Biomedical subjects

K Kurashima

Publications and source records attributed to K Kurashima.

At least 37 records · Page 2Linked to original sources

PubMed2000

Effect of losartan, a type 1 angiotensin II receptor antagonist, on bronchial hyperresponsiveness to methacholine in patients with bronchial asthma.

It is unclear whether angiotensin II receptors are involved in bronchial hyperresponsiveness in asthmatic patients. We examined the effect of losartan, a specific angiotensin II type 1 (AT1) receptor antagonist, on bronchial responsiveness to inhaled methacholine in eight patients with stable asthma. Bronchial responsiveness to methacholine, assessed as the concentration of methacholine producing a 20% fall in FEV(1) (PC(20)-FEV(1)) and a 35% fall in standardized partial expiratory flow at 40% of FVC (PC(35)-PEF(40)), was measured on two occasions 2 wk apart. Losartan (50 mg once a day) or a placebo was orally administered for 1 wk before methacholine provocation test in a double-blind, randomized, crossover fashion. Although the PC(20)-FEV(1) values after placebo (2.037 [geometric standard error of the mean, GSEM = 0.210] mg/ml) and losartan (2.098 [GSEM, 0.239] mg/ml) were identical (p = 0.840), the geometric mean PC(35)-PEF(40) values significantly (p = 0.034) increased from 0.258 (GSEM, 0.156) mg/ml with placebo to 0.456 (GSEM, 0.186) mg/ml with losartan. We conclude that AT1 receptors are involved in bronchial hyperresponsiveness in asthmatic patients. This is the first report demonstrating the involvement of AT1 receptors in bronchial asthma.

Adult↗

PubMed2000

Type 1 angiotensin II receptor antagonism reduces antigen-induced airway reactions.

Although the renin-angiotensin system is activated in patients with asthma during severe acute attacks and angiotensin II has been shown to cause bronchoconstriction in patients with asthma, the role of angiotensin II in patients with asthma is unclear. We investigated the effects of two specific antagonists at type 1 and type 2 angiotensin II receptors, candesartan cilexetil (TCV-116) and PD123319, on antigen-induced airway reactions in guinea pigs. Sixty minutes after intraperitoneal administration of candesartan cilexetil (0.1, 1.0, or 10 mg/kg) or PD123319 (30 mg/kg), animals received an antigen challenge. Airway responsiveness to inhaled methacholine was assessed as the dose of methacholine required to produce a 200% increase in the pressure at the airway opening (PC(200)). Differential cell counts in bronchoalveolar lavage fluids (BALF) were measured 24 h after antigen challenge. Candesartan cilexetil did not inhibit antigen-induced bronchoconstriction in sensitized guinea pigs or alter PC(200) in nonsensitized guinea pigs. Antigen inhalation significantly increased bronchoconstrictor responses to methacholine and increased airway accumulation of eosinophils; both responses showed dose-dependent prevention by candesartan but not by PD123319. These results indicate that endogenous angiotensin II promotes antigen-induced airway hyperresponsiveness and eosinophil accumulation by acting at type 1 receptors.

Angiotensin Receptor Antagonists↗

PubMed1999

The apical Na(+)/H(+) exchanger isoform NHE3 is regulated by the actin cytoskeleton.

The epithelial isoform of the Na(+)/H(+) exchanger, NHE3, associates with at least two related regulatory factors called NHERF1/EBP50 and NHERF2/TKA-1/E3KARP. These factors in addition interact with the cytoskeletal protein ezrin, which in turn binds to actin. The possible linkage of NHE3 with the cytoskeleton prompted us to test the effect of actin-modifying agents on NHE3 activity. Cytochalasins B and D and latrunculin B, which interfere with actin polymerization, induced a profound inhibition of NHE3 activity. The effect was isoform-specific inasmuch as the "housekeeping" exchanger NHE1 was virtually unaffected. Cytoskeletal disorganization was associated with a subcellular redistribution of NHE3, which accumulated at sites where actin aggregated, suggesting a physical interaction of exchangers with the cytoskeleton. An interaction was further suggested by the co-sedimentation of a detergent-insoluble fraction of NHE3 with the actin cytoskeleton. Inhibition of transport was not due to diminution in the number of transporters at the plasmalemma. Functional analyses of NHE1/NHE3 chimeras revealed that the cytoplasmic domain of NHE3 conferred sensitivity to cytochalasin B. Progressive carboxyl-terminal and internal deletions of the cytoplasmic region of NHE3 indicated that the region between residues 650 and 684 is critical for this response. This region overlaps with the domain reported to interact with NHERF and also contains a putative ezrin-binding site; hence, it likely plays a role in interactions with the cytoskeleton.

Actins↗

PubMed1999

Structure analysis of new homologous compounds Ga2O3(ZnO)m (m = integer) by high-resolution analytical transmission electron microscopy.

The crystal structure of a new homologous compound series, Ga(2)O(3)(ZnO)(m) (m = integer), is determined by high-resolution lattice imaging and high spatial resolution energy-dispersive X-ray spectroscopy (EDS) analysis in a field-emission analytical transmission electron microscope. This work was carried out mainly on the compound with m = 9 (digallium nonazinc dodecaoxide), which belongs to the orthorhombic system and has lattice constants a(o) = 0.33, b(o) = 2.0 and c(o) = 3.4 nm. From the extinction rules three possible space groups are selected and from them a unique space group is assigned as noncentrosymmetric Cmc2(1) (No. 36) on the basis of structural requirements. Ga(2)O(3)(ZnO)(m) is a layered structure consisting of Ga-O and m + 1 Ga/Zn-O layers stacked alternately along the c axis. It is shown that the structure of Ga(2)O(3)(ZnO)(m) differs from that of M(2)O(3)(ZnO)(m) (M = In, Fe; m = integer) reported previously. In Ga(2)O(3)(ZnO)(m) the Ga atoms occupy the tetrahedral sites in the Ga-O layers, whereas the M atoms in the M-O layers occupy the octahedral sites in M(2)O(3)(ZnO)(m) (M = In, Fe).

Journal Article↗

PubMed1999

Boron nitride cones: structure determination by transmission electron microscopy.

A form of turbostratic boron nitride containing a large percentage of micrometre-size conically shaped particles was investigated by transmission electron microscopy. Electron diffraction patterns revealed the presence of correlations between adjacent boron nitride layers. The diffraction patterns were also used to determine the cone apex angle of many cone-like objects. The apex angles exhibited a broad distribution from 84 to 130 degrees with an ill defined peak in the 92-95 degrees region. These results could be accounted for by a structure model in which conical boron nitride layers are helically wound about a disclination axis, according to overlap angles that correspond to high densities of coincidental lattice sites between successive layers. The clustering of the apex angles near 93 degrees was attributed to the formation of a square ring during the nucleation stage. This appears to be specific to the boron nitride system.

Journal Article↗

PubMed1999

Enzymatic synthesis of (-)- and (+)-acetoxyhexamides and (-)- and (+)-hydroxyhexamides.

The enantioselective hydrolysis of (+/-)-4-(1-acetoxyethyl)-N-(cyclohexylcarbamoyl)-benzenesulfona mides 3 with lipase Amano P from Pseudomonas sp. in a water-saturated solvent gave (R)-4-(1-hydroxyethyl)-N-(cyclohexylcarbamoyl)benzenesulfonamide 2 (39%, > 99% ee) and unchanged (S)-3 (50%, 62% ee). On the other hand, enantioselective esterification of (+/-)-2 with lipase Amano P in the presence of vinyl acetate provided (R)-3 (41%, > 99% ee) and unchanged (S)-2 (46%, 78% ee).

Acetohexamide↗

PubMed1998

Endosomal recycling of the Na+/H+ exchanger NHE3 isoform is regulated by the phosphatidylinositol 3-kinase pathway.

The NHE3 isoform of the Na+/H+ exchanger localizes to both the plasmalemmal and endosomal compartments in polarized epithelial and transfected Chinese hamster ovary (AP-1) cells. It is unclear how the distribution of NHE3 between these compartments is regulated. In this study, we examined the potential involvement of phosphatidylinositol 3'-kinase (PI3-K) in regulating the activity and distribution of NHE3, as this lipid kinase has been implicated in modulating vesicular traffic in the endosomal recycling pathway. Wortmannin and LY294002, both potent inhibitors of PI3-K, markedly inhibited NHE3-mediated H+ extrusion across the plasma membrane in a concentration- and time-dependent manner. The subcellular distribution of the antiporters was monitored by transfecting epitope-tagged NHE3 into AP-1 cells. In parallel with the inhibition of transport, PI3-K antagonists induced a pronounced loss of NHE3 from the cell surface and its accumulation in an intracellular compartment, as assessed by immunofluorescence microscopy and enzyme-linked immunosorbent assays. Further analysis using cells transfected with antiporters bearing an external epitope tag revealed that the redistribution reflected primarily a decrease in the rate of recycling of intracellular NHE3 to the cell surface. The wortmannin-induced inhibition and redistribution of NHE3 were prevented when cells were incubated at 4 degreesC, consistent with the known temperature dependence of the endocytic process. These observations demonstrate that NHE3 activity is controlled by dynamic endocytic and recycling events that are modulated by PI3-K.

Amino Acid Sequence↗

PubMed1998

Bafilomycin A1 induces apoptosis in the human pancreatic cancer cell line Capan-1.

Bafilomycin A1, a specific inhibitor of vacuolar type H(+)-ATPase, can inhibit the growth of a variety of cultured cells in a dose-dependent manner, but its mechanism is unclear. The aim of this study was to examine whether bafilomycin A1 inhibits the growth of Capan-1 human pancreatic cancer cells through apoptosis. The effect of bafilomycin A1 on tumour growth in vitro and in vivo was examined using an MTT assay and an in vivo tumour model. The presence or absence of apoptosis was determined by morphology and DNA analysis of tumour cells. The concentration of bafilomycin A1 for 50 per cent inhibition of cell viability during 72 h by the MTT assay was 5 nm. In DNA analysis, a ladder of fragmented DNA was detected in Capan-1 cells treated with bafilomycin A1 at concentrations greater than 10 nm for 24 h. Nude mice bearing a xenografted Capan-1 cell line tumour received 4 weeks of bafilomycin A1 (1.0 mg/kg per day). This treatment significantly inhibited tumour growth compared with controls after 21 days (P < 0.05). Histopathological examination of tumour cells in the treated group demonstrated signs of apoptosis with chromatin condensation and cell shrinkage. These observations suggest that bafilomycin A1 inhibits the growth of Capan-1 human pancreatic cancer cells through apoptosis.

Adenocarcinoma↗

PubMed1997

Identification of sites required for down-regulation of Na+/H+ exchanger NHE3 activity by cAMP-dependent protein kinase. phosphorylation-dependent and -independent mechanisms.

We recently identified a region within the cytoplasmic C-terminal tail of the Na+/H+ exchanger NHE3 isoform (residues 579 to 684) which is essential for inhibition of transport activity by cAMP-dependent protein kinase (PKA) (Cabado, A. G., Yu, F. H., Kapus, A., Gergely, L., Grinstein, S., and Orlowski, J. (1996) J. Biol. Chem. 271, 3590-3599). To further define determinants of PKA regulation, six serine residues located in potential recognition sequences for PKA within, or adjacent to, this region (positions 552, 605, 634, 661, 690, and 691) were altered either independently or in various combinations using site-directed mutagenesis. Wild type and mutant NHE3s tagged with the influenza virus hemagglutinin epitope were stably expressed in exchanger-deficient Chinese hamster ovary cells (AP-1) for functional studies. Of the individual mutations examined, only substitutions at Ser605 or Ser634 affected sensitivity to forskolin, an activator of adenylate cyclase, although partial inhibition of NHE3 activity by forskolin remained. By contrast, simultaneous mutation of both these serines completely abolished cAMP-mediated inhibition of NHE3 without greatly affecting basal transport activity. Two-dimensional analysis of tryptic digests of immunoprecipitated NHE3 labeled in vivo with [32P]orthophosphate revealed several phosphopeptides under basal conditions. Phosphorylation was increased approximately 3-fold in one of these peptides following forskolin treatment, and this change was eliminated by mutation of residue Ser605. Thus, phosphorylation of Ser605 is essential for cAMP-mediated inhibition of NHE3. In addition, Ser634 is also required for the effect of cAMP, even though this residue does not become phosphorylated upon activation of PKA.

Amiloride↗

PubMed1997

Surface activity of sputum from acute asthmatic patients.

The cause of airway resistance that develops during an asthma attack is not fully understood. Besides bronchospasm and airway edema, a surfactant dysfunction has been suggested as a reason for an increase in airway resistance. This study sought to determine if surface activity of airway fluid is altered in acute asthmatic attacks. Sputa were collected serially from seven patients with acute asthmatic attacks and compared with that of normal subjects (n = 15) and subjects with stable asthma (n = 14). Sputa were analyzed for minimal surface tension (STmin), phospholipids (PL), and total proteins (TP). There were no significant differences in STmin, PL, and TP between sputa from normal subjects and subjects with stable asthma. In the acute phase of the attack, TP, TP/PL ratio, and STmin significantly increased. However, in the recovery phase of the attack, TP/PL ratio and STmin significantly decreased. These data demonstrated that surface properties of airway fluid deteriorated in the early phase of the asthma attack, but were ameliorated in the recovery phase possibly by the surfactant recruitment. The drastic changes in surface activity of sputum suggest that changes in airway surfactant may be partly involved in the pathogenesis of the airway obstruction during asthmatic attack.

Acute Disease↗

PubMed1997

Elevated chemokine levels in bronchoalveolar lavage fluid of tuberculosis patients.

In pulmonary tuberculosis, the proportion of lymphocytes, particularly that of CD4+ T lymphocytes, was increased in bronchoalveolar lavage fluid (BALF), reflecting their protective role against mycobacterial infections. In order to elucidate the mechanisms of lymphocyte accumulation in lungs, we measured the levels of chemokines with potent lymphocyte chemotactic activities, including interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and, regulated on activation, normal T-cell expressed and secreted (RANTES) present in BALF from patients with pulmonary tuberculosis in acute (n = 10) and convalescent phases (n = 6), as well as normal subjects (n = 10). During the acute phase of the disease, the proportions of lymphocytes and neutrophils were increased, as reported in previous studies. The levels of IL-8, MCP-1, and RANTES in the acute phase of pulmonary tuberculosis were also markedly elevated as compared with those of normal subjects. MCP-1 and RANTES, but not IL-8 levels present in BALF, decreased in the convalescent phase. Moreover, the concentration of RANTES correlated significantly with the absolute number of CD4+ cells in BALF. These data suggest that chemotactic cytokines are differentially produced and participate in the host response to Mycobacterium tuberculosis infection.

Bronchoalveolar Lavage Fluid↗

PubMed1997

A specific elevation of RANTES in bronchoalveolar lavage fluids of patients with chronic eosinophilic pneumonia.

Chronic eosinophilic pneumonia (CEP) is a rare, idiopathic lung disorder characterized pathologically by massive eosinophil infiltration into lung. In the bronchoalveolar lavage fluid (BALF) of patients with CEP, eosinophil numbers were markedly increased but returned to normal-levels upon the resolution of clinical symptoms, which suggests the crucial role of eosinophils in the pathogenesis of CEP. To clarify the mechanism of eosinophil accumulation in CEP, we determined the BALF levels of RANTES and macrophage inflammatory protein-1 alpha, two chemokines that predominantly exhibit in vitro eosinophil chemotactic activities. RANTES (106.7 +/- 27.2 pg/mg albumin; n = 16) concentrations in BALF from patients with CEP were significantly elevated in comparison with those of normal control subjects (1.4 pg/mg albumin; n = 13), whereas BALF macrophage inflammatory protein-1 alpha levels were not. In addition, eosinophils, lymphocytes, and macrophages in BALF were positively stained with a specific anti-RANTES antibody, which suggests that RANTES was produced locally in the lungs of CEP patients. Moreover, BALF-RANTES levels correlated significantly with the proportion of eosinophils in BALF. Furthermore, nearly half of the eosinophil chemotactic activities in BALF were abrogated by the anti-RANTES antibody in vitro. Collectively, these data suggest that locally produced RANTES is involved in eosinophil accumulation in the pulmonary alveolus and interstitium of patients with CEP.

Bronchoalveolar Lavage↗

PubMed1997

Effect of surfactant inhalation on allergic bronchoconstriction in guinea pigs.

BACKGROUND: In the small airway, surfactant reduces surface tension, prevents liquid filling of bronchioles, thereby maintaining patency in the small airways. Recent reports demonstrated that surfactant dysfunction develops in experimental asthma in immunized guinea pigs. However, there are few reports concerning surfactant and lung function in an experimental asthma model. OBJECTIVE: To examine whether inhaled surfactant improves lung mechanics in antigen-induced bronchoconstriction in guinea pigs. METHOD: We developed a passively immunized guinea pig model for allergic bronchoconstriction induced by antigen inhalation. Using this model, we investigated the effect of inhaled exogenous surfactant, surfactant TA, on the airway opening pressure (Pao) after antigen challenge. RESULTS: Aerosol antigen challenge produced a gradual and long-lasting increase in Pao. Twenty minutes after antigen challenge, aerosolized surfactant TA, 20 mg/ml, was inhaled for 90 s, and it significantly reduced the Pao by 32.8% in 12 min, while a 10.2% reduction was observed in a control group in the same period. When surfactant TA was administered by 90-s inhalation before antigen challenge, it inhibited the Pao increase in a dose-dependent manner: mean inhibitory rates of Pao were 33.6% in surfactant TA 10 mg/ml and 61.9% in surfactant TA 20 mg/ml, respectively. CONCLUSION: Inhaled surfactant showed preventive and recovery effects on antigeninduced bronchoconstriction in an immunized guinea pig model.

Administration, Inhalation↗

PubMed1996

Increase of chemokine levels in sputum precedes exacerbation of acute asthma attacks.

Basophils and eosinophils can be activated in vitro by several chemokines such as RANTES, monocyte chemotactic and activating factor (MCAF/MCP-1), macrophage inflammatory peptide-1 alpha (MIP-1 alpha), and interleukin-8 (IL-8). To explore the clinical relevance of the in vitro observations, we measured here the concentrations of these chemokines in sputa from asthmatic patients during acute attacks. Before the onset of a late-phase exacerbation, sputum MCAF/MCP-1, MIP-1 alpha, and IL-8 levels transiently but markedly increased from the basal levels in all of the patients with exacerbation, whereas the sputum levels of these chemokines remained unchanged during the course in the patients without a late-phase exacerbation. These results suggest the involvement of these chemokines in the late-phase exacerbation of asthma.

Acute Disease↗

PubMed1996

The role of vacuolar H(+)-ATPase in the control of intragranular pH and exocytosis in eosinophils.

The presence of vacuolar H(+)-ATPase (V-ATPase) on exocytotic granules in eosinophils and the role of this enzyme in exocytosis were explored in this study. Antibody against 116-kd subunit of V-ATPase positively stained eosinophil granules in immunofluorescence analysis. When eosinophil lysate was extracted immunomagnetically with the same antibody, the extracted fraction contained a considerable amount of eosinophil peroxidase, a marker of eosinophil-specific granules, which indicates that V-ATPase was present on the membranes of eosinophil exsosomal granules. The pH of the eosinophil granules, measured fluorometrically with acridine orange as a delta pH-sensitive dye, was estimated to be 5.1. The acidity of the eosinophil granules was perturbed by bafilomycin A1, a potent selective inhibitor of V-ATPase, which indicates that the low pH of these granules is maintained by V-ATPase activity. BafilomycinA1 and NH4Cl, both of which raise the intragranular pH to neutral, inhibited the eosinophil peroxidase exocytosis induced by platelet-activating factor. These agents did not, however, affect the changes in cytosolic free calcium concentration [Ca2+]i induced by platelet-activating factor. These observations suggest that bafilomycin A1 inhibited a delta pH-requiring step in eosinophil exocytosis that was preceded by the [Ca2+]i transient in the signal transduction pathway, and, hence, the findings suggest the pivotal role of V-ATPase in maintaining intragranular pH and its function of eosinophil exosomal granules.

Ammonium Chloride↗

PubMed1995

Effect of thromboxane A2 synthetase inhibitor, OKY-046, on sputum in chronic bronchitis and diffuse panbronchiolitis.

The mechanisms of excessive sputum production are only partially understood. We speculated that a selective thromboxane (Tx) A2 synthetase inhibitor, OKY-046, now used in the treatment of asthma in Japan, could decrease excess sputum production in patients with chronic airways disease. To test this hypothesis, we carried out a double-blind, placebo-controlled study of the effects of OKY-046, administered orally at 400 mg.day-1, on the sputum of patients with chronic bronchitis and patients with diffuse panbronchiolitis. Patients treated with OKY-046 showed a significant decrease (22%) in sputum volume after 1 month, and a 39% decrease after 3 months. Although the rheological properties of the sputum and the concentrations of fucose and immunoglobulin (Ig) A in the sputum remained unchanged, significant decreases were observed in the concentrations of total protein, albumin, sialic acid and phospholipid. Since albumin and fucose are chemical markers of plasma exudation and mucus secretion, respectively, whilst sialic acid and phospholipid are derived both from serum and mucus, our results indicate that this TxA2 synthetase inhibitor reduced sputum volume by inhibiting microvascular leakage in the airway. OKY-046 may, therefore, be of value in the treatment of chronic bronchitis and diffuse panbronchiolitis.

Adult↗

PubMed1994

Peripheral neuroectodermal tumor presenting pleural effusion.

Pleural effusion is a common finding of peripheral neuroectodermal tumor (PNET) of the chest wall (Askin's tumor), but little is known about the characteristics. A case of Askin's tumor with pleural effusion is reported. Repeated cytologies were negative for malignancy, but levels of lactic dehydrogenase (LDH) and neuron-specific enolase (NSE) in the pleural effusion were increased. Surgical biopsy was performed and immunohistochemical study of the tumor revealed the diagnosis.

Adult↗

PubMed1994

Inhibitory effect of aerosol administration of a sulfidopeptide leukotriene antagonist on bronchoconstriction induced by antigen inhalation in guinea pigs.

The effects of inhalation of the sulfidopeptide leukotriene antagonist AS-35 (9-[(4-Acetyl-3-hydroxy-2-n-propyl-phenoxy)methyl]-3-(1H- tetrazol-5-yl)-4H-pyrido [1,2-a]pyrimidin-4-one, CAS 108427-72-1) on bronchoconstriction induced by aerosol antigen, histamine, and leukotriene C4, D4 (LTC4, D4) were investigated in anesthetized and artificially ventilated guinea pigs. The increase in pressure at the airway opening (P(ao)) was measured as an index representing the grade of bronchial response. The bronchoconstriction induced by aerosol antigen was suppressed dose-dependently by pretreatment with inhaled AS-35 (0.1 mg and 1 mg) through a pressurized meter-dosed inhaler in the passively sensitized animals pretreated with diphenhydramine hydrochloride. But the histamine-induced bronchoconstriction was not altered by the pretreatment with AS-35 inhalation. On the other hand, LTC4- and LTD4-induced bronchoconstriction was inhibited by the pretreatment with aerosol AS-35 in a dose-dependent manner. The deposited dose of inhaled AS-35 in the peripheral airways was 3.5 micrograms and 6.5 micrograms when 0.1 mg and 1 mg of the drug was inhaled, respectively. These results suggest that sulfidopeptide leukotrienes (s-LTs) play an important role in the allergic bronchoconstriction in guinea pigs pretreated with antihistamine, and AS-35 inhalation may be beneficial in the treatment of asthma.

Administration, Inhalation↗