Biomedical subjects

A Hayashi

Publications and source records attributed to A Hayashi.

At least 163 records · Page 9Linked to original sources

PubMed1997

Caspase-dependent apoptosis of COS-7 cells induced by Bax overexpression: differential effects of Bcl-2 and Bcl-xL on Bax-induced caspase activation and apoptosis.

Bcl-2 family proteins and ICE/CED-3 family proteases (caspases) are regarded as the basic regulators of apoptotic cell death. They are evolutionarily conserved and implicated in a variety of apoptosis. However, the precise mechanism by which these two families interact to regulate cell death is not yet known. In this study, we found that the overexpression of the Bcl-2 family member Bax induced apoptotic cell death in COS-7 cells through the activation of CPP32 (caspase-3)-like proteases that cleaved the DEVD tetrapeptide. This apoptotic cell death was suppressed by the viral proteins CrmA and p35, as well as by the chemically synthesized caspase inhibitors Z-Asp-CH2-DCB and zVAD-fmk. We also found that the Bax-induced apoptosis of COS-7 cells was suppressed by Bcl-xL and Bcl-2, though both Bcl-xL and Bcl-2 similarly prevented etoposide-induced apoptosis in COS-7 cells. In addition, Bcl-xL inhibited the activation of caspase-3-like proteases accompanying Bax-induced COS-7 cell death but Bcl-2 did not. These results indicate that the caspase activation is essential for Bax-induced apoptosis, and that the ability of Bcl-2 and Bcl-xL to prevent the Bax-induced caspase activation and apoptosis in COS-7 cells could be differentially regulated. Our results also suggest that Bcl-2 family proteins function upstream of caspase activation and control apoptosis through the regulation of caspase activity.

Animals↗

PubMed1997

Complementary DNA cloning and chromosomal mapping of a novel phosphatidylinositol kinase gene.

A cDNA for a putative new member for phosphatidylinositol kinase family was cloned from an adult human whole brain cDNA library. The predicted translation product was composed of 961 amino acid residues and contained a sequence feature characteristic for lipid/protein kinases. The messenger RNA was ubiquitously expressed in various tissues, while relatively higher expression was observed in heart, skeletal muscle and testis. The chromosomal location of the gene was determined by fluorescence in situ hybridization and PCR-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel.

1-Phosphatidylinositol 4-Kinase↗

PubMed1997

HLA-A locus-restricted and tumor-specific CTLs in tumor-infiltrating lymphocytes of patients with non-small cell lung cancer.

HLA class I restriction and tumor specificity of cytotoxicity in the IL-2-activated tumor-infiltrating lymphocytes from 16 patients with non-small cell lung cancer were investigated. Six HLA class I-restricted and tumor-specific CTL lines were established: (i) HLA A2-restricted and adenocarcinoma-specific CTLs in three (two A0201+ and one A0206+) patients with adenocarcinoma, (ii) HLA A3101- and A3302-restricted and adenocarcinoma-specific CTLs in an HLA A3101/3302+ patient with adenocarcinoma, and (iii) HLA A3302-restricted CTLs and (iv) HLA A2402-restricted CTLs recognizing tumors with different types of histology in an HLA A3302+ patient with adenocarcinoma and an HLA A2402+ patient with squamous cell carcinoma (SCC), respectively. The three HLA A2-restricted CTL lines recognized 4, 4, or 6 of 15 HLA A2+ adenocarcinoma cell lines that originated from lung, stomach, colon, and breast with different subtypes (HLA A0201, A0206, and A0207), respectively. Furthermore, the CTLs of an HLA A0206+ patient recognized five different fractions of peptides eluted from an HLA A0201+ adenocarcinoma cell line. These results showed evidence of the existence of HLA class I-restricted and tumor-specific CTLs recognizing peptide antigens on HLA-A alleles of adenocarcinoma or SCC in tumor sites of a substantial number of patients with non-small cell lung cancer.

Adenocarcinoma↗

PubMed1997

Role of protein tyrosine phosphorylation in rat corneal neovascularization.

BACKGROUND: Recent studies have suggested that tyrosine kinase pathways that are activated by angiogenic growth factors may play a role in corneal neovascularization. METHODS: Corneal neovascularization was induced in rat corneas by chemical cauterization. At 6, 24, 48, 96, and 168 h after chemical cauterization the rat corneas without the corneal epithelium were prepared for gel electrophoresis. Total protein profiles of the corneal samples were examined by staining gels with Coomassie brilliant blue. Tyrosine-phosphorylated proteins, three angiogenic growth factors (basic fibroblast growth factor, vascular endothelial growth factor, and platelet-derived growth factor-B chain), and three intracellular signal proteins in the tyrosine kinase pathways (phospholipase C gamma, SHC, and mitogen-activated protein kinase) in the corneal samples were examined by western blot analysis. A topical treatment of genistein eye drop (5 mg/ml) was used for inhibition of corneal neovascularization after chemical cauterization in rats. RESULTS: In total protein profiles, three bands in the corneal samples were increased after cauterization. Overall tyrosine-phosphorylated proteins and all three angiogenic growth factors increased with progression of corneal neovascularization. The tyrosine-phosphorylated forms of three intracellular signal proteins were also increased after cauterization. Treatment with topical genistein was effective in inhibiting corneal neovascularization in rats. CONCLUSION: Protein tyrosine phosphorylation was involved in inflammation-induced corneal neovascularization. Tyrosine kinase inhibitors may have utility as inhibitors of corneal neovascularization.

Animals↗

PubMed1997

Mediastinal parathyroid adenoma detected by 99mTc-methoxyisobutylisonitrile: report of a case.

We report herein the case of a 30-year-old man in whom ectopic mediastinal parathyroid adenoma was detected by 99mTc-methoxyisobutylisonitrile scintigraphy (99mTc-MIBI). The patient presented with a history of recurrent renal stones and was diagnosed as having primary hyperparathyroidism due to elevated serum levels of calcium and intact parathyroid hormone (i-PTH) levels. On admission, his serum calcium value was 12.1 mg/dl and the i-PTH level was 137 pg/ml. Ultrasonography, computed tomography (CT), and 201TlCl-99mTcO4- subtraction scintigraphy (Tl-Tc) were performed but none of these imaging techniques was able to detect an enlarged parathyroid gland. Selective venous sampling revealed that the serum i-PTH level was highest at 422 pg/ml in the left brachiocephalic vein. Exploration of the neck also proved unsuccessful. Finally, we performed 99mTc-MIBI which revealed and delineated an enlarged parathyroid gland in the upper mediastinum. Tumor extirpation was subsequently performed through a left thoracotomy. Pathologically, the tumor was confirmed to be a parathyroid adenoma and the serum calcium and i-PTH levels returned to within the normal ranges postoperatively.

Adenoma↗

PubMed1997

Cervical cord ependymoma with numerous microrosettes.

"Microrosette ependymoma," which is ependymoma with numerous microrosettes throughout the tumor, has rarely been reported. We describe an autopsy case of cervical cord ependymoma with two unusual features: the presence of numerous microrosettes and the formation of trabecular architecture. The tumor originated in the C2 segment of a man aged 23 years and gradually expanded over the following 15 years and 10 months until the entire cervical cord was involved. Beside the low grade of malignancy, the tumor cells exhibited a strong tendency to form microrosettes and trabecular architecture, which formed many perivascular pseudorosettes. The microrosettes mostly consisted of only two or a few more cells, in the absence of large rosettes. Thus the constituent cells were those forming perivascular pseudorosettes. Electron microscopy and immunohistochemistry characterized the ependymal properties of the microrosettes, whose lumina frequently contained fibril bundles similar to those of the Reissner's fiber fibrils, in addition to cilia and microvilli. The pathogenesis of the occurrence of numerous microrosettes is unknown; however, a defect in the mechanism of regulation of rosette formation and enlargement is the most likely explanation.

Adult↗

PubMed1997

Stepladder V-Y advancement flap for repair of postero-plantar heel ulcers.

Random fasciocutaneous flaps with bilateral adipofascial pedicles were elevated from the posterior heel and transferred distally to defects of the postero-plantar heel in 4 patients, using a stepped incision technique, in combination with the V-Y advancement principle. In all cases the flap was successfully transferred without any linear scar or scar contracture in the posterior heel. The absence of recurrence of ulcers during the postoperative follow-up between 1.5 and 4 years indicated the reliability and durability of the flaps. Application of this procedure permits rapid resurfacing and excellent recontouring of small to moderate-sized defects of the heel with minimal donor site morbidity.

Aged↗

PubMed1997

Induction of autoimmune-like hepatic and ductal lesions by administration of lipopolysaccharide in mice undergoing graft-versus-host reaction across MHC class I difference.

In this paper, we examined the induction of autoimmune-like histologic changes in the liver and other organs of mice undergoing graft-versus-host reaction (GVHR) with MHC class I disparity by the administration of bacterial lipopolysaccharide (LPS), on the assumption that stimulation with LPS could be an exacerbating factor. Spleen cells of C57BL/6 (B6) mice were injected twice into (B6 x bml) F1 recipient mice at an interval of 7 days to induce MHC class I GVHR and then challenged with 1 microg of LPS intravenously on the next day of the cell transfer. The hepatic lesions of the group of MHC class I GVHR mice challenged with LPS showed marked cellular infiltration at the portal area and focal necrosis was observed in the hepatic lobule. The major infiltrating cells were CD8+, and others including CD4+ cells being of minor populations. In addition, ductal lesions in extrahepatic organs, including the pancreas and salivary glands also showed marked cellular infiltration. Thus, we have demonstrated that LPS induced ductal lesions in mice with MHC class I disparity. CD8+ cells were detected at the destructive hepatic lesions, which might be effector cells. These findings indicate that LPS might be one of the potential factors which augment autoimmune-like lesions.

Alanine Transaminase↗

PubMed1997

Elastin expression is up-regulated by retinoic acid but not by retinol in chick embryonic skin fibroblasts.

Effects of retinoid derivatives (retinol and retinoic acid) on elastin expression and cell proliferation in chick embryonic skin fibroblasts were studied. Retinoic acid inhibited cell proliferation one half of control at the concentration 10(-5) M. Retinoic acid exhibited stimulatory effect on elastin synthesis with a maximum stimulation of 2.0-fold at the concentration of 10(-6) M for 24 h treatment. Elastin level detected by Western blot analysis in the pooled conditioned medium was compatible with the increase of elastin synthesis by retinoic acid treatment. Comparable increase in elastin mRNA level was observed by retinoic acid treatment. Retinol showed no significant effects on either cell proliferation or elastin expression. The results indicate that retinoic acid was potentially active and retinol was inactive for modulation of cell proliferation and elastin expression.

Animals↗

PubMed1997

Japanese families with autosomal dominant pure cerebellar ataxia map to chromosome 19p13.1-p13.2 and are strongly associated with mild CAG expansions in the spinocerebellar ataxia type 6 gene in chromosome 19p13.1.

Autosomal dominant cerebellar ataxia is a group of clinically and genetically heterogeneous disorders. We carried out genomewide linkage analysis in 15 families with autosomal dominant pure cerebellar ataxia (ADPCA). Evidence for linkage to chromosome 19p markers was found in nine families, and combined multipoint analysis refined the candidate region to a 13.3-cM interval in 19p13.1-p13.2. The remaining six families were excluded for this region. Analysis of CAG-repeat expansion in the alpha1A-voltage-dependent calcium channel (CACNL1A4) gene lying in 19p13.1, recently identified among 8 small American kindreds with ADPCA (spinocerebellar ataxia type 6 [SCA6]), revealed that 8 of the 15 families studied had similar, very small expansion in this gene: all affected individuals had larger alleles (range of CAG repeats 21-25), compared with alleles observed in neurologically normal Japanese (range 5-20 repeats). Inverse correlation between the CAG-repeat number and the age at onset was found in affected individuals with expansion. The number of CAG repeats in expanded chromosomes was completely stable within each family, which was consistent with the fact that anticipation was not statistically proved in the SCA6 families that we studied. We conclude that more than half of Japanese cases of ADPCA map to 19p13.1-p13.2 and are strongly associated with the mild CAG expansion in the SCA6/CACNL1A4 gene.

Adult↗

PubMed1997

Experience of expandable metallic stents for central airway obstruction.

The aim of this study was to investigate retrospectively the efficacy of expandable metallic stents (EMSs) for severe respiratory distress in patients with central airway obstruction. Twenty patients with central airway obstructions were treated with an EMS. An intraluminal stricture was present in 15 and an extraluminal stricture in 5 patients. Of the 15 patients with intraluminal stenosis, 11 exhibited symptomatic improvement. All 11 patients had tumor infiltration occupying less than 50% of the endoluminal diameter. The other four patients with intraluminal stenosis had tumor infiltration occupying > 50% of the endoluminal diameter and demonstrated no improvement. All five patients with extraluminal stenosis were improved. EMS is useful for an extraluminal stricture in the central airway and the effect of EMS for intraluminal stenosis is related to the degree of infiltration and of tumor progression itself.

Adult↗

PubMed1997

Role of Helicobacter pylori infection in perforation of peptic ulcer: an age- and gender-matched case-control study.

Evidence showed a marked decrease in recurrence rate of peptic ulcer after eradication of Helicobacter pylori infection. However, whether H. pylori infection is etiologically related to perforation of peptic ulcer remains to be clarified. We therefore conducted an age- and gender-matched case-control study between perforated and nonsurgical peptic ulcers in H. pylori infection and examined differences in the cytotoxin genes cagA and vacA. Serum H. pylori IgG antibody (ELISA) was positive in 20/21 (95%) of perforated vs. 37/40 (93%) of nonsurgical duodenal ulcers and in 5/5 (100%) of perforated vs. 24/28 (86%) of nonsurgical gastric ulcer patients. Positivity of H. pylori DNA in gastric juice, which was amplified by PCR and identified by Southern blot hybridization, was 17/23 (74%) of perforated vs. 32/45 (71%) in the nonsurgical duodenal ulcer group. Positivity of the cytotoxin genes cagA and vacA in H. pylori DNA-positive gastric juice was as follows: perforated vs. nonsurgical duodenal ulcer, cagA 11/ 13 (85%) vs. 24/27 (89%); vacA1: 9/13 (69%) vs. 22/27 (82%); vacA2 8/13 (62%) vs. 21/27 (78%). There were no significant differences between the perforated and nonsurgical peptic ulcer groups for these H. pylori serum and gene markers. It is assumed that H. pylori infection is not etiologically related to perforation of peptic ulcer.

Antigens, Bacterial↗

PubMed1997

Detection of MAGE-4 protein in sera of lung cancer patients.

We investigated the level of MAGE-4 protein in sera of patients with primary lung cancer to understand better the biological roles of the MAGE proteins. MAGE-4 protein was detected as a non-degraded form in both the supernatant of a MAGE-4+ tumor cell line and in a patient's serum. Serum level of the MAGE-4 protein in lung cancer patients (n=100, mean=1.17 ng/ml) was significantly higher than that in either patients with benign pulmonary diseases (n=80, 0.33 ng/ml) or healthy donors (n=68, 0.32 ng/ml). It was higher than the cutoff level (1.15 ng/ml) in 34 of 100 cancer patients, but not in anyone in the other groups.

Adult↗

PubMed1997

Cytotoxin genes of Helicobacter pylori in chronic gastritis, gastroduodenal ulcer and gastric cancer: an age and gender matched case-control study.

Helicobacter pylori (H. pylori) infection is involved in many gastrointestinal diseases, such as chronic gastritis (CAG), peptic ulcer and gastric cancer (GCA). Both host factors and H. pylori strain differences may contribute to differences in the diseases. Thus, we conducted an age and gender matched case-control study of 35 patients each with CAG, gastric ulcer (GUL), duodenal ulcer (DUL) and gastric cancer (GCA) to examine the role of strain differences of the H. pylori cytotoxin genes cagA and vacA in these diseases. We employed polymerase chain reaction to examine the gastric juice for H. pylori DNA. The test was positive for 26 (74.3%) CAG, 29 (82.9%) GUL, 28 (80.0%) DUL and 27 (77.1%) GCA patients, showing no statistically significant difference among the diseases (P = 0.84). cagA and vacA genes (picked up by using a vacA1 + vacA2 primer pair which detected non-variable regions of the vacA gene) were detected by PCR in the H. pylori DNA-positive cases as follows: CAG, 92.3% and 76.9%; GUL, 100% and 86.2%; DUL, 89.3% and 89.3%; GCA, 92.6% and 85.2%, respectively. No statistically significant differences were found in the frequencies of these cytotoxin genes in H. pylori-positive cases among the various gastric diseases (P = 0.39 for cagA and P = 0.64 for vacA).

Adult↗

PubMed1997

Frequent occurrence of hepatocytic apoptosis in acute rejection of the grafted rat liver.

The present study was conducted to examine the histopathological changes of rat liver grafts in the early post-transplantation period. A total of 44 orthotopic liver transplantations were performed using cuff techniques without anastomosis of the hepatic artery. They were divided into four groups: (i) group 1 (syngeneic, Lewis-->Lewis; n = 10); (ii) group 2 (allogeneic, ACI-->Lewis; n = 20); (iii) group 3 (allogeneic with immunosuppression, ACI-->Lewis; n = 12); and (iv) long-surviving grafts (PVG-->Lewis; n = 2). The histological findings were classified into four categories: (i) mild dilatation of Disse's spaces due to ischemia; (ii) bile duct proliferation; (iii) acute rejection; and (iv) zonal necrosis. Bile duct proliferation occurred on day 6 in all three groups and in one of the two long-surviving grafts (120 days). Acute rejection occurred on day 3 and progressed in groups 2 and 3. Zonal necrosis developed in group 2 after day 6, while acute rejection subsided after day 4 in group 3 receiving 15-deoxyspergualin. A few single cell deaths or acidophilic bodies were variably noted. Most of these cells showed signals by in situ nick end-labeling in their nuclei, implying the cells were undergoing apoptosis. The number of apoptotic hepatocytes increased as the progress and decreased with the regression of acute rejection in group 3. Thus, the extent of hepatocytic apoptosis may reflect the magnitude of acute rejection. More frequent distribution of apoptotic hepatocytes in periportal areas suggests that apoptosis may be induced by a variety of pathological stimuli, including the direct cytotoxic effects of lymphocytes, various cytokines and local circulatory disturbances.

Animals↗

PubMed1997

[Serotype determination of enteroviruses that cause hand-foot-mouth disease; identification of enterovirus 71 and coxsackievirus A16 from clinical specimens by using specific probe].

Coxsackievirus A16 (CA16) and enterovirus 71 (EV71) are known to be major causative agents of hand-foot-and-mouth disease prevalent in summer in Japan. Discrimination and identification of these viruses were often hampered by a nonneutralizable or nontypable virus. Therefore, a Southern blot hybridization that utilizes mixed probes specific to serotype was developed. Firstly, an approximately 650 bases spanning 5'-noncoding region to one third of VP2 including entire VP4 was amplified with a set of primers containing enterovirus common sequences and a genomic RNA as template. Secondary, the nucleotide sequences were determined using seven CA16 and eighteen EV71 strains including the standard strains, and the deduced amino acid sequences of VP4 were searched to find residues which are conserved in the same serotypes but diverged among different serotypes. Candidate positions for the mixed probes were defined at the carboxyl terminus of VP4. Thirdly, Southern blot analyses were carried out using thirty-nine enterovirus standard strains, seven CA16 isolates and sixty-six EV71 isolates previously identified by the neutralization test. The results revealed that each mixed probe exclusively bound to the homologous DNAs but not to the heterologous ones. In an attempt to determine serotypes without virus isolation, clinical specimens from hand-foot-and-mouth disease were examined. Of 78 throat swabs and 15 vesicular fluids, 71 (91.0%) and 13 (86.7%) specimens were clearly identified, indicating that the method described here offer advantages over the traditional neutralization assay: It is rapid, specific and less labor-consuming.

Amino Acid Sequence↗

PubMed1997

Enzymes involved in DNA synthesis in the testes are regulated by temperature in vitro.

OBJECTIVE: Temperature is considered to be an important local regulating factor of testicular function. We investigated whether a high temperature would affect the activity of DNA polymerases alpha, beta, and gamma (EC2.7.7.7) and DNA topoisomerase I in the testes which are required for germ cell differentiation. METHODS: An in vitro rat testis tissue culture system was used to evaluate the activity of DNA polymerases alpha, beta, and gamma and DNA topoisomerase I after incubation at 31 and 37 degrees C. The expression of proliferating cell nuclear antigen was evaluated immunohistochemically in the testis. RESULTS: Incubation at 37 degrees C significantly reduced the activity of all four enzymes as compared with incubation at 31 degrees C. Reducing the temperature to 31 degrees C after prior incubation at 37 degrees C partially restored DNA polymerase alpha activity. Expression of proliferating cell nuclear antigen demonstrated a reduction in DNA synthesis in the seminiferous tubules after incubation at 37 degrees C CONCLUSION: These findings indicate that a high temperature of 37 degrees C reduces the activity of the enzymes involved in the testicular synthesis of DNA which may cause the impairment of spermatogenesis.

Animals↗

PubMed1997

Inhibitory mechanism of mizoribine on the antibody production of mouse B cells stimulated with lipopolysaccharide.

It has been reported that the immunosuppressant mizoribine (MZR) inhibits T cell proliferation by depleting intracellular guanine nucleotides via competitive inhibition of inosine 5'-monophosphate (IMP) dehydrogenase in the purine metabolism pathway. This study was performed to determine if the mechanism by which MZR suppresses the proliferation of mouse B cells and antibody production by these cells is dependent on the depletion of intracellular guanine nucleotides. Stimulation of purified splenic B cells of mice with lipopolysaccharide (LPS), a mitogen to B cells, increased both proliferation and antibody production. MZR suppressed both of these functions in a dose-dependent fashion. MZR also caused a decrease in the amount of intracellular guanosine 5'-triphosphate (GTP). When the cultures were grown on plates containing guanosine plus 8-aminoguanosine, the amount of intracellular GTP, which had been reduced by MZR, was restored. Furthermore, the repletion of GTP pools restored both proliferation and antibody production almost to their previous levels. These results suggest that MZR suppresses antibody production and proliferation of B cells by acting directly on B cells. Furthermore, it is suggested that the inhibitory effect of MZR on antibody production, as well as on T cell proliferation, is dependent on the decrease in intracellular guanine nucleotide pools of mouse B cells.

Animals↗