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Biomedical subjects

S Ishihara

Publications and source records attributed to S Ishihara.

At least 37 records · Page 2Linked to original sources

A role of glial fibrillary acidic protein in hippocampal degeneration after cerebral trauma or kainate-induced seizure.

Astrocytes perform a variety of functions in the adult central nervous system (CNS). Recent evidence suggests the robust upregulation of glial fibrillary acidic protein (GFAP) after CNS insult. However, little is known about the role of GFAP in the hippocampal degeneration after brain injury. We herein compared the GFAP knockout (KO) and wild type (WT) mice on the histological and behavioral outcome in response to cerebral trauma or kainic acid (KA)-induced seizure. Although all KO mice showed hippocampal CA3 neuronal degeneration. WT mice did not show any neuronal degeneration in CA3 subfield at 72 hrs after trauma. Thereafter, KO mice showed a higher susceptibility to KA-induced seizures and an increased number of pyknotic CA3 neurons 72 hrs after KA administration. These results indicate that GFAP plays a crucial role in the hippocampal neurodegeneration after CNS insult.

Animals↗

Expression of midkine and receptor-like protein tyrosine phosphatase (RPTP)-beta genes in the rat stomach and the influence of rebamipide.

BACKGROUND: Midkine has been reported to bind to receptor-like protein tyrosine phosphatase (RPTP)-beta and to play important roles in growth and differentiation of various cells. Midkine is expressed in rat stomach during experimental ulcer healing, suggesting that the midkine-RPTP-beta system has some physiological functions in the stomach. Rebamipide is a mucoprotective drug used for the treatment of gastric ulcers. We have tested the hypothesis that the ulcer healing mechanism stimulated by rebamipide is linked physiologically to the gastric midkine-RPTP-beta system. MATERIALS AND METHODS: Seven-week-old-male Wistar rats were used. Midkine and RPTP-beta gene expression in rat stomach was investigated by laser capture microdissection coupled with the reverse transcription-polymerase chain reaction (RT-PCR). The effects of rebamipide on midkine and RPTP-beta expression in rat stomach and the gastric epithelial cell line RGM1 were evaluated by RT-PCR and Northern blot analyses. RESULTS: Midkine and RPTP-beta expression was detected in the gastric mucosal, submucosal and muscle layers. Rebamipide stimulated both midkine and RPTP-beta expression in rat stomach and RGM1 cells. CONCLUSION: Rebamipide may protect the gastric mucosa by regulating midkine and RPTP-beta expression.

Alanine↗

Prediction of tumor radiosensitivity in rectal carcinoma based on p53 and Ku70 expression.

The identification of predictive indicators of radiosensitivity is extremely useful in selecting patients suited for preoperative radiotherapy and avoiding unnecessary preoperative treatment. In this study, we evaluated the possible role of the immunohistochemical expression pattern of p53 and Ku70 protein in determining tumor radiosensitivity in rectal cancer before preoperative irradiation. We examined pretreatment biopsy materials from 111 patients by immunohistochemistry. The expression pattern of p53 and Ku70 was evaluated for association with tumor radiosensitivity, which was defined according to the criteria of the Japanese Research Society for Cancer of the Colon and Rectum. There was a significant correlation between the expression pattern of p53 and tumor radiosensitivity (P = 0.045); Ku70 and tumor radiosensitivity (P < 0.001); and the combination of p53 and Ku70, and tumor radiosensitivity (P < 0.001). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in both p53 and Ku70-positive cases for radioresistance were all superior to those of the group positive for p53 alone. In conclusion the examination of the combination of p53 and Ku70 may predict the radiosensitivity of rectal cancer before preoperative irradiation.

Adult↗

Female gender as a risk factor for hypokalemia and QT prolongation after subarachnoid hemorrhage.

Subarachnoid hemorrhage (SAH) often causes hypokalemia and QT prolongation. The sex disparities among 73 patients with SAH and 96 control subjects were examined. On the first day of SAH, the mean serum potassium level was significantly lower and the mean corrected QT interval was significantly longer in the female SAH group compared with either the female control or male SAH group. These findings suggest female susceptibility to hypokalemia and QT prolongation after SAH.

Acute Disease↗

The static and cyclic strength of a bone-cement bond.

Four-point bending static and fatigue tests were carried out on bone-cement bonds. The effects of the pressurization and the washing of the bone joint face on the bond strength were investigated. The results are summarized as follows. When the bond surface of cancellous bone is washed prior to the application of the bone cement, both the static and fatigue strengths of the bond are increased relative to the corresponding properties of unwashed bone-cement bonds. From observations of bone-cement interfaces as well as the fracture surfaces of bone-cement specimens, it has been determined that bone cement was able to infiltrate into fine holes present in washed cancellous bone. However, such infiltration occurred to a much lesser degree in the case of unwashed cancellous bone. Increasing the molding pressure during the time of cement application to the bone from 39200 to 117600 Pa had a beneficial effect on the bending strength and fatigue properties, particularly in the case of washed bone cement specimens. An increase in molding pressure also resulted in a reduction in the amount of scatter in test results.

Journal Article↗

Time-course changes of ECL cell markers in acetic acid-induced gastric ulcers in rats.

BACKGROUND AND AIM: Enterochromaffin-like (ECL) cells are the major source of histamine for the regulation of gastric acid secretion, and also contain histidine decarboxylase (HDC), vesicular monoamine transporter 2 (VMAT2), and chromogranin A (CgA). Although gastric acid secretion is suppressed during ulcer healing, the role of ECL cells in that process is not yet fully understood. In the present study, we investigated the changes in ECL cell number during healing of experimental ulcers in rats. MATERIALS AND METHODS: Seven-week-old male Wistar rats were used. Acetic acid-induced ulcers were caused by an application of 100% acetic acid to the serosal surface of the rat stomachs. At different time points following the induction (12 h-15 days), time-course changes of HDC, VMAT2, and CgA mRNA expression were investigated by Northern blot analysis. The expressions of HDC, VMAT2, and CgA were immunostained on gastric mucosal sections with ulcers. RESULTS: HDC, VMAT2, and CgA mRNA in gastric mucosa each showed an initial marked transient decrease, followed by an increase on day 10 back to the initial value. HDC, VMAT2, and CgA-immunoreactive cells at the ulcer margin were reduced in number on day 3, compared with those in distant areas. On day 10, however, they returned to levels similar to those in distant areas. CONCLUSION: The present study revealed a local down-regulation of HDC, VMAT2, and CgA in ECL cells at the ulcer margin. As a result, we concluded that a suppression of ECL cell activity during ulcer healing may be involved in suppressed gastric acid secretion.

Acetic Acid↗

Activation of prostaglandin E2-receptor EP2 and EP4 pathways induces growth inhibition in human gastric carcinoma cell lines.

The effect of prostaglandin E2 (PGE2) on the proliferation of gastric cancer cells is still unclear. PGE2 receptors are divided into four subtypes - EP1, EP2, EP3, and EP4 - which are coupled to three different intracellular signal-transduction systems. Stimulation of EP2 and EP4 is linked with cyclic adenosine 3', 5'-monophosphate (cAMP)-dependent protein kinase A (PKA). In some human gastric cancer cells, PGE2 has been suggested to have an antiproliferative effect by way of increased cAMP production. Expression of EP2 and EP4 in human gastric carcinoma cells, however, has not been examined. We examined the expression of EP2 and EP4 and the antiproliferative effects of specific EP2 and EP4 agonists on four different human gastric cancer cell lines. Our data clarified that all the cell lines investigated in this study expressed EP2 and EP4 and that the specific agonists of these receptors induced growth inhibition with an accompanying increase in cAMP production. In summary, gastric cancer cells have EP2 and EP4 receptors, and their selective activation is linked with the decreased cell proliferation.

Cell Division↗

Peripheral blood dendritic cells, but not monocyte-derived dendritic cells, can augment human NK cell function.

Dendritic cells (DCs) are essential antigen-presenting cells with a wide variety of functions relating to both adaptive and innate immunity. Recently, interactions of DCs with natural killer (NK) cells and NK1.1-positive T cells have been reported in mice. However, in humans, this interaction is not well understood. Here we report the use of a coculture method to analyze the modulation of NK cell function in antitumor immunity by DCs. We found that peripheral blood DCs (PDCs) enhanced NK cell activity in cytotoxicity assay, even without direct contact between DC and NK cells. In contrast, neither monocyte-derived DCs (MoDCs), nor TNF-alpha-treated MoDCs, stimulated NK lytic activity. Secretion of IL-12 and TNF-alpha into the PDC-NK coculture supernatant was increased. However, blocking antibodies against these cytokines could not completely abolish the upregulation of NK activity, suggesting the presence of other soluble factor(s) that affect DC-NK cell interaction. To summarize, this study demonstrates for the first time the direct activation of human NK cells by DC-NK cell interaction in vitro, suggesting that DCs may have a central role linking the innate and adaptive immune responses. Moreover, in stimulating NK cell function, PDCs appear to have a different potential from MoDCs.

Cells, Cultured↗

In vitro anti-tumour activity of alpha-galactosylceramide-stimulated human invariant Valpha24+NKT cells against melanoma.

alpha-galactosylceramide (KRN 7000, alpha-GalCer) has shown potent in vivo anti-tumour activity in mice, including against melanoma and the highly specific effect of inducing proliferation and activation of human Valpha24+NKT-cells. We hypothesized that human Valpha24+NKT-cells activated by alpha-GalCer might exhibit anti-tumour activity against human melanoma. To investigate this, Valpha24+NKT-cells were generated from the peripheral blood of patients with melanoma after stimulation with alpha-GalCer pulsed monocyte-derived dendritic cells (Mo-DCs). Valpha24+NKT-cells did not exhibit cytolytic activity against the primary autologous or allogeneic melanoma cell lines tested. However, proliferation of the melanoma cell lines was markedly suppressed by co-culture with activated Valpha24+NKT-cells (mean +/- SD inhibition of proliferation 63.9 +/- 1.3%). Culture supernatants of activated Valpha24+NKT-cell cultures stimulated with alpha-GalCer pulsed Mo-DCs exhibited similar antiproliferative activities against melanoma cells, indicating that the majority of the inhibitory effects were due to soluble mediators rather than direct cell-to-cell interactions. This effect was predominantly due to release of IFN-gamma, and to a lesser extent IL-12. Other cytokines, including IL-4 and IL-10, were released but these cytokines had less antiproliferative effects. These in vitro results show that Valpha24+NKT-cells stimulated by alpha-GalCer-pulsed Mo-DCs have anti-tumour activities against human melanoma through antiproliferative effects exerted by soluble mediators rather than cytolytic effects as observed against some other tumours. Induction of local cytokine release by activated Valpha24+NKT-cells may contribute to clinical anti-tumour effects of alpha-GalCer.

Adjuvants, Immunologic↗

Peroxisome proliferator-activated receptor gamma ligand-induced growth inhibition of human hepatocellular carcinoma.

Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands have been implicated in the growth inhibition and differentiation of certain human cancers with diverse tissue origin. In this study, expression of PPARgamma in human hepatocellular carcinoma (HCC) and the effect of PPARgamma ligands on HCC cells were investigated in vitro using Hep G2, HuH-7, KYN-1 and KYN-2 cell lines. All cell lines were found to express functionally active PPARgamma and a marked growth inhibition was induced by thiazolidinedione ligands troglitazone, and pioglitazone as well as with its natural ligand 15-deoxy-Delta(12,14)-prostaglandin J(2). The growth inhibitory effect was associated with a dose-dependent inhibition of DNA synthesis, cell cycle progression and alpha fetoprotein expression.

Carcinoma, Hepatocellular↗

Stimulatory effects of centrally injected kappa-opioid receptor agonists on gastric acid secretion in urethane-anesthetized rats.

Gastric acid secretion has been proposed to be regulated by opioid receptors in the central nervous system (CNS). However, whether the effect of morphine is stimulatory or inhibitory, and the role of type specificity of opioid receptors have not been established. We investigated the effects of centrally injected opioid receptor agonists on gastric acid secretion in the perfused stomach of urethane-anesthetized rats. Injection of morphine (1-30 microg/rat, mu-opioid receptor agonist) into the fourth cerebroventricle inhibited the secretion stimulated by i.v. injection of 2-deoxy-D-glucose. Morphine itself did not show an inhibitory effect. In contrast, injection of kappa(1)-opioid receptor agonists such as (5alpha,7alpha,8beta)-(+)-N-methyl-N-(7-[1-pyrrolidinyl]-1-oxaspiro[4.5]dec-8-yl)benzeneacetamide (U59593, 0.3-3 microg) and (trans)-(+/-)-3,4-dichloro-N-methyl-N-(2-[1-pyrrolidinyl]cyclohexyl) benzeneacetamide hydrochloride (U50488H, 10 microg) and the kappa(2)-opioid receptor agonist, bremazocine (3 microg), into the lateral cerebroventricle markedly stimulated secretion. The effect of U59593 was inhibited by naloxone and norbinaltorphimine (an antagonist of kappa-opioid receptors) and in vagotomized rats. [D-Pen(2)-D-Pen(5)]enkephalin (10microg, delta-opioid receptor agonist) had no effect on secretion. The dual roles of the opioid system in the CNS in gastric acid secretion are discussed.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

Observation of orbital waves as elementary excitations in a solid.

A basic concept in solid-state physics is that when some kind of symmetry in a solid is spontaneously broken, collective excitations will arise. For example, phonons are the collective excitations corresponding to lattice vibrations in a crystal, and magnons correspond to spin waves in a magnetically ordered compound. Modulations in the relative shape of the electronic clouds in an orbitally ordered state could in principle give rise to orbital waves, or 'orbitons', but this type of elementary excitation has yet to be observed experimentally. Systems in which the electrons are strongly correlated-such as high-temperature superconductors and manganites exhibiting colossal magnetoresistivity-are promising candidates for supporting orbital waves, because they contain transition-metal ions in which the orbital degree of freedom is important. Orbitally ordered states have been found in several transition-metal compounds, and orbitons have been predicted theoretically for LaMnO3. Here we report experimental evidence for orbitons in LaMnO3, using Raman scattering measurements. We perform a model calculation of orbiton resonances which provides a good fit to the experimental data.

Journal Article↗

Analysis of gastrin receptor gene expression in proliferating cells in the neck zone of gastric fundic glands using laser capture microdissection.

Gastrin stimulates proliferation of progenitor cells in the neck zone of gastric fundic mucosa. However, whether it directly enhances this proliferation through its receptors remains unclear. We investigated the expression of gastrin receptors in neck zone proliferating cells in rat gastric fundic glands using a reverse transcription polymerase chain reaction (RT-PCR) coupled with laser capture microdissection and in situ RT-PCR. Gastrin receptor expression was identified in c-fos-expressing cells located in the neck zone, and results of the RT-PCR analysis argued against contamination by other cells, such as enterochromaffin-like, parietal or D cells. Supporting this finding, gastrin receptor gene expression was identified in the neck zone as well as base glands by in situ RT-PCR. Therefore, it is suggested that proliferating cells in the neck zone are stimulated directly by gastrin via their gastrin receptors.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Correlation between microsatellite instability and metachronous disease recurrence after endoscopic mucosal resection in patients with early stage gastric carcinoma.

BACKGROUND: Since endoscopic treatment was first evaluated and established as a treatment for patients with early stage gastric carcinoma, metachronous disease recurrence at other sites in the stomach after endoscopic treatment has become a major problem. METHODS: A retrospective case-control study was conducted on 10 patients with metachronous recurrence of gastric carcinoma after undergoing successful endoscopic mucosal resection (EMR) therapy for early stage gastric carcinoma and on 14 patients without recurrence. Gastric mucosal tissues obtained during the initial EMR were dissected, and DNA samples from the tumor tissue and surrounding nonneoplastic mucosa were extracted separately. Microsatellite instability (MSI) was tested in five microsatellite markers (D2S137, D3S1067, TP53, TGFbetaRII, and BAX). The authors also looked for K-ras codon 12 point mutations in the tumor tissues. In addition, immunohistochemical staining was done to test for the presence of proliferating cell nuclear antigen (PCNA), p53, hMSH2, and hMLH1 in the mucosal tissues. Finally, the correlation between the presence or absence of metachronous recurrence and the characteristics of the primary tumor (MSI, K-ras, p53, etc.) were investigated. RESULTS: Three of 10 patients with recurrent disease showed MSI in more than two microsatellite markers among 3-5 investigated site (MSI-H), whereas none of the patients with nonrecurrent disease did so. There was no significant correlation between metachronous recurrence after EMR and immunohistochemical staining reactions, including those for PCNA, p53, hMSH2, and hMLH1. None of the patients showed K-ras mutations. CONCLUSIONS: Thirty percent of patients with recurrent disease showed MSI-H, whereas none of the patients with nonrecurrent disease did so.

Adaptor Proteins, Signal Transducing↗

Co-infection by serologically-silent hepatitis B virus may contribute to poor interferon response in patients with chronic hepatitis C by down-regulation of type-I interferon receptor gene expression in the liver.

Intrahepatic mRNA levels of type-I interferon (IFN) receptor genes have been shown to correlate with the clinical efficacy of IFN therapy in patients with chronic hepatitis C. Recently, co-infection by serologically-silent hepatitis B virus (HBV) has been assumed to be associated with the poor IFN response in patients with chronic hepatitis C. The aim of this study was to investigate the relationship between the co-infection of serologically-silent HBV and type-I IFN receptor gene expression in the liver of patients with chronic hepatitis C. The intrahepatic mRNA levels of IFNAR2, one of the two subunits of the type-I IFN receptor, were quantified and compared with both the prevalence of HBV DNA and the hepatitis C virus (HCV) genotype in 45 patients with chronic hepatitis C, who were negative for hepatitis B surface antigen. Co-infection, as evaluated by a nested polymerase chain reaction, was present in 22 patients (48.9%), with dominance of the HCV genotype 1b (65.2%) over genotype 2a (31.8%). Co-infection was associated with lower IFNAR2 mRNA levels, higher levels of serum HCV RNA, and a poor IFN response, regardless of the HCV genotype. The findings suggest the possibility that co-infection by serologically-silent HBV is one of the factors that can lead to an unfavorable IFN response in chronic hepatitis C by down-regulation of IFN receptor gene expression in the liver.

Adult↗

A mutation in SPC42, which encodes a component of the spindle pole body, results in production of two-spored asci in Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, SPC42 is an essential gene, which encodes one of the major components of the spindle pole body (SPB). We report on a mutation in the SPC42 gene (spc42-102) that results in a sporulation-specific defect. Mitotic growth of haploid and diploid spc42-102 strains is normal and both exhibit the same growth rates as the isogenic wild-type strains. Many diploid spc42-102/spc42-102 cells undergo normal meiotic nuclear divisions, producing four haploid nuclei. However, a significant fraction of meiotic spc42-102/spc42-102 cells contain two immature SPBs and aberrant nuclei that are not surrounded by a prospore membrane. Some 40% of the resultant asci contain only two spores, while wild-type diploid cells almost always produce four-spored asci. Segregation of auxotrophic markers that are tightly linked to the centromere reveals that two-spore asci formed from spc42-102/spc42-102 diploid cells exclusively contain nonsister haploid spores. Western analysis and measurements of the fluorescent signal from an Spc42p-GFP (green fluorescent protein) fusion reveal that the mutant strain fails to accumulate Spc42p at meiosis. Thus, our results suggest that insufficiency of Spc42p during meiosis results in a pair of immature nonsister SPBs that are not enclosed by prospore membrane.

Blotting, Western↗

Nonrotation of the midgut with appendiceal mucocele in an adult.

Nonrotation of the midgut in adults and appendiceal mucocele are both rare pathological conditions. We report here the first case of nonrotation of the midgut associated with appendiceal mucocele. The patient was a 51-year-old man admitted to hospital with ileus. An upper gastrointestinal series and a barium enema revealed nonrotation of the midgut without midgut volvulus and with a mass in the ileocecal area. Laparotomy revealed an appendiceal mucocele adhering to the urinary bladder and the rectum. The mucocele had partly ruptured; yellowish mucinous material had entered the abdominal cavity, resulting in pseudomyxoma peritonei. Histological examination revealed a mucinous tumor of the appendix with borderline malignancy. In this patient, the ileus was caused by the appendiceal mucocele rather than being caused by nonrotation of the midgut.

Appendix↗