Search PubMed⌕ Search

Biomedical subjects

Shinichi Hayashi

Publications and source records attributed to Shinichi Hayashi.

15 recordsLinked to original sources
PubMed2026

Differential regulation of CYP46A1 in ischemic core and peri-infarct regions of male mouse brain after permanent middle cerebral artery occlusion.

Cholesterol 24-hydroxylase (CYP46A1) regulates brain cholesterol homeostasis and synaptic plasticity, playing a crucial role in ischemic stroke. Although previous studies have reported post-ischemic CYP46A1 upregulation, its spatiotemporal dynamics remain poorly defined. To elucidate these dynamics, we investigated the expression of CYP46A1 and other essential cholesterol homeostasis-related genes from 6 h to 3 days after permanent middle cerebral artery occlusion (pMCAO) in CB-17 mice. We utilized single-cell and single-nucleus transcriptomics, regional quantitative PCR, and high-resolution immunohistochemistry. CYP46A1 is predominantly expressed in neurons. Following ischemia, the cholesterol network exhibited a dynamic spatiotemporal divergence. Acutely (6 h post-ischemia), surviving regions transiently upregulated cell-autonomous cholesterol synthesis genes and CYP46A1. Subacutely (3 days), this response shifted toward a widespread upregulation of glia-dependent cholesterol transport genes and general CYP46A1 downregulation. At 24 h, CYP46A1 protein was substantially reduced in the necrotic core and superficial layer II/III of the peri-infarct cortex, but upregulated in deeper layer V, hippocampus, and lateral striatum. Notably, this localized upregulation spatially coincided with reactive microglial hypertrophy. These findings indicate that CYP46A1 is dynamically modulated in viable tissues following ischemic stress. This spatial divergence likely reflects a synergistic interaction between inflammatory propagation and neural circuit-mediated oxidative stress. Resolving these spatiotemporal profiles provides a rigorous foundation for evaluating CYP46A1 functionality and developing stage-specific therapeutic interventions.

Cholesterol 24-hydroxylase↗
PubMed2006

An inhibitory effect of A20 on NF-kappaB activation in airway epithelium upon influenza virus infection.

Influenza is a major disease in humans. The reemergence of avian influenza A viruses has indicated that hyperinflammatory responses are closely related to the severity of disease. Influenza virus infection induces nuclear transcription factor kappaB (NF-kappaB) activation. NF-kappaB and NF-kappaB-dependent gene products promote lung inflammation and injury. Therefore, it is important to investigate the means to attenuate NF-kappaB activation. A20 is a cytoplasmic zinc finger protein that inhibits NF-kappaB activity, However, little is known about the role of A20 in influenza virus infection. Here, we have examined the role of A20 in influenza virus infection-induced NF-kappaB promoter activation in human bronchial epithelial cells. The results showed that (1) A20 protein and mRNA are inducible and expressed in the lung from mice and human bronchial epithelial cells upon influenza virus infection; (2) NF-kappaB promoter activation was induced in bronchial epithelial cells upon influenza virus infection; and (3) overexpression by transient transfection of A20 attenuated NF-kappaB promoter activation in bronchial epithelial cells. These results indicate that A20 may function as a negative regulator of NF-kappaB-mediated lung inflammation and injury upon influenza virus infection, thereby protecting the host against inflammatory response to influenza virus infection.

Animals↗
PubMed2006

Postchemotherapy characteristics of hepatic colorectal metastases: remnants of uncertain malignant potential.

Accepted management for colorectal cancer (CRC) involves resection of the primary neoplasm and chemotherapy; the debate continues over the most beneficial order of these components. Preoperative chemotherapy aimed at liver metastases may result in complete pathologic response and replacement of the malignancy with scar. The McGill University liver diseases database was retrospectively reviewed. Forty-one patients receiving treatment between December 2003 and August 2004 were identified, their medical records examined, and liver histology reviewed. The histology of the remnants was linked to the appearance of the lesions on preresection imaging and to the primary colorectal neoplasms. Twenty-seven of the 41 patients (66%) received preoperative chemotherapy (oxaliplatin or irinotecan). Features of the primary neoplasm that predicted resolution of the metastases were absence of tumor budding (P = 0.04), absence of a diffusely infiltrative tumor margin (P = 0.02), and loss of expression of the DNA repair gene O6-methylguanine-DNA methyltransferase (P = 0.08). Oxaliplatin and irinotecan demonstrate beneficial effects in treating hepatic colorectal metastases and should be considered in such patients before resection. We propose the acronym RUMP to denote the remnants of uncertain malignant potential remaining. Further investigation is required to determine any correlation between the drug received and the resulting lesion.

Adult↗
PubMed2005

Correlation between DNA alterations and p53 and p16 protein expression in cancer cell lines.

To investigate the interaction between DNA abnormalities, p53 and p16 gene mutations, and methylation and protein expression, 20 cancer cell lines were examined by Western blotting. A clear relation was found to exist between p53 accumulation and mutation status. Of 20 cell lines examined, 14 demonstrated p53 homozygous mutations in exons 3-10, including 12 missense mutations, one nonsense mutation, and one frameshift mutation. Overexpression of p53 was always linked to missense mutations in exons 6-8. Intermediate expression of p53 was noted in cells with missense mutations or polymorphism to proline at codon 72 in exons 4-5, whereas there was slight or no visible expression in wild type cells and in cells with nonsense and frameshift mutations. DNA aberration in the p16 promoter gene correlated significantly with protein expression of the p16 suppressor gene. Overexpression was noted in six cell lines, intermediate expression in two, and slight or no visible expression in 12. Methylation-caused disappearance of p16 protein was noted in 40% (8/20) of the cell lines. Of six cell lines overexpressing p16 protein, two could be amplified with primers for both unmethylated and methylated forms in a methylation-specific RCP analysis. One cell line with no visible expression could also be amplified with both primers. Overexpression or disappearance of p16 protein may readily occur when one of two alleles has been methylated.

Biomarkers, Tumor↗
PubMed2005

Cyclooxygenase-2 in sporadic colorectal polyps: immunohistochemical study and its importance in the early stages of colorectal tumorigenesis.

Cyclooxygenase-2 (COX-2) is one of the important targets for the chemoprevention of colorectal cancer by non-steroidal anti-inflammatory drugs (NSAIDs). To evaluate the role of COX-2 in early stages of colorectal tumorigenesis, we immunohistochemically investigated the frequency and localization of COX-2 in sporadic colorectal polyps that showed various histology using a commercially available monoclonal antibody. A total of 105 colorectal polyps were examined. These included 33 low-grade adenomas (LGAs), 28 high-grade adenomas (HGAs), 32 HGAs with p53 overexpression (HGAs-p53), and 12 cases of carcinoma in adenoma (CIA). Regarding the immunohistochemical expression of p53, MIB-1, and CD63, histological classification was made for each case. COX-2 was expressed in neoplastic epithelial cells and interstitial macrophages that were distributed mainly in the superficial areas of polyps. COX-2 labeling indices (LIs) were 8.2% in LGAs, 6.3% in HGAs, 0.9% in HGAs-p53, and 0.6% in the carcinomatous components of CIAs. COX-2 LIs were significantly higher in adenomas, including LGAs and HGAs, than in HGAs-p53 and CIAs (p < 0.001). Within CIAs, significantly higher COX-2 LIs were obtained in the adenomatous components than in the carcinomatous components (p < 0.05). The size of polyps was not correlated with COX-2 expression irrespective of their histology. The results show that COX-2 might be involved in early stages of colorectal tumorigenesis. Colorectal adenomas could be a target for the chemopreventive strategy irrespective of their sizes.

Adenocarcinoma↗
PubMed2005

Fos plays no role in apoptosis of epithelia in the mouse male accessory sex organs and uterus.

Roles of Fos in apoptosis of epithelia in the mouse male accessory sex organs and uterus were investigated using Fos-deficient mice. Normal 30- and 50-day-old and Fos-deficient 50-day-old male and female mice were castrated, and testosterone propionate and estradiol-17 beta were daily injected into male and female mice, respectively, for 5 days. An apoptotic index (a percentage of apoptotic cells) in the epithelium was examined from the day following the last injection (day 1) to day 8. The body weights and the weights of the ventral prostate (VP), coagulating gland (C), seminal vesicle (SV) and epididymis (Ep) and uterus of 50-day-castrated Fos-deficient mice on day 1 suggested that the development of these mice corresponded to that of 30-day-castrated normal mice at the most. The extents of apoptosis estimated by an apoptotic index in the VP, C, SV, Ep and uterus in 50-day-castrated Fos-deficient mice were comparable to those in 30-day-castrated normal mice. The extents of apoptosis in the SV, Ep and uterus in 30-day-castrated normal and 50-day-castrated Fos-deficient mice were similar to those in 50-day-castrated normal mice, while the extents of apoptosis in the VP and C in the former two groups of mice were less than those in the latter mice. The present results show that Fos-deficiency does not affect apoptosis in the SV, EP and uterus. However, the extents of apoptosis in the VP and C were less in 50-day-castrated Fos-deficient mice than in 50-day-castrated normal mice. This seems to be due to the retarded development of 50-day-castrated Fos-deficient mice, but not to a role of Fos in apoptosis.

Animals↗
PubMed2005

[A case of metastatic submandibular lymphnode treated successfully with palliative oral (5-FU + PSK) chemotherapy in the elderly].

The patient was a 87-year-old woman diagnosed as type 2 advanced colon cancer in the ascending colon. The patient underwent right hemicolectomy. The pathological diagnosis showed poorly-differentiated adenocarcinoma, si, ly2, v1, n0 (0/41) and Stage IIIa. The postoperative course was uneventful, and she was discharged on POD 23. But a left submandibular lymph node enlarged rapidly within two months after the operation. Aspiration cytology of the lymph node indicated poorly-differentiated adenocarcinoma, and she was diagnosed as recurrent colon cancer. Combined chemotherapy of 5-FU (200 mg/day/po) and PSK (3.0 g/day/po) was started as palliative chemotherapy. The metastatic lymph nodes were reduced in size within two months after the treatment. Oral administration of 5-FU+PSK succeeded without serious adverse effects or worsening of quality of life. Ten months later, no recurrence was detected on physical examination or computed tomography. We conclude that palliative oral (5-FU+PSK) chemotherapy is useful for recurrent colon cancer in the elderly because of its excellent safety and effectiveness.

Adenocarcinoma↗
PubMed2004

FDG-PET imaging in duodenal cancer.

F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) imaging is a useful modality in the detection of various tumors, including colon cancer and gastric cancer. We evaluated a patient with duodenal cancer with multiple metastases including brain metastases using FDG-PET imaging. It revealed multiple tumor uptake in the brain, clavicular fossa, mediastinum, right adrenal gland and duodenum. These results suggest that FDG-PET imaging may be useful in detecting the primary and metastatic lesions of duodenal adenocarcinoma.

Adenocarcinoma↗
PubMed2004

Differential response of human lung epithelial cells to fas-induced apoptosis.

The Fas (CD95)/Fas ligand (CD178) system plays an important role in epithelial damage during the acute respiratory distress syndrome. The goal of this study was to determine whether proximal and distal human lung epithelial cells differ in their sensitivity to Fas ligand (rh-sFasL), and whether the response of lung epithelium to Fas ligation is modulated by proinflammatory cytokines. Although the expression of both Fas message and protein was similar in proximal and distal lung epithelial cells, only distal cells became apoptotic when exposed to serial dilutions of rh-sFasL. Stimulation with tumor necrosis factor-alpha, interleukin-1beta, or interferon-gamma significantly increased the sensitivity of proximal cells to rh-sFasL, and exposure to either tumor necrosis factor-alpha or interferon-gamma enhanced the sensitivity of distal cells to Fas ligation. These findings suggest that in normal human lungs, the responses of the epithelium to Fas ligation become more pronounced from proximal to distal locations. Furthermore, proinflammatory cytokines sensitize lung epithelium to Fas-induced death. These findings are relevant for understanding the role of the Fas/FasL system in acute lung injury, in which epithelial damage occurs primarily in distal airway and alveolar epithelium, whereas sFasL is present throughout the airspaces.

Antigens, CD↗
PubMed2004

Using a customized DNA microarray for expression profiling of the estrogen-responsive genes to evaluate estrogen activity among natural estrogens and industrial chemicals.

We developed a DNA microarray to evaluate the estrogen activity of natural estrogens and industrial chemicals. Using MCF-7 cells, we conducted a comprehensive analysis of estrogen-responsive genes among approximately 20,000 human genes. On the basis of reproducible and reliable responses of the genes to estrogen, we selected 172 genes to be used for developing a customized DNA microarray. Using this DNA microarray, we examined estrogen activity among natural estrogens (17beta-estradiol, estriol, estrone, genistein), industrial chemicals (diethylstilbestrol, bisphenol A, nonylphenol, methoxychlor), and dioxin. We obtained results identical to those for other bioassays that are used for detecting estrogen activity. On the basis of statistical correlations analysis, these bioassays have shown more sensitivity for dioxin and methoxychlor.

Biological Assay↗
PubMed2003

Apoptosis signal-regulating kinase 1-mediated signaling pathway regulates hydrogen peroxide-induced apoptosis in human pulmonary vascular endothelial cells.

OBJECTIVE: Reactive oxygen species initiate pulmonary vascular endothelial cell damage leading to an increase in endothelial permeability resulting in the production of pulmonary edema. Apoptosis signal-regulating kinase (ASK)-1 is a ubiquitously expressed mitogen-activated protein kinase kinase kinase (MAPKKK) that activates the MKK3/MKK6-p38 MAPK and the SEK1-c-Jun N-terminal kinase (JNK) signaling cascade. ASK1 has been implicated in cytokine- and stress-induced apoptosis. However, little is known about the role of ASK1 in apoptosis in hydrogen peroxide (H2O2)-stimulated pulmonary vascular endothelial cells and how ASK1-mediated apoptosis is executed. To clarify this issue, we examined the role of ASK1-p38 MAPK/JNK cascade in apoptosis and caspase-3 activation in H2O2-stimulated pulmonary vascular endothelial cells. DESIGN: Experimental laboratory study. SETTING: University laboratory. SUBJECTS: Normal human pulmonary artery endothelial cells. INTERVENTIONS: Western blot analysis and quantification of apoptosis in cells. MEASUREMENTS AND MAIN RESULTS: The results showed that H2O2 induced ASK1 phosphorylation and concomitantly p38 MAPK and JNK phosphorylation as well as induced caspase-3 activation in pulmonary vascular endothelial cells. To further characterize the role of ASK1 cascade in H2O2-induced apoptosis of pulmonary vascular endothelial cells, the dominant negative form of ASK1-stably transfected porcine artery endothelial cells was used. p38 MAPK and JNK phosphorylation, caspase-3 activation, and apoptosis in the dominant negative form of ASK1-stably transfected porcine artery endothelial cells were depressed compared with those in the parental porcine artery endothelial cells. CONCLUSION: ASK1-p38 MAPK/JNK cascade regulates apoptosis of H2O2-stimulated human pulmonary vascular endothelial cells.

Animals↗
PubMed2003

[A study of pharmacists' consciousness toward separation of medical practice from pharmaceutical dispensing].

Separation of medical practice from pharmaceutical dispensing had been a system with just the notation without the reality even after Showa era, although its concept was brought into Japan along with European medicine in Meiji era. Since 1970's, its concept has been prevailing gradually, and the so-called separation rate currently has reached to 47 percent. Increasingly keen attention has been paid recently by the public toward medicine in general. Taking into consideration these facts, a questionnaire survey was conducted for dispensing pharmacists in order to find out where pharmacists stand at present. Most of pharmacists (98.3%) answered that they practice patient counseling either by documents or by verbal communication, and that they take patient profiles. They said that patients could enjoy a proper medical treatment (32.9%) as well as a rational prescribing (14.3%). For a question asking what pharmacists should do to promote the separation of medical practice from pharmaceutical dispensing, 40.3% of them answered that they should make progress in their knowledge on drug therapy.

Adult↗
PubMed2002

Modified and simplified western blotting protocol: use of intermittent microwave irradiation (IMWI) and 5% skim milk to improve binding specificity.

A modified western blotting protocol was developed to increase the binding specificity of antigens and antibodies, using intermittent microwave irradiation (IMWI) with seven antibodies and two cell lines. The method was based on IMWI of the blotting membrane in the immunoblotting step using 5% skim milk as the diluting buffer. For some antibodies against p53, CDK4 and cyclinE, there were no distinct differences between the IMWI(+) and IMWI(-) counterparts; but improvement over the standard protocol was noted in both. For some antibodies, such as the polyclonal antibody against tubulin and the monoclonal antibodies against beta-tubulin, cyclinA and cyclinB1 (which were otherwise difficult to obtain good results with), IMWI was extremely effective, resulting in clear, specifically binding bands and a clean background. Moreover, the times were reduced from 8 to 3 h. Both the IMWI(+) and IMWI(-) protocols can be applied as simple, rapid and highly specific detection techniques for applications with various antigens, reducing background 'noise' to a minimum.

Animals↗