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Biomedical subjects

T Murata

Publications and source records attributed to T Murata.

At least 181 records · Page 10Linked to original sources

Peroxisome proliferator-activated receptor-gamma ligands inhibit choroidal neovascularization.

PURPOSE: To determine the antiangiogenic effects of peroxisome proliferator-activated receptor (PPAR)-gamma agonists on ocular cells involved in the pathogenesis of choroidal neovascularization (CNV) in vitro and on experimental laser photocoagulation-induced CNV in vivo. METHODS: PPAR-gamma expression in human retinal pigment epithelial (RPE) cells and bovine choroidal endothelial cells (CECs) was determined using an RNase protection assay and Western blot analysis. Two PPAR-gamma ligands, troglitazone (TRO) and rosiglitazone (RSG; 0.1-20 microM), were used to assess effects on RPE and CEC proliferation and migration and CEC tube formation in response to vascular endothelial growth factor (VEGF). The effects of intravitreal injection of TRO on laser photocoagulation-induced CNV lesions in rat eyes (15 experimental, 15 control, nine burns per eye) and cynomolgus monkey eyes (two experimental, two control, seven paramacular burns per eye) was assessed by fluorescein angiography and histologic evaluation. RESULTS. PPAR-gamma1 was expressed in both RPE and CEC. PPAR-gamma ligands significantly inhibited VEGF-induced migration and proliferation in both cell types and tube formation of CEC in a dose-response manner. CNV in rats was markedly inhibited by intravitreous injection of TRO (P < 0.001). Lesions showed significantly less fluorescein leakage and were histologically thinner in the TRO-treated animals. Similar findings were present in the TRO-treated lesions in two monkey eyes. The drug showed no apparent adverse effects in the adjacent retina or in control eyes. CONCLUSIONS: The inhibition of VEGF-induced choroidal angiogenesis in vitro, and CNV in vivo by PPAR-gamma ligands suggests the potential application of these agents in the large group of patients with age-related macular degeneration complicated by CNV.

Animals↗

[Video-assisted thoracoscopic surgery for an intrapulmonary bronchogenic cyst: a case report].

A 23-year-old female was referred to us for an abnormal shadow at the right pulmonary hilum on chest X-ray film. CT scan of the thorax showed a cystic lesion measuring 4 x 3.5 cm with an air-fluid level in S3 of the right upper lobe. We performed video-assisted thoracoscopic S3 segmentectomy for this cyst. The pathological diagnosis was intrapulmonary bronchogenic cyst lined ciliated epithelium and contained bronchial glands, cartilage and smooth muscle.

Adult↗

Chromosome of the enterohemorrhagic Escherichia coli O157:H7; comparative analysis with K-12 MG1655 revealed the acquisition of a large amount of foreign DNAs.

A complete Xba I and Bln I cleavage map was constructed for the chromosome of an enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain isolated from an outbreak in Sakai City, Japan, in 1996. A comparative chromosome analysis with E. coli K-12 strain MG1655 was made. The EHEC chromosome was approximately 5600 kb in length, 1 Mb larger than that of MG1655. Despite the marked difference in chromosome length, the location and direction of seven rRNA operons of the EHEC strain were similar to those for MG1655. Overall organization of genes common in both strains is also highly conserved. Chromosome expansion was observed throughout the EHEC chromosome, albeit in an uneven manner. A large portion of the chromosome enlargement was observed in the region surrounding the replication terminus, particularly in a segment containing the terA locus. Sample sequencing of 3627 random shotgun clones suggested the presence of approximately 1550 kb strain-specific DNAs on the EHEC chromosome, most of which are likely to be of foreign origin.

Bacterial Toxins↗

An Application of Percolation Theory to the Electrolyte Penetration through Porous Water-Swollen Cellulose Triacetate Membrane.

Permeability coefficients P of KCl through porous cellulose triacetate (CTA) membranes were measured as a function of the water volume fraction V(W) and diffusion coefficients D were determined using solubility parameters K and a membrane thickness d from the relationship of P = KD/d. D increased with an increase in V(W). D especially increases abruptly around V(W) = 0.5, which corresponds to 2% triethylene glycol (TEG) content. The percolation theory was applied to the experimental results under the conditions D(A) = D (V(W) = 1) = 1.8 x 10(-5) cm(2) s(-1), D(B) = D(V(W) = 0) = 1.8 x 10(-8) cm(2) s(-1), coordination number (Z) = 2.5, 3, 3.5, and 4, and packing fraction f = 1.0. A good fit was obtained at Z = 3.5 because the experimental and calculated results also shifted at the same V(W) below V(W) = 0.5. It is suggested that a phase inversion, that is, change of a discontinuous water phase to a continuous water phase, occurs around V(W) = 0.5. Above V(W) = 0.5, the experimental results agree well with the calculated line for Z = 3 or Z = 2.5 which means that the coordination numbers decrease with an increase in water content. It is thought that V(W) is overestimated because it is hard to completely wipe off the excess water quickly from the membrane surface. Z = 3.5 means that a pore can connect in 3.5 directions. Copyright 1999 Academic Press.

Journal Article↗

Prevention of leukostasis and vascular leakage in streptozotocin-induced diabetic retinopathy via intercellular adhesion molecule-1 inhibition.

Diabetic retinopathy is a leading cause of adult vision loss and blindness. Much of the retinal damage that characterizes the disease results from retinal vascular leakage and nonperfusion. This study shows that diabetic retinal vascular leakage and nonperfusion are temporally and spatially associated with retinal leukocyte stasis (leukostasis) in the rat model of streptozotocin-induced diabetes. Retinal leukostasis increases within days of developing diabetes and correlates with the increased expression of retinal intercellular adhesion molecule-1 (ICAM-1). ICAM-1 blockade with a mAb prevents diabetic retinal leukostasis and vascular leakage by 48.5% and 85.6%, respectively. These data identify the causal role of leukocytes in the pathogenesis of diabetic retinopathy and establish the potential utility of ICAM-1 inhibition as a therapeutic strategy for the prevention of diabetic retinopathy.

Acridine Orange↗

The DNA-binding and transcriptional activities of MAZ, a myc-associated zinc finger protein, are regulated by casein kinase II.

Myc-associated zinc finger protein (MAZ) is a transcription factor that contains proline-rich, alanine repeats and six C(2)H(2)-type zinc finger motifs, as well as five putative sites of phosphorylation by casein kinase II (CKII). Site-specific mutagenesis of MAZ revealed that the serine residue at position 480 was the major site of phosphorylation by CKII both in vitro and in vivo. Phosphorylation of MAZ by CKII at this serine residue was required for maximum binding of MAZ to the pyrimidine-rich DNA of the nuclease-hypersensitive element (NHE) in the 5'-end promoter region of the c-myc gene. Mutation of serine at position 480 to alanine eliminated the DNA-binding activity of MAZ to this element. Moreover, the mutated MAZ was unable to enhance the expression of luciferase encoded by a c-myc promoter/luciferase reporter gene in HeLa cells in the presence of CKII. These results suggest that phosphorylation of the serine residue at position 480 of MAZ by CKII can control the function of MAZ by altering its DNA-binding activity.

Amino Acid Substitution↗

Enzymatic synthesis of alpha-L-fucosyl-N-acetyllactosamines and 3'-O-alpha-L-fucosyllactose utilizing alpha-L-fucosidases.

An alpha-L-fucosidase from porcine liver produced alpha-L-Fuc-(1-->2)-beta-D-Gal-(1-->4)-D-GlcNAc (2'-O-alpha-L-fucosyl-N-acetyllactosamine, 1) together with its isomers alpha-L-Fuc-(1-->3)-beta-D-Gal-(1-->4)-D-GlcNAc (2) and alpha-L-Fuc-(1-->6)-beta-D-Gal-(1-->4)-D-GlcNAc (3) through a transglycosylation reaction from p-nitrophenyl alpha-L-fucopyranoside and beta-D-Gal-(1-->4)-D-GlcNAc. The enzyme formed the trisaccharides 1-3 in 13% overall yield based on the donor, and in the ratio of 40:37:23. In contrast, transglycosylation by Alcaligenes sp. alpha-L-fucosidase led to the regioselective synthesis of trisaccharides containing a (1-->3)-linked alpha-L-fucosyl residue. When beta-D-Gal-(1-->4)-D-GlcNAc and lactose were acceptors, the enzyme formed regioselectively compound 2 and alpha-L-Fuc-(1-->3)-beta-D-Gal-(1-->4)-D-Glc (3'-O-alpha-L-fucosyllactose, 4), respectively, in 54 and 34% yields, based on the donor.

Alcaligenes↗

Alterations of 3-nitrotyrosine concentration in the cerebrospinal fluid during aging and in patients with Alzheimer's disease.

To investigate the significance of nitric oxide (NO)-mediated neuron death in aging and Alzheimer's disease (AD), the 3-nitrotyrosine concentration in the cerebrospinal fluid (CSF) was investigated in neurologically normal controls and patients with AD. The 3-nitrotyrosine concentration and the 3-nitrotyrosine/tyrosine ratio significantly increased with advancing age, whereas the tyrosine concentration was unaltered. In patients with AD, the 3-nitrotyrosine concentration and the 3-nitrotyrosine/tyrosine ratio increased significantly (>six-fold) compared with controls of similar age, and increased significantly with decreasing cognitive functions, whereas the tyrosine concentration did not change. These findings suggest that an activation of tyrosine nitration, increase in nitrated tyrosine-containing proteins, and/or its degradation may be involved in brain aging and play an important role in the pathogenesis of AD.

Aged↗

Dynamic computed tomography predicts tumor temperature and response to thermoradiotherapy in superficial and subsurface tumors.

BACKGROUND: Dynamic computed tomography (CT) was performed on patients undergoing thermoradiotherapy for superficial or subsurface tumors, and the correlation between tumor enhancement and tumor temperature during hyperthermia was evaluated. The authors further investigated whether tumor enhancement by dynamic CT is predictive of tumor response to thermoradiotherapy. METHODS: Thermoradiotherapy was given to 26 patients. Radiotherapy consisted of 40-70 gray. Hyperthermia was conducted over 3-5 sessions, and tumor temperature was measured at each session. Dynamic CT was performed prehyperthermia (within 1 week before the initiation of hyperthermia) and midtherapy (within 1 week after 2 sessions of hyperthermia). RESULTS: A complete response (CR) was obtained in 11 patients (42%) and either a partial response or no response (non-CR) in 15 (58%). There was no correlation between tumor enhancement obtained by prehyperthermia CT and tumor temperature parameters or response. However, the deltaCTmax (maximum increased enhancement) by prehyperthermia and midtherapy CT was 39.0 +/- 18.9 HU and 26.1 +/- 14.2 HU, respectively, in CR patients, and 46.4 +/- 21.1 HU and 49.6 +/- 19.1 HU, respectively, in non-CR patients. This change in deltaCTmax at midtherapy was significantly different between groups (P < 0.01). The deltaCTmax ratio for prehyperthermia and midtherapy CT studies correlated with the average tumor temperature (P < 0.05). CONCLUSIONS: Tumor enhancement by prehyperthermia and midtherapy dynamic CT predicted tumor temperature during hyperthermia and response to thermoradiotherapy for superficial or subsurface malignancies.

Adult↗

Dynamic changes in glucose metabolism induced by thiamine deficiency and its replenishment as revealed by a positron autoradiography technique using rat living brain slices.

Dynamic changes in the cerebral glucose metabolic rate (CMRglc) before and after thiamine replenishment were investigated in living brain slices obtained from pyrithiamine-treated (PT) and pair-fed control rats by use of a positron autoradiography technique. Fresh rat brain slices (300 microm thick) were incubated with [18F]2-fluoro-2-deoxy-D-glucose ([18F]FDG) in oxygenated Krebs-Ringer solution at 36 degrees C, during which serial two-dimensional images of [18F]FDG uptake in the slices were constructed on the imaging plates. The net influx constant (=K) of [18F]FDG was determined by a Patlak graphical method of the image data. Prior to thiamine pyrophosphate (TPP)-loading, the K value in the neurologically symptomatic PT was higher in all brain regions except the thalamus and mammillary body than the control, suggesting compensatory enhanced glycolysis. The rapid decrease in this heightened net influx constant immediately after TPP-loading was surmised to be due to activation of pyruvate oxidation with lactate as the substrate, with this inhibiting the glycolysis. From > or = 150 min after TPP-loading, the K value continued to show low values in the thalamus and mammillary body, which are regarded as the responsible sites for Korsakoff syndrome, whereas in all other sites recovery to control values was observed. These findings suggest that using this technique the quantitative evaluation of serial local changes in CMRglc from thiamine deficiency to after its replenishment may be useful in elucidating the pathophysiology and prognosis of Wernicke's encephalopathy.

Animals↗

Rhophilin, a small GTPase Rho-binding protein, is abundantly expressed in the mouse testis and localized in the principal piece of the sperm tail.

Tissue distribution and cellular localization of rhophilin, a 71 kDa Rho-binding protein, were examined in mice. Rhophilin mRNA was highly expressed in adult testis, but was absent in the testis of W/WV mice deficient in germ cells. An anti-rhophilin antibody detected a band of an expected size in sperm extracts, which was enriched in the tail fraction. Immunofluorescence analysis revealed two lines of striated staining running in parallel in the principal piece of the sperm tail. These results suggest that rhophilin is expressed in germ cells and localized in the fibrous sheath of the sperm tail.

Adaptor Proteins, Signal Transducing↗

Increase in oxidized NO products and reduction in oxidized glutathione in cerebrospinal fluid from patients with sporadic form of amyotrophic lateral sclerosis.

To determine the role of free radical mechanisms in the pathogenesis of amyotrophic lateral sclerosis (ALS), cerebrospinal fluid concentrations of oxidized nitric oxide (NO) products (nitrite and nitrate) and reduced and oxidized forms of glutathione (GSH and GSSG, respectively) were compared between patients with the sporadic form of ALS (SALS) and controls. In the SALS patients, the nitrate levels were significantly higher (by 73%) in contrast to remarkably lower GSSG/GSH ratio, approximately 3-fold, compared to controls. These results suggest that NO production or oxidation is activated in SALS patients, leading to a decrease in superoxide radicals to oxidize GSH. The subsequent generation of a highly reactive anion, peroxynitrite, may play a causal role in the pathogenesis of SALS.

Aged↗

A Case of Esophageal Squamous Cell Carcinoma Metastatic to the Breast.

Metastasis to the breast from extramammary malignancies is rare. There are especially few reports of metastasis from esophageal cancer. We report the pathological and autopsy findings of a 44-year-old man with advanced esophageal cancer and a left breast tumor. Squamous cell carcinoma invading the mammary glands wasdemonstrated histologically. Immunostains for ER, PgR, and ErbB-2 were negative. At autopsy, metastatic lesions were found in lung, liver, diaphragm, peritoneum, spine, and mediastinal lymph nodes, with no evidence of metastasis to the skin. While metastatic breast tumors are rarely the initial sign of malignancy, it isimportant to distinguish a metastasis from primary breast cancer to avoid unnecessary conflicting treatments.

Journal Article↗

Ion Transport across a Polyelectrolyte-Adsorbed Cellulose Triacetate Membrane in the Multicomponent Ionic Systems.

The effects of polyelectrolyte adsorption by cellulose triacetate (CTA) membrane on ionic transport are investigated in two systems: the three-ionic-component system and the multicomponent-ionic system. In the three-ionic-component system, the permeabilities of two anions are affected by the competitive ion. Especially in the case of the albumin-adsorbed CTA membrane, there exists much greater specificity for the permeability of SO2-4 than in the case of the lysozyme-adsorbed membrane. On the other hand, in the case of the PAS-H(10L)(polydiallyldimethylammonium chloride)-adsorbed membrane, the permeability coefficient of HPO2-4 increases, though there exists the effect of a competitive ion. In a multicomponent-ionic system, the logarithmic permeability coefficient ratios (rP) of each ion in an adsorbed membrane to that in a nonadsorbed membrane decreased by PAS-H(10L) adsorption for all cations. The rP of bivalent cations decreased more than those of univalent cations because of the rejection from the positively charged adsorbed layer. On the other hand, the permeabilities slightly increase because of the attraction from the PAS-H(10L)-adsorbed layer when competitive anions exist among them. Furthermore, the increase in the HPO2-4 permeability is confirmed by PAS-H(10L)-adsorption on a CTA membrane for a case very similar to the actual anion multicomponent system. These are the most important results in the application for an approach to phosphate extraction from blood across an artificial kidney membrane. Copyright 1999 Academic Press.

Journal Article↗

Promoter characterization of the rat gene for Ca2+-binding protein regucalcin. Transcriptional regulation by signaling factors.

To understand the mechanism underlying the regulation of the Ca2+-binding protein regucalcin gene expression, we characterized the 5'-flanking region of the rat regucalcin gene. The transcriptional start site of the rat regucalcin gene was determined by the cap site hunting method with rat liver cap site cDNA. The 5'-flanking region of the rat regucalcin gene ligated to a luciferase reporter gene possessed functional promoter activity in rat H4-II-E hepatoma cells. 3'- and 5'-deletion analyses indicated the sequence required for basal functional promoter activity of the rat regucalcin gene. The promoter activity of the rat regucalcin gene was enhanced by treatment with Bay K 8644, dibutyryl cAMP, phorbol esters, insulin, and dexamethasone. Using gel mobility shift assays, we found that nuclear proteins from H4-II-E cells specifically bind to the 5'-flanking region of the rat regucalcin gene. Moreover, gel mobility shift assays revealed that Bay K 8644, dibutyryl cAMP, phorbol esters, and insulin stimulated the binding of nuclear factors to the 5'-flanking region of the rat regucalcin gene in H4-II-E cells. These results suggest that Bay K 8644-, dibutyryl cAMP-, phorbol ester-, and insulin-inducible nuclear factors mediate the stimulatory effect of each regulator on promoter activity of the rat regucalcin gene.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Mannosylerythritol lipid is a potent inducer of apoptosis and differentiation of mouse melanoma cells in culture.

Malignant melanomas are tumors that are well known to respond poorly to treatment with chemotherapeutic reagents. We report here that mannosylerythritol lipid (MEL), an extracellular glycolipid from yeast, markedly inhibited the growth of mouse melanoma B16 cells in a dose-dependent manner. Exposure of B16 cells to MEL at 10 microM and higher concentrations caused the condensation of chromatin, DNA fragmentation, and sub-G1 arrest, all of which are hallmarks of cells that are undergoing apoptosis. Analysis of the cell cycle also suggested that both the MEL-mediated inhibition of growth and apoptosis were closely associated with growth arrest in the G1 phase. Moreover, MEL exposure stimulated the expression of differentiation markers of melanoma cells, such as tyrosinase activity and the enhanced production of melanin, which is an indication that MEL triggered both apoptotic and cell differentiation programs. Forced expression of Bcl-2 protein in stably transformed B16 cells had a dual effect: it interfered with MEL-induced apoptosis but increased both tyrosinase activity and the production of melanin as compared with these phenomena in vector-transfected MEL-treated control B16 cells. These results provide the first evidence that growth arrest, apoptosis, and the differentiation of mouse malignant melanoma cells can be induced by a microbial extracellular glycolipid.

Animals↗