Biomedical subjects

K Shirato

Publications and source records attributed to K Shirato.

At least 109 records · Page 6Linked to original sources

PubMed1998

Arg506Gln mutation of the coagulation factor V gene not detected in Japanese pulmonary thromboembolism.

The incidence of pulmonary thromboembolism (PTE) is lower in Japanese than in Caucasians. The basis for the different incidence has not been clarified. A poor anticoagulant response to activated protein C based on a single point mutation of the factor V gene (Arg506Gln) was found to be a pathogenetic factor for venous thrombosis and PTE in North America and Europe. We investigated whether the Arg506Gln mutation of factor V is responsible for the occurrence of PTE among Japanese. We analyzed genomic DNA prepared from fresh peripheral blood of 25 patients with PTE of unknown etiology (12 of acute type and 13 of chronic type) and that of 110 controls without respiratory or circulatory disorders. To detect the Arg506Gln mutation, 267 bp DNA fragments of the factor V gene including the Arg506Gln region were amplified by PCR, digested by MnlI and electrophoresed. After digestion of PCR products with MnlI, DNA fragments of 163 bp length, but not DNA fragments of 200 bp length, were identified in all samples, indicating the absence of the Arg506Gln mutation in the patients with PTE and control subjects. These results suggest that the Arg506Gln mutation is absent or very rare and not an important pathogenetic factor for PTE in Japanese.

Aged↗

PubMed1998

Use of balloon-occluded retrograde transvenous obliteration with ethanolamine oleate for the treatment of hepatic encephalopathy in a cirrhotic patient with a large spontaneous splenorenal shunt.

We describe a technique for the treatment of hepatic encephalopathy in a cirrhotic patient with a large spontaneous splenorenal shunt. This large shunt was successfully occluded without severe complications by retrograde transrenal venous obliteration with ethanolamine oleate after balloon occlusion. This procedure may be an effective method to occlude a large spontaneous splenorenal shunt for the treatment of hepatic encephalopathy.

Aged↗

PubMed1998

Laminin inhibits Cl- -secretion across canine tracheal epithelium.

We examined the effect of laminin (a major component of the basement membrane) on potential difference (PD) and short circuit current (SCC) in both posterior epithelial membranes (native tissue) and cultured epithelial cell layers from canine trachea using an Ussing chamber. Although laminin itself did not alter the baseline values of SCC or PD, it significantly inhibited in a dose-dependent manner the isoproterenol induced SCC and PD rises of the epithelial membrane. In the cultured epithelial layer treated with amiloride, laminin did not alter the baseline values of SCC, PD or resistance (R) but it significantly inhibited the isoproterenol induced SCC rises. However, laminin did not significantly inhibit the bradykinin induced SCC rise in the cultured epithelial layer treated with amiloride. These findings indicate that laminin is a determinant in ion transport (mainly Cl- secretion) across canine tracheal epithelium, inhibiting the beta-agonist induced Cl- transport.

Adrenergic beta-Agonists↗

PubMed1998

Dexamethasone suppresses gene expression and production of IL-13 by human mast cell line and lung mast cells.

BACKGROUND: IL-13 has been shown to induce IgE production in B cells by promoting class switching to IgE. Mast cells are known to play an important role in the pathogenesis of allergic diseases. We evaluated the ability of human mast cells to produce IL-13 using human mast cell line HMC-1 and freshly isolated lung mast cells and then examined the effect of dexamethasone on the gene expression and production of IL-13 by these cells. METHODS: HMC-1 cells and lung mast cells were cultured with 10 ng/ml phorbol 12-myristate 13-acetate (PMA) and 1 micromol/L ionomycin and with 5 microg/ml phytohemagglutinin (PHA) and 10 ng/ml PMA, respectively, in the presence of dexamethasone. The gene expression of IL-13 at 3 hours (HMC-1 cells) or 12 hours (human lung mast cells) after stimulation was assessed semiquantitatively by sequential reverse transcription-polymerase chain reaction and Southern blot analysis. IL-13 production at 12 hours after stimulation was assayed by ELISA. RESULTS: The gene expression of IL-13 by HMC-1 cells and human lung mast cells, which was detected at a low level in an unstimulated condition, was increased by PMA/ionomycin and suppressed by dexamethasone. The supernatant of HMC-1 cells and human lung mast cells showed a low level of IL-13, which was increased by the stimulation and suppressed by dexamethasone. CONCLUSION: These findings indicate that HMC-1 cells and human lung mast cells produce IL-13 and that dexamethasone suppresses the production of IL-13 by these cells through an inhibitory action on the gene expression.

Cell Line↗

PubMed1998

Effects of L-NAME and L-arginine on diaphragm contraction in a septic animal model.

The effects of nitric oxide on diaphragm contraction after endotoxin administration were studied in Wistar rats. The animals were divided into seven treatments: a saline-injected group as control, three groups injected with L-NAME (0.01, 0.1, 1 mg/kg) and three groups injected with L-arginine (1, 10, 100 mg/kg). Escherichia coli endotoxin was injected into the peritoneal cavity 15 min later. Twitch kinetics and force-frequency curves were measured 0, 2, and 4 hr after endotoxin injection. In the control group, the force-frequency curves significantly decreased from 0 hr to 4 hr. In the L-NAME group, the force-frequency curves at 4 hr showed significant increases in a dose-dependent manner. In the L-arginine group, the force-frequency curve with 100 mg/kg at 4 hr showed a significant increase. There was no consistent change in the contraction time, half relaxation time, or fatiguability. NADPH diaphorase histochemistry performed on diaphragm muscle samples 4 hr after endotoxin injection showed positive in the control and L-arginine group, but was only weakly observed in L-NAME group. These data suggest that nitric oxide contributes to the endotoxin induced diaphragm contractile deterioration.

Animals↗

PubMed1998

Non-invasive estimation of human left ventricular end-diastolic pressure.

Sato et al. (Electronic Letters 32, 949-950, 1996) reported that one can obtain a non-invasive estimate of left ventricular (LV) pressure at around end-diastole in an isolated canine preparation. In this study we examined whether this method can be applied to humans. Using the method proposed by Kanai et al. (IEEE. Trans. UFFC, 43, 791-810,1996), we detected small amplitude LV vibration from an ultrasonic pulse Doppler signal reflected from the interventricular septum in five patients (44-63 y.o., male;4, female;1). We measured the oscillation frequency of the LV wall through the wavelet transform of small amplitude LV vibration, and calculated LV pressure at around end-diastole from the values of oscillation frequency, internal radius and wall thickness using Mirsky's equation. The estimated LV pressures at around end-diastole were similar to end-diastolic pressure measured directly by cardiac catheterization. These results show the possibility that this method allows for the non-invasive estimate of LV pressure at around end-diastole, and furthermore provides the basis for future clinical applicability of this technique.

Adult↗

PubMed1998

Heterogeneous fatty acid uptake early after reperfusion in rat hearts.

We determined whether spatial distributions of substrate uptake are heterogeneous within the area at risk during reperfusion. Quantitative autoradiography with imaging plates and two long-lived radioisotopes was applied to 15 open-chest, anesthetized rats subjected to 30 min of coronary artery ligation and 30 min of reperfusion. Regions showing increased beta-methyl-[1-14C]heptadecanoic acid ([14C]BMHDA) uptake (166 +/- 17% of that in the nonischemic area) appeared at the lateral borders and subepicardial layer within the area at risk, and 2-deoxy-D-[1-3H]glucose ([3H]DG) uptake was 103 +/- 24% in these regions. Regions with decreased [14C]BMHDA uptake (28 +/- 11%) occupied the midmyocardial layer except at the lateral borders within the area at risk, and [3H]DG uptake was 62 +/- 18% in these regions. The percentage interregional coefficients of variation (index of heterogeneity) in [14C]BMHDA uptake, [3H]DG uptake, and blood flow were higher in the area at risk than in the nonischemic area (76 +/- 23 vs. 21 +/- 7%, 39 +/- 10 vs. 21 +/- 7%, and 49 +/- 19 vs. 14 +/- 4%, respectively). Heterogeneous distributions of substrate uptake may explain the conflicting results concerning substrate metabolism during reperfusion.

Animals↗

PubMed1998

Pulmonary function and regional distribution of emphysema as determined by high-resolution computed tomography.

In patients with pulmonary emphysema, emphysematous changes are not uniform and vary from minimum alveolar destruction to advanced bullous formation, depending on the lobe or site in the lungs. However, we have little knowledge on whether or how this nonuniformity or localization affects pulmonary function in PE patients. Therefore, we measured the computed tomography (CT) density of divided sites in lungs with high-resolution CT images from 25 PE patients (FEV1.0%, mean +/- SD 36 +/- 9%, %DLCO 48 +/- 16%, all men, 68 +/- 4 years) and compared them to various parameters of pulmonary function. The mean CT density of whole lungs correlated with 12 pulmonary function parameters including FEV1.0 and diffusion capacity. When both lung fields were divided into peripheral, intermediate and central portions, the CT density of the central portion correlated with all pulmonary function parameters with which CT density of whole lungs correlated. In contrast, the CT density of the peripheral portion significantly correlated with only 7 parameters with smaller correlation coefficient values than those of the central portion. When divided into upper, middle and lower portions, the CT densities of upper, middle and lower portions correlated with 6, 8 and 10 of the 12 pulmonary function parameters which correlated with the density of whole lungs, respectively. The delta value of CT densities between the upper and lower portions or between the lateral and medial portions correlated with obstructive impairment (FEV1.0 and FEV1.0%). These findings suggest that (1) central rather than peripheral emphysematous changes affect pulmonary function, and (2) uniformity of emphysematous change correlates with the severity of airway obstruction in PE patients.

Adult↗

PubMed1998

Methacholine bronchial hyperresponsiveness in chronic sinusitis.

The coexistence of chronic sinusitis (CS) may deteriorate the clinical condition of lower airway diseases such as bronchial asthma (BA) or chronic bronchitis (CB). However, the bronchial hyperresponsiveness (BH) in CS without any apparent lower airway disease is not fully understood nor are the effects of treatment. We examined lower airway hyperresponsiveness to methacholine (MCh) in 42 subjects with CS but without allergic rhinitis (AR) who had normal lung functions without any pulmonary symptoms, comparing it with that of 50 subjects with stable BA, 50 subjects with simple CB and 40 subjects with AR, and further examined the effect of endoscopic sinus surgery in 7 CS subjects with BH. The BH to MCh was measured in terms of the minimum dose (Dmin), defined as the cumulative dose at the point where respiratory conductance began to decrease. A Dmin <50 units was defined as BH. Seventy-one percent of CS subjects showed BH without relation to the severity or duration of CS, or atopic status. BH in CS subjects, which was less than that in BA subjects, was similar to that in simple CB or AR in both its prevalence and degree. After the surgical treatment of CS, BH significantly decreased (p < 0.01) with improvements in both nasal symptoms and sinus lesions. These findings suggest that CS itself induces BH to a degree similar to simple CB and AR without any relationship to the clinical background, and that adequate treatment of CS reduces BH.

Adolescent↗

PubMed1998

Tazanolast inhibits ozone-induced airway hyperresponsiveness in guinea pigs.

We studied the inhibitory effect of tazanolast, a selective mast-cell-stabilizing drug, on ozone-induced airway hyperresponsiveness in guinea pigs. Airway responsiveness to methacholine peaked at 2 h after ozone exposure (2.0 ppm for 2 h) and the number of neutrophils in bronchoalveolar lavage (BAL) fluid continued to increase until 6 h. Tazanolast administered before ozone exposure at doses of 30, 100, or 300 mg/kg inhibited ozone-induced airway hyperresponsiveness in a dose-dependent manner. However, tazanolast administered after ozone exposure did not inhibit the airway hyperresponsiveness. Tazanolast did not significantly change the cell distribution of BAL cells at 2 h after the exposure. We conclude that tazanolast significantly inhibits ozone-induced airway hyperresponsiveness in guinea pigs. This result suggests that mast cells may play an important role in its development.

Airway Resistance↗

PubMed1998

Surfactant protein A2 gene expression by human airway submucosal gland cells.

To determine whether human airway submucosal glands produce and secrete surfactant proteins, we examined their protein and gene expression in submucosal glands from trachea and bronchi obtained from operated and autopsied lungs within 4 h of death. Using a monoclonal antibody (PE-10) against surfactant protein A (SP-A), a positive immunoperoxidase stain was observed over serous cells of submucosal glands in histologic sections of airway walls. Measurement of SP-A in culture medium samples using single-step enzyme-linked immunosorbent assay showed a significant secretion of SP-A by isolated submucosal glands (1.2 +/- 0.08 ng/ml/h, SEM, n = 40). In gene expression experiments by reverse transciption-polymerase chain reaction, the SP-A complementary DNA (cDNA) segment was amplified from isolated submucosal glands, indicating the presence of SP-A messenger RNA (mRNA) in airway submucosal glands. Bronchial superficial epithelial cells failed to show the presence of SP-A mRNA. No cDNA segment of SP-B, SP-C, or SP-D cDNA was amplified from isolated submucosal glands or superficial epithelial cells, whereas all were amplified from alveolar tissue. Furthermore, in contrast to the control alveolar tissue, which expressed both SP-A1 and SP-A2 genes, SP-A2 gene transcript alone was detected in isolated submucosal glands by Southern analysis that included the digestion of the amplified SP-A cDNA fragment with the restriction enzyme Apa I. These findings indicate that human airway submucosal gland cells can transcribe the SP-A2 gene and produce SP-A protein in a manner different from peripheral airways and alveoli, playing a role in the airway defense mechanism.

Adult↗

PubMed1998

Effects of histamine and endothelin-1 on membrane potentials and ion currents in bovine tracheal smooth-muscle cells.

We tested the effects of tetraethylammonium (TEA), acetylcholine (ACh), histamine, and endothelin-1 on single airway smooth-muscle cells from bovine trachea, using the patch-clamp technique. Resting membrane potential was -48 +/- 1 mV (n = 47). Both TEA and ACh significantly depolarized the membrane, by +28 +/- 4 mV (P < 0.001, n = 12) and +21 +/- 2 mV (P < 0.01, n = 7), respectively, in the whole-cell configuration. In contrast, both histamine and endothelin-1 hyperpolarized the membrane, by -21 +/- 6 mV (P < 0.01, n = 8) and -15 +/- 2 mV (P < 0.01, n = 8), respectively. Calcium-dependent large-conductance K+-channels (127 pS) and small-conductance K+ channels (21 pS) were identified in excised patches. The small-conductance K+ channel was inhibited by 4-aminopyridine and activated by both histamine and endothelin-1. Furthermore, TEA did not alter the membrane hyperpolarization by these agonists, suggesting that the small-conductance K+ channel or delayed-rectifier K+ channel was involved in the membrane hyperpolarization. Membrane hyperpolarization by histamine and endothelin-1 suggests that activation of voltage-dependent calcium channels (VDCCs) or of calcium influx does not contribute substantially to the contractile response of airway smooth-muscle contraction to these agonists.

Acetylcholine↗

PubMed1998

Relationship between cholinergic airway tone and serum immunoglobulin E in human subjects.

It has recently been shown that immunoglobulin (Ig)E facilitates the cholinergic bronchoconstrictor pathway in human tissue in vitro. However, whether this occurs in humans in vivo has not been clarified. In this study, the bronchodilator responses were examined to inhalation of a submaximal dose of the anticholinergic agent oxitropium bromide (600 microg) in normal and allergic subjects with various levels of total serum IgE. Values of the forced expiratory volume in one second (FEV1) for all subjects were greater than 80% of predicted, but were negatively correlated with serum IgE levels (p<0.01). Oxitropium bromide inhalation induced an increase in FEV1 that was significantly greater in allergic rhinitis patients with high serum IgE (155+/-20 mL (mean+/-SEM), p<0.05) than in healthy subjects (64+/-21 mL) or those with allergic rhinitis but low serum IgE (82+/-21 mL, p<0.05). In contrast, the effects of the inhaled beta2-adrenergic agent orciprenaline sulphate (2.25 mg) were not significantly different among the three groups. In conclusion, higher serum immunoglobulin E levels were correlated with lower values of the forced expiratory volume in one second, and anticholinergic agents, but not beta2-adrenergic agents, caused more pronounced bronchodilation in subjects with high than in those with low immunoglobulin E levels. These data suggest that serum immunoglobulin E may be one of the factors that determine the airway tone, possibly via cholinergic mechanisms.

Administration, Inhalation↗

PubMed1998

Induction of nitric oxide synthase by lipopolysaccharide inhalation enhances substance P-induced microvascular leakage in guinea-pigs.

Inducible nitric oxide (NO) synthase (iNOS)-mediated hyperproduction of NO in airways has been reported in asthmatic patients. However, the role of NO in the pathogenesis of asthma has not yet been fully elucidated. The aim of this study was to examine whether the iNOS-derived NO affects airway microvascular leakage, one of the characteristic features of asthmatic airway inflammation. Guinea-pigs were exposed to lipopolysaccharide (LPS) (1 mg x mL(-1)) by inhalation in order to induce iNOS in the airways, and the histochemical staining of reduced nicotinamide-adenine dinucleotide phosphate (NADPH)-diaphorase activity was determined 5 h after the inhalation to confirm the iNOS induction. Airway microvascular leakage to subthreshold doses of substance P (0.3 microg x kg(-1), i.v.) was also examined in the absence and presence of an iNOS inhibitor (aminoguanidine) in LPS- or saline-exposed (control) animals using Evans blue dye and Monastral blue dye. In the LPS-exposed animals, increased NADPH-diaphorase activity was observed in the airway microvasculature compared with the control animals. Substance P caused significant airway microvascular leakage assessed by Evans blue dye in all airway levels in the LPS-exposed animals but not in the control group. This was also confirmed by Monastral blue dye extravasation. Aminoguanidine abolished this LPS-induced enhancement of plasma leakage to substance P without changing the systemic blood pressure. These results may suggest that inducible nitric oxide synthase-derived nitric oxide is capable of potentiating neurogenic plasma leakage in airways.

Administration, Inhalation↗

PubMed1998

Suppression of gene expression and production of interleukin 13 by dexamethasone in human peripheral blood mononuclear cells.

We examined the effect of dexamethasone on the gene expression and production of interleukin (IL)-13 by human peripheral blood mononuclear cells from healthy controls. The gene expression was assessed semiquantitatively by sequential transcription polymerase chain reaction and Southern blot analysis, and the production of this cytokine was assessed by enzyme-linked immunosorbent assay (ELISA). Dexamethasone suppressed IL-13 gene expression induced by stimulation with phytohemagglutinin and phorbol 12-myristate 13-acetate in a dose-dependent manner, with 96% suppression at 10(-6) M, and also suppressed the increased production of IL-13. This is suggested to be one of the mechanisms by which glucocorticoids suppress allergic inflammation.

Anti-Inflammatory Agents↗

PubMed1998

Protective effects of interleukin-10 on diaphragm muscle in a septic animal model.

The effects of Interleukin (IL)-10 intravenous injection after endotoxin administration on diaphragm muscle were studied using Wistar rats. The animals were divided into two treatment groups: A saline + endotoxin group as control and an IL-10 + endotoxin group. E. coli endotoxin (10 mg/kg) was injected intraperitoneally 5 minutes after saline or IL-10 (1250 U, 0.25 mg) injection. The force-frequency curves, twitch kinetics and fatigability were measured at 0 and 4 hours after endotoxin injection. In the saline + endotoxin group, the force-frequency curves and half relaxation time were significantly decreased at 4 hours (p < 0.001 and p < 0.05, respectively) compared to those at 0 hour. In the IL-10 + endotoxin group, the decrement in the force-frequency curves by endotoxin was prevented at 4 hours compared to that at 0 hour. Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry showed positive staining in the saline + endotoxin at 4 hours, but there was no significant staining at 0 or 4 hours in the IL-10 + endotoxin group. These data suggest that IL-10 prevents the deterioration of contraction induced by endotoxin by inhibiting nitric oxide production in the diaphragm muscle.

Animals↗

PubMed1998

Influence of sustained hypoxia on the sensation of dyspnea.

We assessed the effect of sustained isocapnic hypoxia (PCO2 = 40 Torr, SaO2 = 80%) on the sensation of dyspnea in 16 normal healthy males. Subjects rated the sensation of dyspnea (c) on 15 cm visual analog scales during 20 min of sustained hypoxia. Following this hypoxic period, 8 subjects undertook mild exercise (10-50 W on a bicycle ergometer for 3 min) under the continuation of hypoxia. During sustained hypoxia, psi increased initially with ventilation from 0.6 +/- 0.2 (n = 16, mean +/- SE) to 2.9 +/- 0.6 at peak ventilation, but it decreased with ventilatory depression to 1.6 +/- 0.4. Dyspnea intensity during hypoxic exercise was significantly smaller than that at peak ventilation in the resting hypoxic period (2.3 +/- 0.7 vs. 3.9 +/- 1.0), although the ventilation was greater during exercise (24.0 +/- 3.0 vs. 19.7 +/- 1.4 l/min). These results indicate that sustained hypoxia has a biphasic, i.e., initial stimulatory and delayed depressant, effect on dyspnea and on ventilation. It is suggested that the dyspnea sensing mechanism is suppressed during mild exercise under sustained hypoxia.

Adult↗

PubMed1998

[SMANCS-TAE combined with PEI in the treatment for hepatocellular carcinoma].

From January 1996 to August 1997, 24 patients with advanced hepatocellular carcinoma (HCC) equal to or more than 2 cm (mean +/- SD; 4.1 +/- 3.0 cm) in main tumor diameter were treated by SMANCS-TAE (20 cases) or SMANCS-TAI (4 cases) combined with PEI. Six cases had solitary lesion, 16 cases had multiple lesions, and 2 cases had massive lesions. After this combination therapy, 21 of 24 cases had complete tumor necrosis. During 3 to 19 months follow up period, 12 cases had cancer-free survival (SMANCS-TAI; 3 cases), and 9 cases had tumor recurrences (3 cases were local recurrences and 6 cases involved new lesions). Two cases died of hepatic infarction and cancer death, however, the remaining 22 cases were surviving. SMANCS-TAE combined with PEI is useful treatment for advanced large or multiple HCC lesions in patients who are poor surgical risks.

Aged↗