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Biomedical subjects

Y Murakami

Publications and source records attributed to Y Murakami.

At least 37 records · Page 2Linked to original sources

DARPP-32 expression arises after a phase of dysplasia in oesophageal squamous cell carcinoma.

This is the first report to correlate DARPP-32 immunoreactivity (dopamine and cAMP-regulated phosphoprotein, M(r) 32 000) to clinicopathological status in human cancer. DARPP-32 is recognised as a neuronal protein. A recent study demonstrated that DARPP-32, and a truncated isoform t-DARPP, are overexpressed in gastric carcinoma during the process of carcinogenesis. The biological function of DARPP-32, however, is still unclear. The purpose of this study was to clarify the roles of DARPP-32 and t-DARPP in oesophageal squamous cell carcinoma (OSCC). Initially, we investigated DARPP-32 and t-DARPP expression in OSCC cell lines by Reverse transcription-polymerase chain reaction and Western blot. DARPP-32 expression was observed in four out of seven (57.1%) cell lines, but t-DARPP expression was not observed in any cell lines. In oesophageal tissue sample, DARPP-32 expression was observed in four out of seven (57.1%) tumour tissues, while t-DARPP was not observed in any tissues. Subsequently, DARPP expression was assessed by immunohistochemistry, using a polyclonal antibody, in tissue sections from 122 patients with primary OSCC. DARPP immunoreactivity was not observed in any normal oesophageal mucous membranes. On the other hand, positive DARPP immunostaining was detected in 37 patients (30.3%) and correlated inversely with pathologic stage (P=0.0284), pT (P=0.0438), pN (P=0.0303) and tumour size (P=0.012). The overall survival rate was worse in patients with DARPP-negative tumours than in patients with DARPP-positive tumours (P=0.0453). Interestingly, DARPP expression was observed in only one out of 45 cases of dysplasia. These observations suggest that DARPP-32 (rather than t-DARPP) expression arises after a phase of dysplasia in OSCC, and that tumours expressing DARPP-32 progress less rapidly than DARPP-32-negative tumours.

Adult↗

Commensurate-incommensurate crossover of charge stripe in La2-xSrxNiO4 (x approximately 1/3).

The temperature (T) dependence of the charge-stripe order in La2-xSrxNiO4 has been investigated in the vicinity of x approximately 1/3 by synchrotron radiation x-ray diffraction measurements. With decreasing T, a prominent commensurate-incommensurate (C-IC) crossover is observed in the x<1/3 region, while for the x>1/3 region the IC order is dominant over the whole T range. Such a C-IC crossover is interpreted as the entropy-driven self-doping of the charge stripes, and its x dependence indicates the clear electron-hole asymmetry with the x=1/3 compound as the Mott insulator.

Journal Article↗

Effect of erythropoietin on nitric oxide production in the rat hippocampus using in vivo brain microdialysis.

In order to investigate the role of erythropoietin (EPO) in the hippocampus, we studied the effect of EPO on nitric oxide (NO) production in the rat hippocampus using brain microdialysis. The dialysis probe was stereotaxically inserted into the rat hippocampus 24 h before the dialysis experiment. The perfusion fluid (Krebs-HEPES buffer, pH 7.4) was collected at 15-min intervals under freely moving conditions and NO metabolites (NOx) in the perfusate were immediately measured using a NOx-analyzing high performance liquid chromatography (HPLC)system. Following the collection of four fractions, 1 microl of EPO (10(-10) M, 10(-8) M and 10(-6) M) or vehicle (saline) was gently injected into the hippocampal tissue. The perfusion fluid was collected for 3 h after the injection. The NOx levels were unchanged by the injection of vehicle alone. After the injection of EPO, NOx levels gradually increased. The EPO-induced increase in NOx levels was significant at 10(-6) M EPO. The EPO-induced increases in NOx levels were eliminated in the presence of anti-EPO antibody. The increase in NOx levels induced by EPO was blunted by nicardipine, a Ca2+ channel blocker, but not by MK-801, an antagonist of N-methyl-D-aspartate (NMDA) receptors. These findings, taken together, suggest that EPO increased NO production in the rat hippocampus by activating voltage-gated Ca2+ channels, but not through NMDA receptors.

Animals↗

Over-expression of E2F-1 in esophageal squamous cell carcinoma correlates with tumor progression.

The transcription factor E2F-1, a downstream regulator of the p16-cyclinD-Rb pathway, is required for cell cycle progression. Evidence shows that overexpression of E2F-1 can either promote or inhibit the development of tumors, depending on tissue or experimental conditions. However, the clinical impact of E2F-1 expression on esophageal squamous cell carcinoma (ESCC) remains unknown. To analyze E2F-1 expression in ESCC, we investigated the immunoreactivity of E2F-1 and its correlation with clinicopathological features in 122 patients who underwent surgical resection for ESCC. Positive E2F-1 immunostaining was detected in 73 patients (59.8%). Positive E2F-1 immunostaining correlated positively with pathologic stage (P = 0.0103), p-Grade (P = 0.0014) and pT (P = 0.0192). The overall survival rate was worse in patients with E2F-1-positive tumors than in patients with E2F-1-negative tumors (P = 0.0290). Over-expression of E2F-1 is associated with tumor progression and a worse prognosis after surgery in ESCC.

Adult↗

[Extrapleural pneumonectomy for malignant pleural mesothelioma].

Standard treatment for malignant pleural mesothelioma (MPM) has not been proved yet. However, it has been recognized that extrapleural pneumonectomy (EPP) is a treatment of choice for epithelial MPM when combined with adjuvant therapies though EPP may frequently cause fetal complications. We report 5 cases of MPM with EPP, including 1 with good prognosis. Sixteen patients with MPM were admitted to our hospital between 1988 and 2003. Five patients underwent EPP, among which 4 were male and 1 female with ages from 46 to 61 years old. Histologically, 3 of them were epithelial and 2 were biphasic. Those with biphasic experienced acute respiratory failure and empyema, and died 81 days and 8 months after the surgery respectively. Among those with epithelial MPM, 2 are alive with no recurrence at 129 and 29 months after the surgery, and the other, followed by postoperative radiotherapy, died at 12 months. More cases with EPP or randomized controlled trials regarding EPP are necessary to evaluate efficacy of EPP for MPM.

Female↗

Magnetization distribution in the mixed-phase state of hole-doped manganites.

The effect of 'colossal magnetoresistance' (CMR) in hole-doped manganites--an abnormal decrease of resistivity when a magnetic field is applied--has attracted significant interest from researchers in the past decade. But the underlying mechanism for the CMR phenomenon is not yet fully understood. It has become clear that a phase-separated state, where magnetic and non-magnetic phases coexist, is important, but the detailed magnetic microstructure of this mixed-phase state is so far unclear. Here we use electron microscopy to study the magnetic microstructure and development of ferromagnetic domains in the mixed-phase state of La(1-x)Sr(x)MnO3 (x = 0.54, 0.56). Our measurements show that, in the absence of a magnetic field, the magnetic flux is closed within ferromagnetic regions, indicating a negligible magnetic interaction between separated ferromagnetic domains. However, we also find that the domains start to combine with only very small changes in temperature. We propose that the delicate nature of the magnetic microstructure in the mixed-phase state of hole-doped manganites is responsible for the CMR effect, in which significant conduction paths form between the ferromagnetic domains upon application of a magnetic field.

Journal Article↗

Expression of hypothalamic corticotropin-releasing hormone-like immunoreactivity in isolated ACTH deficiency: a report of an autopsied case.

A 77-yr-old man died of metastising adenocarcinoma of the lung. He had been treated with hydrocortisone for isolated ACTH deficiency for 9 yr. At autopsy, the pituitary and the adrenal glands were atrophic. Lymphocyte infiltration was not observed in the pituitary, the adrenal and the thyroid glands. Immunohistochemistry revealed selective loss of ACTH-like immunoreactivity in the pituitary gland whereas CRH-immunoreactive cells were found in the paraventricular nucleus of the hypothalamus. This is the first report demonstrating immunohistochemical examinations of the pituitary and the hypothalamus in isolated ACTH deficiency.

Adrenal Glands↗

A study on surface roughness and microleakage test in cavities prepared by Er:YAG laser irradiation and etched bur cavities.

The purposes of this study were to investigate the surface morphology, suface roughness of cavities prepared by Er:YAG laser irradiation, and compared the microleakage degree after composite resin restoration with etched bur cavities, in vitro. In each of the 30 human extracted teeth, two shallow cavities were prepared; one prepared with the Er:YAG laser system on the buccal surface, and one produced on the lingual (palatal) surface with a high-speed turbine. Five cavities from each group were investigated by scanning electron microscopy (SEM), and five were subjected to surface roughness analysis by a colour laser three-dimensional (3D) microscope. The remaining cavities were filled with a composite resin and subjected to a microleakage test under thermocycling. Only bur cavities were acid-etched before filling. Statistical analysis was performed using the Mann-Whitney U test; a value of p <0.01 was considered significant. Morphologically, the prepared cavities showed an irregular surface with the absence of a debris-like smear layer; enamel prisms and opening of dentinal tubules were recognised. Surface roughness was significantly increased with the laser system. Microleakage test revealed no significant differences between the laser and bur cavities. Crosscut sections of the cavities with no microleakage showed no gap at the interface. Laser cavity may facilitate good adaptation of composite resin with enamel and dentine, because an increase of surface roughness and the openings of dentinal tubules may facilitate the formation of a hybrid zone, since a primer and an adhesive can penetrate the surface better when the smear layer is removed. It can be concluded that shallow cavity prepared by Er:YAG laser is capable of decreasing microleakage of composite resin restorations, and its efficiency is similar to etched bur cavities.

Acrylic Resins↗

Effects of dexamethasone on the pathogenesis of porcine circovirus type 2 infection in piglets.

To investigate the effect of immunosuppression on porcine circovirus type 2 (PCV2) infection, hysterectomy-produced, colostrum-deprived piglets were inoculated with the virus by the intranasal or intraperitoneal route, with or without dexamethasone (DEX) treatment. Eleven piglets aged 8 days were divided into four groups, namely group A (four animals given PCV2), B (three given PCV2 with DEX), C (two given sterile medium with DEX) and D (two given sterile medium). No significant clinical signs were observed. Histopathological and immunohistochemical examination revealed granulomatous inflammation and PCV2 antigen in the lymphoid tissues of group B piglets, but not in the other three groups. Flow cytometric analysis of peripheral blood lymphocytes showed a reduced number of CD4+ T cells in DEX-treated piglets (groups A and C). No differences between groups were observed in respect of the number of B cells, serum IgG concentration, or PCV2 antibody titre. These results indicate that DEX influenced the pathogenic effects of PCV2 infection in lymphoid organs, and that suppression of cell-mediated immunity may play a role in the aetiology of postweaning multisystemic wasting syndrome.

Animals↗

Excited state processes in a four-component photosensitive system based on a bisimidazole derivative.

The four component system consisting of bis[2-(o-chlorophenyl)-4,5-diphenylimidazole] (Cl-HABI), N,N'-bis(diethylamino)benzophenone (EAB), N-phenylglycine (NPG) and an iodonium salt (BIP-T) can be used for the photoinitiation of radical polymerization reactions. The excited state processes involved are reported and all the transient species, including the triplet state and ketyl radical of EAB as well as the lophyl radical, are characterized. Rate constants of interaction between the different partners in the four component system are determined. Redox potentials of the different compounds are evaluated enabling the free energies for electron transfer reactions to be calculated. An overall scheme for the evolution of the excited states and a general discussion on the role played by the different components in photoinitiating polymerization are then provided.

Journal Article↗

Atomic analysis and knoop hardness measurement of the cavity floor prepared by Er,Cr:YSGG laser irradiation in vitro.

In the present study, the compositional changes and knoop hardness of the cavity floor prepared by Er,Cr:YSGG laser irradiation was compared with that of the conventional bur cavity. Fifteen laser and 15 bur cavities were cross-sectioned, and subjected to atomic analysis by SEM-EDX and knoop hardness test. Statistical analyses were performed using the Mann-Whitney U-test; a value of P < 0.01 was considered significant. Surface characteristics of the prepared cavities were also investigated by light microscopy and scanning electron microscopy (SEM). The results showed that the quantities of Ca (Ca weight %) and P (P weight %) were increased significantly in the laser cavity floor but no significant differences were found between the Ca/P ratio and knoop hardness number of laser and bur cavities. The SEM observation revealed that the lased cavity surface was irregular and there was also the absence of a smear layer; the orifice of dentinal tubules was exposed. Er,Cr:YSGG laser device is considered as one of the most effective and safe devices for cavity preparation because of its many advantages. This includes easy delivery system, minimal thermal damage to the surrounding tissues, minimal thermal-induced changes of dental hard tissue compositions, and favourable surface characteristic.

Calcium↗

A morphological and immunolabeling study of freeze-substituted Bacteroides forsythus.

We used a rapid freezing and freeze-substitution technique without glutaraldehyde and OsO4 fixation for the electron microscopic immunocytochemical demonstration of the surface structure of Bacteroides forsythus, an anaerobic Gram-negative periodontopathogen. Cells were applied to pieces of filter paper and freeze-substituted by plunge-freezing in liquid propane, substituted in methanol containing 0.5% uranyl acetate, and infiltrated with LR White resin. The membrane ultrastructure of B. forsythus was preserved well, and the labeling density of the freeze-submitted cells was compared to a conventional processing method. Our results show the usefulness of the freeze-substitution method for immunohistochemical studies of B. forsythus.

Bacteroides↗

Expression of two kinds of recombinant glutamate dehydrogenase from Aeropyrum pernix with different N-terminal sequence length in Escherichia coli.

Two recombinant Aeropyrum pernix glutamate dehydrogenase (GDH) enzymes with different length N-termini were cloned and expressed in Escherichia coli: sGDH begins with the amino acid sequence of the extracted native enzyme (M-Q-P-T-D-P-L-E-E), whereas lGDH begins with the sequence of the predicted ORF (M-E-V-L-A-L-Q-P-T-D) and is longer than sGDH by five amino acids (M-E-V-L-A). Purified recombinant lGDH was more stable than sGDH, indicating that the N-terminal extension, containing mostly hydrophobic residues, affected the overall stability of recombinant lGDH. This stabilising effect of extending the N-terminal sequence on an oligomeric enzyme such as GDH is novel; factors affecting stabilisation have previously only been discussed in the context of the contribution of internal amino acids.

Amino Acid Sequence↗

Inhibitory effect of synthetic histatin 5 on leukotoxin from Actinobacillus actinomycetemcomitans.

Actinobacillus actinomycetemcomitans is a gram-negative bacterium strongly implicated in the pathogenesis of juvenile periodontitis. This periodontal pathogen synthesizes a leukotoxin that destroys human polymorphonuclear leukocytes (PMNs), and this toxin is thought to be responsible for the virulence of A. actinomycetemcomitans. It was therefore of interest to assess whether major virulence factors of periodontal pathogens were neutralized by salivary components. This study focuses on the effect of histatins, components of the nonimmune oral defense system, on leukotoxin activity. Leukotoxin was extracted with polymyxin B from freshly grown anaerobic cultures of A. actinomycetemcomitans strain Y4. PMNs isolated from blood of healthy human volunteers were incubated in a cytotoxicity assay containing PMNs (10(7) cells/ml) and leukotoxin preparation (0-500 microg/ml) in Hanks' balanced salt solution at 37 degrees C for 0-120 min with or without synthetic histatin 5 (0-500 microM). Cytotoxicity was measured by release of lactate dehydrogenase (LDH) at different time intervals. Histatin 5 neutralized the toxic effect of the leukotoxin preparation in a concentration-dependent manner, with an IC(50) value of 150 microM. When PMNs were preincubated with histatin 5 (300 microM), washed and subsequently exposed to leukotoxin, no protective effect was observed. This observation suggests a mechanism of inhibition whereby histatin 5 either directly neutralizes the leukotoxin or interferes with the leukotoxin-PMN interaction. The inhibitory effect of histatin 5 on leukotoxic activity may suggest a new biological function of histatins in the oral cavity as a naturally occurring secondary antibiotic.

Adult↗

Bacteroides forsythus hemagglutinin is inhibited by N-acetylneuraminyllactose.

Bacteroides forsythus, which has been recognized as a pathogen associated with periodontitis, produces a hemagglutinin. The hemagglutinin was localized in the envelope of B. forsythus. The hemagglutinating activity was inhibited by lactose at concentrations as low as 1 mM, and by L-arginine and L-lysine at concentrations of 100 mM. N-Acetylneuraminyllactose (NeuAc-lactose) was at least 100 times more potent an inhibitor than lactose, as it completely inhibited the hemagglutination at concentrations below 10 microM. This is similar to the Helicobacter pylori hemagglutinin. The hemagglutinin was heat-labile, and resistant to treatment with proteases such as trypsin. A specific antibody raised against one of the S-layer proteins that are major species-specific proteins had no inhibitory effect on the hemmaglutination. These results suggest that the NeuAc-lactose-sensitive adhesin of B. forsythus may play an important role in colonization in the oral cavity.

Adhesins, Bacterial↗

LPS-stimulated human gingival fibroblasts inhibit the differentiation of monocytes into osteoclasts through the production of osteoprotegerin.

Periodontitis is an inflammatory bone disease caused by Gram-negative anaerobic bacteria, but the precise mechanism of bone destruction remains unknown. Activated T lymphocytes secrete receptor activator of NF-kappaB ligand (RANKL) and support the differentiation of monocytes into mature osteoclasts. The purpose of this study was to examine the expression of RANKL and its inhibitor, osteoprotegerin (OPG), in inflamed gingival tissue and to clarify the role of human gingival fibroblasts (HGFs) in osteoclastogenesis regulated by RANKL. HGFs and gingival mononuclear cells (GMCs) were obtained from chronic periodontitis patients during routine periodontal surgery. Expression of OPG and RANKL mRNA in gingival tissue and HGFs was examined with RT-PCR. OPG production was measured using ELISA. Expression of RANKL, CD4, CD8 and CD69 on GMCs was determined by flow-cytometry using RANK-Fc fusion protein and the respective monoclonal antibodies. Osteoclastogenesis by RANKL was assayed by counting the number of tartarate-resistant acid phosphatase (TRAP)-positive cells after culturing human peripheral blood monocytes with recombinant human RANKL and macrophage-colony stimulating factor (M-CSF) for 10 days. OPG and RANKL mRNA were expressed in 80% (16/20) and 25% (5/20) of periodontitis lesions, respectively. OPG, but not RANKL, mRNA was expressed within HGFs. OPG mRNA expression and production by HGFs was augmented by LPS stimulation. All GMC samples expressed CD69, and two of five GMC samples expressed RANKL. The culture supernatant of LPS-stimulated gingival fibroblasts significantly reduced the number of TRAP positive cells generated by culturing monocytes with RANKL and M-CSF. The present study suggests that LPS-stimulated HGFs inhibit monocyte differentiation into osteoclasts through the production of OPG.

Antigens, CD↗