Biomedical subjects

H Hayashi

Publications and source records attributed to H Hayashi.

At least 145 records · Page 8Linked to original sources

PubMed2001

Possible involvement of Rho kinase in Ca2+ sensitization and mobilization by MCh in tracheal smooth muscle.

We examined the effects of Rho kinase on contraction and intracellular Ca2+ concentration ([Ca2+](i)) in guinea pig trachealis by measuring isometric force and the fura 2 signal [340- to 380-nm fluorescence ratio (F340/F380)]. A Rho kinase inhibitor, Y-27632 (1-1,000 microM), inhibited methacholine (MCh)-induced contraction, with a reduction in F340/F380 in a concentration-dependent manner. The values of EC(50) for contraction and F340/F380 induced by 1 microM MCh with Y-27632 were 27.3 +/- 5.1 and 524.1 +/- 31.0 microM, respectively. With 0.1 microM MCh, the values for these parameters were decreased to 1.0 +/- 0.1 and 98.2 +/- 6.2 microM, respectively. Tension-F340/F380 curves for MCh indicated that Y-27632 caused an ~50% inhibition of MCh-induced contraction, without a reduction in F340/F380. These effects of Y-27632 were not inhibited by a protein kinase C inhibitor, GF-109203X. Our results indicate that inhibition of Rho kinase attenuates both Ca2+ sensitization and [Ca2+](i).

Amides↗

PubMed2001

Effect of suprachiasmatic nucleus lesion on circadian dentin increment in rats.

Mammalian dentin universally shows circadian increments. However, little is known about the mechanism of this phenomenon. The purpose of the present study was to investigate the role of the suprachiasmatic nucleus (SCN) in the generation of circadian rhythm in dentin increment. Rats underwent lesion of the SCN by electrodes and were maintained under constant light to examine whether the circadian increment free runs. The rats were injected with nitrilotriacetato lead to chronologically label the growing dentin. Two weeks after the operation, maxillary incisors and the locations of lesions in the brain were examined histologically. A harmonic (Fourier) analysis was performed to examine the densitometric pattern of the dentin increments to determine their periodicity. In rats with a completely lesioned SCN, ultradian increments, but no circadian increments, were observed in the dentin. Alternatively, in rats with an intact or only partially lesioned SCN, circadian increments persisted or were only temporarily disturbed. These results suggest that the SCN plays an important role in the generation of the circadian dentin increment in rats.

Activity Cycles↗

PubMed2001

Sialyl Lewis X and anti-P-selectin antibody attenuate lipopolysaccharide-induced acute renal failure in rabbits.

BACKGROUND/AIM: Acute renal failure (ARF) is one of the common problems associated with sepsis or multiple organ dysfunction syndrome (MODS). We have investigated the effects of inhibiting selectin-mediated cell adhesion on lipopolysaccharide-induced ARF in rabbits, using sialyl Lewis X oligosaccharide and PB1.3, an anti-human P-selectin monoclonal antibody, as inhibitors. METHODS: ARF was induced by intravenous administration of lipopolysaccharide (0.3 mg/kg, i.v. bolus injection) to New Zealand White rabbits. Induction of ARF was characterized by increases in blood urea nitrogen (BUN), creatinine, and the number of polymorphonuclear leukocytes infiltrating glomeruli, and by fibrin deposition in glomeruli, and tubular dilatation in the kidney. Sialyl Lewis X oligosaccharide (14 mg/kg, i.v. bolus injection immediately before lipopolysaccharide administration and 9 mg/kg/h, i.v. infusion) or PB1.3 (5 mg/kg, i.v. bolus injection before lipopolysaccharide administration), anti-P-selectin antibody, were treated. RESULTS: Treatment with sialyl Lewis X oligosaccharide inhibited the increases in BUN, creatinine, and the number of infiltrating polymorphonuclear leukocytes, and attenuated histopathological impairments. Similarly, treatment with PB1.3 prevented some of the characteristics associated with lipopolysaccharide-induced ARF, not but the increase in creatinine. CONCLUSION: These results suggest that selectin inhibitors, including sialyl Lewis X oligosaccharide and PB1.3, may provide clinical benefits in the prevention of ARF associated with sepsis and MODS. To our knowledge, this is the first report that P-selectin is directly involved in lipopolysaccharide-induced ARF in rabbits.

Acute Kidney Injury↗

PubMed2001

Treatment of steroid-resistant idiopathic thrombocytopenic purpura in pregnancy with repeated high-dose intravenous immunoglobulin.

High-dose intravenous immunoglobulin (HD-IVIG) has temporary but reliable efficacy in idiopathic thrombocytopenic purpura (ITP). HD-IVIG has been described as the representative management in pregnant cases of refractory to corticosteroid or immunosuppressants. There have been few cases treated with repeated HD-IVIG to sustain pregnancy from the early phase of pregnancy. This case report describes a pregnant case of steroid-refractory ITP, treated with six times repeated HD-IVIG, resulting in the successful delivery of a healthy newborn with a normal platelet count. No adverse effects were observed.

Adolescent↗

PubMed2001

Immunocytochemical localization of secretory phospholipase A(2)-like protein in the pituitary gland and surrounding tissue of the bullfrog, Rana catesbeiana.

Previously, we obtained a protein that has considerable amino acid sequence homology with secretory phospholipase A(2) (PLA(2)) from a bullfrog pituitary fraction obtained during the purification of thyrotropin (TSH). Subsequently, partial amino acid sequence (N-terminal 45 amino acid residues) analysis revealed this protein to be identical to the N-terminal amino acid sequence of otoconin-22, the major protein of aragonitic otoconia in the Xenopus saccule. In this study we developed an antibody against the N-terminal peptide of the bullfrog protein and applied it for immunocytochemical study of the pituitary and its surrounding tissue. Western blotting analysis showed that this antibody recognizes a 20.4-kD protein that has a molecular mass close to that of otoconin-22. Immunohistochemical reaction with the antibody was not found in any anterior pituitary cells but was intense in the monolayer epithelial cells of the endolymphatic sac surrounding the pituitary gland, which is a major storage site of calcium carbonate in amphibians. An electron microscopic study revealed that the cuboidal cells in the endolymphatic sac contained large, polymorphic secretory granules in their apical cytoplasm. Immunogold particles indicating the presence of a PLA(2)-like protein were observed predominately in these secretory granules. These findings support the view that this PLA(2)-like protein obtained during purification of TSH was derived from the endolymphatic sac adhering to the pituitary and that this protein is a bullfrog otoconin. (J Histochem Cytochem 49:631-637, 2001)

Animals↗

PubMed2001

Acyclic carotenoids and their oxidation mixtures inhibit the growth of HL-60 human promyelocytic leukemia cells.

Lycopene has been known as a potential food component for cancer prevention, since tomato consumption was shown to be associated with reduced risk of certain cancers. We used HL-60 cells as a model of cancer cells to investigate whether acyclic carotenoids, such as phytoene, phytofluene, and zeta-carotene present in tomatoes, other than lycopene, as well as oxidation mixtures of these carotenoids, are potentially involved in the cancer-preventive action of tomatoes. When HL-60 cells were grown in the carotenoid-supplemented medium for 120 hours, zeta-carotene and phytofluene at 10 microM inhibited cell growth to 3.7% and 22.6% of the growth in control culture, respectively, although they were extremely unstable in the culture medium. The oxidation mixture of each carotenoid, which was prepared by incubation in toluene at 37 degrees C for 24 hours, more strongly inhibited cell growth than each intact carotenoid. The growth inhibition by lycopene was remarkably enhanced by its oxidation before supplementation to the medium. Phytofluene, zeta-carotene, and the oxidation mixture of lycopene induced apoptosis in HL-60 cells during incubation for 24 hours. The addition of alpha-tocopherol to the medium did not eliminate growth inhibition by the oxidation mixture of lycopene. These results suggest that the acyclic carotenoids inhibit cell growth through apoptosis induction and that oxidation products of the carotenoids participate in the growth inhibition.

Anticarcinogenic Agents↗

PubMed2001

BMP-2 augments FGF-induced differentiation of PC12 cells through upregulation of FGF receptor-1 expression.

When exposed to various neurotrophic factors, including fibroblast growth factors (FGF)-1 and -2, rat pheochromocytoma-derived PC12 cells differentiate into sympathetic neuron-like cells possessing elongated neurites. We found that while bone morphogenetic protein-2 (BMP-2) exerted little effect by itself on the differentiation of PC12 cells, in combination with FGF it strongly induced neurite outgrowth, even at subthreshold concentrations of FGF. Analysis of gene expression revealed that FGF receptor-1 (FGFR-1) mRNA was abundantly expressed in PC12 cells and that its expression was upregulated by pretreating the cells with BMP-2. Crosslinking the receptors with (125)I-FGF-2 and then immunoprecipitating them confirmed that expression of FGFR-1, but not other FGF receptor types, was enhanced by BMP-2. Furthermore, Scatchard analyses revealed that the numbers of FGF-2 binding sites were increased by approximately 40% after BMP-2 treatment. Pretreatment with BMP-2 also enhanced peak and sustained levels of FGF-induced ERK1/2 phosphorylation in PC12 cells. Finally, the augmentation of neurotrophic activity by BMP-2 was inhibited by SU5402, an FGFR-1 inhibitor. These findings indicate that BMP-2 augments FGF-induced differentiation of PC12 cells through selective upregulation of FGFR-1 expression, and suggest that BMP-2 and FGF act in concert to regulate cell differentiation in the nervous system.

Animals↗

PubMed2001

Effects of San'o-shashin-to and the constituent herbal medicines on theophylline-induced increase in arterial blood pressure of rats.

San'o-shashin-to, composed of Scutellariae Radix, Coptidis Rhizoma and Rhei Rhizoma (volume ratio = 1:1:1), reduced an increase in arterial blood pressure of anesthetized rats induced by theophylline (5 mg/kg, i.v.). The hypotensive effect of San'o-shashin-to was produced in a dose dependent manner and was maximum at its 0.5 g/kg. Then the constituent herbal medicines were examined for their possible hypotensive effect. Scutellariae Radix of 0.2 g/kg slightly decreased in the blood presure. Rhei Rhizoma of 0.2 g/kg decreased in the blood pressure and the hypotensive effect was significantly produced even at the dose of 0.05 g/kg, while Coptidis Rhizoma had little effect. Among fractions of San'o-shashin-to separated by Diaion HP-20 column chromatography, the 50% methanol-eluted fraction had a large hypotensive effect. The 50% methanol-eluted fraction of Scutellariae Radix and Rhei Rhizoma were also effective and, especially, that of Rhei Rhizoma had a large hypotensive effect. In isometric tension study, Scutellariae Radix and Rhei Rhizoma (10-30 microg/ml) slightly exerted contractile and relaxant effects, respectively, on the phenylephrine-contracted endothelium-intact rat thoracic aorta. Coptidis Rhizoma (1-10 microg/ml) caused both endothelium-dependent and -independent relaxantion. These results suggest that the hypotensive effect of San'o-shashin-to is not mediated by the direct action on blood vessel but by other actions. Some components in Scutellariae Radix and Rhei Rhizoma, especially in the latter may play a main role in the hypotensive effect.

Animals↗

PubMed2001

Selective nonpeptidic inhibitors of herpes simplex virus type 1 and human cytomegalovirus proteases.

The proteases encoded by herpesviruses including herpes simplex virus type 1 (HSV-1) and human cytomegalovirus (HCMV) are attractive targets for antiviral drug development because of their important roles in viral replication. We randomly screened a chemical compound library for inhibitory activity against HSV-1 protease. 1,4-Dihydroxynaphthalene and three naphthoquinones were found to be potent inhibitors of HSV-1 protease with IC50 values of 6.4 to 16.9 microM. Inhibitory mode analysis of the compounds against HSV-1 protease suggested that, in spite of structural similarities, only 1,4-dihydroxynaphthalene was a competitive inhibitor, whereas the three naphthoquinones were noncompetitive inhibitors. Among all assayed dihydroxynaphthalene derivatives in the chemical compound library, 1,4-dihydroxynaphthalene proved to be the most potent inhibitor of HSV-1 protease. Therefore, the two hydroxyl groups located at positions 1 and 4 on the naphthalene structure seemed essential for exertion of a potent inhibitory activity against HSV-1 protease. In addition, we have found that these compounds are also potent inhibitors of HCMV protease with extremely low micromolar IC50 values. This differed from the results of inhibitory mode analysis of HSV-1 protease, 1,4-dihydroxynaphthalene was a noncompetitive inhibitor of HCMV protease, and three naphthoquinones were competitive inhibitors. These compounds showed no effective inhibitory activity against several mammalian serine proteases (trypsin, chymotrypsin, kallikrein, plasmin, thrombin and Factor Xa) at 100 microM. These results suggest that 1,4-dihydroxynaphthalene and three naphthoquinones may be useful in the development of nonpeptidic antiherpesvirus agents.

Amino Acid Sequence↗

PubMed2001

Accumulation of medium chain acyl-CoAs during beta-oxidation of long chain fatty acid by isolated peroxisomes from rat liver.

We have reported fatty alcohol synthesis accompanied by chain elongation in liver peroxisomes (Biochim. Biophys. Acta, 1346, 38 (1997)). In the present experiment, we studied what kind of acyl-CoA(s) destined to be utilized as primer for fatty alcohol synthesis accumulate(s) during peroxisomal beta-oxidation. Peroxisomes were prepared from rat liver treated with clofibrate, a peroxisome proliferator, and incubated with [U-14C]palmitate, in order to investigate acyl-CoAs after beta-oxidation. At 1 mM concentration, MgATP activated beta-oxidation, but inhibited beta-oxidation at concentrations higher than 1 mM. After incubation of peroxisomes with palmitate, various acyl-CoAs were formed. Among medium-chain labelled acyl-CoAs, octanoyl-CoA was mainly detected. These results suggest that octanoyl-CoA accumulates during beta-oxidation of palmitate. When peroxisomes were incubated with [9,10-(3)H]palmitate and [9,10-(3)H]stearate, among medium-chain acyl-CoAs, octanoyl-CoA and decanoyl-CoA were primarily detected, respectively, suggesting the occurrence of at least 4 cycles of beta-oxidation of both fatty acids by peroxisomes.

Acyl Coenzyme A↗

PubMed2001

Cloning and characterization of a cDNA encoding beta-amyrin synthase involved in glycyrrhizin and soyasaponin biosyntheses in licorice.

An oxidosqualene cyclase cDNA, termed GgbAS1, was isolated from cultured cells of licorice (Glycyrrhiza glabra) by heterologous hybridization with cDNA of Arabidopsis thaliana LUP1 lupeol synthase. The yeast transformed with an expression vector containing the open reading frame of GgbAS1 produced beta-amyrin, indicating that GgbAS1 encodes beta-amyrin synthase involved in the glycyrrhizin and soyasaponin biosyntheses in licorice. Northern blot analysis showed that the level of beta-amyrin synthase mRNA was drastically changed in the cultured licorice cells, whereas the mRNA level of cycloartenol synthase was relatively constant.

Amino Acid Sequence↗

PubMed2001

Radiofrequency ablation at the coronary sinus ostium interrupts the vagal efferent input to the atrioventricular node in the canine heart.

The fat pad at the junction of the inferior vena cava and inferior left atrium is the area of convergence of vagal projections into the atrioventricular node (AVN) region. The present study investigated whether radiofrequency (RF) ablation applied to the area around the coronary sinus (CS) ostium would impair vagal input to the AVN in the canine heart. Twenty-four dogs were anesthetized by sodium pentobarbital and RF energy was delivered at 20W for 5-10s. In the baseline state without vagal stimulation (10Hz, 2ms), the electrophysiological variables did not change significantly after RF ablation. Vagally induced changes in the sinus cycle length and effective refractory period of the right atrium and left ventricle did not differ after RF ablation. However, the effects of vagal stimulation on the AVN function were impaired after RF ablation to the CS area from the ostium to 10mm within the ostium. After ablation was applied to the fast pathway area, the vagally induced changes in the AVN function decreased, but these changes were not affected after RF ablation in the slow pathway area. RF ablation in the vicinity of the CS would attenuate vagal input to the AVN.

Animals↗

PubMed2001

Transient prevention of ethanol-induced gastric lesion by capsaicin due to release of endogenous calcitonin gene-related peptide in rats.

Pre-exposure of the rat gastric mucosa to capsaicin reduced the mucosal lesion by 50% ethanol to 1/4. Treatment with an antagonist of calcitonin gene-related peptide (CGRP), CGRP (8-37), nullified the effect of capsaicin. During constant perfusion of the gastric lumen with physiological saline + pepstatin, the CGRP level was not increased by 50% ethanol, but it showed a peak (802.5 +/- 145.7 pg/2 min) after 1.6 mM capsaicin. Four minutes after capsaicin, the CGRP level was kept at a high level and the gastric lesion was markedly reduced by re-exposure of the mucosa to 50% ethanol. At 20-30 min after capsaicin, the CGRP levels returned to the resting level and the reddened area by 50% ethanol was not reduced. It was concluded that capsaicin transiently prevented the mucosal lesion through CGRP release.

Animals↗

PubMed2001

Importance of the carbohydrate-binding module of Clostridium stercorarium Xyn10B to xylan hydrolysis.

The Clostridium stercorarium xylanase Xyn10B is a modular enzyme comprising two thermostabilizing domains, a family 10 catalytic domain of glycosyl hydrolases, a family 9 carbohydrate-binding module (CBM), and two S-layer homologous (SLH) domains [Biosci. Biotechnol. Biochem., 63, 1596-1604 (1999)]. To investigate the role of this CBM, we constructed two derivatives of Xyn10B and compared their hydrolytic activity toward xylan and some preparations of plant cell walls; Xyn10BdeltaCBM consists of a catalytic domain only, and Xyn10B-CBM comprises a catalytic domain and a CBM. Xyn10B-CBM bound to various insoluble polysaccharides including Avicel, acid-swollen cellulose, ball-milled chitin, Sephadex G-25, and amylose-resin. A cellulose binding assay in the presence of soluble saccharides suggested that the CBM of Xyn10B had an affinity for even monosaccharides such as glucose, galactose, xylose, mannose and ribose. Removal of the CBM from the enzyme negated its cellulose- and xylan-binding abilities and severely reduced its enzyme activity toward insoluble xylan and plant cell walls but not soluble xylan. These findings clearly indicated that the CBM of Xyn10B is important in the hydrolysis of insoluble xylan. This is the first report of a family 9 CBM with an affinity for insoluble xylan in addition to crystalline cellulose and the ability to increase hydrolytic activity toward insoluble xylan.

Catalysis↗

PubMed2001

Increases of secondary metabolite production in various plant cell cultures by co-cultivation with cork.

Cork tissues increased secondary metabolite production of various plant cell cultures in a different manner from those of conventional elicitors. In Sophora flavescens and Glycyrrhiza glabra cultured cells, cork tissues increased the amounts of both lipophilic and hydrophilic flavonoids without affecting the cell growth, although elicitors such as copper ion and yeast extracts showed a clear inhibition of cell growth with the increasing amount of these lipophilic ones. The validity of this effect of cork tissues covered a wide range of aromatic compounds produced by suspension cell cultures derived from diverse plant species. Woody tissues of Japanese cypress had a very similar effect to that of cork. Partial purification of cork tissues suggested that the production-stimulating factor was present in the hemicellulose B fraction that was not included in the dedifferentiated cultured tissues.

Cell Wall↗

PubMed2001

Anti-tumor promoting activity of bufadienolides from Kalanchoe pinnata and K. daigremontiana x tubiflora.

Five bufadienolides (1-5) isolated from the leaves of Kalanchoe pinnata and K. daigremontiana x tubiflora (Crassulaceae) were examined for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation in Raji cells induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. All bufadienolides showed inhibitory activity, and bryophyllin A (1) exhibited the most marked inhibition (IC50 = 0.4 microM) among the tested compounds. Bryophyllin C (2), a reduction analogue of 1, and bersaldegenin-3-acetate (3) lacking the orthoacetate moiety were less active. These results strongly suggest that bufadienolides are potential cancer chemopreventive agents.

Animals↗

PubMed2001

Assessment of abdominal aortic aneurysms using a cone-beam CT system: an experimental phantom study and an initial clinical evaluation before and after stent-graft treatment in patients with an abdominal aortic aneurysm.

The aim of this study is to conduct a quantitative analysis of cone-beam CT (CBCT) images using a phantom, and then to evaluate the clinical usefulness of CBCT in the assessment of abdominal aortic aneurysms (AAA) before and after stent-grafting, both qualitatively as well as quantitatively. The phantom used in this study was a rectangular plate made of an acrylic resin, which contained eight through-holes to mimic blood vessels. Each columnar cavity was filled with contrast media and the diameter of each was then measured using a cone-beam multiplanar reformation/curved planar reformation (CB-MPR/CPR) technique, and the results were compared with the corresponding results obtained by actual measurement. In the clinical assessment, nine patients with AAA (consisting only of males with an average age of 68 years old: 56 approximately 80) were enrolled. The clinical qualitative analysis of CBCT consisted of: 1) for the pre-operative state, the shape of the aortic aneurysm, the relationship between the aneurysm and the aortic branches, and 2) for the post-operative state, the shape of the stent and any endoleakage present. The clinical quantitative analysis of CBCT included, for the aneurysm, its inflection angle, its maximum diameter, the diameter of the proximal and distal necks, and the distance of these two necks from specific reference points. The quantitative analysis using the phantom showed no significant differences between the results based on CB-MPR/CPR and those obtained by actual measurement. In the clinical qualitative analysis three-dimensional CBCT (3D-CBCT) depicted the anatomical relationship between the aneurysm and the aortic branches well, an accomplishment that was not possible by conventional angiography. Cone-beam maximum intensity projection (CB-MIP) was as good in tracing the migration and deformation of the stent following endovascular intervention as plain radiograms and conventional angiograms. CB-MPR/CPR enabled us to obtain any cross-sectional image of the aorta desired, including a curved, longitudinal cross-section of the aorta. Thus, with the CB-MPR/CPR technique it is easy to determine the distance of the proximal and distal necks of the aneurysm, and the inflection angle, and those results were not significantly different from those obtained by angiography. The diameter of the aneurysm, and the diameter of the proximal and distal necks as measured by CB-MPR/CPR images were significantly different from those obtained by conventional contrast enhanced-CT (p<0.05). This suggests that CB-MPR/CPR yields a cross-sectional view that is more perpendicular to the longitudinal direction of the aorta than that given by conventional contrast enhanced-CT, and thus provides a more accurate cross-sectional image of the aneurysm than the latter. We conclude that, in the experimental phantom study CBCT had a high quantitative reliability, and that, in the clinical study CBCT provided useful information for both qualitatively and quantitatively evaluating AAA before and after stent-grafting.

Aged↗

PubMed2001

Association of angiotensinogen gene polymorphism with erythropoietin-induced hypertension: a preliminary report.

The association of the angiotensinogen (AGT) gene variation at codon 235, the T235 variant, with hypertension induced by erythropoietin (Epo) was investigated in patients with progressive renal disease requiring treatment for renal anemia with Epo. The subjects for the study were patients with renal diseases with serum creatinine concentration exceeding 2 mg/dl and a hematocrit (Ht) of less than 30%. During the run-in period, blood pressure was well controlled with an appropriate salt restricted diet and/or antihypertensive treatment. The patients were then given 6,000 IU of Epo once a week until the Ht rose by 5%. For the overall patient group, AGT gene polymorphism analysis revealed T235T (T/T) in 31 cases (61%), M235T (M/T) in 19 cases (37%), and M235M (M/M) in 1 case (2%). In response to treatment with Epo, hypertension (defined as an increase in mean blood pressure greater than 10 mmHg) was found in 11 cases (22%), all of who carried the homozygous T allele (T/T). On the other hand, the frequency of T/T in patients who did not develop hypertension was 50% (T/T:T/M=20:19 cases), indicating a significant difference (p=0.003 by Chi-square). Variables estimated to be associated with Epo-induced hypertension were the T allele, gender (male), and the degree of increase in Ht, in descending order. Our preliminary research indicates that individuals who carry two copies of the T allele, i.e., who are homozygous for T, are highly susceptible to development of hypertension when subjected to Epo. These results suggest that the AGT T235 variant may be the primary gene responsible for the development of Epo-induced hypertension.

Adult↗