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Biomedical subjects

M Iwata

Publications and source records attributed to M Iwata.

At least 199 records · Page 11Linked to original sources
PubMed1997

Regulation of thymocyte lineage commitment by the level of classical protein kinase C activity.

Thymocyte-positive selection involves signaling through TCR and accessory molecules, and the signaling intensity appears to be critical for this event. The specific inhibitor of classical Ca2+-dependent protein kinase C (cPKC), Gö 6976, inhibited positive selection in fetal thymus organ culture, indicating that cPKC activation is essential for positive selection. The major protein kinase C isoforms in CD4+ CD8+ thymocytes are cPKC-alpha, cPKC-beta, and the novel Ca2+-independent protein kinase C, nPKC-epsilon. To analyze the effect of cPKC activation level on positive selection, we used thymocytes from TCR transgenic mice with nonselecting and RAG-2 -/- backgrounds as they were developmentally arrested at the CD4+ CD8+ stage without positive selection signals. These thymocytes survived and acquired CD4/CD8 lineage commitment in suspension culture upon transient stimulation with limited concentrations of the selective activator of cPKC-alpha and -beta, thymeleatoxin, and the calcium ionophore, ionomycin. However, neither 12-deoxyphorbol 13-phenylacetate 20-acetate, which selectively activates cPKC-beta, nor ingenol 3,20-dibenzoate, which selectively activates nPKC-epsilon, exerted such an effect. The thymeleatoxin/ionomycin concentrations corresponded to those that inhibit glucocorticoid-induced apoptosis in thymocytes and were lower than those that induce proliferation of mature T cells. The CD4 lineage commitment required a higher level of cPKC activity than the CD8 lineage commitment. CD8alpha or CD4 mRNA expression was down-regulated. Functional helper and killer T cells were induced from the CD4 and CD8 lineage-committed cells, respectively, by additional stimulation. These results suggest that thymocyte lineage commitment in positive selection is regulated by the level of cPKC-alpha activity or by the levels of cPKC-alpha and -beta activities.

Animals↗
PubMed1997

El Tor hemolysin of Vibrio cholerae O1 forms channels in planar lipid bilayer membranes.

We investigated the channel formation by El Tor hemolysin (molecular mass, 65 kDa) of Vibrio cholerae O1 biotype El Tor in planar lipid bilayers. The El Tor hemolysin channel exhibited asymmetric and hyperbolic membrane current with increasing membrane potential, meaning that the channel is voltage dependent. The zero-current membrane potential measured in KCI solution showed that permeability ratio PK+/PCl- was 0.16, indicating that the channel is 6-fold more anion selective over cation. The hemolysin channel frequently flickered in the presence of divalent cations, suggesting that the channel spontaneously opens and closes. These data imply that the El Tor hemolysin damages target cells by the formation of transmembrane channels and, consequently, is the cause of osmotic cytolysis.

Bacterial Toxins↗
PubMed1997

Pattern reversal visual evoked potentials in classic and common migraine.

Pattern reversal visual evoked potentials (PVEPs) to transient checkerboard were recorded in 19 patients with migraine with visual aura (i.e., classic migraine), 14 patients with migraine without aura (i.e., common migraine) in the interictal period and 43 normal subjects. Latencies and amplitudes of PVEPs in each group were analyzed. In classic migraine patients, P100 amplitude was significantly higher than in normal subjects (p < 0.01), whereas latencies of PVEPs did not significantly differ. There were no significant differences between the common migraine and normal subjects, nor within the classic and common migraine groups in latencies and amplitudes of PVEP. Four patients with classic migraine underwent PVEPs during or 1-2 h immediately after their migraine attacks. Two of these patients who underwent PVEPs 1.5-2 h after their attacks showed abnormally increased PVEP amplitudes. These results suggest that there are different pathophysiologies in the visual pathway between classic and common migraine and furthermore, classic migraine patients in interictal periods may have hyperexcitability in the visual pathway and that the increased amplitude of PVEPs after attacks may be due to cortical spreading depression.

Adolescent↗
PubMed1997

Development of a time-resolved fluoroimmunoassay (TR-FIA) for testosterone: measurement of serum testosterone concentrations after testosterone treatment in the rainbow trout (Oncorhynchus mykiss).

A sensitive time-resolved fluoroimmunoassay (TR-FIA) for testosterone was developed, and the assay system was used for measuring serum testosterone concentrations in rainbow trout. Testosterone-3-(O-carboxymethyl)oxime-bovine serum albumin (T-3-CMO-BSA) was immobilized by physical adsorption to the wells of microtiter plates. A competitive assay using two antibodies was performed among T-3-CMO-BSA in the solid-phase, unknown amounts of testosterone, testosterone antibodies, and europium labeled secondary antibodies, followed by measurements using a time-resolved fluorometer (DELFIA system). The TR-FIA had a sensitivity of 0.075 pg/50 microliters sample (1.5 pg/ml), and the range of the assay system was between 1.5 pg/ml and 25 ng/ml. The intra- and interassay coefficients of variation for the testosterone TR-FIA were satisfactorily low, and were between 1.62 and 6.38% and 2.96 and 8.29%, respectively. The assay system was applied to measure the serum testosterone concentrations after an injection of testosterone dissolved in saline, propyleneglycol, or coconut oil. Among the three solvents, the coconut oil group showed continuously high serum testosterone level. In contrast, the saline and propyleneglycol groups had maximum concentrations 24 hr after the injection, but their levels were significantly lower than that of the coconut oil group. The testosterone TR-FIA method is sensitive, repeatable, and is as accurate as conventional RIAs. It is very good for measuring serum testosterone concentrations.

Animals↗
PubMed1997

Hypogenesis of right hepatic lobe accompanied by portal hypertension: case report and review of 31 Japanese cases.

We report a rare case of hypogenesis of the right lobe of the liver with portal hypertension and a review of 31 cases of agenesis or hypogenesis of the right hepatic lobe reported in Japan. A 74-year-old man consulted our hospital for further examination after a mass screening for gastric cancer. On physical examination liver enlargement was palpable, but liver function tests were normal. Abdominal ultrasonography, computed tomography, technetium-99m liver scintigraphy, and endoscopic retrograde cholangiopancreatography revealed a small right hepatic lobe and moderate splenomegaly, in contrast to a hypertrophic lateral segment of the left hepatic lobe, as well as ectopic dislocation of the gallbladder. Endoscopic examination revealed esophageal varices, indicating portal hypertension. Abdominal angiography demonstrated mild shunt flow between the hepatic artery feeding from the gastroduodenal artery and the portal vein. A biopsy specimen taken from both lobes of the liver showed normal liver tissue histologically. Based on these findings, we made a definite diagnosis of hypogenesis of the right lobe of the liver with portal hypertension. The present case appears to be the first such case accompanied by portal hypertension reported in Japan.

Aged↗
PubMed1997

Immunocytochemical and ultrastructural study of pericapillary rosettes in amyotrophic lateral sclerosis.

This report concerns a comparative immunocytochemical and ultrastructural investigation on pericapillary rosettes (PR) in the lumbar spinal cords of 21 patients with amyotrophic lateral sclerosis (ALS) and 18 age-matched neurologically normal individuals. The purpose of the study was to determine the alteration of PR in relation to the neuronal loss in ALS. The PR were almost always positively immunostained for phosphorylated neurofilament, and some PR immunoreacted with antibodies to synaptophysin and beta-amyloid precursor protein. This finding suggests that axonal transport, whether fast or slow, is impaired in the terminal portion of the axon that reaches the capillaries. Some PR were also positively immunostained by the antibody against ubiquitin, anti-calbindin-D 28 K antibody, anti-parvalbumin antibody and the antibody to superoxide dismutase 1. Morphometrically, the number of PR in the anterior horns and lateral column was markedly diminished in ALS compared with controls. At the ultrastructural level, the PR consisted mostly of unmyelinated degenerated axons, and were frequently found outside the basal laminae of the endothelial cell and of the astrocytic foot processes on the opposite side of the capillary, and less often in the space between the two basal laminae. The data indicate that the fate of PR is intimately associated with the neuronal loss of the anterior horn cells and with degenerative change of nerve fibers extending from their mother neurons to the capillaries.

Adult↗
PubMed1997

A de novo recombination in the ABO blood group gene and evidence for the occurence of recombination products.

We have encountered a paternity case where exclusion of the putative father was only observed in the ABO blood group (mother, B; child, A1; putative father, O), among the many polymorphic markers tested, including DNA fingerprints and microsatellite markers. Cloning a part of the ABO gene, PCR-amplified from the trio's genomes, followed by sequencing the cloned fragments, showed that one allele of the child had a hybrid nature, comprising exon 6 of the B allele and exon 7 of the O1 allele. Based on the evidence that exon 7 is crucial for the sugar-nucleotide specificity of A1 and B transferases and that the O1 allele is only specified by the 261G deletion in exon 6 of the consensus sequence of the A1 allele, we concluded that the hybrid allele encodes a transferase with A1 specificity, resulting, presumably, from de novo recombination between the B and O1 alleles of the mother during meiosis. Screening of random populations demonstrated the occurrence of four other hybrid alleles. Sequencing of intron VI from the five hybrid alleles showed that the junctions of the hybrid alleles were located within intron VI, the intron VI-exon 7 boundaries, or exon 7. Recombinational events seem to be partly involved in the genesis of sequence diversities of the ABO gene.

ABO Blood-Group System↗
PubMed1997

Serum vitamin K level and bone mineral density in post-menopausal women.

OBJECTIVE: Vitamin K is known to influence bone metabolism by facilitating the synthesis of osteocalcin (BGP). The bone mineral density decreases drastically after menopause. We investigated the relationship of bone mineral density, vitamin K levels and other biological parameters of bone metabolism in post-menopausal women. METHODS: Serum levels of vitamin K, BGP and other markers of bone metabolism were measured in 71 post-menopausal women (19 with reduced bone mineral density and 52 with normal bone density), and 24 women with climacteric symptoms receiving hormone replacement therapy (HRT), (6 with reduced bone mineral density and 18 with normal density). RESULTS: In the first group, women with reduced bone mineral density showed lower levels of vitamin K1 and K2 than those with normal bone mineral density. In the other group, the level of BGP decreased but levels of vitamin K showed no increase during HRT. CONCLUSION: The present findings suggested that vitamin K was related to post-menopausal bone mineral loss.

Bone Density↗
PubMed1997

cDNA for ribosomal protein S2 in sockeye salmon, Oncorhynchus nerka.

We isolated a cDNA encoding ribosomal protein S2 in sockeye salmon, Oncorhynchus nerka, using a reverse transcriptase-polymerase chain reaction (RT-PCR) method. The cDNA encoding ribosomal protein S2 is composed of 933 nucleotides, and has a 5'-noncoding sequence of 9 bases, a 885 base open reading frame coding for a 294 amino acid polypeptide, and a 39 base 3'noncoding sequence. The amino acid sequence of sockeye salmon S2 protein deduced from the nucleotide sequence is highly homologous to those from the rat (86.1%) and Drosophila melanogaster (73.6%). The N-terminal region of S2 protein is rich in arginine-glycine sites, including eight tandem repeats, and has two consecutive copies of the RGGF motif. The sequences are considered to be requisites for nucleolar localization and binding to RNA for nucleolar proteins. Southern blot analysis indicates that there may be only a single copy of the S2 gene, which is a multiple copy gene in the rat and the fruit fly. Northern blot analysis shows that the S2 gene is expressed in the brain, pituitary, heart, liver kidney, muscle, testis and ovary of sockeye salmon.

Amino Acid Sequence↗
PubMed1997

Epigallocatechin gallate-induced histamine release in patients with green tea-induced asthma.

BACKGROUND AND OBJECTIVE: Epigallocatechin gallate is the causative agent of green tea-induced asthma. To determine whether an IgE-mediated mechanism plays a pathogenetic role in this disorder, we measured histamine release after in vitro exposure to epigallocatechin gallate. METHODS: Subjects included eight patients (four men and four women) with green tea-induced asthma, who had been diagnosed by skin test and inhalation challenge, and eight controls (four asthmatic subjects with no previous exposure tea dust and four healthy volunteers). Heparinized whole blood samples were taken and incubated with epigallocatechin gallate at various concentrations (final concentration range, 0.003 to 300 micrograms/mL) for 30 minutes at 37 degrees C. After centrifugation, histamine was measured in the cell-free supernatants by radioimmunoassay. Histamine release was expressed as a percentage of total histamine. A result higher than 10% was considered positive. RESULTS: In one of the tea-sensitive patients, epigallocatechin gallate did not cause histamine release. Five of the other seven patients (71%) demonstrated a positive, dose-dependent histamine release to epigallocatechin gallate. In asthmatic and normal controls, histamine release was not observed at any epigallocatechin gallate concentration. Furthermore, a significant correlation was noted between the maximum percentage histamine release and the threshold epigallocatechin gallate concentration for intradermal skin testing. CONCLUSION: These results indicate that an IgE-mediated response is the basis for green tea-induced asthma.

Adult↗
PubMed1997

Altered ceramide and sphingosine expression during the induction phase of ischemic acute renal failure.

UNLABELLED: Recent evidence indicates that a "sphingomyelin signaling pathway" exists: in response to heterogeneous influences, sphingomyelin is hydrolyzed, liberating ceramide, and subsequently its sphingoid base, sphingosine. Ceramide and sphingosine can influence diverse cellular processes, including cell differentiation, proliferation, protein trafficking, and apoptosis. Each of these processes have important implications for post-ischemic acute renal failure (ARF). However, sphingosine and ceramide expression during the induction of ischemic/reperfusion injury have not been previously assessed. To this end, CD-1 mice were subjected to 45 minutes of unilateral renal ischemia +/- reperfusion, followed by cortical sphingosine, ceramide, and sphingomyelin assessments. Contralateral kidneys served as controls. Ischemia caused approximately 50% sphingosine and ceramide decrements. During reperfusion, sphingosine rebounded to normal values. Conversely, ceramide rose to, and was maintained at, supranormal levels (approximately 175% of controls). Subsequent studies performed with hypoxic or oxygenated isolated proximal tubules suggested that these changes: (1) had a multifactorial basis; (2) were partially simulated by enhanced PLA2 activity; (3) and were dissociated from alterations in net sphingomyelin content. To assess the potential pathogenic relevance of the documented ceramide increments, cultured human proximal tubule (HK-2) cells were subjected to ATP depletion/Ca2+ ionophore- or PLA2-induced attack with or without exogenous C2 ceramide loading. Ceramide worsened both forms of injury without exerting an independent lethal effect. Conversely, ceramide markedly attenuated arachidonic acid cytotoxicity. This occurred without any decrease in arachidonate uptake, suggesting a direct cytoprotective effect. IN CONCLUSION: (1) sphingosine and ceramide fluxes are hallmarks of early ischemic/reperfusion injury; (2) these changes occur via divergent metabolic pathways; and (3) that ceramide increments can affect divergent injury pathways, and that sphingosine and ceramide have potent cell signaling effects, suggest that the currently documented sphingosine/ ceramide fluxes could have important implications for the induction phase and evolution of post-ischemic ARF.

Acute Kidney Injury↗
PubMed1997

Simultaneous recording of pattern reversal electroretinograms and visual evoked potentials in migraine.

We recorded full-field pattern reversal electroretinograms (PERGs) and visual evoked potentials (PVEPs) simultaneously in 15 migraine with aura, 14 migraine without aura patients during the interictal period, and in 23 sex- and age-matched normal subjects. All subjects had normal visual fields. The visual aura in all patients was hemianopsia or fortification spectra. Neither migraine group showed significant differences from normal in latency and amplitude of PERGs. In migraine with aura, the amplitudes of PVEPs in classic migraine at the mid-occipital electrode were significantly (p < 0.01) higher than normal. PVEP amplitudes were significantly (p < 0.01) higher on the contralateral side of the aura than the ipsilateral side in both visual aura and normal subjects, but there was no significant difference in latency. This high amplitude and asymmetry of PVEPs may contribute to defective inhibition between interhemispheric visual occipital areas or striate and peristriate areas.

Adult↗
PubMed1997

Alterations of platelet, coagulation, and fibrinolysis markers in patients with acute ischemic stroke.

Alterations of platelet, coagulation, and fibrinolysis markers were investigated to determine the indications for antithrombotic therapy in patients with different clinical categories of acute cerebral infarction. Marked platelet activation was observed in platelet function tests, including measurements of platelet-specific proteins and platelet survival, platelet scintigraphy in the brain, and platelet fibrinogen binding assay, in patients with atherothrombotic stroke. Among patients with atherothrombotic stroke, increases of thrombin-antithrombin III complex (TAT) and D-dimer were frequent in addition to the findings of platelet activation in patients showing progressing stroke. Patients with cardioembolic stroke demonstrated marked elevation of coagulation markers, TAT and fibrinopeptide A, and fibrinolysis markers, D-dimer and plasmin-alpha2-plasmin inhibitor complex, as well as platelet activation. In contrast, neither activation was seen in patients with lacunar stroke. On the basis of these findings, antiplatelet therapy is indicated for stable or improving stroke, whereas anticoagulant therapy is indicated for progressing stroke among patients with atherothrombotic stroke. In patients with cardioembolic stroke, anticoagulant therapy should be started as soon as possible, or should be followed by thrombolytic therapy in the hyperacute phase. However, further investigation appears necessary for recommendations for patients with lacunar stroke.

Acute Disease↗
PubMed1997

Mechanisms of lymphocyte adhesion to cultured human corneal epithelial cells.

PURPOSE: The authors studied the adhesion mechanisms between peripheral blood lymphocytes (PBL) and cultured human corneal epithelial (HCE) cells to investigate the lymphocyte interaction with corneal epithelial cells in the corneal immune response. METHODS: First, the authors examined the expression of intercellular adhesion molecule (ICAM)-1 or lymphocyte function-associated antigen (LFA)-3 on the normal human corneal epithelium and cultured HCE cells by an immunostaining technique and flow cytometry. Effects of inflammatory cytokines such as interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha, on ICAM-1 or LFA-3 expression on cultured HCE cells were also examined. Second, the authors performed an adhesion assay with 51Cr-labeled monocyte-depleted PBL from normal, healthy volunteers and cultured HCE cells, with and without treatment of IFN-gamma or TNF-alpha in 96-well-plates for 1 hour at 37 degrees C in 5% CO2. After unbound PBL were removed, the radioactivity of the sample in each well was counted with a scintillation counter. In addition, the authors evaluated the blocking effects of monoclonal antibodies (mAbs) on the adhesion of PBL to the cultured HCE cells. RESULTS: ICAM-1 expression was not detected in the normal human corneal epithelium. However, the expression of ICAM-1 was detected on the cultured HCE cells with Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum. In addition, both IFN-gamma and TNF-alpha increased ICAM-1 expression on the cultured HCE cells dramatically. LFA-3 expression was detected in all cell layers of the normal human corneal epithelia. Neither IFN-gamma nor TNF-alpha had any effect on LFA-3 expression on the cultured HCE cells. The PBL adhesion to the HCE cells with and without treatment of IFN-gamma or TNF-alpha was blocked dominantly by anti-ICAM-1 or anti-LFA-1 alpha mAb. Anti-LFA-3 mAb also blocked the PBL adhesion but had less blocking effect than anti-ICAM-1 or anti-LFA-1 alpha mAb. Anti-very late activation antigen beta, or anti-human leukocyte antigen (HLA)-class I or HLA-class II mAb had no effect on the PBL adhesion to the HCE cells. The adhesion percentile of the PBL applied to the HCE cells pretreated with IFN-gamma or TNF-alpha showed a dose-response curve dependent on the concentration of these cytokines. CONCLUSIONS: The results in the present study demonstrate that (i) adhesion of lymphocytes to HCE cells could be mediated by the LFA-1-ICAM-1 pathway and/or the CD2-LFA-3 pathway; (ii) the LFA-1-ICAM-1 pathway could be crucial in lymphocyte adhesion to HCE cells; (iii) IFN-gamma or TNF-alpha exerts an enhancing effect not only on the ICAM-1 expression on HCE cells but also on the adhesion of lymphocytes to HCE cells.

Antibodies, Monoclonal↗
PubMed1997

Effect of 6-fluoro-8-methoxy quinolone (AM-1155) against chronic airway infection with Pseudomonas aeruginosa in a rat model.

We studied the effect of AM-1155, a newly developed quinolone, against chronic airway infection with Pseudomonas aeruginosa using a previously described rat model. AM-1155 (25 mg/kg) or ciprofloxacin (25 mg/kg) or saline (controls) were injected s.c. for 14 days (from day 4 to day 17) after the inoculation of agar beads containing P. aeruginosa. The number of viable cells of intrapulmonary P. aeruginosa, histological findings of the lungs and immunoglobulin levels of serum and bronchoalveolar lavage fluid were examined in rats 11 and 18 days after the treatment. The findings indicated that the number of viable cells of P. aeruginosa in lungs was significantly decreased in the AM-1155- or ciprofloxacin-treated group compared with the non-treated control group. Histological examination in the non-treated control group showed hyperplasia of bronchus-associated lymphoid tissue as well as cellular infiltration in airways, but not prominently in the AM-1155- or ciprofloxacin-treated group. The IgG and IgA levels in serum and bronchoalveolar lavage fluid were significantly lower in the AM-1155- and ciprofloxacin-treated groups than in the control group. These in-vivo effects of AM-1155 were comparable to those of ciprofloxacin. These findings suggest that treatment with AM-1155 and ciprofloxacin suppressed excessive immune responses, preventing progression of airway damage in the chronic infectious state.

Animals↗
PubMed1997

Isolated collagen IV retains the potential to form an 18-nm sided polygonal meshwork of the lamina densa.

Polygonal meshworks were revealed in the aggregates reconstituted from isolated collagen IV as well as in the lamina densa of mouse pancreas by the quick-freezing and deep-etching technique. Collagen IV solution with acetic acid from the bovine lens capsules and that isolated from the pepsin-solubilized fraction of human placenta reconstituted polygonal meshworks, which were dimensionally similar to the meshwork seen in the lamina densa of mouse pancreas. The average lengths between branching points in the lamina densa, and the meshworks of collagen IV derived from the bovine lens capsules and from the human placenta were 20.1 +/- 7.7 nm (mode = 18 nm), 21.6 +/- 11.3 nm (mode = 14 nm), and 21.6 +/- 9.0 nm (mode = 18 nm), respectively. These observations suggest that collagen IV can, in the absence of other macromolecules including laminins, comprise a skeletal meshwork similar in dimensions to that of the lamina densa.

Animals↗