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Biomedical subjects

Y Hata

Publications and source records attributed to Y Hata.

At least 127 records · Page 7Linked to original sources
PubMed1998

Neurabin-II/spinophilin. An actin filament-binding protein with one pdz domain localized at cadherin-based cell-cell adhesion sites.

In a preceding paper, we reported a novel actin filament (F-actin)-binding protein, named neurabin, which was specifically expressed in neural tissue and implicated in neurite formation. We purified from rat brain another F-actin-binding protein, which had a domain organization similar to that of neurabin but was ubiquitously expressed, and named it neurabin-II. The original neurabin, renamed neurabin-I, had 1095 amino acids and a calculated Mr of 122,729, whereas neurabin-II had 817 amino acids and a calculated Mr of 89, 642. Both neurabin-I and -II had one F-actin-binding domain at the N-terminal region, one PDZ domain at the middle region, a domain known to interact with transmembrane proteins, and domains predicted to form coiled-coil structures at the C-terminal region. Both neurabin-I and -II bound along the sides of F-actin and showed F-actin-cross-linking activity. The subcellular distribution analysis indicated that neurabin-II was enriched at the postsynaptic density fraction in rat brain and the adherens junction fraction in rat liver. Immunofluorescence microscopic analysis revealed that neurabin-II was highly concentrated at the synapse in primary cultured rat hippocampal neurons and at the cadherin-based cell-cell adhesion sites in Madin-Darby canine kidney cells. Neurabin-II turned out to be the same as a recently reported protein phosphatase 1-binding protein named spinophilin. These results suggest that neurabin-II/spinophilin plays an important role in linking the actin cytoskeleton to the plasma membrane.

Actins↗
PubMed1998

Nucleotide sequence of an alternative glucoamylase-encoding gene (glaB) expressed in solid-state culture of Aspergillus oryzae.

The DNA (glaB) and a cDNA-encoding glucoamylase produced in solid-state culture of Aspergillus oryzae were cloned using oligodeoxyribonucleotide probes derived from internal amino acid sequences of the enzyme. Comparison of the nucleotide sequences of a genomic DNA fragment with its cDNA showed the glaB gene carried three exons interrupted by two introns and had an open reading frame encoding 493 aa residues. The 5'-flanking region had a TATA box at nt -87 from the start codon and two putative CAAT sequences at nt -276 and -288. The glaB gene shared 57% homology at the aa level with the glaA gene which was cloned previously from A. oryzae. Interestingly, the glucoamylase encoded by the glaB gene had no C-terminal domain such as that proposed to have starch binding activity in Aspergillus glucoamylases. Introduction of cDNA of the glaB gene to Saccharomyces cerevisiae caused the secretion of active glucoamylase to culture medium and introduction of the glaB gene to A. oryzae increased glucoamylase productivity in solid-state culture. Northern blot analysis showed the glaB gene was expressed in solid-state culture, but not in submerged culture.

Amino Acid Sequence↗
PubMed1998

Effects of changes in the criteria for nationally shared kidney transplants for HLA-matched patients.

BACKGROUND: Nine years ago, a prospective trial began in all U.S. transplant centers to determine whether the results of renal transplantation would improve with the nationwide shipment of kidneys from cadaveric donors to HLA-matched patients. Since then, the stringency of criteria for HLA matching have been liberalized twice, from sharing only those kidneys that matched at all six HLA-A, -B, -DR antigens, to sharing phenotypically HLA-matched kidneys, and most recently to sharing zero HLA-mismatched kidneys. METHODS: Data reported to the United Network for Organ Sharing Scientific Renal Transplant Registry from October 1987 to December 1996 were analyzed to examine the transplant results of nationally shared HLA-matched kidneys and the effects of changes to the HLA matching criteria on graft survival and the distribution of HLA-matched kidneys. RESULTS: The overall 1-year graft survival rate of 5102 HLA-matched transplants was 88% compared with 81% for 58,207 recipients of kidneys with at least one HLA mismatch (P < 0.001). HLA-matched kidneys had a projected 12-year graft half-life, 50% higher than the 8-year half-life of mismatched grafts (P < 0.01). After the first change in the match criteria in August 1990, 1365 phenotypically matched kidneys with fewer than six HLA antigens identified had an 89% 1-year graft survival rate compared with 84% for 466 six antigen-matched kidneys transplanted before the change. After March 1995, 1067 zero HLA-mismatched kidneys that were not phenotypically identical nor six antigen matched, had a 1-year graft survival rate of 88%. Graft survival has not decreased as a result of these changes in the criteria for national sharing, despite an increase in the percentage of matched transplants from 2.5% during the six antigen-match era to 15.5% during the zero antigen-mismatch era. CONCLUSIONS: Changes to the United Network for Organ Sharing policy for national sharing of HLA-matched kidneys have increased the number of patients, and especially minority patients, who can benefit by receiving a well-matched graft without compromising the high graft survival rates provided by an HLA-matched kidney.

Cadaver↗
PubMed1998

Effects of thapsigargin and KCl on the O2 use of the excised blood-perfused rat heart.

We have already reported a curvilinear end-systolic pressure-volume relation and a linear oxygen consumption per beat (Vo2)-systolic pressure-volume area (PVA) relation of the left ventricle in the rat blood-perfused whole heart preparation. Recently, we have proposed that a PVA at an appropriate left-ventricular end-diastolic volume (0.15 ml/g), such as PVA0.15 and the Vo2 intercept (minimally loaded Vo2) of the Vo2-PVA relation, can be good indexes for assessing rat left-ventricular mechanoenergetics. The minimally-loaded Vo2 would mainly consist of Vo2 for Ca 2+ handling in the excitation-contraction (E-C) coupling and basal metabolism. However, the fraction of the Vo2 by the sarcoplasmic reticulum (SR) Ca2+-ATPase in the total Ca2+ handling Vo2 has not yet been examined. To study this for the first time, we investigated the effects of thapsigargin and KCl on left-ventricular mechanoenergetics. Blockade of the SR Ca2+ pump by thapsigargin (2.5 micromol/l) decreased PVA 0. 15 by 65% and decreased the Vo2 intercept by 40% without a change in the slope of the Vo2-PVA relation. We measured the basal metabolic Vo2 by intracoronary KCl and found a 75% decrease in the Vo2 intercept. We conclude that the minimally-loaded Vo2 mainly consists of Vo2 for Ca2+ handling in the E-C coupling (at least 40% is consumed by the SR Ca2+ ATPase) and basal metabolism (25%).

Animals↗
PubMed1998

Synergistic effect of electric pulses and bleomycin on cultured rabbit subconjunctival fibroblasts.

BACKGROUND: The combined effect of electric pulses (EP) and antiproliferative agents on the proliferation of rabbit Tenon's capsule fibroblasts was investigated. METHODS: Rabbit Tenon's capsule fibroblasts were cultured. Some of these cells were exposed to various intensities of EP alone (500-2500 V/cm). Other cells were then exposed for 30 min to an antiproliferative agent: bleomycin (BLM; 0.0005-50 mumol/l), mitomycin C (MMC; 0.0005-50 mumol/l), 5-fluorouracil (5-FU; 0.05-5000 mumol/l), or streptomycin (SM; 0.0005-50 mumol/l) with or without EP (2000 V/cm, 99 mus, eight pulses). Cell proliferation was assessed by cell counting on day 3 and by a 3H-thymidine uptake assay. DNA fragmentation was assessed by flow-cytometric analysis and agarose gel electrophoresis. RESULTS: A significant reduction in the cell number was observed only at 2500 V/cm (P < 0.05). BLM, MMC and 5-FU treatment inhibited cell proliferation in a dose-dependent manner either with or without EP (ID50: BLM alone, 0.029 mumol/l; BLM and EP, 0.00022 mumol/l; MMC alone, 41.6 mumol/l; MMC and EP, 27.5 mumol/l; 5-FU alone, 1045 mumol/l; 5-FU and EP, 690.2 mumol/l; P < 0.05). EP treatment induced an inhibitory effect of BLM on cell proliferation which was 100 times more prominent than BLM alone (0.0005 mumol/l of BLM alone 103.4 +/- 4.4%, 0.0005 mumol/l of BLM and EP 26.0 +/- 4.4%; P = 0.021). BLM treatment with EP also augmented the apoptotic-like DNA fragmentation in both a flow-cytometric DNA histogram and agarose gel-electrophoresis. CONCLUSION: EP treatment enhanced the inhibitory effect of BLM on the cell proliferation of Tenon's capsule fibroblasts of rabbits. The combination of electric pulses and antiproliferative drug treatments may therefore reduce the necessary dose of antiproliferative agents in filtering surgery.

Animals↗
PubMed1998

Role of NMDA receptors in the propagation of excitation in rat visual cortex as studied by optical imaging.

To examine the role of the N-methyl-D-aspartate (NMDA) type of glutamate receptors in the propagation of information in visual cortex, optical imaging with high spatial and temporal resolution of neuronal activity was used in cortical slices of rats. Single-shock stimulation of the white matter elicited a vertical propagation of excitation toward the cortical surface simultaneously with a horizontal spread of excitation in lower layers. The horizontal spread in upper layers occurred subsequent to the vertical spread reaching these layers. The results from perfusion of Ca2+-free medium and application of an antagonist of non-NMDA receptors indicated that this intracortical propagation of signals is due mostly, if not exclusively, to the postsynaptic excitation of cortical neurons. Blockade of NMDA receptors attenuated the rising and peak phases of the upper horizontal spread, but did not affect those of the lower horizontal or vertical propagation of excitation. Perfusion with Mg2+-free solution enhanced the upper horizontal spread, but in most cases did not significantly change the spread of excitation in the other pathways. These results indicate that NMDA receptors are involved in the flow of information in the upper layers of visual cortex, and further suggest that this propagation of activity is mediated mainly by horizontal connections intrinsic to the upper layers.

2-Amino-5-phosphonovalerate↗
PubMed1998

Synaptic PDZ domain-containing proteins.

Synaptic junctions accumulate various molecules involved in neurotransmissions and synaptic plasticity. The proper organizations of these molecules at synaptic junctions sustain the interneuronal signal transmissions and the subsequent signal cascades underling learning and memory. Recent studies have revealed that a protein-interacting module named a PDZ domain plays important roles in the interactions among the components of synaptic junctions. In this article, we summarize synaptic PDZ domain-containing proteins and discussed their functions.

Adaptor Proteins, Signal Transducing↗
PubMed1998

Fluorophotometric measurement of the buffering action of human tears in vivo.

PURPOSE: The buffering action of human tears is thought to be important to keep its pH constant. We measured the change in pH in the precorneal tear film in vivo when the acidic solution is challenged, using a fluorophotometric technique. METHODS: Twelve eyes from 6 healthy subjects were entered in this study. Each subject was pretreated with either one drop of 0.4% oxybuprocaine for once (light anesthesia), three times (deep anesthesia), or none (controls). The measurement was initiated by instilling 20 microl of 0.067 M phosphate buffer at pH 5.5 containing 2 mM bis-carboxyethyl-carboxyfluorescein free acid, a pH sensitive dye, into the subject's eye. The pH was determined by the ratio of fluorescent intensities at two excitation wavelengths (490 and 430 nm). pH recovery time (PHRT) as defined by the time required for pH to reach 95% of pH at equilibrium was used for the marker of tear buffering action. Tear turnover rate was also determined using the fluorescent decay curve at 430 nm, which was independent of pH, but dependent on dye concentration. RESULTS: Immediately after the instillation, the pH value in the tear film was around 6.0-6.5 in all cases. The tear film rapidly became more alkaline, reaching its normal value in 2.3 +/- 0.5 min in untreated eyes. The pretreatment with 0.4% oxybuprocaine retarded the neutralization process. A single regression analysis revealed that the PHRT had a significant negative correlation with the tear turnover rate (r = -0.78). CONCLUSIONS: Our results suggest that the neutralization process of tears largely depends on the tear turnover rate. The buffering action of tears in vivo consists of the tear turnover as well as its chemical buffering capacity.

Acid-Base Equilibrium↗
PubMed1998

Long-range effect of mutation of calcium binding aspartates [correction of asparates] on the catalytic activity of alkaline protease from Pseudomonas aeruginosa.

Apart from a catalytic domain, the alkaline protease of Pseudomonas aeruginosa has a novel parallel beta-helix domain stabilized through Ca2+ binding. In order to clarify the importance of the beta-helix structure in maturation of the enzyme, aspartic residue D-356 or D-365 in the Ca2+ binding sequence motif was replaced with L-alanine, and the catalytic activity of each mutant was assayed. These mutants did not show any proteolytic activity, although the composition of their polypeptide chains was the same as that of the wild type except for the mutated alanine residue. These results suggest that D-356 and D-365 are important in control of the beta-helix folding induced by Ca2+ binding and that incomplete beta-helix folding due to the lack of their side-chains affects the maturation of the enzyme in the long-range order.

Aspartic Acid↗
PubMed1998

X-ray structure of a reaction intermediate of L-2-haloacid dehalogenase with L-2-chloropropionamide.

The crystal structure of a complex prepared by soaking a single crystal of the S175A mutant of L-2-haloacid dehalogenase from Pseudomonas sp. YL in a solution containing a poor substrate, L-2-chloropropionamide, has been determined by X-ray analysis at 2. 15 A resolution with a crystallographic R factor of 19.8%. The present analysis has revealed the structure of the reaction intermediate trapped in the crystal. In the intermediate, the substrate moiety lacking chlorine is covalently bound to the carboxyl group of D10, and adopts the pro-D-configuration at the C2 atom. The amide group of the substrate is hydrogen-bonded with the hydroxy group of S118. The methyl group bound to the C2 atom exists in a hydrophobic pocket which is important for recognition of the alkyl group of the substrate. The guanidino group of R41 has reasonable orientation for halogen-abstraction.

Amides↗
PubMed1998

Cathepsin D expression in skin metastasis of breast cancer.

Cathepsin D, an aspartic proteinase, correlates with invasion and metastasis in breast cancer and with poor prognosis. In the present study, we examined the immunohistological expression of cathepsin D in both primary (5 cases) and skin-metastatic breast cancers (13 cases) and compared it to those in gastric (2 cases) and lung (4 cases), and primary eccrine cancers (3 cases). All breast and gastric cancers were adenocarcinomas. The 2 gastric cancers were poorly differentiated, while the 4 lung cancers consisted of 2 poorly differentiated adenocarcinomas, 1 poorly differentiated large cell carcinoma, and 1 moderately to poorly differentiated squamous cell carcinoma. We also surveyed the immunohistological distribution of cathepsin B, carcinoembryonic antigen, gross cystic disease fluid protein-15, c-erbB-2, and estrogen receptor. In almost all breast cancer samples, the cancer cells demonstrated strong expression of cathepsin D in the cytoplasm, but weak staining patterns with other antibodies. Gastric and lung cancer cells did not respond with cathepsin D (except one metastatic lung cancer) or the other immunohistological markers. Since cathepsin D is strongly expressed in primary and metastatic lesions of breast cancer, cathepsin D could be useful as an adjunct to a panel of immunohistochemical stains in determining the primary site of origin of metastatic cancer in the skin.

Adenocarcinoma↗
PubMed1998

Frontotemporal dementia and Alzheimer disease: evaluation of cortical atrophy with automated hemispheric surface display generated with MR images.

PURPOSE: To determine the features of cortical atrophy in frontotemporal dementia (FTD) and Alzheimer disease by using a hemispheric surface display generated with magnetic resonance (MR) images. MATERIALS AND METHODS: The extent of cortical atrophy was evaluated with automated MR hemispheric surface display and volumetry in 18 patients with FTD and in 18 matched patients with Alzheimer disease. Results were compared with those in 18 healthy, matched control subjects. RESULTS: Most cortical regions were significantly atrophic in FTD and Alzheimer disease. The frontal and anterior temporal lobes were significantly more atrophic in FTD than in Alzheimer disease. The mean hemispheric-to-intracranial volume ratio in patients with FTD (56.2%) and those with Alzheimer disease (58.4%) was significantly smaller than the ratio in the control subjects (66.0%). Asymmetry of hemispheric volume was significantly larger in the FTD group than in the Alzheimer disease and control groups. CONCLUSION: Cortical atrophy in FTD is more widespread than was previously thought. Asymmetric frontal and anterior temporal atrophy is a distinctive feature of FTD and distinguishes it from Alzheimer disease. Hemispheric surface display is a useful complement to tomographic images and is useful for the evaluation of focal cortical atrophy in degenerative dementias, especially FTD.

Aged↗
PubMed1998

The role of NF-kappaB in retinal neovascularization in the rat. Possible involvement of cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin-8 family.

Hypoxia precedes neovascularization in many retinal diseases that can lead to irreversible vision loss. The transcription factor NF-kappaB is activated by hypoxia and regulates the expression of many genes, including angiogenic factors. The relation between the NF-kappaB activation and the cytokine-induced neutrophil chemoattractant (CINC), a member of the interleukin-8 (IL-8) family, was investigated by immunohistochemistry in a rat model of proliferative retinopathy presumably caused by relative hypoxia. Activated NF-kappaB and CINC immunoreactivity was detected in retinal glial cells in the nonperfused retina and in neovascular cells. Activated NF-kappaB was detected before the CINC staining, and both of these events occurred before the development of neovascularization. The intensity of both activated NF-kappaB and CINC staining remained increased during the development of neovascularization and then declined as neovascularization regressed. In rat retinal glial cells in vitro, dexamethasone, an inhibitor of NF-kappaB activation, prevented the hypoxia-induced increase in the amount of CINC mRNA. Furthermore, CINC induced neovascularization in a rat corneal pocket model. These results suggest that hypoxia-induced activation of NF-kappaB results in CINC production and participates in the induction of retinal neovascularization.

Actins↗
PubMed1998

Endothelium-dependent potentiation of prostaglandin F2alpha-induced contractions by (+/-)-[6]-gingerol is inhibited by cyclooxygenase- but not lipoxygenase-inhibitors in mouse mesenteric veins.

The mechanism of potentiation of prostaglandin (PG) F2alpha-induced contraction of mouse mesenteric veins by (+/-)-[6-gingerol was investigated in vitro. (+/-)-[6]-Gingerol (0.3mM) potentiated the maximal contraction response elicited by PGF2alpha (0.28 mm) in the presence of intact vascular endothelium, but not in its absence (de-endothelialized preparations). The potentiating effect was completely inhibited by cyclooxygenase inhibitors (0.2 mm aspirin and 0.2 mm indomethacin) and partly by calcium antagonists (2 microM verapamil, 8 nM nitrendipine and 1 microM ryanodine), but not inhibited by nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor and ONO-3708, a thromboxane (TX) A2 antagonist. The potentiation by (+/-)-[6]-gingerol is also observed in mesenteric veins of streptozotocin-diabetic mice where the enhancement of PGF2alpha-induced contraction is caused mainly by activation of lipoxygenase. The potentiation of PGF2alpha-induced contraction by (+/-)-[6]-gingerol may be caused by a cyclooxygenase-dependent release of vasoconstrictors, other than PGF2alpha and TXA2, or by inhibiting vasorelaxants released from endothelial cells of mouse mesenteric veins.

Animals↗
PubMed1998

Linear O2 use-pressure-volume area relation from curved end-systolic pressure-volume relation of the blood-perfused rat left ventricle.

We measured rat left ventricular pressure, volume, and oxygen consumption ( = arteriovenous oxygen content differencexcoronary flow) to establish a new evaluation of its mechanoenergetics in the whole heart preparation by using the cross-circulation method. We obtained a curved end-systolic pressure-volume relation in contrast to a linear end-systolic pressure-volume relation in dogs, rabbits, and humans. However, we obtained a linear oxygen consumption per beat (VO2)-systolic pressure-volume area (PVA, a measure of left ventricular total mechanical energy per beat) relation as in other species. Thus PVA can be a good index for assessing rat left ventricular mechanoenergetics. The VO2 intercept and slope of the linear VO2-PVA relation correspond to those in other species. Intracoronary calcium elevated the curved end-systolic pressure-volume relation and significantly increased PVA at 0.15 ml/g of left ventricular end-diastolic volume (PVA0.15) by 50%. Calcium also significantly increased the VO2 intercept of the VO2-PVA relation by 30% without a change in its slope. We conclude that the rat left ventricular end-systolic pressure-volume relation is curved, but the VO2-PVA relation is linear, and that the VO2 intercept is mainly composed of PVA-independent VO2, presumably VO2 for Ca2+ handling in the excitation-contraction coupling and basal metabolism. Therefore we propose PVA at an appropriate left ventricular volume and the VO2 intercept as good rat left ventricular mechanoenergetic indexes despite the nonlinearity of the end-systolic pressure-volume relation.

Animals↗