The thiaminase, which is produced by Bacillus aneurinolyticus Kimura et Aoyama differs entirely in enzymic properties from that of other sources, which have hitherto been studied. Aniline, pyridine, quinoline and many other amines, which markedly activate the thiaminases of various sources, are completely inactive or rather inhibitive. Thiamine is decomposed by the enzyme into its pyrimidine and thiazole moieties contrary to the known enzymes, by which the thiazole moiety of thiamine is replaced by an amine. The only exception is aniline, which can replace the thiazole moiety of thiamine. The enzyme synthesizes pyrimidinemethylaniline from pyrimidinemethanol and aniline. Its reverse reaction is also possible. The nature of this reaction was discussed.

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First, 5-bromo-3, 6-dinitropseudocumene was prepared follows: Coal tar cumene→pseudocumene-5-sulfonic acid→5-bromopseudocumene→5-bromo-3, 6-dinitropseudocumeme. The latter compound gave by hydrogenation in the presence of palladium charcoal 3, 6-diaminopseudocumene, which was then oxidized with ferric salt to pseudocumoquinone.

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Allithiamine was previously prepared by the reaction between thiamine and an ingredient of garlic, and its structure was established as S-allylmercaptothiamine. In the present paper are reported the syntheses of allithiamine and its homologues by the reaction of thiamine with thiolsulfinates, thiolsulfonates, thiosulfates, thiothiocyanates, sulfenyl chlorides, etc.

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Addition of pyridine, quinoline, niacin, niacinamide, aniline, p-aminobenzoic acid or indole to anaqueous solution of thiamine hydrochloride in the presence of sulphurous acid accelerated the degradation of the thiamine and at the same time caused replacement of the thiazole-portion of the thiamine with the amine. This reaction was affected by pH, and an equilibrium was found when a tertiary amine was employed. It is very interesting to observe that sulphurous acid acts as if it were a thiamine destructive enzyme.

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1. The presence of vitamin B₁₂-forming strains of microbe was positively confirmed in the alimentary tract of ducks. Actinomycetes indicated strong B₁₂-producing ability, but other microbes were rather weak in this ability.
2. There was a fair quantity of vitamin B₁₂ in the contents of digestive tract. Caecum was found to be maximum in gram per cent and small intestine to be highest in total quantity.
3. Further feeding experiments with Co⁶⁰Cl₂ were repeatedly performed and the biosynthesis de novo of Co⁶⁰-labeled vitamin B₁₂ was positively confirmed in the digestive tract of ducks. Moreover, it was also found that the form of Co⁶⁰ in the liver may be Co⁶⁰-labeled vitamin B₁₂.

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In patients suffering from thiaminase disease, thiaminase was found in the lower half of the alimentary canal, i.e. from the lower part of the small intestine to the rectum, and thiamine was present less in amount there. A very small portion of the ingested thiamine could be found in feces. The content of ammonia-like nitrogen in the feces was much higher in the patients, compared with that in normal subjects. Particular subjects only can be afflicted temporarily with thiaminase disease by oral administration of Bacillus thiaminolyticus.

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Asakusanori, the air-dried Porphyra tenera, used as a foodstuff in Japan, was found to be relatively rich in vitamin B₁₂ and to possess a growth-promoting activity on the young chicks fed an all plant protein ration. Over thirty species of other seaweeds were tested and the vitamin contents were found to vary from 0.1 to 4.0γ/100g on a fresh basis. Several zoo-and phytoplanktons and bottom muds of pond were also analysed and proved to be very rich in this vitamin. The author's view on the significance of the vitamin in algae is proposed, especially on the utility of marine and freshwater algae as APF sources.

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1. Guinea pig maintained on a vitamin C and tryptophan deficient diet dies of scurvy earlier (12-15 days).
2. In an insufficient vitamin C feeding, the appearance of scurvy can be delayed by administrating anthranilic acid.
3. Experimentally induced alcaptonuria disappears by giving anthranilic acid. The effects of glutathione and Na₂S₂O₃ are not apparent. Vitamin C and anthranilic acid have no effect on congenital alcaptonuria.
4. The oxidation by liver homogenate of p-hydroxyphenylpyruvate and homogentisic acid is accelerated by 5-hydroxyanthranilic acid, which has no effect on the oxidation of 2, 5-dihydroxyphenylpyruvate. Consequently, the efficacy of anthranilic acid is ascribable to the action of 5-hydroxyanthranilic acid.
5. Indolelactic acid appears in the urine of the guinea-pig fed a vitamin C deficient diet when tryptophan is administered to the animal.
6. A high casein (40%) diet causes an earlier induction of scurvy.
7. The content of vitamin C in the guinea-pig liver rapidly decreases by the administration of tyrosine, cystine and histidine, but is little influenced by alanine, glutamic acid and arginine.
8. The length of survival of the guinea-pig on an insufficient vitamin C diet is prolonged by more than one week by the addition of homogentisic acid.
9. 5-Hydroxyanthranilic and homogentisic acid are therefore capable of sparing vitamin C.
10. A new modification for the estimation of benzoquinoneacetic acid was proposed.
11. When butyric acid and tyrosine are simultaneously injected to the guinea pig maintained on a vitamin C deficient diet, benzoquinoneacetic acid is excreted in the urine and cataract appears in the eye lens.
12. By using the 2, 6-dichlorophenolindophenol, 2, 4-dinitrophenylhydrazine, and ascorbic oxidase methods, it was proved that 2, 3-diketogulonic acid is converted to ascorbic acid by the rat and rabbit liver, but not by the liver slice and homogenate of the guinea pig tissues.
13. Diketogulonic acid was estimated in tissues and organs of the rat, rabbit, guinea pig and hatching eggs.
14. It was established by the use of the rabbit lens extract that diketogulonic acid increases following the formation of ascorbic acid from mannose, whereas in the traumatic cataractal lens the diketogulonic acid but not ascorbic acid is formed from mannose indicating that in the normal lens ascorbic acid is produced through diketogulonic acid, contrary to the cataraetal lens.
15. It was discussed that the metabolisms of ascorbic acid and protein (amino acids) interact on each other and that diketogulonic acid is an intermediate metabolite of ascorbic acid.

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