VITAMINS

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Print ISSN : 0006-386X

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Volume 33, Issue 1

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EXPERIMENTAL STUDIES ON THE INFLUENCE OF VITAMIN K DEFICIENCY UPON THE FEMALE SEXUAL FUNCTION : (II) CHANGES IN WEIGHTS AND HISTOLOGICAL FINDINGS OF THE HYPOPHYSIS, ADRENAL, OVARY AND UTERUS

Yuichi ABE

Article type: Article
1966 Volume 33 Issue 1 Pages 1-14
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_1

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The adult rats fed on vitamin K free diet were sacrificed in various stage of the experiment and their hypophysis, adrenal, ovary and uterus were weighted and investigated histologically and histochemically. 1) In an early stage of the experiment an apparent weight reduction of hypophysis and uterus was noted and with progress of vitamin K-deficiency the ovarian weight reduction was added. 2) In hypophysis, it was observed the decrease of the acidophilic and basophilic cells in number, contrary to the increase of the chromophobic cells. But when the basophiles are divided into two types of the gonadotrophs and thyrotrophs, the increase of the former is seen throughout the gland and conversely, the latter mostly diminished from the gland. 3) Adrenal cortex was in the state of hyperfunction on the 20th day of vitamin K-deficiency. But about on the 30th day the adrenal cortex was fallen in the state of hypofunction. 4) In ovary, a slight decrease of growing follicles in number was observed on the 20th day of vitamin K-deficiency. 5) In uterus, with the progress of vitamin K deficiency an atrophy in all layers became more obvious gradually.

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EFFECTS OF LARGE AMOUNTS OF THIAMINE AND ITS DERIVATIVES ON GROWTH OF HUMAN FETAL KIDNEY CELLS IN TISSUE CULTURE

Makoto INABA, Shigetane MORI, Susumu INOUE

Article type: Article
1966 Volume 33 Issue 1 Pages 15-18
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_15

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Human fetal kidney cells were grown in TC Med. 199 containing thiamine and its derivatives, such as TPD, TATD, BTMP, BTDS and TTFD at high concentration equivalent to 50μg per ml of thiamine hydrochloride. The effects of these compounds on the cell growth were studied. As a result, it was recognized that TPD and TATD at high concentration had remarkable inhibitory effects on the cell growth, especially in the former, however, TTFD, BTDS and BTMP had no such significant effect. On the other hand, thiamine showed no such effect on the cell growth.

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COMPLEX COMPOUNDS OF THIAMINE AND HEAVY METALS : (II) AN IMPROVED METHOD OF CRYSTALLIZATION OF THIAMINE-COPPER-COMPOUND AND SOME PROPERTIES OF THE CRYSTAL

Akira TANAKA

Article type: Article
1966 Volume 33 Issue 1 Pages 19-24
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_19

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An improved method of crystallization of a new type of thiamine-copper-complex, having a composition of C_<12>H_<16-17>O_2N_4SCu and mp. 152-157℃ (decomp.) is described. The crystal is easily soluble in 25% KCl-HCl solution or in diluted HCl at pH 1 and its thiamine content is about 78.6%. From the results of crystallizing processes, ultimate analysis, copper and thiamine contents, ultraviolet and infrared absorption spectra, it is assumed that this compound is the complex of bivalent copper and thiamine of thiol-type, and is constituted by two components of different copper content.

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TRANSPORT OF VITAMIN B_6 IN HUMAN ERYTHROCYTE : (I) AMOUNT OF VITAMIN B_6 TRANSPORTED IN HUMAN ERYTHROCYTE IN VARIOUS CONDITIONS

Kozo YAMADA, Minoru TSUJI

Article type: Article
1966 Volume 33 Issue 1 Pages 25-30
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_25

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The transport of vitamin B_6 in human erythrocyte was investigated, in vitro. The reaction mixture was consisted with the suspension of human erythrocyte in phosphate buffer containing glucose and various types of vitamin B_6,such as pyridoxine, pyridoxal, pyridoxamine and their phosphates. The amount transported in erythrocyte after incubation was determined by bioassay with Saccharomyces carlsbergensis. The rate and total amount of transportation in the erythrocyte increased according to the concentration of vitamin B_6 in the medium, linearly between 1-4μg/ml of pyridoxine and 1-160μg/ml of pyridoxal. The rate of transportation lowered in acidic pH range in the cases of pyridoxine and pyridoxamine, while in pyridoxal the effect of pH was scarcely observed. The concentration of erythrocyte in the suspension did not influenced on the transport. In the cases of vitamin B_6 phosphates, the transported amounts in erythrocyte were very small compared with the free types. These results lead to the conclusion that vitamin B_6 is transported into erythrocyte against a concentration gradient by energy-dependent mediated by a membrane carrier.

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TRANSPORT OF VITAMIN B_6 IN HUMAN ERYTHROCYTE : (II) AMOUNT OF PYRIDOXAL PHOSPHATE IN ERYTHROCYTE AFTER INCUBATION WITH VARIOUS FORMS OF VITAMIN B_6 AND RELEASE OF PYRIDOXAL FROM ITS PHOSPHATE BY GHOST CELL

Kozo YAMADA, Minoru TSUJI

Article type: Article
1966 Volume 33 Issue 1 Pages 31-34
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_31

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Studies were made on the pyridoxal phosphate content in erythrocyte after an incubation in phosphate buffer, containing glucose with free or phosphorylated forms of vitamin B_6. Pyridoxal phosphate did not increased whatever the concentrations of pyridoxine, pyridoxal or pyridoxamine in the reaction mixture. However, when pyridoxal phosphate was added, its content in erythrocyte increased moderately with the increase of given pyridoxal phosphate amount. With regard to the binding of pyridoxine or pyridoxal with ghost cells, the authors could not recognized the binding. The releasing of pyridoxal from pyridoxal phosphate by ghost cells to the extent of about 24%, and the reaction was inhibited by an addition of ATP.

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TRANSPORT OF VITAMIN B_6 IN HUMAN ERYTHROCYTE : (III) THE EFFECT OF PHOSPHATE ON TRANSPORT OF VITAMIN B_6 IN HUMAN ERYTHROCYTE

Kozo YAMADA, Minoru TSUJI

Article type: Article
1966 Volume 33 Issue 1 Pages 35-38
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_35

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Studies were made on the effect of phosphate on the transport of pyridoxine, pyridoxal and pyridoxamine in human erythrocyte. The rate of transfer of pyridoxine into erythrocyte markedly decreased in parallel with the amount of phosphate in the medium. As for pyridoxamine, it slightly decreased, while pyridoxal did not. Further, the decrease of phosphate concentration in the medium resulted in the diminution of pyridoxal phosphate in erythrocyte after an incubation with various forms of vitamin B_6. The transport of phosphate in human erythrocyte was investigated. Incubation of erythrocyte with glucose in phosphate medium resulted in accumulation of inorganic and organic phosphate along with lactic acid in erythrocyte. Lactic acid formed in erythrocyte was increased in parallel with the amount of phosphate in the medium. These results lead to the conclusion that the amount of vitamin B_6 transported in erythrocyte increases in accordance with the acceleration of glycolysis induced by phosphate.

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STUDIES ON THE MICROBIOLOGICAL ASSAY FOR VITAMIN B_6 : (I) THE REQUIREMENT OF AMINO ACIDS AND THEIR UTILIZATION ON SACCHAROMYCES CARLSBERGENSIS 4228

Keiichi OSHIBA, Hyozo KAWAKITA

Article type: Article
1966 Volume 33 Issue 1 Pages 39-42
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_39

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A study was made on the requirement of individual amino acid and on the relation between the utilization of the amino acids, vitamin B_6 and thiamine in basal medium for the growth of Saccharomyces carlsbergensis. Using 18 amino acids found in casein hydrolysate, the yeast was cultivated under following conditions. (1) Absence of both thiamine and vitamin B_6,(2) presence of thiamine, (3) presence of vitamin B_6 and, (4) presence of both thiamine and vitamin B_6. The amino acids were able to classify into 4 groups by their effects on growth. The requirement of individual amino acid in assay medium for vitamin B_6 was examined by the removal of each amino acid, it was supposed that glutamic acid, histidine, methionine, isoleucine and tryptophan are necessary for the growth of the yeast as nitrogen sources.

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STUDIES ON THE MICROBIOLOGICAL ASSAY FOR VITAMIN B_6 : (II) THE EFFECT OF NITROGEN SOURCES IN PLACE OF CASEIN HYDROLYSATE IN ATKIN'S BASAL MEDIUM ON THE GROWTH OF SACCHAROMYCES CARLSBERGENSIS 4228

Keiichi OSHIBA, Hyozo KAWAKITA

Article type: Article
1966 Volume 33 Issue 1 Pages 43-46
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_43

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Based on the results of the previous report, nitrogen sources in place of casamino acid in vitamin B_6 assay medium was studied. Any combination of amino acids and inorganic nitrogen compounds did not show so excellent response as casamino acid. But when the mixture of glutamic acid, methionine, histidine, isoleucine, tryptophan and ammonium sulfate or of glutamic acid, aspartic acid, asparagine, and arginine was used as nitrogen sources in vitamin B_6 assay medium, it was found the highest growth response of all combinations, so these mixture may be applied for the assay of the sample of simple composition.

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STUDIES ON THE MICROBIOLOGICAL ASSAY FOR VITAMIN B_6 : (III) EFFECT OF THE CORRELATION OF VITAMINS IN THE ASSAY MEDIUM UPON THE GROWTH OF SACCHAROMYCES CARLSBERGENSIS 4228

Keiichi OSHIBA, Hyozo KAWAKITA

Article type: Article
1966 Volume 33 Issue 1 Pages 47-51
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_47

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The effect of the ratios of thiamine, vitamin B_6 and other vitamins in vitamin B_6 assay medium on the growth of Saccharomyces carlsbergensis 4228 was examined, and it was tried to make clear the mechanism of the antagonism between thiamine and vitamin B_6. The results show that this antagonism was not characteristic to this organisms, but common to a part of Saccharomyces. This antagonism was remarkably affected by balance of pantothenic acid, thiamine and vitamin B_6 in the medium, and it seems to be occurred when the organism required pantothenic acid for the growth but not thiamine and vitamin B_6 cultured in medium containing a small amount of pantothenic acid.

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STUDIES ON THE QUANTITATIVE DETERMINATION OF β-CAROTENE AND VITAMIN A IN MARGARINE : (I) SEPARATION AND DETERMINATION OF β-CAROTENE

Hiromu KANEMATSU, Isao NIIYA, Masao IMAMURA, Hyozo KAWAKITA

Article type: Article
1966 Volume 33 Issue 1 Pages 52-56
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_52

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For the purification of β-carotene from other pigments and vitamin A, column partition chromatographic procedure was investigated. As an adsorbent, activated alumina was suitable, which was dried for 6 hours at 110℃, or heated for 5-7 hours at 400-600℃ and then added with 0.5% of water. β-Carotene on the column was eluted with 10ml of acetone・petroleum ether (1 : 49) and the other pigments were eluted with 15ml of acetone・petroleum ether (1 : 9), while vitamin A remained on the column. About 93% of β-carotene added in margarine was recovered by this method.

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STUDIES ON THE QUANTITATIVE DETERMINATION OF β-CAROTENE AND VITAMIN A IN MARGARINE : (II) SEPARATION AND DETERMINATION OF VITAMIN A

Hiromu KANEMATSU, Isao NIIYA, Masao IMAMURA, Hyozo KAWAKITA

Article type: Article
1966 Volume 33 Issue 1 Pages 57-60
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_57

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The separation and determination of vitamin A in margarine by column chromatography were studied. By the zinc reduction method of Yamashita et al., the decolorization of β-carotene is imcomplete and the remaining carotene causes an error in vitamin A value. After the removal of β-carotene and other coloring matters from alumina column as shown in previous paper, vitamin A is eluted by ether. The recovery of added vitamin A in margarine was about 99% by zinc reduction method, while that by column chromatographic method was about 89%. The corrected values were about 100% in the latter and 106% in the former.

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STUDIES ON S-CARBALKOXYTHIAMINE : (VII) METABOLISM OF O, S-DICARBETHOXYTHIAMINE IN THE INTESTINAL ABSORPTION PROCESS

Minoru MORITA, Tetuo MINESITA

Article type: Article
1966 Volume 33 Issue 1 Pages 61-66
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_61

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As the oral administration of O, S-dicarbethoxythiamine (DCET) caused a remarkable increase of thiamine content in erythrocytes, which could not be attained by oral or parenteral administration of thiamine hydrochloride, it is suggested that DCET was absorbed from the intestine without being converted to thiamine in the epithelial cells and further penetrated into erythrocytes where it was metabolized to thiamine. To obtain evidence for this hypothesis, perfusion experiments in situ have been carried out. Fifty mg of DCET was administrated into a ligated sac of rabbit jejunum and portal vein blood which flowed out from the ligated part of the intestine was collected for the analysis of metabolites of DCET. About 12 % of the absorbed DCET was found in erythrocytes as thiamine and no other metabolites were demonstrated. In the plasma, on the other hand, thiamine, O-carbethoxythiamine (OCET), S-carbethoxythiamine (CET) and DCET were found in percentages of 7,11,69,13,respectively. In substitution of perfusing blood by Locke-Ringer's solution, conversion to thiamine or OCET was markedly diminished but the rate of CET formation was not affected. These results showed that DCET is mainly metabolized to CET, which still possesses a high permeability to the cells, in the intestinal absorption process.

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STUDIES ON S-CARBALKOXYTHIAMINE : (VIII) PENETRATION OF O, S-DICARBETHOXYTHIAMINE INTO ERYTHROCYTES

Minoru MORITA, Tetuo MINESITA

Article type: Article
1966 Volume 33 Issue 1 Pages 67-71
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_67

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The mechanism of penetration into erythrocytes and metabolism of O, S-dicarbethoxythiamine (DCET) were investigated in vitro, using rabbit erythrocyte suspension. DCET readily entered into erythrocytes untill the concentration in the cell became equal with that in the medium, and this penetration was not affected by metabolic inhibitors such as monoiodoacetic acid or 2,4-dinitrophenol. After penetration into the cell, DCET was gradually converted to thiazole type metabolites in which thiamine and O-carbethoxythiamine were included, and accumulated in the cells because of low permeability of these metabolites. This metabolic reaction was suppressed when the erythrocytes were denaturated by heat treatment, or when the incubation was carried out at a low temperature. In addition, the relationship between the sub-strate concentration and the rate of metabolism was shown to satisfy the equation of Michaelis and Menten. These results suggested that DCET penetrates the erythrocyte membrane by a simple diffusion mechanism and is metabolized to thiamine by an enzyme present in the erythrocytes.

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STUDIES ON S-CARBALKOXYTHIAMINE : (IX) PARTIAL PURIFICATION OF S-CARBALKOXYTHIAMINE METABOLIZING FACTOR

Minoru MORITA, Tsuyoshi IWATA, Tetuo MINESITA

Article type: Article
1966 Volume 33 Issue 1 Pages 72-75
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_72

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The factor which metabolizes S-carbalkoxythiamine (CAT) to thiamine is widely distributed in animal tissues, but in different degrees. The highest activity was found in liver, especially in microsomal fraction of the cell. Among other tissues, fairly high activities were found in erythrocyte, blood plasma, kidney and intestine, on the other hand, the activities in nervous and muscular tissues were relatively low. In order to elucidate the characters of this factor, its partial purification from rabbit liver has been attempted. CAT metabolizing factor in the liver was extracted by treating the homogenate with ethylene dichloride, which was found to solubilize and stabilize the factor, and the extract was successively fractionated with basic lead acetate and ammonium sulfate, then further purified by removing inactive protein with calcium phosphate gel and finally lyophilized after dialyzed the extract against water. The specific activity of this partially purified preparation was about 65-fold that of the homogenate.

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STUDIES ON S-CARBALKOXYTHIAMINE : (X) ENZYMATIC PROPERTIES OF S-CARBALKOXYTHIAMINE METABOLIZING FACTOR

Minoru MORITA, Tetuo MINESITA

Article type: Article
1966 Volume 33 Issue 1 Pages 76-80
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_76

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Enzymatic properties of S-carbalkoxythiamine (CAT) metabolizing factor partially purified from rabbit liver has been investigated. The factor was stable for at least 20 minutes at 50℃, while it was completely inactivated within 2 minutes at 100℃. Optimal pH for S-carbobutoxythiamine metabolism was around 7.5,but in case of O, S-dicarbethoxythiamine, the higher the pH, the more the thiamine produced. The activity was inhibited by eserine, neostigmine and heavy metal ions such as Hg^<2+> and Cu^<2+>. The final reaction products of CAT with this factor were identified as thiamine, carbon dioxide and alcohol corresponded to the alkoxy radical of CAT, so that this metabolic reaction was considered to be a hydrolytic reaction accompanied by decarboxylation, and the catalytic factor for this reaction should be classified as a hydrolase.

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STUDIES ON S-CARBALKOXYTHIAMINE : (XI) COMPARISON OF S-CARBALKOXYTHIAMINE HYDROLYZING ENZYME WITH LIVER ESTERASES

Minoru MORITA, Tsuyoshi IWATA, Tetuo MINESITA

Article type: Article
1966 Volume 33 Issue 1 Pages 81-86
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_81

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In order to investigate a possibility that S-carbalkoxythiamine (CAT) hydrolyzing enzyme may be identical with a known non-specific esterase such as aliesterase or cholinesterase, both activities being found in the partially purified liver extract, the characters of these three hydrolase activities have been compared with one another. As each hydrolase activity in the crude extract was increased to different degrees after purification by a same procedure, and effects of neostigmine, cupric ion and rivanol treatment on each activity were not similar, it was suggested that CAT hydrolyzing enzyme is different from such esterases. CAT hydrolyzing enzyme was shown to be able to hydrolyze S-acyl-thiamine derivatives, but it was also demonstrated that liver contained an another S-acylthiamine hydrolyzing enzyme which could be separated from CAT hydrolyzing enzyme by treating the homogenate with ethylene dichloride and showed no activity toward CAT.

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Article type: Article
1966 Volume 33 Issue 1 Pages 87-
Published: January 25, 1966
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DOIhttps://doi.org/10.20632/vso.33.1_87_1

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Article type: Article
1966 Volume 33 Issue 1 Pages 87-88
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DOIhttps://doi.org/10.20632/vso.33.1_87_2

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Article type: Article
1966 Volume 33 Issue 1 Pages 88-
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Article type: Article
1966 Volume 33 Issue 1 Pages 88-89
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DOIhttps://doi.org/10.20632/vso.33.1_88_2

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1966 Volume 33 Issue 1 Pages 89-
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1966 Volume 33 Issue 1 Pages 89-
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1966 Volume 33 Issue 1 Pages 89-
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1966 Volume 33 Issue 1 Pages 89-90
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1966 Volume 33 Issue 1 Pages 90-
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1966 Volume 33 Issue 1 Pages 91-
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Article type: Article
1966 Volume 33 Issue 1 Pages 91-
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1966 Volume 33 Issue 1 Pages 91-92
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Article type: Article
1966 Volume 33 Issue 1 Pages 92-
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Article type: Article
1966 Volume 33 Issue 1 Pages 92-
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Article type: Article
1966 Volume 33 Issue 1 Pages 92-93
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Article type: Article
1966 Volume 33 Issue 1 Pages 93-
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Article type: Article
1966 Volume 33 Issue 1 Pages 93-94
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Article type: Article
1966 Volume 33 Issue 1 Pages 94-95
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DOIhttps://doi.org/10.20632/vso.33.1_94

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Article type: Article
1966 Volume 33 Issue 1 Pages 95-
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DOIhttps://doi.org/10.20632/vso.33.1_95_1

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Article type: Article
1966 Volume 33 Issue 1 Pages 95-96
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Article type: Article
1966 Volume 33 Issue 1 Pages 96-97
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Article type: Article
1966 Volume 33 Issue 1 Pages 97-98
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Article type: Article
1966 Volume 33 Issue 1 Pages 98-
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Article type: Article
1966 Volume 33 Issue 1 Pages 98-99
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Article type: Article
1966 Volume 33 Issue 1 Pages 99-100
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[in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 100-
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_100_1

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[title in Japanese]

[in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 100-101
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_100_2

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[title in Japanese]

[in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 101-
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_101_1

JOURNAL FREE ACCESS

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[title in Japanese]

[in Japanese], [in Japanese], [in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 101-102
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_101_2

JOURNAL FREE ACCESS

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[title in Japanese]

[in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 102-
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_102_1

JOURNAL FREE ACCESS

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[title in Japanese]

[in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 102-103
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_102_2

JOURNAL FREE ACCESS

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[title in Japanese]

[in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 103-104
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_103

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[title in Japanese]

[in Japanese], [in Japanese], [in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 104-
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_104_1

JOURNAL FREE ACCESS

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[title in Japanese]

[in Japanese], [in Japanese]

Article type: Article
1966 Volume 33 Issue 1 Pages 105-
Published: January 25, 1966
Released on J-STAGE: February 09, 2018

DOIhttps://doi.org/10.20632/vso.33.1_105_1

JOURNAL FREE ACCESS

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