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R Kido

Publications and source records attributed to R Kido.

At least 55 records · Page 3Linked to original sources
PubMed1990

Purification and characterization of kynurenine-pyruvate aminotransferase from rat kidney and brain.

Kynurenine-pyruvate aminotransferase (KPT), the enzyme responsible for the biosynthesis of the endogenous excitatory amino acid receptor antagonist kynurenic acid, was purified to homogeneity from rat kidney, as judged by polyacrylamide and sodium dodecyl sulfate electrophoresis. The protein appeared to consist of 2 identical subunits of approximately 48 kDa. Kinetic analysis showed Km values of 2.8 mM (kynurenine) and 3.8 mM (pyruvate), respectively. KPT was also partially purified from rat brain. Kidney and brain KPT were found to be identical when analyzed by a spectrum of biochemical, physico-chemical and, after production of anti-kidney KPT antibodies, immunological methods. Partially purified anti-KPT antiserum was used for first immunohistochemical studies, which revealed the presence of the enzyme in astrocyte-like cells throughout the brain. Less frequently, KPT was also found in discretely arranged neurons. The availability of pure KPT and specific anti-KPT antibodies can be expected to be of value for the further examination of the neurobiology of kynurenic acid.

Animals↗
PubMed1990

The effects of caffeic acid and its related catechols on hydroxyl radical formation by 3-hydroxyanthranilic acid, ferric chloride, and hydrogen peroxide.

The effect of caffeic acid on hydroxyl radical formation through a reaction, which contained 0.22 M carbonate buffer (pH 7.4), 0.22 mM 3-hydroxyanthranilic acid, 87 mM 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), 2.9 mM hydrogen peroxide, and 14 microM FeCl3, was investigated. The addition of 30 microM caffeic acid resulted in the decrease of hydroxyl radical formation in the reaction mixture. Chlorogenic acid, 3,4-dihydroxy-phenylalanine noradrenaline, gallic acid, dopamine, epicatechin, and D-(+)-catechin also suppressed the hydroxyl radical formation. In regard to the positional isomers of benzenediol, o-benzenediol inhibited the hydroxyl radical formation, but m and p-benzenediol did not. The inhibitory effect of the hydroxyl radical formation seems to be due to the chelation of iron ions by the catechols. Supporting evidence includes the diminished effect of catechols in the presence of EDTA (a potent iron ion chelator) and the observation of a visible band at 450 nm caused by the interaction between caffeic acid and iron ions. Additionally, the visible band (506 nm) was observed in the solution of o-benzenediol and ferric chloride but not in the solution of m- or p-benzenediol and ferric chloride. Thus compounds with adjacent hydroxyl groups on aromatic rings might inhibit hydroxyl radical formation.

3-Hydroxyanthranilic Acid↗
PubMed1990

Superoxide dismutases enhance the rate of autoxidation of 3-hydroxyanthranilic acid.

The autoxidation of 3-hydroxyanthranilate to cinnabarinate at 37 degrees C and at pH 7.4 is hastened by superoxide dismutase (SOD). The Cu,Zn-containing enzyme from bovine erythrocytes and the Mn-containing enzyme from Escherichia coli were equally effective in this regard; whereas the H2O2-inactivated Cu,Zn enzyme was ineffective. Catalase appears to augment the effect of superoxide dismutase, because it prevents the bleaching of cinnabarinate by H2O2. It follows that O2-, which is a product of the autoxidation, slows the net autoxidation by engaging in back reactions and that SOD increases the rate of autoxidation by removal of O2- and thus by prevention of these back reactions.

3-Hydroxyanthranilic Acid↗
PubMed1990

Effects of some insecticides on several enzymes of tryptophan metabolism in rats.

The activities of tryptophan 2,3-dioxygenase (EC 1.13.11.11), indoleamine 2,3-dioxygenase (EC 1.13.11.17), kynurenine 3-hydroxylase (EC 1.14.13.9), kynureninase (EC 3.7.1.3), kynurenine transaminases, and pyridoxal phosphokinase (EC 2.7.1.35) in the liver, kidney and lung rats were measured after administration of a single dose and repeated doses of dimethoate, carbaryl and fenvalerate, respectively. Ten percent LD50 of each insecticide was orally administered to a rat for a single dose, while 5% LD50 was orally given for five consecutive days as repeated doses. The control animals received the same volume of vehicle (polyethylene glycol 300). Body weight and organs weight losses were recognized only after repeated doses of dimethoate, while protein content remained constant compared to control animals. Repeated administration of dimethoate caused significant decrease in the activity of kynurenine 3-hydroxylase (28.3% decrease in liver, and 32.5% in kidney), kynurenine-pyruvate transaminase (EC 2.6.1.7) (40% in liver, and 24.2% in kidney), kynurenine- pyruvate transaminase (EC 2.6.1-) (24.5% in kidney) and pyridoxal phosphokinase (36.1% in liver). Repeated doses of carbaryl resulted in a significant decrease in the activity of apo-tryptophan 2,3-dioxygenase (42.8%), kynurenine-2-oxoglutarate transaminase (40% in liver), kynurenine-pyruvate transaminase (30.6% in liver), and serine-glyoxylate transaminase (EC 2.6.1.51) (47.9% in liver). Externally added insecticides at different concentrations to the incubation mixture resulted in an inhibition to tryptophan 2,3-dioxygenase, while the other enzymes examined showed no change in their activities.

Animals↗
PubMed1989

Separation of polyunsaturated fatty acid radicals by high-performance liquid chromatography with electron spin resonance and ultraviolet detection.

In the reaction of soybean lipoxygenase (EC 1.13.11.12) with polyunsaturated fatty acids such as linoleic, linolenic and arachidonic acids, some radical species were detected using the electron spin resonance (ESR) spin-trapping technique. The radical species derived from the three polyunsaturated fatty acids were not distinguishable because the ESR spectra of the spin adducts of nitrosobenzene with their three radical species showed no difference in their hyperfine splittings. To overcome this defect of the spin-trapping technique, these spin-adducts were separated by employing high-performance liquid chromatography (HPLC) combined with ESR spectroscopy. The spin adducts were eluted from a C18 reversed-phase column in the order linolenic acid, linoleic acid and arachidonic acid. The half-lives of the spin adducts separated by HPLC-ESR were determined as linoleic acid 600 min, linolenic acid 360 min and arachidonic acid 160 min. The use of an ultraviolet detector together with the HPLC-ESR technique resulted in a 500-fold increase in sensitivity in the detection of the radical species.

Chromatography, High Pressure Liquid↗
PubMed1989

Organ distribution, purification and characterization of kynureninase in Suncus murinus (Insectivora) and anthranilic acid level in the serum.

1. The highest kynureninase activity was measured in Suncus liver compared with other organs tested. The holo-enzyme activity detected in liver was 25%. 2. The enzyme mainly localized in liver cytosol was purified to a single protein band by heat treatment, ammonium sulfate fractionation, DEAE-sepharose, hydroxyapatite and phenyl-sepharose column chromatography. 3. The purified enzyme was effective for both kynurenine and 3-hydroxykynurenine, and showed the optimum pH at 8.5 against both substrates. 4. Km values were 250 microM, 18 microM and 70 microM for kynurenine, 3-hydroxykynurenine and pyridoxal 5'-phosphate, respectively. Various compounds such as histidine, aspartate and nicotinamide enhanced the enzyme activities. 5. Anthranilate, the product of kynureninase, was present at a concentration in Suncus serum of 1.96 microM. This value was higher than those of rat and human.

Animals↗
PubMed1989

Enhancement by catechols of hydroxyl-radical formation in the presence of ferric ions and hydrogen peroxide.

The effect of caffeic acid, a kind of catechol, on the Fenton reaction was examined by using the ESR spin trapping technique. Caffeic acid enhanced the formation of hydroxyl radicals in the reaction mixture, which contained caffeic acid, hydrogen peroxide, ferric chloride, EDTA, and potassium phosphate buffer. Chlorogenic acid, which is an ester of caffeic acid with quinic acid, also stimulated the formation of the hydroxyl radicals. Quinic acid did not stimulate the reaction, suggesting that the catechol moiety in chlorogenic acid is essential to the enhancement of the hydroxyl-radical formation. Indeed, other catechols and related compounds such as pyrocatechol, gallic acid, dopamine, and noradrenaline effectively stimulated the formation of the hydroxyl radicals. The above results confirm the idea that the catechol moiety is essential to the enhancement. Ferulic acid, 4-hydroxy-3-methoxybenzoic acid, and salicylic acid had no effect on the formation of the hydroxyl radicals. The results indicate that the enhancement by the catechols of the formation of hydroxyl radicals is diminished if a methyl ester is formed at the position of the hydroxyl group of the catechol. In the absence of iron chelators such as EDTA, DETAPAC, desferrioxamine, citrate, and ADP, formation of hydroxyl radicals was not detected, suggesting that chelators are essential to the reaction. The enhancement of the formation of hydroxyl radicals is presumably due to the reduction of ferric ions by the catechols. Thus, the catechols may exert deleterious effects on biological systems if chelators such as EDTA, DETAPAC, desferrioxamine, citrate, and ADP are present.

Caffeic Acids↗
PubMed1988

Superoxide dismutase enhances the formation of hydroxyl radicals in the reaction of 3-hydroxyanthranilic acid with molecular oxygen.

Superoxide dismutase (SOD) enhanced the formation of hydroxyl radicals, which were detected by using the e.s.r. spin-trapping technique, in a reaction mixture containing 3-hydroxyanthranilic acid (or p-aminophenol), Fe3+ ions, EDTA and potassium phosphate buffer, pH 7.4. The hydroxyl-radical formation enhanced by SOD was inhibited by catalase and desferrioxamine, and stimulated by EDTA and diethylenetriaminepenta-acetic acid, suggesting that both hydrogen peroxide and iron ions participate in the reaction. The hydroxyl-radical formation enhanced by SOD may be considered to proceed via the following steps. First, 3-hydroxyanthranilic acid is spontaneously auto-oxidized in a process that requires molecular oxygen and yields superoxide anions and anthranilyl radicals. This reaction seems to be reversible. Secondly, the superoxide anions formed in the first step are dismuted by SOD to generate hydrogen peroxide and molecular oxygen, and hence the equilibrium in the first step is displaced in favour of the formation of superoxide anions. Thirdly, hydroxyl radicals are generated from hydrogen peroxide through the Fenton reaction. In this Fenton reaction Fe2+ ions are available since Fe3+ ions are readily reduced by 3-hydroxyanthranilic acid. The superoxide anions do not seem to participate in the reduction of Fe3+ ions, since superoxide anions are rapidly dismuted by SOD present in the reaction mixture.

3-Hydroxyanthranilic Acid↗
PubMed1988

Mechanism of interferon-gamma action. Characterization of indoleamine 2,3-dioxygenase in cultured human cells induced by interferon-gamma and evaluation of the enzyme-mediated tryptophan degradation in its anticellular activity.

Induction by interferon-gamma of indoleamine 2,3-dioxygenase (a tryptophan degradation enzyme) was examined with 11 human cell lines. The enzyme induction was demonstrated in 7 of the 11 cell lines. The induced enzyme in each of the 7 cell lines was identical to the enzyme purified from human placenta, as evidenced by immunoblot analysis with a monoclonal antibody specific to the placental one. The extent of the induction varied largely with the cell line; a relatively high induction was observed with HEL (lung fibroblasts), NY (osteosarcoma), and A-431 (epidermoid carcinoma). The enzyme induction was dependent on the concentration of interferon-gamma and occurred 12-18 h after addition of interferon-gamma to the cultures. Interferon-alpha or -beta was completely ineffective in this induction. Interferon-gamma inhibited the growth of the 7 cell lines observed with the enzyme induction, and this growth inhibition was accompanied with a complete deletion of tryptophan (less than 1 microM) in the culture medium by the induction of the enzyme. For two of these cell lines, the inhibition was partially reversed by an addition of exogenous tryptophan to the medium not to be depleted. These findings indicated that the growth inhibition by interferon-gamma was in part explained by the tryptophan depletion in the medium caused by the enzyme induction.

Ascorbic Acid↗
PubMed1988

Enzymatic studies on tryptophan metabolism disorder in rats chronically exposed to carbon disulfide.

Eight-week-old female Wistar rats were exposed to carbon disulfide for 6 hr a day, 5 days a week, for 12 weeks in inhalation chambers. Then the activities of the main enzymes of tryptophan metabolism (i.e., L-tryptophan 2,3-dioxygenase, indoleamine 2,3-dioxygenase, kynurenine 3-hydroxylase, kynureninase, and kynurenine aminotransferases) in their tissues were determined. The results showed that exposure to carbon disulfide caused a significant increase in the activities of kynureninase and kynurenine-2-oxoglutarate aminotransferase in the kidneys, but only a slight increase in their activities in the liver. The activities of L-tryptophan 2,3-dioxygenase and kynurenine 3-hydroxylase also tended to increase, but the increases were not statistically significant. These results suggests that the kynurenine pathway of tryptophan metabolism in the kidneys of rats exposed to carbon disulfide is activated and that the increased activities of kynurenine-2-oxoglutarate aminotransferase in the kidneys may cause the increased excretion of tryptophan metabolites after tryptophan loading as shown in an earlier study (A. Okayama, L. Fun, A. Yamatodani, Y. Ogawa, H. Wada, and S. Goto, 1987, Arch. Toxicol. 60, 460-463.

Animals↗
PubMed1988

Alanine:glyoxylate aminotransferase activities in liver of Suncus murinus (insectivora).

1. The distribution of L-alanine:glyoxylate aminotransferase (AGT) activities were found in Suncus liver, 55% in particulate fraction and 45% in supernatant. 2. 65% of AGT activities in particulate were dependent on AGT isoenzyme 2 (AGT 2) having molecular weight 210,000, the remainder (35%) of AGT activities were dependent on AGT isoenzyme 1 (AGT 1) which have aminotransferase activity for serine. AGT activities in supernatant were dependent on AGT 1, AGT 2 and alanine:2-oxoglutarate aminotransferase (GPT), and their activity ratios were 10, 15 and 75%, respectively. 3. Km values for alanine were 0.52 mM; AGT 1, 3.3 mM; AGT 2, 0.88 mM; GPT measuring with AGT activity. AGT activity of GPT was inhibited by addition of glutamate and its Ki value was 1.8 mM. 4. Some other properties of AGT 1, AGT 2 and GPT are described.

Alanine Transaminase↗
PubMed1988

Decreased membrane fluidity of erythrocytes and cultured vascular smooth muscle cells in spontaneously hypertensive rats: an electron spin resonance study.

1. Membrane fluidity of erythrocytes and cultured vascular smooth muscle cells was studied in spontaneously hypertensive rats (SHR; Okamoto and Aoki strains) by means of an electron spin resonance (e.s.r.) and a spin-label technique. 2. The values of outer hyperfine splitting (2T') and of the order parameter (S) determined from e.s.r. spectra for a fatty acid spin-label agent (5-nitroxy stearate) were significantly higher in erythrocytes of SHR (4 and 10-13 weeks of age) than in those of age-matched Wistar-Kyoto rats (WKY). This finding suggests that the membrane fluidity of erythrocytes might be decreased in SHR. 3. The fluidity of cultured vascular smooth muscle cells obtained from 10-13-week-old SHR was also reduced compared with that from WKY. 4. These results suggest that the decreased membrane fluidity might be a genetically determined abnormality of the cell membranes in SHR.

Animals↗
PubMed1988

Effects of calcium antagonists on membrane fluidity in hypertension--an electron spin resonance study.

We examined alterations in membrane fluidity of hypertension and the effects of calcium antagonists on these changes by means of an electron spin resonance (ESR) and spin-label technique. Washed erythrocytes from spontaneously hypertensive rats (SHRs; aged 4 and 10-13 weeks) and from patients with essential hypertension, or cultured vascular smooth muscle cells from SHRs were examined and compared with those from normotensive controls. Electron spin resonance spectra for a fatty acid spin label-agent (5-nitroxy stearate) in the membranes were obtained. The values of hyperfine splitting and other parameters of the spectra were significantly higher in erythrocytes from hypertensive rats than in normotensive controls. Similar results were obtained in cultured vascular smooth muscles. These findings indicate that the membrane fluidity might be decreased in hypertension. When calcium was loaded to erythrocytes with Ca-ionophore A23187, the fluidity was decreased. The alternative degrees were greater in hypertensive rats than in controls, and these hypertensive changes were antagonized by diltiazem or verapamil. The results revealed that abnormalities of membrane fluidity in hypertension might be more prominent in the presence of calcium. Further, it is suggested that calcium antagonists could correct these membrane abnormalities in hypertension.

Adult↗
PubMed1988

Detection of nucleic acid-derived radicals formed by alloxan.

Pyrimidine base-derived radical spin adducts were detected in reaction mixtures containing pyrimidine bases, glutathione, and alloxan by the ESR spin trapping technique with a spin trap, alpha-phenyl-N-tert-butyl nitrone (PBN). Pyrimidine nucleoside- and nucleotide-, and ribose- and deoxyribose-derived radical spin adducts of PBN were also observed. However, purine base- and nucleoside-derived radical spin adducts of PBN were not detected. A cytosine-derived radical spin adduct of PBN was not generated under anaerobic conditions. Catalase and mannitol inhibited the formation of the cytosine-derived radical spin adduct of PBN but superoxide dismutase (SOD) did not. EDTA stimulated it and desferrioxamine suppressed it nearly completely. From these results it is presumed that the hydroxyl radical is involved in the formation of the cytosine-derived radical spin adduct of PBN generated by alloxan.

Alloxan↗
PubMed1988

Organ distribution of rat kynureninase and changes of its activity during development.

Kynureninase activity was measured in various organs of the rat by a sensitive assay method based on the use of an HPLC system. High activities were detected in liver, kidney and spleen, and much lower activities in adrenals, intestine, lung, heart, brain, skeletal muscle and pancreas. Kynureninase of liver, kidney and spleen showed the same molecular weight (110,000) and the same isoelectric point (pI 5.4). They also showed the same properties of heat stability and apparent optimum pH. The enzymes in kidney and spleen were localized in the cytosol. The developmental study showed increasing activities of the enzyme after birth and a maximum on the 60th day in both liver and spleen. The activity in kidney increased after birth and reached a plateau on the 30th day.

Aging↗