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Akiko NAKAMURA, Keiko SANADA, Eisuke KATSURA
Article type: Article
1968 Volume 37 Issue 1 Pages
1-4
Published: January 25, 1968
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The study was made on the separation and the determination of thiamine and its phosphates in animal tissues. It was confirmed that thiamine and its phosphates could be separated completely with success by means of a stepwise DEAE・Sephadex A-25 column chromatography in which elution was made with 0.1,0.2 and 0.3M formic acid solution. Thiamine is determined by thiochrome method. It was demonstrated that this procedure is rather simple, very ready to perform and the recovery rate of thiamine and its phosphates added to the column was about 100 per cent.
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Akiko NAKAMURA, Eisuke KATSURA
Article type: Article
1968 Volume 37 Issue 1 Pages
5-11
Published: January 25, 1968
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The distributions of thiamine and its phosphates in rat's tissues, such as liver, heart and blood were determined, 30 minutes after the intravenous injection of either thiamine-^<14>C or thiamine propyldisulfide (TPD)-^<14>C at a dose of 1.5mg per 100g body weight. Thiamine, its di- and tri-phosphates (TDP and TTP) were separated by DEA・Sephadex A-25 column chromatography. As a result, free thiamine was found in largest amunt in tissues, followed by TDP, and TTP was found in small amount and TMP was not able to determined because of little amount. The injected TPD was taken up by heart tissue as thiamine and its phosphates more rapidly and in greater amount than in the case of thiamine injection.
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Yasuyuki SHIMIZU
Article type: Article
1968 Volume 37 Issue 1 Pages
12-18
Published: January 25, 1968
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Riboflavin-tetrabutyrate is said to have antioxydative property in vivo, and to stay in animal body longer than riboflavin. These properties are expected to be advantageous for supplementation to fish diet. The author studied at first on its determination. One g of fish tissue is homogenized under ice cooling and is extracted with chloroform methanol (3 : 1). The extract is dried up in vacuo, again dissolved in chloroform, washed with phosphate buffer, and centrifuged, the chloroform is dried up in vacuo, added with 5ml of ethanol and 1ml of 0.5N NaOH in a dark room and shake for 10 minutes at 30℃, by these treatment, the riboflavin-tetrabutyrate is hydrolyzed into free form. The ethanolic solution is dried up in vacuo, dissolved in water, then measured by the lumiflavin method. Multiplication the value of free riboflavin by 1.745 gives the riboflavin tetrabutyrate. By this method at least 1μg of riboflavin-tetrabutyrate may be determined in the presence of a large amount of free riboflavin.
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Toshio ANMO, Mitsunori WASHITAKE, Yasuji TAKASHIMA, Seiichi SATO
Article type: Article
1968 Volume 37 Issue 1 Pages
19-24
Published: January 25, 1968
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The influence of water on the change of vitamin A acetate was investigated without using surface active agents, by comparing it with the changes in anhydrous ethanol and in aqueous ethanolic solution (90,80,70,60,50,and 45%). By ultraviolet spectrometric method (Morton・Stubbs correction) and Carr・Price reaction method, the change of vitamin A acetate in aqueous ethanolic solution was examined. Supposing that the value determined by the former method represents almost correctly unchanged vitamin A itself, and comparing it with the value determined by the latter method, we examined the total amount of the intermediately changed substance (Carr・Price reaction positive). By Al_2O_3 column chromatography, we examined each amount of the intermediately changed substances, respectively, and studied the process of these changes. In ethanolic solution the change process of vitamin A acetate seems to be as follows : Vitamin A acetate→intermediately changed substances (Carr・Price reaction positive substances, mainly anhydro-vitamin A)→degradated substances (Carr・Price reaction negative). Increasing of water content accelerates these reactions and seems to accelerate especially the second step reaction.
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Tamotsu DANNO, Yuzo MATSUOKA, Seiichi NURIMOTO, Goro HAYASHI
Article type: Article
1968 Volume 37 Issue 1 Pages
25-30
Published: January 25, 1968
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The recovery of serum GOT and GPT activities which were lowered in vitamin B_6-deficient rat, was more prompt and retained high value for longer time in pyridoxal-homocysteine administration than that in pyridoxal or its phosphate administration. The blood pyridoxal phosphate level was also higher in the former than that in pyridoxal phosphate administration. Pyridoxal-homocysteine did not show any marked effect on cardiovascular system or gastrointestinal tract as well as the latter two.
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Hideo IKEHATA, Yoshiko SUGIYAMA
Article type: Article
1968 Volume 37 Issue 1 Pages
31-36
Published: January 25, 1968
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Rose's hemolysis test using dialuric acid was simplified and the following facts were observed by the method. Dialuric acid, H_2O_2 and peroxidized ethyl linoleate specifically hemolyzed the vitamin E-deficient erythrocytes under certain conditions, although HgCl_2,KMnO_4 and Tween 80 equally hemolyzed the erythrocytes of vitamin E-deficient and normal rats. Vitamin K_3,K_2CrO_4 and K_2Cr_2O_7 had the antihemolytic effect on in vitro dialuric acid-induced hemolysis of vitamin E-deficient erythrocytes. The method was expected to be used as a simple biological assay procedure of vitamin E.
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Hideo IKEHATA, Kiyoko KOYANAGI, Michiko WAKAIZUMI
Article type: Article
1968 Volume 37 Issue 1 Pages
37-43
Published: January 25, 1968
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Horwitt's H_2O_2-induced hemolysis test was observed to be dependent on the temperature of reaction mixture, and a revised method was proposed. High percentages of hemolysis of fourteen adults were all lowered by the supplementation of 150 mg of DL-α-tocopherol acetate. Supplementation of 10g per day of vitamin E-depleted sufflower oil to three adults raised hemolysis in 11-18 days. Average hemolysis percent of sick children was significantly higher than that of normal children.
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Masahiro KAWASAKI, Tadayoshi ONO
Article type: Article
1968 Volume 37 Issue 1 Pages
44-49
Published: January 25, 1968
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It was demonstrated that thiaminase and thermostable factor existed in the fruit body of Lentinus edodes. The prescence of thiaminase in L. edodes was confirmed by chemical and microbiological assay method. The optimum pH of the enzyme was 6.5,and its optimum temperature 20℃. It was inactivated at 45℃ in 10 minutes. The organic bases such as pyridine, aniline, quinoline and p-aminobenzoic acid acted as activators, as in the case of thiaminase I. L. edodes contains not only thiaminase, but also thermostable thiamine-decomposing factor. The activity of the latter was found at pH greater than 6.0 and at 60℃ or more. The enzyme activity in L. edodes was most potent in the gillus, less in the stipe and the pileus. Thiamine was found to contain mostly in the gillus, followed by the pileus and slightly in the stipe. Of the total thiamine, the ester form was about 70 percent.
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Motohiko UEDA, Shoichi MATSUMURA, Masaru KAWAKAMI, Tetuo MINESITA
Article type: Article
1968 Volume 37 Issue 1 Pages
50-59
Published: January 25, 1968
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It is very important to know the pharmacodynamic effects of a new thiamine derivatives, because the so-called "mass therapy" of thiamine derivative is being carried out without proven action mechanisms. As O, S-dicarbethoxythiamine (DCET) had an quinidine-like antiar-rhythmic effects on myocardium as well as striated muscle, it was studied whether DCET had an quinidine-like antiarrhythmic effects in situ preparations or not. Methacholine induced A-V block in cats was effectively antagonized by the DCET administration as quinidine. These effects, however, were not specific to DCET, for the similar antiarrhythmic effects were observed by the thiamine administration. Only slight antiarrhythmic effects were observed by the DCET administration in the epinephrine or epinephrine-petroleum ether induced ventricular arrhythmia in cats, while quinidine effectively antagonized to the arrhythmias at the same dose range as in the methacholine arrhythmias. In ouabain arrhythmias, β-adrenergic blocker propranolol was the only compound which markedly increased the ouabain doses required for various arrhythmias. Threshold voltages in the ventricular fibrillation of rabbit were elevated in the following order : quinidine>procaine amide>DCET. It may be concluded that vagotonic atrial arrhythmias are effectively suppressed by the DCET administration.
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Motohiko UEDA, Shoichi MATSUMURA, Tetuo MINASITA
Article type: Article
1968 Volume 37 Issue 1 Pages
60-68
Published: January 25, 1968
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In the preceeding paper it was observed that the atrial arrhymias like A-V block by methacholine were effectively antagonized by O, S-dicarbethoxythiamine (DCET) administration but the ventricular arrhythmias by epinephrine or ouabain were only slightly antagonized. In this paper the effects of DCET on the atrial fibrillation in cats were compared with quinidine, pronethalol, and atropine, because an atrial fibrillation was one of the common arrhythmias observed in clinic. DCET, in a dose 20-40 mg/kg, prevented the incidence of atrial fibrillation or decreased the duration of atrial fibrillation caused by methacholine-electrical stimulation. As a moderate shortening of atrial fibrillation was also observed by thiamine administration, antifibrillatory effects of DCET could not be considered as a specific effects of DCET but one of the common properties of thiamine derivatives. But the antifibrillatory effects of thiamine were inferior to those of DCET. Atropine, at a dose of 1mg/kg, completely suppressed the incidence of methacholine induced atrial fibrillation. While quinidine or pronethalol prevented the methacholine induced atrial fibrillation at the same dose range as in the prevention of epinephrine arrhythmias. Antiarrhythmic effects of DCET to methacholine induced atrial fibrillation may provide some reasonable evidence in the so-called "mass therapy" of thiamine derivatives.
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Chikataro KAWASAKI, Yoshio ITO
Article type: Article
1968 Volume 37 Issue 1 Pages
69-72
Published: January 25, 1968
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The chemical nature of a colored product obtained from the reaction of retinoic acid with 74% sulfuric acid was studied. The colored product in its sulfuric acid layer, when it was diluted with water, was extracted by chloroform and the extracted material was soluble in KOH or NaOH. Thin layer chromatogram of the extracted material suggested that the reaction product was a single substance. UV-, IR-, and NMR-patterns of the material suggested that the chemical structure of the colored reaction product will be simillar with retinoic acid but hydrated at one of the conjugated double bonds in the side chain.
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Chikataro KAWASAKI, Yoshio ITO, Katsunori TANINO
Article type: Article
1968 Volume 37 Issue 1 Pages
73-77
Published: January 25, 1968
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Retinoic acid, when dissolved in 0.1N KOH and diluted with 0.1M phosphate buffer (pH 8), was proved to be stable on heating or standing for days at 37℃, if light was intercepted. Retinoic acid added in the homogenate of animal tissues or in urine or feces, can be extracted by 70% ethanol. The extract after the centrifugation was diluted by 0.1M phosphate buffer and extracted by ligroin. Retinoic acid in the ligroin layer was purified by the transfer to 0.1N KOH in 50% ethanol and the re-extraction by ligroin then retinoic acid was estimated by E value at 355mμ or 74% sulfuric acid color reaction. Retinoic acid when 3 mg were orayll given to rats, was found in the feces for 24 hours to the extent of 3-15% of the dosage but was unable to be detected in the urine. Retinoic acid, when 1.2mg were infused in ligated small intestine of rats, was recovered from the intestine after one hour only to the extent of 33% of the dosage and the retinoic acid in livers and kidneys was estimated as 46 and 17μg even at the highest peak after 2 hours. It was concluded from the repeated infusion experiments into ligated intestines of rats that retinoic acid was so quickly metabolized and decomposed in animals after absorption from the intestine that it was hardly detected in animal tissues and feces.
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Chikataro KAWASAKI, Yoshio ITO, Masaharu OGAWA
Article type: Article
1968 Volume 37 Issue 1 Pages
78-84
Published: January 25, 1968
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2.5mg of retinoic acid-6,7^<14>C_2 were infused into the midportions of ligated intestines of rats and estimation of both radioactivity and retinoic acid in organs and tissues of the animals was undertaken at 1 and 6 hours after the infusion. Both radioactivity and retinoic acid were detected in liver, kidney, bone, muscle and skin after 1 hour, but after 6 hours, only trace of or no radioactivity was detected in them, except liver. Radioactivity in the upper portions of the intestines and the urine after 6 hours was 32 and 18% of the total given activity. Excretion of metabolites of retinoic acid was shown to have two ways to urine through kidney and to upper intestine through gall-bladder. If one way was shut down by ligation, the excretion of activity was shown to be exclusive to the urine or to the upper intestine. Radioautography of the urine and the upper intestine content demonstrated 4 metabolites (Rf 0.05,0.3,0.55,0.95) in the urine and 4 metabolites (Rf 0.05,0.15,0.75,1.0) in the upper intestine. Retinoic acid and its ester was recovered from the upper intestine but no tetinoic acid was found in the urine.
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Chikataro KAWASAKI, Takeo KISHI, Masahiro NISHII
Article type: Article
1968 Volume 37 Issue 1 Pages
85-90
Published: January 25, 1968
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Modified thiamine compounds are resistant to thiaminase but some of them, such as thiamine propyldisulfide (TPD) can be reduced to thiamine on the incubation with Bacillus thiaminolyticus or Escherichia coli. When thiamine-HCl, TPD, dicarbethoxythiamine (DCET) and cyclocarbothiamine (CCT) were incubated with the culture of B. thiaminolyticus in the presence of pyridine, thiamine-HCl was easily decomposed but DCET and CCT were proved to be stable and resistant to degradation, while TPD was decomposed in lower concentrations but resistant to degradation in higher concentrations of the substrate. TPD, when incubated with the saline suspension of B. thiaminolyticus, was reduced to thiamine but poorly decomposed because of the lack of replacing basic compounds. O-Benzoylthiamine disulfide (BTDS) and dibenzoylthiamine (DBT) were proved to be more resistant than TPD but more easily decomposed than DCET or CCT on incubation with the culture of B. thiaminolyticus. TPD, BTDS and DBT were decomposed by the culture of B. thiaminolyticus even on anaerobic incubation in nitrogen.
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Isamu UTSUMI, Keiichi KOHNO, Kazuo NODA, Yoshikazu TAKEUCHI
Article type: Article
1968 Volume 37 Issue 1 Pages
91-94
Published: January 25, 1968
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Comparative study was made on acyl-TDS metabolizing ability of animals. OBT- and OBuT- deacylating activities were found to widely distribute in rabbit and rat tissues, such as liver, kidney, heart, intestine, brain, spleen and muscle. But the activities in rat tissues were much lower than in rabbit ones, especially in liver, kidney and brain. For the OBT esterase activity in blood, profound differences were found among man, dog, rabbit and rat. 100 ml of human blood was able to deacylate 25mg OBT to thiamine for ten minutes under the assay condition, and was 20- or 200-fold active than that of rabbit or rat blood, respectively.
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Isamu UTSUMI, Keiichi KOHNO, Kazuo NODA, Sigeo MATSUO, Toshio UMETANI
Article type: Article
1968 Volume 37 Issue 1 Pages
95-98
Published: January 25, 1968
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O-Acylthiamine esterase activities in the blood of in-patients of liver cirrhosis and chronic hepatitis were compared with that of healthy subjects. OBT esterase activities (expressed by an amount of thiamine produced per 100ml blood for 10 minutes) ranged from 11.8 to 23.8mg in the patients (9 cases) and from 17.4 to 31.2mg in healthy man (10 cases). OBuT esterase activities ranged from 147 to 239mg and from 126 to 256mg, respectively, in the patients and the normal subjects. It was concluded that there was no practically significant difference in the acylthiamine metabolizing ability between them.
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Isamu UTSUMI, Keiichi KOHNO, Yoshikazu TAKEUCHI
Article type: Article
1968 Volume 37 Issue 1 Pages
99-104
Published: January 25, 1968
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Kinetic study was made on the metabolic fate of the benzoyl group of BTDS in man. First order rate constants for absorption, metabolism and excretion were calculated from data of urinary hippuric acid elimination after an oral administration of BTDS or benzoic acid. There could be found no appreciable difference on the rate constant in each process between the benzoyl group and benzoic acid. It was concluded that excretion of the benzoyl group as hippuric acid was rapid and complete as that of benzoic acid.
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Kunio YAGI, Ikuo NISHIGAKI, Hirobumi OHAMA
Article type: Article
1968 Volume 37 Issue 1 Pages
105-112
Published: January 25, 1968
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Article type: Article
1968 Volume 37 Issue 1 Pages
113-
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Article type: Article
1968 Volume 37 Issue 1 Pages
113-
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[in Japanese], [in Japanese], [in Japanese], [in Japanese]
Article type: Article
1968 Volume 37 Issue 1 Pages
113-114
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Article type: Article
1968 Volume 37 Issue 1 Pages
114-
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Article type: Article
1968 Volume 37 Issue 1 Pages
114-115
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Article type: Article
1968 Volume 37 Issue 1 Pages
115-
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Article type: Article
1968 Volume 37 Issue 1 Pages
115-116
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Article type: Article
1968 Volume 37 Issue 1 Pages
116-
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Article type: Article
1968 Volume 37 Issue 1 Pages
116-117
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Article type: Article
1968 Volume 37 Issue 1 Pages
117-
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Article type: Article
1968 Volume 37 Issue 1 Pages
118-
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Article type: Article
1968 Volume 37 Issue 1 Pages
118-119
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Article type: Article
1968 Volume 37 Issue 1 Pages
119-
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Article type: Article
1968 Volume 37 Issue 1 Pages
119-120
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Article type: Article
1968 Volume 37 Issue 1 Pages
120-
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Article type: Article
1968 Volume 37 Issue 1 Pages
120-121
Published: January 25, 1968
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Article type: Article
1968 Volume 37 Issue 1 Pages
121-122
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Article type: Article
1968 Volume 37 Issue 1 Pages
122-
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1968 Volume 37 Issue 1 Pages
122-123
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Article type: Article
1968 Volume 37 Issue 1 Pages
123-
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1968 Volume 37 Issue 1 Pages
123-124
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Article type: Article
1968 Volume 37 Issue 1 Pages
124-
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1968 Volume 37 Issue 1 Pages
124-125
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Article type: Article
1968 Volume 37 Issue 1 Pages
125-
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1968 Volume 37 Issue 1 Pages
125-126
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1968 Volume 37 Issue 1 Pages
126-
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1968 Volume 37 Issue 1 Pages
126-127
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1968 Volume 37 Issue 1 Pages
127-
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1968 Volume 37 Issue 1 Pages
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1968 Volume 37 Issue 1 Pages
127-128
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1968 Volume 37 Issue 1 Pages
128-
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1968 Volume 37 Issue 1 Pages
128-
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