In order to study the pantothenic acid metabolism in liver diseases, blood and urinary levels of the vitamin were determined in the patients with various liver diseases and found the following results.
1. The total level of the vitamin in the blood fractionated into free and bound form according to Lipmann's enzymatic treatment.
In 24 patients with liver diseases, there was an increase of the free form and a decrease of the bound form in 34 and 24 cases, respectively; the ratio of the bound form to the total, B/T, was low in 20 cases.
In the patients, whose impaired liver function was improved by treatment, the B/T ratio tended to return to normal.
The level of the free form in the blood had a positive correlation, to some degree, with the icteric index in hepatitis. The ester quotient of cholesterol had a positive correlation with B/T in all the cases having B/T above 0.55.
2. The urinary elimination of the vitamin was low in hepatitis and cirrhosis, whereas it was almost normal in neoplasm.

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1. Studies were made to test whether purine nucleotides other than inosinic acid activated thiamine diphosphate in liver-pyruvate oxidase system and adenosine monophosphate and guanosine monophosphate were found to increase markedly the enzyme activity.
2. In parallel experiments, the metabolic fate of inosinic acid was investigated by paper chromatography, and it was found that there were several pathways for the degradation of the nucleotide linked to the enzyme activation. The results seem to suggest that inosine might accelerate the thiamine diphosphate activity of liver-pyruvate oxidase system.
3. Supplement of inosine or hypoxanthine to D-ribose was found to accelerate the thiamine diphosphate action in liver-pyruvate oxidase system, while no activation was detected after addition of hypoxanthine alone. The final reaction mixture was tested by paper chromatography and the reappearance of inosine after adding both hypoxanthine and D-ribose was confirmed.
These findings lead to the conclusion that the metabolism of inosine may accelerate the thiamine diphsophate activity in liver-pyruvate oxidase system.

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1. V compound was produced non-enzymatically from G compound.
2. The conversion reaction of G compound into V compound proceeds enzymatically in the presence of the enzyme preparation obtained from the spinach.
3. The enzymatic formation of V compound from G compound requires the presence of molecular oxygen, and inhibited completely under anaerobic condition.
4. The enzymatic reaction to form riboflavin from G compound proceeds without molecular oxygen.
5. The stoichiometry of the enzymatic synthesis of riboflavin was established by the reaction under “anaerobic condition”.
6. Formation of V compound was inhibited by KCN and KF.

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1. The effect of S-benzoylthiamine monophosphate (BTMP) on yeast cells was investigated. The compound is known to have a superior affinity to animal tissues and to keep higher blood levels in animal body than thiamine. BTMP showed the thiamine activity on the growth and fermentation of the yeast, but the appearance of its effect was somewhat delayed as compared with thiamine.
Considering from the ineffectiveness of BTMP on lactobacilli, a significant difference on absorbability is supposed between animal tissues and microorganism cell walls.
2. In order to compare exactly the rate of the appearance of the activity of the two compounds, the fermentation test for thiamine assay of Schultz, Atkin and Frey was employed. BTMP was shown to behave similarly to SBT. The appearance of its effect was delayed by a quarter to one hour as compared with that of thiamine, but after the appearance the derivative showed the same accerelating effect on the growth of the yeast as thiamine. In the case of thiamine monophosphate (TMP) the retardation of the appearance of the effect was scarcely noticed. It is therefore assumed that BTMP is incorporated into the cells as TMP after debenzoylation. O-Benzoylthiamine disulfide showed the retardation of 5-6 hours before the appearance of the effect. It is therefore assumed that it takes much time for the production of the adaptive enzyme for cleaving the O-benzoyl group.
3. The investigation of the forms of thiamine in the yeast cells incubated with BTMP by both chemical assay and paper chromatography showed the absence of acylated thiamine (BTMP and SBT), but more esterified thiamine was detected than those incubated with thiamine. It was found to be largely TMP. Thus it was clarified that BTMP is incorporated into the cells as TMP after debenzoylation.

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1. It was confirmed by bioautography that S-benzoyl derivatives of thiamine, i, e., S-benzoylthiamine O-monophosphate (BTMP), S-benzoylthiamine (SBT) and O, S-dibenzoylthiamine (DBT), were converted to thiamine monophosphate (TMP), thiamine and O-benzoylthiamine (OBT), respectively, by de-S-benzoylation with the L-cysteine contained in MaciasR's medium.
2. Both BTMP and SBT are as effective as thiamine on L. fermenti in this medium. However, it is not due to the biological activities of these compounds per se. DBT failed to show the thiamine activity on this organism.
3. BTMP, SBT, DBT and OBT become active even without L-cysteine by autoclaving. Therefore, it must be careful, when the biological activity of the compound is in question.

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S-Benzoyl derivative of thiamine, such as S-benzoylthiamine O-monophosphate (BTMP) is de-S-benzoylated to thiamine monophosphate (TMP) by the SH-compound, e. g., L-homocysteine and thioglycollate as well as L-cysteine in the medium, showing the thiamine activity to L. fermenti. It is not the case with other reducing agents, such as sodium thiosulfate, hydrosulfite and thiourea. So the mechanism of the reaction by the SH-compound is not reduction but possibly transbenzoylation to SH compounds.

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1. Ultraviolet absorption spectra of the aqueous solution of O-benzoylthiamine disulfide (BTDS) was determined at various pH levels. They showed the changes according to pH, the isobestic points being at 239 and 274mμ.
2. The structure of BTDS was definitely proved by infrared absorption spectra.
3. BTDS is soluble in mineral acids, chloroform, glacial acetic acid and methanol, but sparingly soluble in water and other organic solvents.
4. The partition coefficient of BTDS between n-butanol or benzene and 0.1 M phosphate buffer (pH 6.6) is remarkably large as compared with those of related compounds. In O-acyltiamine disulfide homologues, the value increases logarithmically with the rise in the number of carbon atoms of the acyl group.
5. R_F values is as high as 0.85±0.03, when the developing solvent consisted of butanol, acetic acid and water. It can therefore be isolated from other compounds.
6. Aqueous solution of BTDS at a concentration of 2mg/ml is fairly stable even in a weakly alkaline solution.
7. The powder of BTDS mixed with alkaline salts (calcium carbonate or sodium bicarbonate) was proved to be fairly stable even when stored under a hot and humid condition.

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1. O-Benzoylthiamine disulfide (BTDS) showed the same biological activity to both lovebirds and rats as that of thiamine.
2. BTDS gave better effect than thiamine when administered intermittently.
3. BTDS is absorbed very well after oral administration, resulting in high levels of total thiamine and cocarboxylase in the blood and liver and the high levels are retained for relatively a long time.
4. The urinary excretion of BTDS showed a tendency to be retarded even after parenteral administration.

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Reducting conditions of BTDS by cysteine and thiosulfate were investigated and the basal assay procedure was established.
1. The optimum condition for reduction with cysteine is the use of 5mg of cysteine hydrochloride for 1 to 100μg BTDS at pH 8.4 to 8.8 and 37° for 60 minutes. The lower the pH of the solution, the more cysteine is required. At pH 7.0 more than 10mg of cysteine is necessary.
2. O-Benzoylthiamine disulfide (BTDS) is converted into O-benzoylthiamine and thiamine by cysteine reduction, while O-benzoylthiamine alone is produced by thiosulfate reduction. O-Benzoylthiamine thus formed is converted into thlochrome by alkaline oxidation with cyanogen bromide. The fluorescence intensity produced from thiamine was practically the same as that from O-benzoylthiamine.
3. The optimum condition for reduction with thiosulfate is the use of 40mg of sodium thiosulfate for 1 to 100μg of BTDS at pH 4.0 to 5.0, heating at 60° for 20 minutes.
4. Adsorbability of O-benzoylthiamine to permutit is poor. However, it become satisfactory after Takadiastase treatment.
5. The thiamine formed from BTDS by reduction and Takadiastase treatment is well adsorbed on permutit but has a tendency to be eluted less. 25ml of the eluting solution is not enough but 40ml is sufficient for the recovery more than 95 per cent.

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The succinic dehydrogenase and DPNH-diaphorase activities of Ehrlich's ascites tumor cells were studied histochemically and the changes in the nuclei were followed with a phase contrast microscope and the following results were obtained.
1. Succinic dehydrogenase activity is very weak.
2. Strong DPNH-diaphorase activity is found in about 10% of the cells but no activity was detected in the majority of them.
3. In the cells showing no DPNH-diaphorase activity, the nuclear membrane and nucleoli were clearly observed and the cells were in the resting state, whereas in the active cells the nuclear membranes and nucleoli were absent, the chromosomes being coarse and diffusive, sometimes partially gathering in groups and being arranged. The cells were in the mitotic stage.
4. The DPNH-diaphorase activity appears in a part of the cytoplasm, spreads throughout the whole cell reaching a maximum, then gradually weakens and eventually disappears. When the activity appears in a part of the cytoplasm, the nuclear membranes and nucleoli are clearly visible. Thereafter the cells are changed into pro- (highest activity), meta-, ana- and telophase and finally divided into daughter cells.
5. Thus the DPNH-diaphorase is concluded to be closely related to mitosis.

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1. The existence of vitamin A and fat in various parts of the lamprey was histochemically observed under a fluorescence microscope.
2. In the muscular tissue vitamin A was found to exist in the fat droplets in the subcutaneous adipose and the intermuscular tissues as fine granules independently of the fat in the boundary line between the muscular and connective tissues.
3. In the liver most of vitamin A was shown to be much dissolved in the droplets of the fat contained in the cells.
4. In the kidney most of vitamin A was found as minute fluorescent granules in the renal corpuscles and uriniferous tubules. They are seemingly free from fat droplets.
5. The milt showed the vitamin A existing in the fat droplets in the interstitium among the tubuli seminiferi. No vitamin was found in the tubli seminiferi itself.
6. Most of the vitamin A in the ovary was found in the adipose tissue among the follicles. The ovarian cyst had fine fluorescent granules of the vitamin, and the corpus luteum had a diffusive fluorescence. Both kinds of fluorescence disappeared in about the same length of time under ultraviolet-ray irradiation.
7. The alimentary canal showed a majority of vitamin A existing in fine fat droplets in the tela submucosa and lamina propria mucosae. It also had a small part of the vitamin independently of the fat in the cells of the mucous epithelium, serous membrane, and muscular layer. Under ultraviolet irradiation the vitamin in the fat disappeard more slowly than that free from fat.

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