Search PubMed⌕ Search

Biomedical subjects

C Watanabe

Publications and source records attributed to C Watanabe.

At least 19 recordsLinked to original sources
PubMed1998

Functional analysis of peripheral blood B cells in patients with X-linked agammaglobulinemia.

X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disease caused by mutations of Bruton tyrosine kinase (Btk); Btk plays an essential role in the development of mature B cells. However, small numbers of B cells ("leaky B cells") are present in the peripheral blood of most XLA patients. In this study, we analyzed the function of these leaky B cells obtained from XLA patients. Enough numbers of B cells were available for analysis from five of nine XLA patients originally screened. Sequence analysis revealed missense mutations of Btk in four of the five XLA patients. No mutation was found in the coding region of Btk in one patient. Western blotting and/or flow cytometric analysis failed to detect Btk protein in all five patients. B cells isolated from peripheral blood of these XLA patients were CD5-, CD20+, CD19+, and CD21-. If stimulated with anti-CD40 and IL-4, XLA B cells proliferated normally and produced significant amounts of IgE. Anti-CD40 stimulation of XLA B cells resulted in normal expression of CD23. In addition, three of the five XLA patients studied were immunized with bacteriophage phiX174 and produced low but detectable levels of antiphage-specific Ab. Similarly, X-linked immunodeficiency mice, which carry a missense mutation in Btk, produced substantial amounts of antiphage Ab. These results indicate that CD40 signaling is intact in B cells lacking demonstrable Btk, and that leaky B cells in XLA patients can proliferate, undergo isotype switching, and differentiate into specific Ab-producing cells.

Adolescent↗
PubMed1998

Collagen induces tyrosine phosphorylation of Wiskott-Aldrich syndrome protein in human platelets.

Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT) are caused by mutations of the WAS protein (WASP) gene. All hematopoietic stem cell-derived lineages, including platelets, express WASP. Platelets from WAS patients are smaller than their normal counterparts and defects in platelet aggregation and actin polymerization have been reported. To determine if WASP is important for normal platelet function, we examined its role in signal transduction. We found that collagen but not thrombopoietin or thrombin induces a rapid and robust increase in tyrosine phosphorylation of platelet-associated WASP. Collagen-induced tyrosine phosphorylation of WASP was inhibited by cytochalasin D and wortmannin, respectively, suggesting that actin polymerization and phosphatidylinositol 3-kinase (PI3-kinase) play a role in the induction of tyrosine phosphorylation of WASP. Binding of glutathion S-transferase (GST)-Grb2 to WASP was seen in the lysate of resting platelets. The binding was reduced when lysates from collagen-stimulated platelets were incubated with GST-Grb2, suggesting that tyrosine phosphorylation of WASP may directly or indirectly modulate the adapter function of WASP. Although thrombin- and thrombopoietin-induced increase in tyrosine phosphorylation of WASP is negligible or marginal, WASP from thrombin-activated platelets became incorporated into the Triton X-100-insoluble 10, 000g sedimentable residue in an aggregation-dependent manner, suggesting that it may have a regulatory role in platelet cytoskeletal processes during aggregation. Lastly, we found that WASP is cleaved in response to activation of calpain, a protease that may have a role in postaggregation signaling processes. Our data suggest that collagen specifically induces an increase in tyrosine phosphorylation of WASP and that WASP is involved in signaling during thrombin-induced aggregation by its redistribution to the cytoskeleton and its cleavage during aggregation.

Adaptor Proteins, Signal Transducing↗
PubMed1998

Effect of spikelets of Miscanthus sinensis on IgE-mediated biphasic cutaneous reaction in mice.

The effect of spikelets of Miscanthus sinensis Andersson (M. sinensis) on IgE-mediated biphasic cutaneous reactions was investigated in BALB/c mice. Mice were passively sensitized by an intravenous (i.v.) injection of monoclonal antidinitrophenol IgE antibody (anti-DNP IgE mAb), or actively by an intraperitoneal (i.p.) injection of DNP-derivatized ovalbumin (DNP-OVA) plus aluminium hydroxide gel (Alum) as an adjuvant. Skin reactions were elicited by an epicutaneous challenge of dinitrofluorobenzene (DNFB) and occurred biphasically with peak responses at 1 and 24 h in both animal models. The administrations of a nondialysable water extract of M. sinensis within 2 h before or after DNFB challenge via oral or i.p. route significantly inhibited the biphasic cutaneous reactions in passively and actively sensitized mice. The inhibitory effect was much stronger than those of a glucocorticoid, prednisolone, and histamine release inhibitor, amlexanox, as positive controls. The active component(s) was predominantly located in the glycoprotein fraction by gel chromatography. In the ears of DNFB-challenged mice, this fraction suppressed the accumulation of inflammatory cells, including mast cells and neutrophils/macrophages. In addition, the biphasic ear swelling was also improved by an administration of the glycoprotein fraction 24 h before active sensitization. These findings indicate that the glycoprotein fraction of M. sinensis was able to inhibit not only the IgE-mediated allergic inflammatory reaction but also the IgE formation. Thus, this fraction may be a useful antiallergic therapy.

Animals↗
PubMed1998

Argyrophilic structures stimulate glial reactions in neurofibrillary tangles and senile plaques.

Neurofibrillary tangles (NFT) and senile plaques (SP) contain various pathological structures, and the majority of these pathological structures are argyrophilic. To investigate the glial reactions of the argyrophilic substance, we performed immunohistochemistry for microglia or for astroglia after Gallyas-Braak staining, which is one of the most sensitive silver impregnation techniques detecting argyrophilic structures in NFT and SP. We found that extracellular argyrophilic structures in NFT and SP showed glial reactions, and we observed reactive microglia in the center of NFT and SP in contrast to astroglia, which were situated in the periphery. These findings suggest that the exposed argyrophilic components in the extracellular space stimulate both glial reactions, but that there is a striking difference in localization between microglia and astroglia.

Alzheimer Disease↗
PubMed1998

Effects of mild chronic heat exposure on the concentrations of thiobarbituric acid reactive substances, glutathione, and selenium, and glutathione peroxidase activity in the mouse liver.

To determine whether mild and chronic heat stress leads to oxidative stress and to differentiate such effects of different exposure periods, we kept male ICR-mice at an ambient temperature of either 35 degrees C or 25 degrees C for 6 hours, 3 days, or 7 days and measured the concentrations of thiobarbituric acid reactive substances (TBARS), glutathione (GSH), selenium (Se), and glutathione peroxidase (GSH-Px) activities in the liver. Since the food consumption of the heat-exposed group was only half that of the control, we prepared pair-fed groups, which were kept at 25 degrees C and whose food consumption were limited to those of the heat-exposed group for the 3-day and the 7-day exposure. TBARS concentrations of the liver was significantly higher in the heat group than the control after the 3-day exposure, while there was no significant difference among the groups after the 7-day exposure. There was no significant difference in GSH concentrations between the heat-exposed group and the control after the 7-day exposure, when the GSH concentration of the pair-fed group was significantly lower than that of the control. Hepatic cytosolic Se GSH-Px activity in the heat group was significantly less than that in the control group after the 6-hour exposure and it tended to be lower in the heat group than that of the control group after the 7-day exposure, while there was no difference in the total GSH-Px activity among the three groups. Our results showed that mild and chronic heat exposure may cause oxidative damage to organisms and that GSH-related anti-oxidative systems would play an important role to defensive reaction.

Animals↗
PubMed1998

Nonasthmatic case of Churg-Strauss syndrome with rapidly progressive glomerulonephritis.

A 61-year-old man developed mononeuritis multiplex accompanied by eosinophilia in 1993. Approximately 3 years later, acute renal dysfunction, a subendocardial tumor, and a high peripheral anti-neutrophil cytoplasmic antibody titer were also detected. Renal biopsy revealed glomerular crescents and interstitial infiltration of eosinophils, so allergic granulomatosis and angiitis was diagnosed. These clinical abnormalities regressed with steroid therapy. He had no history of asthma. This was therefore considered to be an atypical form of Churg-Strauss syndrome with rapidly progressive glomerulonephritis.

Anti-Inflammatory Agents↗
PubMed1998

High incidence of diabetic nephropathy in early-onset Japanese NIDDM patients. Risk analysis.

OBJECTIVE: Because early-onset Japanese NIDDM patients (diagnosed before age 30 years) can develop diabetic end-stage renal failure (ESRF) in their thirties, this study was performed to elucidate the incidence and determinants for the development of diabetic nephropathy. RESEARCH DESIGN AND METHODS: The incidence of diabetic nephropathy and its relationship to baseline characteristics and long-term metabolic control were determined in 426 early-onset Japanese NIDDM patients who were followed for a mean of 6.8 years. RESULTS: Of these 426 patients, 41 developed diabetic nephropathy manifested by persistent proteinuria (incidence rate [95%CI]/1,000 person-years; 14.1 [10.4-19.1]). Among patients whose mean HbA1c (measured by a high-performance chromatography method that is standardized and comparable to the one used in the Diabetes Control and Complications Trial study) was around 7% or less, few developed nephropathy. The incidence of nephropathy increased with increasing mean HbA1c level in a dose-dependent manner (chi 2 trend = 49.9, P < 0.0001). Diastolic blood pressure and duration of diabetes at entry had significant predictive effects independent of metabolic control. CONCLUSIONS: The incidence rate of diabetic nephropathy in early-onset Japanese NIDDM patients is potentially high, similar to or higher than that in Pima Indian NIDDM or Caucasian IDDM patients of comparable age. Diabetic nephropathy in NIDDM patients aged in their thirties or forties is likely to be an early feature that leads to ESRF, and this would contribute to the marked increase in the number of new patients with diabetic ESRF in Japan. NIDDM is a serious disease if near-normal glycemia is not achieved.

Adult↗
PubMed1997

Carbon tetrachloride does not induce micronucleus in either mouse bone marrow or peripheral blood.

We performed mouse bone marrow and peripheral blood micronucleus tests on carbon tetrachloride (CCl4). In the bone marrow assay, bone marrow cells were collected once after 24 h and twice, with a 24-h interval at a dose of 500, 1000 and 2000 mg/kg. In the peripheral blood assay, blood samples were collected 0, 24, 48 and 72 h after a single intraperitoneal injection at a dose of 1000, 2000 and 3000 mg/kg. As a result, micronucleated polychromatic erythrocytes (MNPCEs) were observed neither in the bone marrow assay nor the peripheral blood assay. We concluded that CCl4 does not induce chromosomal aberrations in the mouse bone marrow cells under these experimental conditions.

Animals↗
PubMed1997

Deficiency of selenium enhances the K+-induced release of dopamine in the striatum of mice.

To determine whether a selenium (Se) deficiency in the brain leads to a functional change in dopaminergic transmission in the striatum, in vivo microdialysis was conducted in mice fed a low-Se diet. After 11-13 weeks of the diet regimen, the activity of glutathione peroxidase (GPx) in the Se-deficient brain was reduced to 60% of the control brain. A high K+ perfusion (100 mM) increased the level of dopamine in the dialysate to 67 +/- 16 times the basal level; the increase was significantly greater than that observed in the control group (28 +/- 4 times). Such a between-group difference was not observed after 4-5 weeks of the Se-diet. These results indicated that prolonged Se deficiency altered the function of striatal dopaminergic neurons in mice. A possible contribution of enhanced oxidative stress due to the reduced GPx activity is discussed.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1997

Decrease of neurons in the medullary arcuate nucleus of multiple system atrophy: quantitative comparison with Parkinson's disease and amyotrophic lateral sclerosis.

The physiological functions of the medullary arcuate nucleus are supposed to be involved in autonomic cardioventilatory regulation, but neuropathological studies on neurodegenerative diseases have rarely reported about the arcuate nucleus. We quantitatively examined the neuronal density of the arcuate nucleus in patients with multiple system atrophy (MSA, n = 3), Parkinson's disease (PD, n = 3), amyotrophic lateral sclerosis (ALS, n = 2), and control subjects (n = 6), and statistically compared the findings in each group. Although the neuronal densities in PD and ALS patients were not different from that in the controls, MSA patients showed a marked depletion of neurons in the arcuate nucleus. The neuronal density (/mm2, mean +/- SEM) in the arcuate nucleus was 9.27 +/- 10.4 in MSA, and was significantly decreased (P < 0.05; Wilcoxon test), compared with that in control subjects (87.1 +/- 12.2). These results suggest that the lesioned arcuate nucleus is related to the pathogenesis of dysatonomia in MSA.

Aged↗
PubMed1997

Wiskott-Aldrich syndrome/X-linked thrombocytopenia: WASP gene mutations, protein expression, and phenotype.

Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT), caused by mutations of the WAS protein (WASP) gene, represent different phenotypes of the same disease. To demonstrate a phenotype/genotype correlation, we determined WASP gene mutations in 48 unrelated WAS families. Mutations included missense (20 families) and nonsense (eight) mutations located mostly in exons 1 to 4, and splice-site mutations (seven) and deletions and insertions (13) located preferentially in exons 7 to 11. Both genomic DNA and cDNA were sequenced and WASP expression was measured in cell lysates using peptide-specific rabbit anti-WASP antibodies. WASP was expressed in hematopoietic cell lines including bone marrow-derived CD34+ cells. Missense mutations located in exons 1 to 3 caused mild disease in all but one family and permitted WASP expression, although frequently at decreased concentration. Missense mutations affecting exon 4 were associated with classic WAS and, with one exception, barely detectable WASP. Nonsense mutations caused classic WAS and lack of protein. Insertions, deletions, and splice-site mutations resulted in classic WAS and absent, unstable, truncated, or multiply spliced protein. Using affinity precipitation, WASP was found to bind to Src SH3-containing proteins Fyn, Lck, PLC-gamma, and Grb2, and mutated WASP, if expressed, was able to bind to Fyn-glutathione S-transferase (GST) fusion protein. We conclude that missense mutations affecting the PH domain (exons 1 to 3) of WASP inhibit less important functions of the protein and result in a mild phenotype, and that missense mutations affecting exon 4 and complex mutations affecting the 3' portion of WASP interfere with crucial functions of the protein and cause classic WAS.

Amino Acid Sequence↗
PubMed1997

Missense mutations affecting a conserved cysteine pair in the TH domain of Btk.

Tec family protein tyrosine kinases have in their N-terminus two domains. The PH domain is followed by Tec homology (TH) domain, which consists of two motifs. The first pattern, Btk motif, is also present in some Ras GAP molecules. C-terminal half of the TH domain, a proline-rich region, has been shown to bind to SH3 domains. Mutations in Bruton's tyrosine kinase (Btk) belonging to the Tec family cause X-linked agammaglobulinemia (XLA) due to developmental arrest of B cells. Here we present the first missense mutations in the TH domain. The substitutions affect a conserved pair of cysteines, residues 154 and 155, involved in Zn2+ binding and thereby the mutations alter protein folding and stability.

Adult↗
PubMed1997

Tissue-specific modification of selenium concentration by acute and chronic dexamethasone administration in mice.

Several clinical reports have shown changes in plasma Se concentration with corticosteroid treatments, but the results have been inconsistent. Few experimental studies have been done on this subject. In the present study the effect of dexamethasone (DEX) treatment on Se concentrations and activities of Se-dependent glutathione peroxidase (EC 1.11.1.9; SeGPx) were examined in adult male ICR mice. In the first experiment, DEX was given via drinking water containing 5 or 50 mg DEX/l. At 1 or 3 weeks of DEX treatment, mice were dissected and the Se concentrations as well as SeGPx activities in various tissues, including plasma, were determined. At 1 week the DEX-treated groups had significantly lower hepatic Se concentrations and significantly higher plasma and cerebral concentrations than the control group. The DEX-treated groups showed lower SeGPx activities in the hepatic cytosol and higher SeGPx activities in the plasma than the saline (9 g NaCl/l)-treated group, in parallel with the changes in Se concentrations. At 3 weeks, neither hepatic nor plasma Se concentrations showed a significant change. In the second experiment, mice were injected subcutaneously with DEX and, thereafter, mice were food-deprived. The DEX-injected groups had higher plasma Se concentrations. A similar finding was obtained also when the DEX- or saline-injected mice were not food-deprived. Thus, the difference between the DEX-treated and control groups was possibly caused by redistribution of tissue Se. These results suggested that the effects of DEX on Se concentrations were tissue dependent and that the higher plasma Se observed in DEX-treated groups might be explained by the release of tissue Se into plasma as plasma SeGPx.

Analysis of Variance↗
PubMed1997

[Sjögren's syndrome with presenting as aseptic meningoencephaloradiculopathy in an elderly woman].

A 76-year-old woman was found to have acute aseptic meningoencephalitis with meningial irritation, disturbance of consciousness, elevation of cell counts in cerebrospinal fluid, and swelling of a right temporal-lobe lesion on a CT scan of the head. Muscle weakness in the lower extremities and urinary dysfunction developed and progressed gradually. The protein content of cerebrospinal fluid was high, and the distal latencies of F waves were prolonged, which suggested that the inflammation extended to the nerve roots. Sjögren's syndrome was diagnosed on the basis of atrophy of the labial salivary glands; invasions of lymphocytes and plasma cells to the intercellular space; and elevation of the titers of serum antinuclear antibody, anti-SS-A antibody, and anti-SS-B antibody. The patient had no xerosis. Aseptic meningoencephalitis was the first manifestation of Sjögren's syndrome. In recent years, several cases in which Sjögren's syndrome was associated with aseptic meningitis have been reported. However, we know of no previous report of such a case in a patient of this age. Aseptic meningoencephalitis can be the first manifestation of Sjögren's syndrome.

Acute Disease↗