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Biomedical subjects

S Kanoh

Publications and source records attributed to S Kanoh.

At least 19 recordsLinked to original sources

Diffuse lung disease caused by cotton fibre inhalation but distinct from byssinosis.

A 66 year old man had inhaled cotton fibre for 50 years at his workplace. He did not have any respiratory symptoms. Chest CT scans revealed diffuse centrilobular and peribronchovascular interstitial thickening. Lung biopsy specimens confirmed the presence of string-like foreign bodies as well as granulomas and fibrosis in the peribronchial region. Infrared spectrophotometry confirmed that the foreign bodies were composed of natural cellulose. This is the first study to show directly by examination of biopsy samples that cotton fibre inhalation can cause diffuse lung disease. The clinical features of the disease were entirely different from those of byssinosis.

Aged↗

Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8.

T cells expressing CD57 (a natural killer cell marker) with interferon-gamma (IFN-gamma) producing capacity increase under various conditions. CD57+ T cells are also present in the bronchoalveolar lavage fluid (BALF) of sarcoidosis, and several phenotypical and functional analyses of these cells have been reported. In the present study, BALF T cells obtained from 52 patients with sarcoidosis were classified further into CD4+CD57+ T cells, CD4+CD57- T cells, CD8+CD57+ T cells and CD8+CD57- T cells and their phenotypes and functional characteristics were assessed. Substantial proportions of these T cell subsets expressed natural killer cell markers CD161 and CD122. The biased expansion of Vbeta2 T cells was observed in both CD4+CD57+ T cells and CD4+CD57- T cells in BALF from most patients, while the expansion of other Vbeta T cells was also observed in some patients. Unexpectedly, the biased expansion of certain Vbeta T cells was also seen in either CD8+CD57+ T cells or CD8+CD57- T cells, while the expanded Vbeta T cells in CD8+ T cells differed substantially among individuals. BALF T cells showed a remarkably lower T cell receptor (TCR) intensity than that of peripheral blood T cells. Both CD8+ T cell subsets in BALF of sarcoidosis expressed the intracellular perforin/granzyme B, while all four subsets expressed intracellular IFN-gamma after in vitro activation, and CD4+ T cells, especially CD4+CD57+ T cells, expressed tumour necrosis factor-alpha. These findings indicate that CD57+ T cells as well as CD57- T cells in the BALF are phenotypically and functionally different from peripheral blood T cells and may play an important role in the Th1 dominant state and inflammation in pulmonary sarcoidosis.

Antigens, CD↗

FK506 inhibits Cl- secretion in airway epithelium via calcineurin-independent mechanism.

FK506 (tacrolimus)-binding protein (FKBP) is associated with intracellular Ca2+ release channel and modulates its function. To elucidate the effect of FK506 on Ca2+ dynamics and Ca2+-mediated Cl- secretion in airway epithelium, we studied intracellular Ca2+ ([Ca2+]i) concentration and Cl(-)-dependent short-circuit current (Isc), in cultured bovine tracheal epithelial cells. Addition of ATP induced an increase in [Ca2+]i, and this response was dose dependently inhibited by FK506. Rapamycin, which binds FKBP with high affinity, likewise inhibited the [Ca2+]i rise, but cyclosporin A, a specific calcineurin inhibitor, did not. In Cl- secretion studies using Ussing chamber, ATP increased Ca2+-mediated Isc in amiloride-treated cells, an effect that was inhibited by FK506 and rapamycin but not by cyclosporin A. Therefore, FK506 inhibits Ca2+ mobilization in airway epithelium via FKBP but not calcineurin-dependent mechanism, which may result in the suppression of Ca2+-activated Cl- secretion.

Adenosine Triphosphate↗

New quinolone, grepafloxacin, inhibits Cl- secretion across bovine airway epithelium in culture.

OBJECTIVE: Transepithelial ion transport plays an important role in the regulation of the amount and the rheological properties of bronchial secretion. The effect of grepafloxacin (GPFX), a new quinolone agent, on bioelectrical properties of airway epithelium was determined. METHODOLOGY: Electrical properties of bovine tracheal epithelium cultured under an air-liquid interface condition were measured by the short-circuit technique. RESULTS: Addition of GPFX (100 microg/mL) to the mucosal side decreased short-circuit current (Isc) from 14.4 +/- 1.3 to 5.6 +/- 0.6 microA/cm2 (P < 0.001), and the response was accompanied by corresponding decreases in transepithelial potential difference and cell conductance. This effect was concentration dependent, and a similar response was also noted when GPFX was added to the submucosal side. The GPFX-induced decrease in Isc was not altered by the Na+ channel blocker amiloride, but was inhibited by the Cl- channel blocker diphenylamine-2-carboxylate or Cl(-)-free medium (P < 0.001, in each case). Furthermore, GPFX reduced Cl- conductance (P < 0.01) without affecting Na+ conductance of the epithelium. CONCLUSIONS: Grepafloxacin selectively inhibits Cl- secretion across tracheal epithelial cells, which may result in the inhibition of water secretion and, hence, the reduction of airway secretion.

Animals↗

Differential regulations between adenosine triphosphate (ATP)- and uridine triphosphate-induced Cl(-) secretion in bovine tracheal epithelium. Direct stimulation of P1-like receptor by ATP.

Adenosine 5'-triphosphate (ATP) stimulates airway epithelial Cl(-) secretion in a complicated manner. We examined the difference between ATP- and uridine 5'-triphosphate (UTP)-induced responses of short-circuit current (Isc) in bovine tracheal epithelium treated with amiloride. Each nucleotide caused an increase in Isc composed of the first and second peaks, where the second peak induced by ATP was higher compared with UTP. The ATP-induced second peak was inhibited by the protein kinase (PK) A inhibitor H89, saturation of P1 receptor with adenosine, and the P1 receptor antagonist 8-p-sulfophenyltheophylline, but not by the Ca(2+) chelator ethyleneglycol-bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid plus the endoplasmic reticulum Ca(2+)-pump inhibitor thapsigargin, the adenosine breakdown enzyme adenosine deaminase, the ectonucleotidase inhibitor alpha,beta-methyleneadenosine 5'-diphosphate, or saturation of P2Y2 receptor with UTP. Thus, the response is associated with PKA-dependent pathway via P1-like receptor but not with Ca(2+)-dependent pathway via P2Y2 receptor, and ATP degradation products do not contribute to this response. Further, stimulation of cells with ATP increased PKA activity. In addition, pretreatment with glybenclamide, an inhibitor of cystic fibrosis transmembrane conductance regulator, reduced the second peak of Isc induced by ATP but was without effect on that induced by UTP. Therefore, ATP stimulates glybenclamide-sensitive Cl(-) secretion, and this action is partly mediated by PKA-dependent pathway via P1-like receptor.

Adenosine↗

Thermal gelation properties of spent hen mince and surimi.

Thermal gelation properties of spent hen mince and surimi were investigated. The mince from 98-wk-old spent hens was washed two times with 0.1% NaCl. A portion of unwashed and washed mince was mixed with 4% sugar, 4% sorbitol, and 0.2% Na-tripolyphosphate to produce surimi and was kept frozen at -20 C. The mince and surimi were ground with 3% NaCl and a small amount of water to adjust the final moisture content to 80%. A 6 to 8% potato starch was mixed with some pastes. The pastes were stuffed into sausage casings and heated by one-step and two-step heating. The effects of washing, heating, and addition of ingredients on the color, composition, and functional properties of the mince and gel were compared. Washed, spent hen mince was lighter and less red in color and higher in collagen, gel strength, water-holding ability, and cooking yield than unwashed mince. The best temperature and time schedule for the gelation of spent hen mince was 90 C for 15 min in one-step heating. Heating at 100 C for 5 min after preheating at 60 to 70 C for 30 min resulted a gel with distinctly improved gel strength. Sucrose (4%), sorbitol (4%), and Na-tripolyphosphate (0.2%) improved the gel quality of nonfrozen mince but showed little cryoprotective effect against the degradation of frozen-stored product. A 6% potato starch improved the gel texture, cooking yield, and water-holding ability compared with 8% starch.

Animals↗

Washing and cryoprotectant effects on frozen storage of spent hen surimi.

Mince of 98-wk-old spent hens was washed two times with 0.1% NaCl. Portions of unwashed and washed mince were mixed with a cryoprotectant (CP) composed of 4% sucrose, 4% sorbitol, and 0.2% Na-tripolyphosphate and were immediately frozen and stored at -20 C. Mince without CP was run as control. Textural properties of the stored mince and surimi were measured at 1-mo intervals for 6 mo, after being thawed at 4 C overnight, ground with 3% NaCl, and heated at 90 C for 15 min. For freeze-thaw stability study, minces were subjected to six freeze-thaw cycles. Each freeze-thaw cycle was carried out at 1 mo of storage. Textural quality parameters (gel strength, breaking strength, deformation, protein solubility, expressible moisture, cooking yield, folding test, drip-loss, and sensory scores) were decreased in both unwashed and washed mince, mostly during the early stages of storage. Washed mince showed significantly better textural properties than unwashed mince. Washing protected the gel quality of the hen mince from degradation during frozen storage. Cryoprotectants could not protect the gel strength or breaking strength, but deformation was slightly improved. Water-retention properties were protected, and folding test and sensory scores were well preserved in the mince with added CP. Cryoprotectants had a beneficial effect on frozen, stored spent hen surimi to protect the elasticity and cohesiveness of the gel.

Animals↗

Effect of FK506 on ATP-induced intracellular calcium oscillations in cow tracheal epithelium.

To elucidate the effect of FK506 on Ca2+ oscillations in airway epithelium, we investigated cultured cow tracheal epithelial cells with a Ca2+ image-analysis system. ATP (1 microM) induced long-lasting Ca2+ oscillations, having nearly constant peak values (300-400 nM) and intervals (20-40 s) in subconfluent cells but not in confluent cells. These responses were gradually attenuated and abolished by the addition of FK506. Rapamycin, which binds the FK506-binding protein (FKBP), likewise inhibited Ca2+ oscillations, whereas cyclosporin A, a calcineurin inhibitor, did not. Treatment of cells with FK506 decreased Ca2+ content in thapsigargin-sensitive stores, suggesting that the partial depletion of the stores causes the inhibition of Ca2+ oscillations. Immunocytochemistry revealed the existence of cytoplasmic FKBP-like immunoreactivities. The expression of a 12-kDa FKBP was greater in subconfluent cells than in confluent cells as determined by Western blotting, suggesting that the 12-kDa FKBP may be one of the factors that regulates Ca2+ oscillations. Therefore, FK506 possesses an inhibitory action on the Ca2+ response via intracellular FKBP but not via calcineurin, which may result in modification of airway epithelial functions.

Adenosine Triphosphate↗

Erythromycin inhibits ATP-induced intracellular calcium responses in bovine tracheal epithelial cells.

Erythromycin (EM) therapy is known to decrease airway secretion in chronic inflammatory airway diseases such as diffuse panbronchiolitis. Airway secretion is regulated by intracellular Ca2+ concentration ([Ca2+]i). To elucidate the intracellular site of action of EM in airway epithelium, we examined the effect of EM on Ca2+ dynamics in cultured bovine tracheal epithelial cells using fura-2. EM per se did not cause any change in [Ca2+]i. Adenosine triphosphate (ATP; 10(-4) M) induced a biphasic [Ca2+]i increase, consisting of a transient response followed by a sustained response. Pretreatment of cells with EM had little effect on the ATP-induced transient Ca2+ response but substantially reduced the sustained response in a dose-dependent manner. Clarithromycin, another 14-membered ring macrolide, likewise showed the inhibitory effect, but ampicillin and cephasolin did not. Uridine triphosphate (UTP; 10(-4) M) induced a biphasic [Ca2+]i increase similar to ATP, and the UTP-induced sustained Ca2+ response was also inhibited by EM. In Ca2+-deficient medium (1 mM ethyleneglycol-bis-(beta-aminoethyl ether)-N, N'-tetraacetic acid [EGTA]) or in the presence of La3+, the sustained Ca2+ response disappeared, suggesting that EM may inhibit Ca2+ influx induced by P2u purinoceptor stimulation. In single-cell Ca2+ image analysis, low concentration of ATP (10(-6) M) induced Ca2+ oscillations, which were also inhibited by EM. The disappearance of [Ca2+]i oscillations after addition of EM was similar to that after addition of EGTA. These results suggest that EM may decrease Ca2+-dependent airway secretion by inhibiting agonist-stimulated Ca2+ influx.

Adenosine Triphosphate↗

Determining specific gravity, specific heat, and surface area of rabbits for a possible thermodynamic approach to body temperature change.

In order to investigate the mechanisms of body temperature change from a thermodynamic aspect, we investigated the specific gravity, specific heat and surface area of rabbit in situ. We obtained the following results. 1. The specific gravity of normal, shorn and ecdysed rabbits is 0.94, 0.95 and 0.97, respectively. 2. The specific heat of normal, shorn and ecdysed rabbits is 0.95, 0.89 and 0.69 cal/g.K, [corrected] respectively. 3. The surface area of the rabbit was also measured by using the weight of aluminum foil which covered the surface.

Animals↗

[Respiratory bronchiolitis-associated interstitial lung disease].

A 48-year-old man was admitted because of persistent dry cough for six months. He had been a smoker for 25 years, averaging a pack a day, and demonstrated clubbing of the fingers. Basilar fine crackles were observed in both lung fields. Chest X-ray films on admission showed diffuse reticulonodular shadows. Chest computed tomograms showed low-attenuation areas mainly in the center of the upper lung field, and ground-glass opacity, air bronchiolograms, and perivascular interstitial thickening of the peripheral vessels mostly in the lower field. A Gascintigram disclosed mild accumulation in both lungs. A transbronchial lung biopsy specimen did not reveal special features. However, a biopsy specimen obtained by thoracoscopy showed evidence of respiratory bronchiolitis, with a mononuclear inflammatory process involving respiratory bronchioles and adjacent air space, associated with mild fibrous thickening of the peribronchiolar interstitium and surrounding alveoli septa. These findings suggested that the patient had respiratory bronchiolitis-associated interstitial lung disease, the second case to be reported in Japan.

Bronchiolitis↗

Increased oxidative metabolism in cow tracheal epithelial cells cultured at air-liquid interface.

Airway epithelial cells cultured at the air-liquid interface possess highly differentiated functions and structures compared with the cells cultured under immersion. We examined the oxidative metabolism and glycolysis in cow tracheal epithelial cells on Days 3, 6, 10, and 13, cultured under three different conditions: (1) immersion culture on porous filters with apical and basolateral feeding (IM), (2) air-exposed culture on porous filters with basolateral feeding, i.e., air-liquid interface culture (AI), and (3) conventional immersion culture in plastic dishes with apical feeding (DI). Lactate production was less in AI than in IM and DI on Day 3 through Day 13, whereas cellular adenosine triphosphate content and basal O2 consumption were greater. Ouabain-sensitive and ouabain-insensitive O2 consumption, and the uncoupled O2 consumption were also greater in AI. Cytosolic lactate dehydrogenase activities on Day 10 were lower in AI, whereas alpha-ketoglutarate dehydrogenase activities were higher. The increased oxidative metabolism in AI was more pronounced at the late phase of culture (Days 10 and 13). In contrast, glycolysis remained elevated during the experiment in IM and DI. These data suggest that (I) AI begins to promote oxidative metabolism from growth phase by the provision of adequate oxygenation, and then further shifts to oxidative metabolism with differentiation; and (2) apical feeding may be responsible for the disturbance of the development of the oxidative metabolism.

Adenosine Triphosphate↗

The two essential light chains of carp fast skeletal myosin, LC1 and LC3, are encoded by distinct genes and change their molar ratio following temperature acclimation.

cDNA libraries were constructed from fast skeletal muscles of carp acclimated to 10 and 30 degrees C for a minimum of 5 weeks and were screened for myosin alkali light chains, LC1 and LC3, using an anti-skipjack LC1 polyclonal antibody. Two types of LC1 cDNA clone were isolated and termed LC1a and LC1b: their nucleotide sequences showed 92% homology. The ratio of LC1a to LC1b cDNA clones isolated was approximately 3:1, showing no apparent changes following temperature acclimation. The occurrence of the two isoforms was further confirmed by N-terminal amino acid sequencing of purified LC1. No isoform was, however, detected for LC3, while homology in the overlapping region between LC1a and LC3 cDNAs was only 65% even after the most probable alignment. Southern blot analyses probed with cDNA clones specific to LC1a and LC3 showed different hybridization patterns from each other, demonstrating that carp LC1 and LC3 are encoded by different genes. These results are in marked contrast to those from higher vertebrates which express LC1 and LC3 from a single gene by alternative RNA transcription and two modes of splicing. Northern blot analysis showed that the ratios of LC3/LC1 mRNAs were significantly higher (3.93) in 30 degrees C-acclimated than in 10 degrees C-acclimated (3.10) carp.

Amino Acid Sequence↗

Polysaccharides as drug carriers: biodisposition of fluorescein-labeled dextrans in mice.

The biodisposition of fluorescein-labeled dextrans (FDs) with different molecular weights (MW = 4-500 kDa) was systematically examined in mice. After intravenous injection of FDs at a dose of 120 mg/kg, the levels of FDs in the blood circulation and in the various organs were measured fluorometrically. FDs with a molecular weight lower than 20 kDa showed poor hepatic distribution (2.1-3.7% of dose/g tissue) due to their rapid elimination from the blood circulation. FDs with higher molecular weights were appreciably distributed in the liver (18.9-24.0% of dose/g tissue) and accumulated there over a long period, whereas the FD levels in the other organs were almost negligible a few days after injection. The hepatic mean residence time of FDs ranged from 22.5 to 28.1 d. Partial depolymerization of FDs which accumulated in the liver was observed within 10 d after administration. The hepatic uptake clearance of FDs was decreased with an increase in molecular weight. A marked molecular weight dependency was also seen in the urinary and fecal excretions of FDs. An appreciable dose-dependency was demonstrated in the hepatic uptake of FDs (MW = 40 kDa), as well. The amount of hepatic uptake as a function of dose showed saturation kinetics and was analyzed by a Michaelis-Menten type equation. The apparent values of K(m) (dose) and Vmax (hepatic level) estimated were 116 +/- 5 mg/kg and 1.10 +/- 0.05 mg/g tissue, respectively.

Animals↗

A possible thermodynamic approach to the change of rectal temperature of rabbits due to forced restricting conditions.

It was found that the rectal temperature of rabbits restricted by a normal neck stock position dropped about 1.2-1.5 degrees C when the restriction position changed from a normal restriction to a supine restriction position. This temperature change was also reversible. We studied the mechanisms of this phenomenon from a thermodynamic point of view and obtained the following results: 1) The rectal temperature of rabbit restricted in the supine position did not increase after the injection of lipopolysaccharide (LPS). 2) In the supine restricted rabbit, endogenous pyrogen which was induced by the injection of LPS to these rabbits appeared the same as the control. 3) Some pharmaceuticals (urethan, iproniazid, atropin and hexamethonium) did not show any influence on the drop of rectal temperature in either normal or supine restrictions. 4) The rectal temperature of shorn rabbits dropped more than that of normal rabbits in both normal and supine restrictions. 5) When the normal restriction position was changed to the supine and prone restriction positions, the mean surface temperature was also decreased. From these results, it was concluded that rabbit rectal temperature was changed with a change in the restriction position which could change the surface skin area.

Animals↗

[Effect of heparin on the increase in intracellular calcium caused by inositol 1,4,5-triphosphate in airway epithelial cells].

We studied the effect of heparin on the inositol 1,4,5-triphosphate (IP3)-dependent increase in intracellular calcium (Ca2+) caused by ATP. The Ca2+ concentration ([Ca2+]i) in cultured epithelial cells from cow tracheas was measured by the fura-2 method. ATP (10(-4) M) stimulated IP3 production and caused a biphasic increase in [Ca2+]i a transient response and a subsequent sustained response. Heparin (l 100 U/ml) dose-dependently inhibited the ATP-induced increase in [Ca2+]i but had no effect on IP3 production. Dextran sulfate, a polysaccharide with negative charge density, had a similar inhibitory effect on the ATP-induced increase in [Ca2+]i. These data suggest that heparin inhibits intracellular Ca2+ mobilization by acting at a site "distal to" IP3 production, and that the mechanism of action of heparin may be related to its negative charge.

Adenosine Triphosphate↗