Biomedical subjects

S Kaneko

Publications and source records attributed to S Kaneko.

At least 325 records · Page 18Linked to original sources

PubMed1997

BDNF prevents NO mediated glutamate cytotoxicity in cultured cortical neurons.

The effects of brain-derived neurotrophic factor (BDNF) on glutamate-induced cytotoxicity were examined using primary cultures of rat cortical neurons. BDNF induced TrkB tyrosine phosphorylation in rat cultured cortical neurons. The cell viability was significantly reduced when cultures were briefly exposed to glutamate and incubated with normal medium for 24 h. Glutamate cytotoxicity was prevented by MK-801, which is a non-competitive blocker of N-methyl-D-aspartate and N(omega)-nitro-L-arginine, which is a blocker of nitric oxide synthetase. Delayed neurotoxicity was also induced by ionomycin, a calcium ionophore, and nitric oxide (NO) donors such as S-nitrosocysteine (SNOC) and 3-morpholinosydnonimine (SIN-1). Incubating cultures with BDNF for 10 min to 24 h protected cortical neurons against glutamate neurotoxicity. The protective effects of BDNF against glutamate cytotoxicity were dependent on both its concentrations and incubation time. BDNF also prevented the ionomycin-, SNOC-, and SIN-1 induced cytotoxicity. These results indicate that BDNF protects cultured cortical neurons from NMDA receptor-mediated glutamate neurotoxicity by reducing cytotoxic action of NO.

Animals↗

PubMed1997

Delineation of regions important for heteromeric association of hepatitis C virus E1 and E2.

Hepatitis C virus (HCV) is the major causative agent of blood-borne non-A non-B hepatitis. The persistence of HCV infection is believed to reflect escape from the host immunosurveillance system by mutations in hypervariable region 1 (HVR1) of the envelope protein 2 (E2). Two envelope proteins of HCV, E1 and E2, have been reported to form a heteromeric complex but the exact organization of the viral envelope proteins remains uncertain. We examined the interaction of E1 and E2 by far- Western blotting using the bacterial recombinant proteins and also by pull-down assay using mammalian expressed proteins. The major E1-interacting site of E2 was mapped within the N-terminal part of E2 (NCD1) (aa 415 to 500 of the polyprotein). Both HVR1 and HVR2, located at the N-terminal part of E2, were dispensable for the interaction. Although several discontinuous regions within NCD1 seemed to contribute to the strong binding to E1, the highly conserved amino acid sequences flanking HVR2 had the most significant effect. The amino acid residues "WHY" from 489 to 491 of E2 played an especially crucial role since the constructs with the internal deletion or substitution of the residues showed severely impaired E1-binding. The N-terminal part of E1 is important for the E2-binding as determined by far-Western blotting using the mammalian- and bacterial-expressed E2 proteins as probes. The mammalian-expressed, glycosylated forms of the E1 and E2 proteins exhibited E1-E2 binding activities similar to those of the bacterial-expressed, nonglycosylated forms in pull-down assays, suggesting that glycosylation is not prerequisite for the heteromeric complex formation of E1 and E2.

Amino Acid Sequence↗

PubMed1997

Overexpression of B cell-specific activator protein (BSAP/Pax-5) in a late B cell is sufficient to suppress differentiation to an Ig high producer cell with plasma cell phenotype.

The B cell-specific activator protein (BSAP) is a DNA-binding transcription factor expressed in pro-B, pre-B, and mature B cells but not in plasma cells. We explored the role of BSAP in B cell function by creating clones in a late B cell and a plasma cell line transfected with a BSAP expression plasmid. We found that the plasma cell line MPC11, which does not produce BSAP, is still permissive to BSAP production driven by heterologous promoter. Overexpression of BSAP in a late B cell line (CH12.LX.A2) and a plasma cell line augmented cell proliferation and led to greater suppression of Ig synthesis in a late B cell line than in the plasma cell line. The reduction was seen mostly in synthesis of a secretory form of Ig. Overexpression of BSAP reduced Blimp-1 expression in CH12.LX.A2 clones but not in MPC11 clones. In addition, overexpression of BSAP in CH12.LX.A2 cells suppressed spontaneous appearance of cells with high Syndecan-1 expression and high amounts of intracytosolic as well as secreted Ig synthesis. To corroborate the above findings, we cloned nontransfected CH12.LX.A2 cells and found reduced BSAP mRNA expression in the high Ig-secreting clones, which produced more Blimp-1 mRNA with greater Syndecan-1 expression than the low Ig-secreting clones. Taken together, these results indicate that BSAP expression is sufficient to reduce Ig production in late B cells; this effect is mediated in part by suppression of differentiation to cells of plasma-cell phenotype.

Animals↗

PubMed1997

Autoreactive T cell clones from patients with systemic lupus erythematosus support polyclonal autoantibody production.

This work examines the functional properties and TCRbeta gene utilization of 15 autoreactive T cell clones derived from five patients with systemic lupus erythematosus. All these clones proliferated and secreted cytokine when stimulated in vitro by autologous (but not allogenic) B cells. Individual T cell clones used diverse TCRbeta genes and did not show skewing toward the preferential usage of anionically charged receptors. Autoreactive T cell clones supported polyclonal B cell activation, as characterized by the production of anti-DNA, anti-Sjögren syndrome A, and anti-tetanus toxoid (anti-TT) Abs. This T cell help was mediated through the production of immunostimulatory cytokines, especially IL-6. Although stimulation of the autoreactive clones was blocked by anti-HLA class II Abs, the T cell clones did not proliferate, nor did they support polyclonal IgG production by HLA class II-matched normal B cells. Unlike the autoreactive clones, TT-specific clones derived from the same patients provided help selectively to B cells secreting anti-TT Abs. These findings suggest that autoreactive T cells from systemic lupus erythematosus patients are triggered to provide help following cognate interactions with self-peptides presented in the context of HLA class II molecules expressed on autologous B cells regardless of their specificities.

Amino Acid Sequence↗

PubMed1997

Effects of B vitamins on glutamate-induced neurotoxicity in retinal cultures.

The effects of B vitamins on glutamate-induced neurotoxicity were examined using primary cultures obtained from the rat retina. Cell viability was markedly reduced by a brief exposure to glutamate followed by incubation with glutamate-free media for 1 h. Glutamate cytotoxicity was reduced in the cultures that had been maintained in thiamine-, pyridoxine- or nicotinamide-containing medium before the exposure to glutamate. Glutamate cytotoxicity was also reduced by chronic application of thiamine pyrophosphate and pyridoxal phosphate, which are active coenzyme forms of thiamine and pyridoxine, respectively. By contrast, chronic application of riboflavin, pantothenate, biotin, folic acid and inositol did not affect glutamate cytotoxicity. None of the B vitamins tested had any effect on glutamate cytotoxicity when added only during the exposure to glutamate. These findings suggest that chronically applied thiamine, pyridoxine and nicotinamide protect retinal neurons against glutamate cytotoxicity.

Animals↗

PubMed1997

Direct evidence for increase in excitatory amino acids release during mossy fiber LTP in rat hippocampal slices as revealed by the patch sensor methods.

We had previously established a patch sensor system for detecting overflow of excitatory amino acids (EAA) from hippocampal slices induced by a single electric stimulus. We performed simultaneous recordings of patch sensor current and whole-cell current in rat hippocampal slices. Tetanus of mossy fibers produced long-term potentiation (LTP) of excitatory postsynaptic current (EPSC) recorded from a CA3 pyramidal cell with an increase in single electric stimulus-evoked patch current amplitude recorded with a patch sensor positioned in the stratum lucidum. In Schaffer collateral-CA1 synapses, however, LTP of EPSC was accompanied by an increase in AMPA-evoked whole-cell current, but not with change in patch current in the stratum radiatum. These results indicate that EAA release is increased during mossy fiber LTP but not CA1 LTP.

Animals↗

PubMed1997

Determination of the effects of caffeine and carbamazepine on striatal dopamine release by in vivo microdialysis.

The effects of carbamazepine and caffeine on adenosine receptor subtypes were determined using in vivo microdialysis in an attempt to elucidate their different psychotropic mechanisms of action. Adenosine and a selective adenosine A1 receptor agonist decreased the striatal extracellular dopamine level, whereas caffeine, carbamazepine and a selective adenosine A1 receptor antagonist increased it, but neither an adenosine A2 receptor agonist nor an antagonist affected it. Under conditions of adenosine A1 receptor blockade, adenosine, carbamazepine and a selective adenosine A2 receptor agonist increased the striatal extracellular dopamine level, whereas caffeine and a selective adenosine A2 receptor antagonist decreased it. These results suggest that adenosine A1 receptor stimulation reduces the striatal extracellular dopamine level, and that adenosine A2 receptor stimulation under conditions of adenosine A1 receptor blockade increases it. Therefore, caffeine is an antagonist of both adenosine A1 and A2 receptor subtypes, and carbamazepine is an adenosine A1 receptor antagonist as well as an adenosine A2 receptor agonist. These properties support the hypothesis that the central actions of both carbamazepine and caffeine result from effects on both adenosine A1 and A2 receptors.

Adenosine↗

PubMed1997

Human hepatitis B virus enhancer 1 is responsive to human interleukin-6.

Serum levels of interleukin-6 (IL-6) are elevated in acute and chronic hepatitis B patients. The effect of IL-6 and its transcription factor of NF-IL6 (a nuclear factor for IL-6) on hepatitis B virus (HBV) enhancer 1 (Enh1), which controls HBV X expression, were investigated in HepG2 cells. Twenty ng/ml of IL-6 increased 4-fold the enhancer activity of Enh1 according to the CAT assay. The IL-6 stimulation was abolished by introducing a mutation either in an AP-1-related site or a C-stretch sequence in the Enh1 sequence, demonstrating that the cis-elements are necessary for the IL-6 response. Co-transfection of NF-IL6 expression plasmid similarly increased the enhancer activity of Enh1 through both binding sites. Further, a specific complex formation of the Enh1 was detected using HepG2 nuclear lysates by electromobility shift assays, and the complex formation was increased in the lysates of cells treated with IL-6 and NF-IL6-transfection. In competition assays, one half of the complex formed was found to remain in the presence of 500-times excess competitor DNA fragment harboring NF-IL2 binding site, suggesting indirect binding of NF-IL6 to the Enh1 sequence. These results indicate that IL-6 increased the enhancer activity of HBV Enh1 through signal transduction pathways, indirectly involving NF-IL6, and may control HBV X expression and viral replication in HBV infected liver.

Base Sequence↗

PubMed1997

In vivo mechanisms for the inhibition of T lymphocyte activation by long-term therapy with tacrolimus (FK-506): experience in patients with Behçet's disease.

OBJECTIVE: To examine the in vivo mechanisms of suppression of T lymphocyte function in patients with Behçet's disease (BD) undergoing long-term treatment with tacrolimus (FK-506). METHODS: Intracellular proteins were analyzed by immunoprecipitation and Western blotting. Messenger RNA expression was studied by a polymerase chain reaction-based technique. RESULTS: Interleukin-2 production was suppressed in patients treated with tacrolimus. This suppression was found to be due to inhibition of interactions between activated calcineurin (Cn) and nuclear factor of activated T cells (NF-AT), inhibition of cleavage of the autoinhibitory domain of the CnA subunit, and inhibition of heterodimer formation by CnA and CnB subunits, resulting in the absence of NF-AT in nuclei of the T cells. We found that T lymphocytes in some BD patients treated with tacrolimus had reduced amounts of FK-506 binding protein (FKBP) in their cytoplasm. CONCLUSION: Tacrolimus reduces the Cn activity of T cells in vivo by the cumulative effects of several distinct mechanisms. It is plausible that reduced amounts of FKBP may be associated with diminished clinical efficacy in some BD patients receiving prolonged treatment with tacrolimus.

Adult↗

PubMed1997

Relationship between the CYP2D6 genotype and the steady-state plasma concentrations of trazodone and its active metabolite m-chlorophenylpiperazine.

The relationship between the cytochrome P450 (CYP) 2D6 genotype and the steady-state plasma concentrations (Css) of trazodone and its active metabolite m-chlorophenylpiperazine (mCPP) was studied in 54 depressed Japanese patients receiving trazodone 150 mg at bedtime. By use of allele-specific PCR analysis, the wild type allele, three mutated alleles causing absent enzyme activity (CYP2D6A, CYP2D6B and CYP2D6D) and one mutated allele causing decreased enzyme activity (CYPZD6 Ch) were identified. The means (ranges) of the Css of trazodone, corrected to the median body weight in 17 cases with no mutated allele, 27 cases with one mutated allele and 10 cases with two mutated alleles, were 556 (281-1115), 643 (302-1362) and 671 (234-1418) ng/ml, respectively, while the values of mCPP were 60 (35-121), 65 (33-99) and 58 (38-112) ng/ml, respectively. Neither the Css of trazodone (F = 0.80, P = 0.45) nor that of mCPP (F = 0.49, P = 0.61) significantly differed among the three groups. The present study thus suggests that the CYP2D6 genotype cannot predict the Css of these compounds.

Adult↗

PubMed1997

Interaction between carbamazepine and bromperidol.

OBJECTIVE: The interaction between carbamazepine and bromperidol was studied in 13 schizophrenic inpatients. METHODS: Before carbamazepine addition, the subjects were taking bromperidol 12-24 mg.day-1 for 1-20 weeks. Carbamazepine 400 mg.day-1 was coadministered for 4 weeks, and blood samplings were performed before carbamazepine addition and at weekly intervals after the addition. Plasma concentrations of bromperidol and its reduced metabolite were measured by high-performance liquid chromatography. RESULTS: Carbamazepine significantly decreased plasma concentrations of both bromperidol and reduced bromperidol for all weeks. On average, the plasma concentrations of bromperidol and reduced bromperidol at 4 weeks were 37% and 23% of the corresponding precarbamazepine values. Despite these decreases in plasma concentration, the Clinical Global Impression scores decreased slightly but significantly after carbamazepine addition. CONCLUSION: The present study suggests that carbamazepine decreases plasma concentrations of bromperidol and its reduced metabolite by inducing the metabolism of these compounds. Nevertheless, adjunctive carbamazepine may be useful for schizophrenic patients treated with bromperidol.

Adult↗

PubMed1997

Metabolic disposition of pantoprazole, a proton pump inhibitor, in relation to S-mephenytoin 4'-hydroxylation phenotype and genotype.

OBJECTIVES: To assess the possible relationship between the metabolic disposition of pantoprazole and genetically determined S-mephenytoin 4'-hydroxylation phenotype and genotype. METHODS: The pharmacokinetic disposition of pantoprazole was investigated in 14 Japanese male volunteers (seven extensive and seven poor metabolizers of S-mephenytoin). All subjects received a single 40 mg oral dose of pantoprazole as the enteric-coated formulation. RESULTS: An interphenotypic difference in the metabolic disposition of pantoprazole was observed: the mean values for area under the concentration-time curve (AUC), elimination half-life (t1/2), and apparent oral clearance were significantly (p < 0.01) greater, longer, and lower, respectively, in the poor metabolizers than in the extensive metabolizers. The mean AUC of pantoprazole sulfone was greater (p < 0.01) in the poor metabolizers than in the extensive metabolizers, whereas the mean AUC of the main demethylated metabolite (M2) was lower (p < 0.01) in the poor metabolizers than in the extensive metabolizers. A significant negative correlation was observed between the individual values for log10% urinary excretion of 4'-hydroxymephenytoin and AUC of pantoprazole (rs = -0.816; p < 0.005). The CYP2C19 genotyping test results were found to be in a complete accordance with the phenotypes. CONCLUSION: These data indicated that the metabolic disposition of pantoprazole is under the pharmacogenetic control of S-mephenytoin 4'-hydroxylase (CYP2C19).

2-Pyridinylmethylsulfinylbenzimidazoles↗

PubMed1997

Soluble E-selectin as a marker of disease activity in atopic dermatitis.

BACKGROUND: Augmentation in the expression of adhesion molecules on endothelial cells can regulate leukocyte migration. These molecules are also shed into the circulation. The level of soluble adhesion molecules in the serum is known to reflect the degree of systemic inflammation, and this level can therefore be used as a marker of inflammation. OBJECTIVE: To elucidate whether soluble adhesion molecules can be used as the marker of disease severity in atopic dermatitis, we examined the levels of soluble E-selectin, soluble intercellular adhesion molecule-1, and soluble vascular cell adhesion molecule-1 and compared these levels with the patients' symptom scores and their total serum IgE levels. METHODS: Fifty-three patients with atopic dermatitis were studied. Soluble adhesion molecules were measured by ELISA. RESULTS: The level of soluble E-selectin was higher in patients with atopic dermatitis than in healthy control subjects (p < 0.01). Moreover, soluble E-selectin is correlated with disease severity (p < 0.01). Soluble E-selectin also reflected the changes in symptom scores. In contrast, there were no differences in soluble intercellular adhesion molecule-1 and soluble vascular cell adhesion molecule-1 between the patients and control subjects. CONCLUSION: Soluble E-selectin is a good marker of disease severity and of its activity in atopic dermatitis.

Adult↗

PubMed1997

Cognitive enhancers and hippocampal long-term potentiation in vitro.

We review our works on the pharmacological modulation of long-term potentiation (LTP) at guinea pig hippocampal mossy fiber-CA3 synapses in vitro. The magnitude of tetanus-induced LTP at the mossy fiber synapse was augmented by perfusion of slices with several cognitive enhancers, such as bifemelane (1 microM). The mossy fiber LTP was enhanced by somatostatin (0.32 microM) and inhibited in somatostatin-depleted slices from cysteamine-treated guinea pigs. An involvement of the 5-HT3 receptor also showed that granisetron (0.1 microM) enhanced the mossy fiber LTP. The above-mentioned enhancements by perfused agents were commonly reversed, at least in part, by muscarinic antagonists. However, the magnitude of mossy fiber LTP was bidirectionally modulated by muscarinic stimulations of slices with physostigmine or carbachol at different concentrations. The enhancing effects of high-concentration carbachol was antagonized by pirenzepine, and in contrast, the inhibition by low-concentration carbachol was antagonized in the presence of AF-DX116. When guinea pigs were preinjected with the cholinotoxin AF64A, the magnitude of LTP was decreased in the slices prepared from AF64A-treated animals. These results suggest that endogenous acetylcholine dominantly plays facilitatory roles through muscarinic M1 receptors in the induction of mossy fiber LTP. The pharmacological characterization of mossy fiber LTP may be of help to the evaluation of cognitive enhancers at a neuronal circuit level.

Animals↗

PubMed1997

Involvement of M2 receptor in an enhancement of long-term potentiation by carbachol in Schaffer collateral-CA1 synapses of hippocampal slices.

We examined effects of carbachol (CCh), muscarinic receptor agonist, on long-term potentiation (LTP) of field excitatory postsynaptic potential (fEPSP) at Schaffer collateral-CA1 synapse of guinea pig hippocampal slices using extracellular recording technique. Application of 0.1 microM CCh to the slices significantly augmented the magnitude of LTP without significant change in the amplitude of pretetanus fEPSP. The enhancement of LTP by 0.1 microM CCh was significantly attenuated by 0.1 microM AF-DX 116, M2 receptor antagonist, but not by 0.1 microM pirenzepine, M1 receptor antagonist. Ten micromolar of carbachol reduced the amplitude of pretetanus fEPSP, while the magnitude of LTP was significantly larger than that in control slices to which tetanus was applied in a stimulus intensity producing pretetanus fEPSPs with an amplitude comparable to those during administration of 10 microM CCh. Neither 0.1 microM pirenzepine nor 0.1 microM AF-DX 116 had significant effect on the enhancement of LTP by 10 microM CCh. These results suggest that the induction of LTP at Schaffer collateral-CA1 synapse was enhanced through the activation of M2 receptors by CCh at a lower concentration.

Animals↗

PubMed1997

Plasma concentrations of trazodone and m-chlorophenylpiperazine at steady state can be predicted from those after an initial dose of trazodone.

1. The authors studied the correlations between plasma concentrations of trazodone and mCPP at steady state and those after an initial dose of trazodone. 2. Fifteen depressed patients received trazodone 150 mg at bedtime for 3 weeks, and blood samplings were taken 12 h after the initial dose and 12 h after the last dose at each week. Plasma concentrations of trazodone and mCPP were measured by high-performance liquid chromatography. 3. Plasma concentration of mCPP, but not trazodone, was significantly higher at each week than after initial dosing. 4. For both trazodone and mCPP, significant linear relationships were found between plasma concentration after initial dosing and the average of 3 weeks' plasma concentrations. 5. The present study thus suggests that plasma concentrations of trazodone and mCPP at steady state can be predicted from those after an initial dose of trazodone.

Adult↗

PubMed1997

Possible interaction between cisapride and bromperidol.

1. The case of a schizophrenic patient taking bromperidol (18 mg/day), whose psychotic symptoms deteriorated markedly after addition of cisapride (7.5 mg/day), is presented. 2. Retrospective determination of drug concentrations revealed that plasma concentrations of bromperidol and its reduced metabolite were increased after cisapride addition. 3. The present study thus suggests that there is an interaction between cisapride and bromperidol.

Adult↗

PubMed1997

Effects of genetically determined S-mephenytoin 4-hydroxylation status and cigarette smoking on the single-dose pharmacokinetics of oral alprazolam.

This study examines the effects of genetically determined S-mephenytoin 4-hydroxylation capacity and cigarette smoking on the single-dose pharmacokinetics of oral alprazolam in 12 healthy male volunteers. Six subjects each were extensive metabolizers (EMs) and poor metabolizers (PMs) of S-mephenytoin 4-hydroxylation. Seven subjects were smokers (> 10 cigarettes/day), and five were nonsmokers, according to their self-reports. Each subject took a single oral dose of 0.8 mg of alprazolam, and blood samples were collected up to 48 hours postdose. Psychomotor function was assessed at times of blood samplings using the Digit Symbol Substitution Test (DSST), Visual Analog Scale (VAS), and UKU Side Effect Rating Scale. Plasma alprazolam concentrations were measured by a high-performance liquid chromatography assay. None of the mean pharmacokinetic parameters was significantly different between the EM and PM phenotype groups. Although the mean elimination half-life was significantly shorter in the smoker group (p < .01) than in the nonsmoker group (13.1 +/- 2.9 vs. 20.0 +/- 2.7 hours, mean +/- SD), other pharmacokinetic parameters did not differ significantly between the two groups. Psychomotor function parameters did not differ significantly either between the EM and PM groups or between the nonsmoker and smoker groups. The present study thus suggests that neither S-mephenytoin 4-hydroxylation status nor self-reports of extensive cigarette smoking has a major impact on the metabolism of alprazolam in humans.

Adult↗