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R Kido

Publications and source records attributed to R Kido.

At least 37 records · Page 2Linked to original sources
PubMed1994

Changes of some serum parameters and amino acids content in rats after chronic sublethal doses of dimethoate.

An intraperitoneal repeated dose of dimethoate (O,O-dimethyl S-[N-methylcarbamoyl methyl] phosphorodithioate) was injected to male Wistar rats for 8 successive days. Body weight and liver weight decreased significantly while liver/body weight ratio and mg protein/g liver remained unchanged. Twenty serum amino acids were assayed in blood samples collected from hepatic vein, portal vein, and aorta. Most of the measured amino acids showed a tendency of decrease compared to control. The decrease was significant only for taurine, serine, threonine, methionine, tyrosine, histidine, and arginine but not for the rest of the amino acids measured. Glucose, 3-hydroxybutyrate, urea, ammonia, and lactate were measured in the same serum samples. These serum parameters showed a tendency of decrease (except urea) of them from control but the decrease was not significant.

3-Hydroxybutyric Acid↗
PubMed1994

Purification and characterization of kynurenine aminotransferase I from human brain.

Two kynurenine aminotransferases (KATs), arbitrarily termed KAT I and KAT II, are capable of producing the neuroinhibitory brain metabolite kynurenic acid from L-kynurenine in human brain tissue. Here we describe the purification of KAT I to homogeneity and the subsequent characterization of the enzyme using physicochemical, biochemical, and immunological methods. KAT I was purified from human brain approximately 2,000-fold with a yield of 2%. Assessed by polyacrylamide gel electrophoresis, KAT I migrated toward the anode as a single protein with a mobility of 0.5. The pure enzyme was found to be a dimer consisting of two identical subunits of approximately 60 kDa. Among several oxo acids tested, KAT I showed highest activity with 2-oxoisocaproate. Kinetic analyses of the pure enzyme revealed an absolute Km of 2.0 mM and 10.0 mM for L-kynurenine and pyruvate, respectively. KAT I activity was substantially inhibited by L-glutamine, L-phenylalanine, and L-tryptophan, using either pyruvate (1 mM) or 2-oxoisocaproate (1 mM) as a cosubstrate. L-Tryptophan inhibited enzyme activity noncompetitively with regard to pyruvate (Ki = 480 microM) and competitively with regard to L-kynurenine (Ki = 200 microM). Anti-KAT I antibodies were produced against pure KAT I and were partially purified by conventional techniques. Immunotitration and immunoblotting analyses confirmed that KAT I is clearly distinct from both human KAT II and rat kynurenine-pyruvate aminotransferase. Pure human KAT I and its antibody will serve as valuable tools in future studies of kynurenic acid production in the human brain under physiological and pathological conditions.

Adult↗
PubMed1994

Purification and amino- and carboxyl-terminal amino acid sequences of alanine-glyoxylate transaminase 1 from human liver.

In order to confirm the amino acid sequence predicted from the nucleotide sequence of cDNA and also to elucidate the intracellular localization and molecular evolution, human liver alanine-glyoxylate transaminase 1 (AGT1) was purified and subjected to partial amino acid sequence determination, with special attention to posttranslational modification. The enzyme was purified to homogeneity from the 10,000 x g supernatant of human liver homogenate. The purified enzyme showed only a single protein band at about 43 kDa on SDS-PAGE, indicating that it is a homodimer of two identical subunits, because the native enzyme has a molecular mass of about 80 kDa. Both the amino- and carboxyl-terminal peptides of the enzyme were isolated from a cyanogen bromide digest of the S-carboxyl-methylated protein and subjected to amino acid sequence determination. The alpha-amino group of the amino-terminal peptide was shown to be blocked by an acetyl group. The carboxyl-terminal sequence contained a putative N-glycosylation sequence (-Asn-Ala-Thr-), the only one present in the whole molecule, but this sequence was normally determined, indicating that the enzyme is not N-glycosylated. Purdue et al. [J. Cell Biol. 111, 2341-2351 (1990)] have reported that Pro-11, Gly-170, and Ile-340 in normal human AGT1 were replaced by Leu, Arg, and Met, respectively, in a patient with primary hyperoxaluria type 1. We confirmed that residue-11 was Pro. Both the amino- and carboxyl-terminal sequences of the enzyme showed extensive similarity with those of rat liver mitochondrial serine-pyruvate aminotransferase and the small chain of hydrogenase from a thermophilic unicellular cyanobacterium, Synechococcus PCC 6716.(ABSTRACT TRUNCATED AT 250 WORDS)

Alanine Transaminase↗
PubMed1993

Autosomal codominant inheritance and Japanese incidence of deficiency of OKT4 epitope with lack of reactivity resulting from conformational change.

A large Japanese family in which some members were homozygous or heterozygous for OKT4 epitope deficiency was studied. Homozygotes, heterozygotes, and normal individuals were identified by differences in the number of OKT4 epitopes on the surfaces of lymphocytes. This deficiency was transmitted as an autosomal codominant trait. The internalization of CD4 molecules and the production of IL-2 by lymphocytes of these subjects were examined. The OKT4 epitope was not needed for internalization of CD4 molecules, and IL-2 was produced in the same amounts by these different kinds of subjects. DNA from four clones lacking OKT4 established from four individuals of this family was sequenced. As reported elsewhere for different subjects, a single nucleotide substitution (CGG-->TGG) was found in all four cell lines. The mutation results in arginine being replaced by tryptophan. Analysis showed different hydrophobicity at positions 239 and 240 from the control, probably giving rise to a conformational change in CD4 accounting for lack of reactivity with the OKT4 monoclonal antibody. The incidence of homozygotes in the Japanese population was found to be 0.47% by examination of 1478 random samples, and on the basis of this value, the incidence of heterozygotes was estimated to be 12.8%.

Amino Acid Sequence↗
PubMed1993

Cytotoxicity of Porphyromonas gingivalis toward cultured human gingival fibroblasts.

Direct cytotoxicity of black-pigmented anaerobic rods was studied on the confluent monolayer of human gingival fibroblasts in vitro. Only strains of Porphyromonas gingivalis caused morphological alteration (cell-rounding) and notable depression of viability of fibroblasts. To determine the location of the cytotoxicity, bacterial surface components, i.e., outer membrane, lipopolysaccharide, fimbriae and outer membrane vesicles were prepared from P. gingivalis and their cytotoxicity was assessed. Among these preparations, only outer membrane vesicles are supposed to have high affinity to human gingival fibroblasts, and the cytotoxicity of outer membrane vesicles was found to be much stronger than that of the other constituents. This cytotoxic factor seemed to consist largely of protein and to be associated with the enzyme activity of outer membrane vesicles. The effects of some protease inhibitors and L-cysteine on the cytotoxicity of outer membrane vesicles suggest that the mechanism of cell-rounding is different from that of cell death.

Adolescent↗
PubMed1993

2-Aminoadipate-2-oxoglutarate aminotransferase isoenzymes in human liver: a plausible physiological role in lysine and tryptophan metabolism.

Two major 2-aminoadipate aminotransferase (AadAT) activities of human liver extract were separated by DEAE-Sepharose column chromatography. The faster eluting enzyme was designated AadAT-I and the other one AadAT-II. AadAT-I had a hgih Km value for aminoadipate, 20 mmol/l, and a low Km value for glutamate, 1.4 mmol/l. In contrast, AadAT-II had a low Km value for aminoadipate, 0.25 mmol/l, and a high Km value for glutamate, 12.5 mmol/l. AadAT-I and AadAT-II were mainly localized in the supernatant and mitochondrial fraction, respectively. AadAT-I demonstrated only glutamate-2-oxoadipate or 2-aminoadipate-2-oxoglutarate aminotransferase activities. AadAT-II further showed the activity of tryptophan and kynurenine. On the basis of Km values and subcellular localization of the isoenzymes, a plausible role was suggested for them involving the metabolism of lysine and tryptophan.

2-Aminoadipate Transaminase↗
PubMed1992

Gene structure of human indoleamine 2,3-dioxygenase.

Two genomic DNA clones that encode human indoleamine 2,3-dioxygenase (IDO) were isolated from the human genomic DNA library using the IDO cDNA as a probe, and their restriction maps and partial nucleotide sequences were determined. The human IDO gene spanned 15 kilobase pairs with ten exons. The 5' terminus of the IDO mRNA was 33 nucleotides upstream of the translation initiation codon ATG. The 5' flanking region contained ISRE, X-box, and Y-box like sequences. Southern blot analysis of the human genomic DNA indicated that the human IDO gene was present in a single copy in the genome.

Alternative Splicing↗
PubMed1992

Localization of kynurenine aminotransferase immunoreactivity in the rat hippocampus.

The localization and distribution of kynurenine aminotransferase (KAT), the biosynthetic enzyme of the excitatory amino acid receptor antagonist, kynurenic acid, was studied in the rat hippocampal formation with immunohistochemical methods. The enzyme was found mainly in glial cells that could be distinguished as 3 types on the basis of their shapes and locations. Typically, these cells shared the morphological features of astrocytes and exhibited glial fibrillary acidic protein immunoreactivity as demonstrated by a double-labeling technique. The distribution of KAT-containing glial cells was heterogeneous throughout the hippocampal formation. In the hippocampus, the stratum lacunosum-moleculare of Ammon's horn and the hilus contained a higher density of KAT-positive glial cells than other regions, whereas the lowest density of KAT glial cells was observed in the granule cell layer of the dentate gyrus and in the stratum radiatum of CA subfields. In the subicular complex, the density of KAT-containing glial cells was generally higher in the superficial than in the deep layer. Hippocampal neurons exhibiting KAT immunoreactivity, distinguished as nonpyramidal cells, were very few in number and mainly distributed in strata oriens and pyramidale of Ammon's horn. Substantially more KAT-positive neurons were observed in layers II and III of the subicular complex. The organization of cellular elements containing KAT may be of relevance for the function and possible dysfunction of kynurenic acid in the rat hippocampal formation.

Animals↗
PubMed1992

Formation of hydroxanthommatin-derived radical in the oxidation of 3-hydroxykynurenine.

Using ESR, a radical (g = 2.004) was detected in the reaction mixture of 3-hydroxykynurenine (3-HKY), H2O2, and horseradish peroxidase. The radical was stable and was detected even after 5 h. On HPLC analysis of the reaction mixture, two radical peaks (Peak-1 and Peak-2) were detected using ESR. The ESR spectra of Peak-1 and Peak-2 radicals were the same and identical with that of the original radical in the reaction mixture. The retention times of Peak-1 and Peak-2 corresponded to those of authentic xanthommatin (XA) and hydroxanthommatin (Hydro-XA), respectively, XA being formed in the oxidation of 3-HKY by potassium ferricyanide and Hydro-XA being formed in the reduction of XA by sodium metabisulfite. The absorbance spectra of Peak-1 and Peak-2 were nearly identical with those of authentic XA and Hydro-XA. The absorbance spectrum of Peak-2 changed from that of Hydro-XA to that of XA, indicating that Hydro-XA auto-oxidized to XA in the air. The ESR signal intensity of the Peak-2 radical developed in accordance with the progress of this auto-oxidation of Hydro-XA to XA. It was supposed that the Peak-2 radical was generated in the auto-oxidation of Hydro-XA after its elution from the HPLC column. Thus, the radical seemed to be the one-electron oxidized form of Hydro-XA. The Peak-1 radical appeared to be the true retention of the radical on the column and to be eluted with a much larger amount of XA. The separation of the radical from XA was impossible on the column. Hemoglobin (Hb) or hematin also induced the same radical in the reaction mixture of 3-KHY, H2O2, and Hb or hematin.

Animals↗
PubMed1992

Oxidation of 3-hydroxykynurenine catalyzed by methemoglobin with hydrogen peroxide.

Methemoglobin (metHb) with H2O2 catalyzed the oxidation of 3-hydroxykynurenine (3-HKY) in the reaction mixture of metHb, 3-HKY, and H2O2. The spectrophotometric experiments suggest the following mechanism for the 3-HKY oxidation by metHb with H2O2. MetHb first reacts with H2O2 to form the ferryl complex of Hb. This species then oxidizes 3-HKY, while it returns to metHb. 3-HKY was more reactive with the ferryl complex than glutathione but less reactive than ascorbic acid. Scavengers of the hydroxyl radical, dimethyl sulfoxide and ethanol, scarcely inhibited the 3-HKY oxidation by metHb with H2O2. Desferrioxamine, a metal chelator, hardly suppressed the 3-HKY oxidation. These results indicate that the hydroxyl radical is not involved in the 3-HKY oxidation by metHb with H2O2.

Free Radicals↗
PubMed1992

In vivo 19F MR spectroscopic study of metabolism of 5-fluorotryptophan in rat liver.

The metabolism of 5-fluorotryptophan in rat liver was examined by in vivo 19F MR spectroscopy. After i.v. injection of 200 mg/kg b.w. of 5-fluorotryptophan the substance was noted immediately as a strong peak, which decreased gradually. Another peak appeared about 40 min after the injection. The chemical shift value of this peak was 1.6 ppm from that of 5-fluorotryptophan. Kynurenine is known to be a major metabolite of tryptophan in the liver. We synthesized 5-fluorokynurenine from 5-fluorotryptophan by ozonolysis. The chemical shift value of 5-fluorokynurenine was confirmed to be coincident with that of the metabolite peak. This strongly suggests that the metabolite peak of 5-fluorotryptophan observed in this study was the 5-fluorokyrurenine signal. We also applied this method for the CCl4-injured liver. In the liver injury group, the metabolite peak appeared slowly and the intensity was low compared to that of the normal group, though the peak of 5-fluorotryptophan decreased similarly as in the normal liver. These results suggest that the decrease of 5-fluorotryptophan is due mainly to the renal excretion, as the injured liver could not metabolize 5-fluorotryptophan.

Animals↗
PubMed1991

Kinetic properties of human liver tryptophan pyrrolase.

Human liver tryptophan pyrrolase (TPO) activity exhibited substrate level regulation. TPO showed biphasic activity to tryptophan when low ascorbate was used as an activator. The high affinity form (Km for tryptophan: 0.05 mM) was promoted by low ascorbate and low tryptophan. The low affinity form (Km for tryptophan: 0.4 mM) was induced by high concentrations of tryptophan or ascorbate. Both high and low affinity forms showed the same affinity to oxygen. The high affinity form was also induced by pyrroloquinoline quinone, but this effect was decreased by catalase, suggesting the participation of H2O2.

Ascorbic Acid↗
PubMed1991

In vitro effect of dimethoate on the activity of tryptophan pyrrolase in rat liver.

Total and holo-enzyme activities of tryptophan 2,3-dioxygenase were measured in vitro in the presence and absence of the organophosphorous insecticide, dimethoate. Addition of dimethoate to the reaction mixture decreased the activities of both total and holo-forms. Total and holo-enzyme activities were decreased by 34% and 26%, respectively, by 1 mM dimethoate. On the other hand, 5 mM dimethoate resulted in 56% and 34% inhibition to total and holo-enzyme activities, respectively. Lineweaver-Burk plot of the total-enzyme activity at different tryptophan concentration in the presence of 2 mM dimethoate gave uncompetitive type of inhibition.

Animals↗
PubMed1991

Superoxide dismutase enhances the toxicity of 3-hydroxyanthranilic acid to bacteria.

Cu,Zn.superoxide dismutase (SOD) enhanced the toxicity of 3-hydroxyanthranilic acid (3-HAT) to Salmonella typhimurium strain TA 102, evaluated as ability to form colonies. MnSOD showed the same effect. Inactivated Cu.ZnSOD had no effect. SODs accelerated the oxidation of 3-HAT, but inactivated Cu.ZnSOD caused little acceleration. It is proposed that the acceleration of 3-HAT oxidation leads to the enhancement of the 3-HAT toxicity. Catalase protected the bacteria from the toxicity of 3-HAT enhanced by Cu,ZnSOD, indicating that hydrogen peroxide generated in the oxidation of 3-HAT is involved in the toxicity. SODs accelerate the oxidation of 3-HAT and generate more hydrogen peroxide, that causes the enhancement of the 3-HAT toxicity to the bacteria. However, hydrogen peroxide alone was not so toxic. Hydrogen peroxide with 3-HAT was more toxic to the bacteria.

3-Hydroxyanthranilic Acid↗