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Keiichi KOHNO
Article type: Article
1970 Volume 41 Issue 2 Pages
57-66
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Studies on the transport into red blood cells of O-acylthiamine disulfide with kinetic and enzymatic viewpoints are reviewed. A mechanism of at least two-steps is revealed ; one is passive diffusion process due to lipophylic nature of the vitamin derivative and the other is accumulation of free thiamine metabolized from the permeated derivative. The metabolic conversion proceeds via reductive cleavage of the disulfide bond with glutathione and glutathione reductase, followed by deacylation of the resultant O-acylthiamine by an endosomal alliesterase. A schematic mechanism is presented which is generally apllied to accumulative uptake of lipophylic thiamine derivatives.
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Kaname KURIAKI, Takuro SADO, Yuichi SHIOBARA, Kenzi SANO, Hirokazu SAS ...
Article type: Article
1970 Volume 41 Issue 2 Pages
67-75
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Distribution and excretion of the radioactivity of cyclocarbothiamine-^<35>S injected intravenously into mice were studied by means of whole body autoradiography, microautoradiography and liquid scintillation spectrometry. The whole body autoradiography showed that the radioactivity was elicited in almost all organs by 3 minutes after the intravenous administration. It was especially prominent in urine, liver salivary gland, kidney, small intestine, pancreas and Harderian gland behind eyeball. Thiamine-^<35>S injected intravenously gave rise to radioactivity in salivary gland, Harderian gland, skeletal muscle and blood in heart, but it was lower than that of cyclocarbothiamine-^<35>S. By 6 hours, the activity became noticeable in the content of the large intestine in both cases. Microautoradiography showed higher activity in the muscular layer of small intestine of mice receiving cyclocarbothiamine, whereas in that of thiamine administered mice, it was diffusely distributed throughout the mucosal, submucosal muscular layers. The result obtained by scintillation counting was consistent with that by autoradiography. Higher level of ^<35>S was detected in muscle, salivary gland, lung and heart of cyclocarbothiamine-injected mice than in those of thiamine-injected ones. The blood level kept on higher in the former, and the excretion into urine was more rapid in the latter.
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Kazushige YAMAZAKI
Article type: Article
1970 Volume 41 Issue 2 Pages
81-88
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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The pentose cycle of the thiamine-deficient rats was studied by tracing the isotope activity in the liver slices, when they were incubated with ^<14>C-glucose. 1) ^<14>CO_2 production was decreased with U-^<14>C-glucose. 2) More ^<14>CO_2 was seen with 1^<14>C-glucose than 6^<14>C-glucose. 3) U-^<14>C-glucose incorporation into RNA-fraction was decreased which was restored by the addition of TDP or TTFD. 4) Transketolase activity was low. The contents of fat or protein in the diets did not influence the incorporation into RNA and transketolase activity. These results suggested that nucleic acid metabolism via transketolase plays a role in thiamine-deficient state.
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Hidehiko KURIMOTO
Article type: Article
1970 Volume 41 Issue 2 Pages
89-100
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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The thymus satrophy in pyridoxine-deficient mice which were produced by dietary vitamin B_6 deficiency or administration of deoxypyridoxine, was histologically studied. The thymus atrophy was characterized by cessation of mitosis, degeneration and destruction of lymphocytes, removal of lymphocytes and replacement with fatty tissue. It was rapidly recovered by an administration of pyridoxine.
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Chikataro KAWASAKI, Tomio NAGAYAMA
Article type: Article
1970 Volume 41 Issue 2 Pages
101-106
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Microbiological activity of CH_3O-BT (O-p-methoxybenzoylthiamine) and NO_2-BT (O-p-nitrobenzoylthiamine) on L. fermenti was compared with OBT (O-benzolthiamine) ; antagonist of thiamine on L. fermenti because of its strong inhibitory effect on thiamine-uptake by the cells. NO_2-BT showed about one-tenth growth-stimulating potency of thiamine in the broth with or without cysteine, while CH_3O-BT had very slight growth-stimulation with cysteine but no potency for growth without cysteine. By means of bioautography in the broth containing thiamine, CH_3O-BT proved to be inhibitory to growth of L. fermenti but NO_2-BT showed no inhibition at the spot. When these 3 compounds were incubated with phosphate buffer solution (pH 6.0) at 50 or 100℃ for 1 hour, NO_2-BT was more easily converted into thiamine than CH_3O-BT or OBT ; while incubation with the broth at 37℃ for 5 hours, conversion to thiamine was observed to a measurable extentonly in the case of NO_2-BT. NO_2-BT, when incubated with thiamine deficient L. fermenti for 2 hours, was more easily absorbed by the cells than other 2 compounds and in the medium with thiamine, it showed weaker inhibition to thiamine-uptake by the cells than CH_3O-BT or OBT. It was concluded that O-acylthiamine was no more antagonistic to thiamine by reducing its anti-thiamine potency on hydrolysis.
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Chikataro KAWASAKI, Takeo KISHI, Tohru NISHIHARA
Article type: Article
1970 Volume 41 Issue 2 Pages
107-112
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Nicotinic acid contents of Sacch. carlsbergensis during the growth were estimated. In the broth with tihamine, biosynthesis of nicotinic acid was greatly inhibited as compared with that in the broth without thiamine. In the broth with thiamine and pyridoxine, the biosynthesis was better than that in the broth with pyridoxine. All the nicotinic acid contents of the cells were increased in parallel with the growth except those in broth containing thiamine ; the inhibition of biosynthesis of nicotinic acid was interpreted as the vitamin B_6 deficiency caused by thiamine, resulting in the blocking the enzymatic system associated with vitamin B_6.
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Kiyoshi HARADA, Yoshiaki KAKIE, Isamu UTSUMI
Article type: Article
1970 Volume 41 Issue 2 Pages
113-119
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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From the estimation of urinary thiamine excretion after intraperitoneal administration of thiamine or O-acylthiamine, it was suggested that O-acylthiamine was much more metabolized than thiamine in rat. Metabolism of O-acylthiamine was investigated in vitro, using various rat tissue homogenates, and the enzyme which metabolizes O-acylthiamine to thiochrome-negative substance was found in liver, but scarcely in other tissues. The enzyme was partially purified from rat liver, and the reaction product of O-acylthiamine with this enzyme was isolated as colorless needle crystal, mp. 231-2℃. It was identified as thiamine thiazolone, which has the structure added oxygen atom to 2C-position of thiazole ring of thiamine, by infrared absortpion spectra, paper and thin layer chromatography and elementary analysis. This enzyme, which produce thiamine thiazolone from O-acylthiamine, was named O-acylthiamine oxidase tentatively.
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Kiyoshi HARADA, Yoshiaki KAKIE, Isamu UTSUMI
Article type: Article
1970 Volume 41 Issue 2 Pages
120-123
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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In the previous paper we reported that O-acylthiamine oxidase, which produce thiamine thiazolone from O-acylthiamine, is present in rat liver. To elucidate one of the properties of O-acylthiamine oxidase, substrate specificities for 26 kinds of thiamine related compounds were investigated and summerized in Table 1. As is evident from the table O-acylthiamine was metabolized in preference to others. Correlation between the metabolizing activity and physical and structural properties of O-acylthiamine was discussed. It was considered that hydrophobic character and plane structure of the acyl residue in O-acylthiamine were important factors for the enzymatic reaction.
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Yoshiaki KAKIE
Article type: Article
1970 Volume 41 Issue 2 Pages
124-128
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Enzymatic properties of partially purified O-acylthiamine oxidase from rat liver were investigated. Optimal pH for this enzyme was about 8.5-9.0. The activity was inhibited by metal chelating agents, such as KCN, diethyldithiocarbamate or o-phenanthroline and sulfhydryl reagents, such as PCMB or NEM. The enzyme reaction proceeds scarcely in anaerobic condition, but do by an addition of DCPI, and the enzyme was considered to be a kind of oxidase which requires molecular oxygen as hydrogen acceptor. The enzyme was supposed to be one of metalloflavoproteins since it was inhibited by FAD antagonist. On the base of above properties of this enzyme, the mechanism was proposed that thiamine thiazolone was formed by enzymatic dehydrogenation from pseudobase type O-acylthiamine. This mechanism was supported by the fact that the maximal activity was exhibited in the region of pH 8.5-9.0,where concentration of the pseudobase type is considered to be maximum.
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Yoshiaki KAKIE
Article type: Article
1970 Volume 41 Issue 2 Pages
129-133
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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A method of biological determination of thiamine thiazolone was devised utilizing thiamine biosynthesis from the compound and MHT by Saccharomyces cerevisiae. By means of the method thiamine thiazolone was detected in urine of the rat administered OBT intraperitoneally. This result suggests that OBT was metabolized by O-acylthiamine oxidase to thiamine thiazolone, in vivo.
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Yasuo KAKIUCHI
Article type: Article
1970 Volume 41 Issue 2 Pages
134-138
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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In order to elucidate the mechanism of growth-inhibition of Saccharomyces carlsbergensis 4228 by thiamine, effects of thiamine on growth and vitamin B_6 content of the yeast cells growing in the media containing alanine, phenylalanine, or glutamic acid and methionine as the nitrogen source were investigated. The transient increase of the vitamin B_6 content at the initial growth phase was depressed by addition of thiamine to the medium in all cases, but at the peak stages vitamin B_6 content of the yeast in the medium containing alanine was much higher than those in the medium containing phenylalanine or glutamate and methionine. The amino acids detected in these media after growth of the yeast were not markedly different from those detected in the media with thiamine, but the consumption rate in the Atkin's medium with thiamine was found to be higher in alanine, aspartic acid, glutamic acid, and arginine, and to be lower in threonine, glycine, valine, proline, and serine than that in the medium without thiamine.
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Yasuo KAKIUCHI
Article type: Article
1970 Volume 41 Issue 2 Pages
139-143
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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The adaptation of Saccharomyces carlsbergensis 4228 to thiamine-and vitamin B_6 free medium containing serine as the single nitrogen source (serine medium) was tried, and the growth of this serine-adapted strain in the presence of thiamine or vitamin B_6 was investigated. It was found that thiamine inhibited the growth of the adapted strain in the serine medium to the later growth phase as well as the growth of the original strain, and the antagonistic recovery by vitamin B_6 of the inhibited growth was demonstrated. In the experiment using this adapted organism heavily inoculated in the serine medium, it was proved that the vitamin B_6 content of the cells at the initial growth phase was greatly depressed by addition of thiamine. Using U-^<14>C-L-serine, it was demonstrated that the incorporation rate of serine into the cells of adapted organism was not inhibited by thiamine.
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
144-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
144-145
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
145-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
145-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
145-146
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
146-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
146-147
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
147-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
148-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
148-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
148-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
149-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
149-150
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
150-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
151-152
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
152-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
152-153
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese], [in Japanese], [in Japane ...
Article type: Article
1970 Volume 41 Issue 2 Pages
153-154
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
154-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
154-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
154-155
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
155-156
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
156-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
156-157
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
157-158
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
158-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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[in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
158-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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Article type: Article
1970 Volume 41 Issue 2 Pages
158-159
Published: February 25, 1970
Released on J-STAGE: March 14, 2018
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[in Japanese], [in Japanese], [in Japanese]
Article type: Article
1970 Volume 41 Issue 2 Pages
159-
Published: February 25, 1970
Released on J-STAGE: February 22, 2018
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