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Biomedical subjects

S Ito

Publications and source records attributed to S Ito.

At least 289 records · Page 16Linked to original sources

Helicobacter pylori CagA protein can be tyrosine phosphorylated in gastric epithelial cells.

Attachment of Helicobacter pylori to gastric epithelial cells induces various cellular responses, including the tyrosine phosphorylation of an unknown 145-kD protein and interleukin 8 production. Here we show that this 145-kD protein is the cagA product of H. pylori, an immunodominant, cytotoxin-associated antigen. Epithelial cells infected with various H. pylori clinical isolates resulted in generation of tyrosine-phosphorylated proteins ranging from 130 to 145 kD in size that were also induced in vitro by mixing host cell lysate with bacterial lysate. When epithelial cells were infected with [(35)S]methionine-labeled H. pylori, a radioactive 145-kD protein was detected in the immunoprecipitates with antiphosphotyrosine antibody or anti-CagA (cytotoxin-associated gene A) antibody. Consistently, the 145-kD protein recognized by the anti-CagA and antiphosphotyrosine antibodies was induced in epithelial cells after infection of wild-type H. pylori but not the cagA::Km mutant. Furthermore, the amino acid sequence of the phosphorylated 145-kD protein induced by H. pylori infection was identical to the H. pylori CagA sequence. These results reveal that the tyrosine-phosphorylated 145-kD protein is H. pylori CagA protein, which may be delivered from attached bacteria into the host cytoplasm. The identification of the tyrosine-phosphorylated protein will thus provide further insights into understanding the precise roles of CagA protein in H. pylori pathogenesis.

Amino Acid Sequence↗

SiO(x) Monolayer Overcoating Effect on the TiO(2) Photocatalytic Oxidation of Cetylpyridinium Bromide.

The effect of SiO(x) monolayer coverage on the rate of TiO(2) photocatalytic oxidation of cetylpyridinium bromide (CPB) in aqueous solutions has been studied. The rate of CPB removal from the solution (5<pH<7) increases with the surface modification at concentrations below 4.5x10(-4) M and its promoting effect is enhanced with decreasing concentration. A modified Langmuir-Hinshelwood model, where the rate of surface reaction is assumed to be proportional to the coverage of CPB at the photostationary state is proposed for dilute reaction systems. As a result of the kinetic analyses, the acceleration of the reaction with the SiO(x) monolayer coverage can be attributed to the increase in the rate of adsorption due to the electrostatic attraction of cetylpyridinium ion. The suppression of Br(-) adsorption is also suggested as a minor contribution. Copyright 2000 Academic Press.

Journal Article↗

Baroreflex dependent and independent roles of the caudal ventrolateral medulla in cardiovascular regulation.

The caudal ventrolateral medulla (CVLM) plays a critical role in cardiovascular regulation. Convincing data now support the hypothesis that inhibition of sympathoexcitatory neurons in the rostral ventrolateral medulla (RVLM) by CVLM neurons constitutes the necessary inhibitory link in baroreceptor reflex mediated control of sympathetic vasomotor outflow. Inhibition or destruction of the CVLM produces severe acute hypertension, consistent with blockade of baroreceptor reflexes and withdrawal of inhibition of RVLM sympathoexcitatory neurons. However, other data indicate that the CVLM also tonically inhibits RVLM sympathoexcitatory neurons in a manner not driven by baroreceptor input. In some studies, inhibition of the CVLM results in an increase in arterial pressure (AP) without inhibiting baroreceptor reflexes, possibly reflecting baroreceptor-independent and baroreceptor-dependent sub-regions of the CVLM. Furthermore, in baroreceptor-denervated rats, inhibition of the CVLM still leads to large increases in AP. In addition, in spontaneously hypertensive rats (SHR) central processing of baroreceptor reflexes appears normal but CVLM-mediated inhibition of the RVLM seems to be attenuated, suggesting that it is specifically a baroreceptor-independent mechanism of cardiovascular regulation in SHR that is altered. Taken together, these findings support an important, tonic, baroreceptor-independent inhibition of RVLM sympathoexcitatory neurons exerted by the CVLM.

Animals↗

Effects of inhibitors for tyrosine kinase and non-selective cation channel on capacitative Ca(2+) entry in rat ileal smooth muscle.

The effects of tyrosine kinase inhibitors and non-selective cation channel blockers on capacitative Ca(2+) entry were examined in the presence of methoxyverapamil in rat ileal smooth muscles. In Ca(2+)-free solution, carbachol or caffeine produced a rapid contraction mediated by Ca(2+) release from the stores (Ca(2+)-release response), and then led to Ca(2+) depletion of the stores. Subsequently, reintroduction of Ca(2+) caused a transient contraction due to capacitative Ca(2+) entry. Tyrosine kinase inhibitors, genistein and tyrphostin 47 but not herbimycin A, suppressed the responses to Ca(2+)-reintroduction much greater than Ca(2+)-release responses to carbachol or caffeine. Similar inhibitory effects on the responses to Ca(2+)-reintroduction were obtained with daidzein and tyrphostin A1, respective inactive analogue of genistein and tyrphostins. After continuous depletion of the stores with thapsigargin, Ca(2+)-reintroduction produced a sustained contraction, which was inhibited by these agents to different extents, but not by herbimycin A. In beta-escin-treated skinned muscles, genistein slightly reduced Ca(2+)-induced contraction. In fura-2-loaded tissues, SK&F 96365 inhibited contractile and [Ca(2+)](i) responses to Ca(2+)-reintroduction but minimally affected Ca(2+)-release responses. Tetrandrine suppressed both responses to Ca(2+)-reintroduction and to Ca(2+)-release. These results suggest that genistein and tyrphostin 47 inhibit capacitative Ca(2+) entry through an inhibition of Ca(2+) entry channels rather than tyrosine kinase. SK&F 96365, but not tetrandrine, seems to selectively inhibit the contractile responses to capacitative Ca(2+) entry in rat ileal smooth muscles.

Alkaloids↗

Epstein-barr virus nuclear antigen-1 binds to nuclear transporter karyopherin alpha1/NPI-1 in addition to karyopherin alpha2/Rch1.

We searched for cellular proteins that interact with Epstein-Barr (EBV) virus nuclear antigen-1, which is a latent EBV origin-binding protein detected in all EBV latently infected cells and essential for maintenance of the latent EBV genome, by a yeast two-hybrid screening of a B lymphocyte cDNA library in this study. Interaction of polypeptides synthesized from three selected cDNA clones with EBNA-1 proteins was confirmed in vitro using their glutathione-S-transferase-fusion polypeptides and by coimmunoprecipitation analyses of B cell extracts with anti-EBNA-1 monoclonal antibodies and monospecific antibodies against cellular proteins of interest. We report the following: (i) Karyopherin alpha (karyopherin alpha1, hSRP1, and NPI-1), an adaptor subunit of nuclear localization signal receptors, which direct proteins to the nuclear pore, interacted with EBNA-1. (ii) EBNA-1 proteins endogenous in the B cell line Raji of Burkitt lymphoma origin bound to another adaptor protein, karyopherin alpha2 (hSRP1alpha, hRch1), interactions of which to recombinant EBNA-1 polypeptides were previously reported. (iii) Nearly 90% of all the cDNA clones examined was p32 (SF2-associated P32, p32/TAP, and gC1q-R), and endogenous EBNA-1 proteins in the Raji cells bound to p32, a potential of which to affect localization of EBNA-1 in transfected Vero cells has been recently suggested. These results suggest that EBNA-1, which has the unique NLS containing Lys-Arg and overlapping with one of the phosphorylation domains, is recognized and transported to the nuclei by these two distinct karyopherin alpha proteins, which are differentially expressed in different cell types, implying a regulatory localization system for EBNA-1.

B-Lymphocytes↗

Physicochemical properties and bioavailability of carbamazepine polymorphs and dihydrate.

The dissolution behaviors of carbamazepine (CZP) polymorphs and pseudopolymorphs (form I, form III and dihydrate) and the bioavailabilities (BA) of each form in dogs after oral administration were investigated. Bioavailability tests were carried out at a dose of either 40 mg/body or 200 mg/body. The results of dissolution tests in JP13 first fluid (pH 1.2) at 37 degrees C indicated that the initial dissolution rate was in the order of form III>form I>dihydrate, while form III was transformed to dihydrate more rapidly than form I, resulting in decrease of the dissolution rate. The solubilities of both anhydrates (form I and form III), calculated from the initial dissolution rate of each anhydrate, were 1.5--1.6 times that of the dihydrate. At the dose of 40 mg/body, there were no significant differences in the area under the curve (AUC) between forms; their AUCs were nearly equal to that of CZP solution using polyethyleneglycol 400. These findings suggested that most crystalline powder of each form administered at the low dose was rapidly dissolved in gastrointestinal (GI) fluid. On the other hand, for the dose of 200 mg/body, significant differences in plasma concentration--time curves of CZP among polymorphic forms and dihydrate were observed. The order of AUC values was form I>form III>dihydrate. The inconsistency between the order of initial dissolution rates and that of AUC values at the high dose may have been due to rapid transformation from form III to dihydrate in GI fluids.

Algorithms↗

Human rhabdomyosarcoma cells retain insulin-regulated glucose transport activity through glucose transporter 1.

We evaluated the expression of glucose transporter (glut) isoforms and its function in RD cells, human rhabdomyosarcoma, which retain the potential to differentiate into muscle. Gluts 1, 3, and 4 were expressed in RD cells, as detected by reverse-transcription polymerase chain reaction and immunocytochemistry. Supraphysiological concentration (1 microM) of insulin treatment increased 2-deoxy glucose transport by up to 1.68-fold together with concomitant tyrosine phosphorylation of the insulin receptor beta subunit and of insulin receptor substrate 1. Suppression of glut 1 mRNA by 38% by antisense oligonucleotide transfection led to a reduction of basal and insulin-stimulated 2-deoxy glucose transport by 38 and 55%, respectively. Suppression of gluts 3 and 4 by antisense oligonucleotide transfection did not affect both basal and insulin-stimulated 2-deoxy glucose transport. Thus, glut 1 accounts for the major part of basal and insulin-stimulated glucose transport in RD cells. Next, we transfected expression vectors carrying human gluts 1 and 4 cDNAs into RD cells to add further support for the role of glut 1 in glucose transport. Overexpression of glut 1 stimulated basal and insulin-stimulated 2-deoxy glucose transport by 1.66- and 1.43-fold, respectively. Glut 4 overexpression did not affect basal and insulin-stimulated 2-deoxy glucose transport. Western blot analysis using glut 1 antibody showed that glut 1 was redistributed from intracellular membrane to plasma membrane. These observations support the notion that RD cells, with the potential to differentiate into muscle, retain insulin responsiveness. As human muscle cell lines are not available at this point, RD cells can serve as a useful alternative to human muscle for studies related to insulin signal transduction and glucose transport.

3-O-Methylglucose↗

Studies on some 3(2H)-pyridazinone derivatives with antinociceptive activity.

Nineteen new [6-(4-methoxyphenyl)-3(2H)-pyridazinone-2-yl]-acetamide (1-10) and 3-[6-(4-methoxyphenyl)-3(2H)-pyridazin-one-2-yl]propanamide (11-19) derivatives have been synthesized in this study. The structures of the compounds have been elucidated by their IR and NMR spectral data and elemental analysis. Antinociceptive activity of the compounds has been investigated by modified Koster's Test in mice, using aspirin as a reference. All the compounds (at 100 mg/kg dose) except 1 and 9 have been found more potent than aspirin. Compound 6 in the group of acetamide derivatives and compound 15 in the group of propanamides exhibited the highest antinociceptive activity. In addition, the propanamides have generally been found more potent than acetamides. In addition to these studies, the quantitative relationships between some structural parameters (such as log P, parachor, molar refractivity, and molecular connectivity indices) and antinociceptive activity of the compounds have been investigated. Statistical regression analysis has shown a close relationship to exist between the first-order molecular connectivity index (1 chi) and the antinociceptive activity.

Acetamides↗

Sandwich ELISA for the measurement of Apo-E4 levels in serum and the estimation of the allelic status of Apo-E4 isoforms.

Apolipoprotein-E (Apo-E) isoforms, especially theApo-E4 isoform, are considered to be genetic risk factors for familial as well as sporadic Alzheimer's disease (AD). Using monoclonal antibodies specific for Apo-E4 (MAb412-1-7) and Apo-E (LZ18), we established a sandwich ELISA for the measurement of Apo-E4 levels in serum. Total Apo-E also was determined by sandwich ELISA with a second pair of monoclonal antibodies, LZ12 and LZ18. Estimation of a patient's homozygous or heterozygous status for the Apo-E4 isoform was subsequently calculated as the ratio of Apo-E4/total Apo-E, respectively. Although the antibody for Apo-E used here does not have equivalent reactivity toward Apo-E2, -E3, or -E4, we were able to attain a fairly reliable assay system to estimate the status of the Apo-E4 isoform through a simple formula for the calculation of total Apo-E versus Apo-E4 isoform.

Alleles↗

Heterogeneity in ornithine cytotoxicity of bovine retinal pigment epithelial cells in primary culture.

Gyrate atrophy of the choroid and retina is a chorioretinal degeneration caused by hyperornithinemia and a deficiency of ornithine-delta-aminotransferase (OAT). We recently showed that ornithine exhibits cytotoxicity to human retinal pigment epithelial (RPE) cell lines treated with the OAT inhibitor, 5-fluoromethylornithine (5-FMOrn), and suggested that this system may be an in vitro model of gyrate atrophy. In the present study, in order to apply this system to primary cultured RPE cells, we freshly prepared RPE cells from bovine eyes and studied the effect of ornithine on cell damage. Two phenotypes, epithelioid and fusiform, which coexisted in the primary culture and epithelioid phenotype cells, but not fusiform ones, were severely damaged and partially detached from the substrate by 10 m m ornithine and 0.5 m m 5-FMOrn. Neither ornithine nor 5-FMOrn alone exhibited such cytotoxicity to both phenotypes of RPE cells. Proline significantly prevented the ornithine-induced cytotoxicity. Epithelioid and fusiform phenotypes isolated from the primary culture showed different distribution of actin filaments. A combination of ornithine and 5-FMOrn time-dependently inhibited [(3)H]thymidine incorporation in the epithelioid, but not fusiform, cells. Proline prevented the inhibition of [(3)H]thymidine incorporation by ornithine in 5-FMOrn-treated epithelioid cells. Furthermore, l -azetidine-2-carboxylic acid, a collagen synthesis inhibitor, reduced [(3)H]thymidine incorporation in epithelioid, but not fusiform, cells, which was reversed by proline. These results demonstrate that the epithelioid phenotype of bovine RPE cells becomes susceptible to ornithine following inactivation of OAT. The phenotypic cells and its prevention by proline may provide insight into biochemical triggers that induce gyrate atrophy.

Animals↗

Abnormal Peripheral Vasodilatory Reserve Without Endothelial Dysfunction in Patients With Aortic Dissection.

Structural changes in the aortic wall have been reported to be present in aortic dissection (AD), while there have been no investigations concerning peripheral vasomotion characteristics. Peripheral arterial stiffness is an important factor in the regulation of the central aortic pressure because it produces excessive wave reflection. The present study investigated in AD patients endothelium-dependent peripheral vasodilation and the reactive hyperemic response which is considered to be altered by the structural abnormality of the peripheral resistance artery. Forearm blood flow (FBF) changes induced by intra-arterial infusion of acetylcholine (ACh), sodium nitroprusside (SNP), and by occlusion-induced reactive hyperemia (RH) were measured plethysmographically in 10 AD patients, 7 healthy volunteers, and 7 patients with uncomplicated hypertension (UHT). There were no significant differences in the peak FBF response to ACh and SNP infusion for the three groups (ACh, 14.2 +/- 1.8 vs 17.2 +/- 4.5 vs 15.7 +/- 2.3 ml/min/dl tissue, NS; SNP 9.5 +/- 1.3 vs 10.6 +/- 1.7 vs 11.9 +/- 0.8 vs ml/min/dl tissue, NS). In the case of RH, however, peak FBF and maximum conductance were significantly lower in AD patients than in healthy volunteers and UHT patients (21.1 +/- 2.6 vs 36.1 +/- 4.7 and 32.4 +/- 2.6 ml/min/dl tissue volume, p < 0.01, and 0.25 +/- 0.03 vs 0.46 +/- 0.06 and 0.37 +/- 0.03 ml/min/dl per mmHg, p < 0.05, respectively). Peripheral vasodilatory function depending on the endothelium and smooth muscle in AD patients was not significantly different from that in healthy volunteers and UHT patients. However, reactive hyperemic vasodilatory reserve, which is a well-established, noninvasive measure of arterial structure, was significantly impaired in AD. These findings indicate that arterial structural abnormalities rather than systemic atherosclerotic changes represented by endothelial dysfunction may be present. We speculated that this aspect of peripheral resistance artery might possibly have an unfavorable effect on the postonset aortic conditions in AD patients.

Journal Article↗

Video-assisted mediastinoscopic resection of a thymic cyst by sternum lifting.

We performed mediastinoscopic surgery on two patients with a thymic cyst using a new instrument by sternum-lifting with tape. The merits of this technique are that a resection of the thymus and cyst can be achieved without a midline sternotomy, and under bilateral ventilation, and in cases with pleural adhesion. This technique can be applicable to most benign thymic lesions including thymectomy for myasthenia gravis and to selected cases with a malignant thymic tumor.

Aged↗

Clinical and histopathological features of colonic stromal tumor in a child.

Neoplasms of the colonic submucosa are rare in children. Gastrointestinal stromal tumors (GISTs) are undifferentiated tumors, usually diagnosed by immunohistochemistry. We report a 4-year-old girl with a submucosal GIST of the ascending colon, which was detected by computed tomography. Diagnosis after ileocecal resection was established by histology. In addition, sections were examined immunohistochemically, using antibodies against vimentin, desmin, alpha-smooth muscle actin, S100, neuron-specific enolase, c-kit, and CD34. Hematoxylin and eosin-stained sections showed interlacing fascicles with occasional palisades of epithelioid and spindle cells. The tumor cells were positive for vimentin and CD34. To our knowledge, this is the first reported case of colonic stromal tumor in a child.

Child, Preschool↗

Quantitative trait loci that modify the sootiness of yellow pigmentation in KK-A(y)/a mice.

Compared with C57BL/6J-A(y)/a, KK-A(y)/a mice have yellow fur that is markedly darker. Furthermore, there is a considerable variation in the tone of color with a continuous range in F(2) progeny produced from C57BL/6J females and KK-A(y)/a males. The aims of this study are to reveal the phenotypic differences between the two A(y) congenic strains and to elucidate the genetic factors responsible for the sooty yellow pigmentation in the KK background. On the basis of a chemical analysis, the sootiness in KK-A(y)/a was the result of increased eumelanin (PTCA) and decreased pheomelanin (AHP). A statistically significant QTL was identified on Chromosome (Chr) 15, responsible for the AHP content. No significant loci responsible for PTCA were identified. On the other hand, on the basis of an optical analysis for color difference and overall sootiness, significant evidence of linkage was identified on the proximal part of Chr 15, in the region similar to AHP QTL. The overall sootiness is thus controlled solely by the locus on Chr 15 in F(2) progeny; however, the KK allele at this locus significantly increased the AHP content.

Alleles↗

A giant urinoma in a neonate without obstructive uropathy.

We report a neonate with a giant urinoma and renal failure. A 7-day-old boy had a giant abdominal mass of 6.5 cm x 8 cm in the right quadrant, gastroesophageal reflux, and renal failure caused by the compression from the mass. Radiological observations revealed a multiseptated cyst and neither kidney could be detected. To relieve these symptoms percutaneous drainage was performed. The contents of the fluid were similar to the patient's urine. The symptoms were improved by the drainage, and we found the left kidney to be absent and the right kidney small. Four prenatal ultrasound scans detected no cystic lesions in his abdomen. Neonatal urinomas are commonly complicated by obstructive uropathy, such as posterior urethral valves or ureteropelvic junction obstruction. These obstructive uropathies were ruled out by retrograde pyelography and voiding cystourethrography. A severely dilated upper pole of a double collecting system was also ruled out by intravenous pyelography and direct observation of the kidney during an open biopsy. The cause of the urinoma is still uncertain, but trauma during delivery and the dysplastic right kidney may be involved.

Cysts↗

Early diagnosis of interferon-induced myocardial disorder in patients with chronic hepatitis C: evaluation by myocardial imaging with 123I-BMIPP.

Interferon (IFN) therapy for chronic hepatitis C is sometimes associated with cardiac complications. In the present study, we performed myocardial imaging with 123I-labeled beta-methyl-p-iodophenylpentadecanoic acid (123I-BMIPP) in order to evaluate myocardial disorders caused by IFN. We studied 40 healthy subjects (H group) and 25 patients with chronic hepatitis C who had been treated with IFN (IFN group). A Holter electrocardiogram (ECG) was performed and the autonomic nervous function was assessed by analyzing the spectral variability and 1/f fluctuation of heart rate. Myocardial planner imaging with 123I-BMIPP was performed to obtain the time activity curve for 20min immediately after administration of 123I-BMIPP (dynamic study). Early and delayed myocardial single photon emission computed tomography (SPECT) images were expressed as Bull's eyes and the myocardium was divided into four segments to calculate the washout rate for each segment on early and late SPECT images (early and late SPECT study). No significant differences in autonomic nervous function were observed between the two groups in heart rate variability. In a dynamic study, the reduction rate from the time activity curve was significantly higher in the IFN group compared with the H group (reduction rate, IFN group, 5.3 +/- 3.7% vs H group, 1.2 +/- 3.3%; P < 0.05). In the early and delayed myocardial SPECT study, the washout rate for the IFN group was significantly increased in all myocardial areas compared to that in the H group. However, the metabolic disorder of fatty acids caused by IFN was reversed on the second 123I-BMIPP myocardial scintigraphy examination several months after IFN therapy. These results indicate that metabolic disorders of fatty acids caused by IFN therapy can be detected before abnormalities are observed by Holter-ECG or echocardiography.

Antiviral Agents↗

Effects of cyclosporin A on water-immersion stress-induced gastric lesion and gastric secretion in rats.

Cyclosporin A is an immunosuppressive agent which is well known as a specific inhibitor of calcineurin (protein phosphatase 2B). In this study, we investigated the effects of cyclosporin A on water-immersion stress-induced gastric ulcer formation and gastric acid secretion in rats. We also examined the localization of calcineurin immunohistochemically. Calcineurin was specifically expressed in gastric parietal cells and chief cells of the gastric mucosa. The intraperitoneal administration of cyclosporin A dose-dependently suppressed the development of gastric mucosal lesions induced by water-immersion stress and inhibited gastric acid secretion, as assessed by pylorus ligation. These results indicated that calcineurin may play an important role in gastric acid secretion.

Animals↗