild cases or animals which survived for a relatively long time, the decrease of free riboflavin was remarkable. It seemed that the above-mentioned changes of the vitamin levels are related to histamine.

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As in the previous report, an anaphylactic shock was caused on guinea pigs by injecting horse serum. The anaphylactic shock was followed by the decrease of riboflavin concentration. The most remarkable decrease was observed in the liver. After 10-20 minutes, the total riboflavin concentration was about 3200-3250 μg% (normal 4780 μg%). The ester ratio decreased gradually after the anaphylactic shock occurred. After 90 minutes, about 23% of the vitamin decreased in the liver. Fourty-one percent decreased in the adrenal glands after 60 minutes. The amount of riboflavin, especially of free form, excreted in the urine during the next 12 hours was slightly higher than that of the control. When the shock occurred, the availability of the vitamin was decreased. It was supposed that a large amount of riboflavin might be transferred to gall bladder than that excreted in the urine.

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Guinea pigs were treated with 100 μg of cortisone per g. of body weight, in order to investigate the effect of cortisone on the riboflavin level in the blood when the anaphylactic shock occurred. Fifteen minutes after the subcutanous injection of cortisone, the anaphylactic shock was caused by the injection of horse serum. Thirty minutes after the shock occurred, the concentration of free riboflavin in blood was slightly higher than that of the animals, to which cortisone had not been administered. The ester forms of riboflavin decreased slightly. In the case of the administration of cortisone and riboflavin, the decrease of riboflavin level was not recognized at 15 minutes after the injection, on the contrary to the administration of riboflavin only. Consequently, the ester ratio was higher than that of the control. It seems therefore, that cortisone is effective on the phosphorylation of riboflavin.

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The riboflavin concentration in blood and in various organs of a guinea pig decreased by the injection which caused an anaphylactic shock. A chromatographical investigation was carried out on the amounts of the riboflavin relating substances in each tissue. In the blood, FAD and FMN decreased 20 minutes after the injection, and 60 minutes after, both increased a little. However, free riboflavin increased from 1.9 μg% to 2.2 μg% and 2.5 μg%, during the same time. In the liver, 20 minutes after the injection, FAD decreased remarkably (about 1400 μg%), while FMN (about 340 μg%) and free riboflavin (about 190 μg%) increased. In the kidneys, 20 minutes after the injection, FAD decreased remarkably (about 1000 μg%), while FMN (about 250 μg%) and free vitamin (about 400 μg%) increased. In the suprarenal capsule, 60 minutes after the injection, the remarkable decrease of FAD (about 1000 μg%) and the conspicuous increase of free vitamin (about 450 μg%) were observed. In the lungs, 20 minutes after the injection the decrease of FAD (about 350 μg%) was observed.

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The riboflavin concentration in the blood of two pharyngeal diphtheria patients with serum disease was examined. In the first case, a wide serum eruption appeared on the knee on the 7th day after the injection of serum. On 5th and 10th day, free and ester forms of riboflavin decreased, especially it was remarkable in case of ester form. In the second case, a slight serum eruption appeared on 5th day after the serum injection. On the same day, a slight increase of the amount of free riboflavin and decrease of that of the ester form were observed. On the 10th day the total riboflavin level was restored to original. The level of ester form, however, was still lower than the original. The results mentioned above suggest that the phosphorylating ability of riboflavin is inhibited when the serum eruption occurs.

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As the case of the investigation on the antagonistic effect of vitamin B_6 relating compounds upon the toxication caused by injection of toxopyrimidine, the preventing action of niacin and some of its derivatives against the toxic action caused by injection of the lethal dose of acetylpyridine to mice was examined. While niacin and niacin diethylamide were found to be inactive, niacinamide was shown to be very effective in accordance with the description of Kaplan et al.. Diphosphopyridine nucleotide was, however, found to be very weakly active contrary to their results. Triphosphopyridine nucleotide was also scarcely effective. Toxopyrimidyl niacinamide was found to inhibit the action of acetylpyridine in an equimolecular amount to that of niacinamide, though the nitrogen of its pyridine nucleus is quaternary like that of DPN or TPN. In order to investigate whether toxopyrimidyl niacinamide could antagonize toxopyrimidyl acetylpyridine, the latter compound was synthesized and investigation on its properties was carried out. The velocity of its toxic action was much higher than that of acetylpyridine itself and the symptom was different from that by toxopyrimidine. The death by the injection of toxopyrimidyl acetylpyridine could not be prevented by the injection of toxopyrimidyl niacinamide.

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Prior to various enzymatic studies on thiaminase II of Bacillus aneurinolyticus, stability of the enzyme was investigated and the following results were obtained. The purified thiaminase II was considerably stable against heating and the reaction catalysed by the enzyme followed Arrhenius's equation below 53℃ and Q_<10> was 2.2 The enzyme was unstable below pH 5.5 but comparatively stable in alkaline solution. Thiaminase II, however, was unstable in solution containing chelating agents which increased the enzymatic activity. Its stability was increased by metallic ions, such as Zn^<..>. Mg^<..> or Mn^<..>, and by several reducing reagents, such as cysteine, Na_2S and Na_2SO_3. On the contrary, Cu^<..> or Co^<..> was strong inhibitor for the enzymatic activity. By the application of the above mentioned results, the inactivated enzyme was reactivated by cysteine, one of the reducing agents and Zn^<..>.

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The activity of thiaminase II was measured by the determination of remained thiamine in case of low concentration of the substrate and of formed oxymethylpyrimidine in case of high concentration of the substrate. The enzymatic reaction in low concentrations of the substrate (below 10^<-4> M/l) followed the first order reaction apparently, but not in fact the first order reaction. The cause of this phenomenon was considered to be brought by the inhibitory effect of the products on the reaction. And then, the reaction was investigated according to an equation induced from the Michaelis-Menten's equation by considering the inhibitory effect of the products. V_<max> was directly determined by the measurement of oxymethylpyrimidine and Mm was calculated from this value. V_<max>=1.05×10^<-9> M/min. protein N'1μg, Km=2.77×10^<-5>M/l (pH 7.0,37.5℃). The velocity of this reaction increased in proportion to OH ion concentration. From this result, it was supposed that the reaction might proceed as follows : [chemical formula]

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