Biomedical subjects

H Ueda

Publications and source records attributed to H Ueda.

At least 415 records · Page 23Linked to original sources

PubMed1996

A novel microsatellite polymorphism in the human OB gene: a highly polymorphic marker for linkage analysis.

The mouse ob gene and its human homologue OB have recently been cloned. The mutations in the ob gene are known to be associated with extreme obesity. The relationship between the human OB gene and disease, however, is largely unknown due to the lack of suitable markers within or adjacent to the OB gene. To obtain informative markers, we searched for simple tandem repeat polymorphisms in the genomic sequence of the human OB gene and identified a novel tetranucleotide repeat in the 3' flanking region. Fifteen alleles were detected in this marker with a heterozygosity of 0.85 and polymorphism information content of 0.83, indicating a highly informative nature of this marker. Two-point linkage mapping in two Centre Etude Polymorphisme Humaine (CEPH) reference families suggested that this marker is located in the interval between D7S514 and D7S530, the same interval where the OB gene is located (recombination fractions with D7S514 and D7S530 were 0.026 and 0.034, respectively). Although allele frequency distributions of this marker did not differ between 84 control subjects and 69 NIDDM patients, there was a tendency to higher body weight in control subjects with class I/class I genotype than in those without this genotype (68.8 +/- 11.1 vs 60.8 +/- 10.3 kg, p = 0.05). The highly polymorphic nature of this marker and its location in the OB gene makes this marker useful for linkage studies of the OB gene with a number of phenotypes, such as obesity, non-insulin-dependent diabetes mellitus, hypertension and the insulin resistance syndrome.

Alleles↗

PubMed1996

Cytophysiology of gonadotropin-releasing-hormone neurons in chum salmon (Oncorhynchus keta) forebrain before and after upstream migration.

Cytophysiology of gonadotropin-releasing-hormone neurons in chum salmon (Oncorhynchus keta) was examined before and after upstream migration by an immunocytochemical technique with a specific antiserum to salmon gonadotropin-releasing hormone and an in situ hybridization technique with an oligonucleotide encoding salmon gonadotropin-releasing hormone precursor (pro-salmon gonadotropin- releasing hormone). In the forebrain (olfactory nerve, olfactory bulb, telencephalon, and preoptic area), salmon gonadotropin-releasing hormone-immunoreactive neurons and neurons showing signals for pro-salmon gonadotropin-releasing hormone mRNA were compared between fish from the coastal sea and those from the spawning ground. Neurons in the dorsal region of the olfactory nerve and in the ventral region of the transitional area between olfactory nerve and olfactory bulb showed strong salmon gonadotropin-releasing-hormone immunoreactivity and strong hybridization signals in fish from the coastal sea, but these activities and signals were not observed or were decreased in number in fish from the spawning ground. The neurons in the olfactory bulb, telencephalon, and preoptic area consistently revealed salmon gonadotropin-releasing-hormone immunoreactivity and hybridization signals, and the hybridization signals of salmon gonadotropin-releasing hormone in the telencephalon and the preoptic area were stronger in fish from the spawning ground than in those from the coastal sea. These findings suggest that salmon gonadotropin-releasing-hormone neurons in the olfactory nerve and the transitional area between olfactory nerve and olfactory bulb have different patterns of hormone production than those in the telencephalon and the preoptic area.

Animals↗

PubMed1996

Surgical atrial septostomy without cardiopulmonary bypass.

Surgical atrial septostomy using a special atriotomy knife without cardiopulmonary bypass in patients with obstruction of the left-sided atrioventricular valve and complex cardiac anomalies is described. This procedure is effective, safe, and economical for patients in the acute stage after intracardiac repair, and available for patients with a closed fossa ovalis.

Cardiac Surgical Procedures↗

PubMed1996

Two different activities of phospholipase A2 in porcine aortic cytosol.

Two different activities of phospholipase A2 (PLA2s, EC 3.1.1.4) which was activated at micro molar concentration of free Ca2+ were identified in cytosolic fraction of porcine aortic tissue. The two enzymes behaved very similarly during purification on ion-exchange, hydrophobic, gel filtration and affinity columns, and finally the active peaks of these enzymes were separated from each other on a hydroxyapatite HPLC column. Both of the enzymes hydrolyzed efficiently phosphatidylcholine (PtdCho) containing arachidonoyl residue as the substrate, but showed the apparent differences in head group specificity of phospholipids and also in the sensitivity against high Ca2+ concentration. This is the first report for the identification and characterization of high molecular weight cytosolic PLA2s in porcine aortic tissue.

Animals↗

PubMed1996

Neurobiology of L-DOPAergic systems.

L-DOPA is proposed to be a neurotransmitter and/or neuromodulator in CNS. It is released probably from neurons, which may contain L-DOPA as an end-product, and/or from some compartment other than catecholamine-containing vesicles. The L-DOPA itself produces presynaptic and postsynaptic responses. All are stereoselective and most are antagonized by competitive antagonist. In striatum, L-DOPA is neuromodulator, mother of catecholamines, not only a precursor for dopamine but also a potentiator of children for presynaptic beta-adrenoceptors to facilitate dopamine release and postsynaptic D2 receptors, and ACh release inhibitor. All may cooperate for Parkinson's disease. Meanwhile, supersensitization of increase in L-glutamate release to nanomolar levodopa was seen in Parkinson's model rats, which may relate to dyskinesia or "on-off" during chronic therapy. In lower brainstem, L-DOPA tonically activates postsynaptic depressor sites of NTS and CVLM and pressor sites of RVLM. L-DOPA is probably a neurotransmitter of primary baroreceptor afferents terminating in NTS. GABA, the inhibitory neuromodulator for baroreflex in NTS, tonically functions to inhibit, via GABAA receptors, L-DOPA release and depressor responses to levodopa. Levodopa inversely releases GABA. L-DOPAergic monosynaptic relay from NTS to CVLM and from PHN to RVLM is suggested. Tonic L-DOPAergic baroreceptor-aortic nerve-NTS-CVLM relay seems to carry baroreflex information. Disturbance of neuronal activity to release L-DOPA in NTS, loss of the activity in CVLM, enhancement of the activity with decreased decarboxylation and increase in sensitivity to levodopa in RVLM may be involved in maintenance of hypertension in SHR. This is a story of "L-DOPAergic receptors" with extremely high affinity and low density.

Afferent Pathways↗

PubMed1996

Ten-year experience with the Carpentier-Edwards pericardial xenograft in the tricuspid position.

We reviewed the cases of 66 patients who underwent 67 tricuspid valve replacements with Carpentier-Edwards pericardial xenografts between April 1985 and January 1994. Average patient age at time of operation was 52 years (range 8 to 71 years). Concomitant mitral or aortic valve replacements were performed in 46 patients. There were 10 operative deaths and 6 late deaths. Actuarial survival at 9 years was 75.4% +/- 5.7%. Prosthetic valve endocarditis occurred twice in one patient. Reoperations for tricuspid regurgitation and for concomitant procedures (maze operation and repair for leak of the mitral prosthesis) were performed in two patients. In both cases, examination of the explanted prostheses showed that the tricuspid regurgitation was the result of nonstructural dysfunction caused by fibrous pannus formation on the cusps of the ventricular side. Among the survivors, 47 patients (92%) were in functional class I or II. Prosthetic valve function was studied by color Doppler echocardiography. Among 38 patients, tricuspid regurgitation more than grade 3/4 or transprosthetic gradient more than 5 mm Hg was found in 11. One patient had right heart failure and the others had no symptoms. In 10 years of experience with the Carpentier-Edwards pericardial xenograft, mortality and morbidity after tricuspid valve replacement were satisfactory. Echocardiographic examination revealed subclinical prosthetic dysfunction in 35% of patients who were followed up for longer than 5 years, however, and we believe that these patients should receive careful follow-up.

Adolescent↗

PubMed1996

Cell biology of kidney glomerulus.

It has been accepted that some artifacts are inevitably produced by the conventional preparation steps for electron microscopy, including fixation, dehydration, embedding, ultrathin sectioning, and staining. Therefore, conventional ultrastructural findings on kidney glomeruli are hardly thought to be correlated with the physiological functions of kidneys in vivo. In this chapter, two preparation techniques, the quick-freezing and deep-etching (QF-DE) method or the quick-freezing and freeze-substitution (QF-FS) method, are presented and shown to be useful for clarifying the ultrastructures of kidney glomeruli more closely to structures in vivo with fewer artifacts. Moreover, the ultrastructures of glomerular capillary loops have been demonstrated by a new "in vivo cryotechnique," that shows that hemodynamic factors should be considered in the morphological study of glomerular functions.

Animals↗

PubMed1996

Open sandwich ELISA: a novel immunoassay based on the interchain interaction of antibody variable region.

We describe an immunoassay that is based on the interchain interaction of separated VL and VH chains from a single chain antibody variable region. In the presence of antigen, the chains reassociate. VL fragments of anti-hen egg lysozyme (HEL) antibody HyHEL-10 were immobilized on microtiter plates. Samples were coincubated with an M13-displayed VH chain, and assayed with peroxidase-labeled anti-M13 antibody. Signal was detected in direct proportion to the amount of HEL in the sample. Wide dynamic range with < 15 ng/ml sensitivity was attained.

Animals↗

PubMed1996

A subtype of kappa-opioid receptor mediates inhibition of high-affinity GTPase inherent in Gi1 in guinea pig cerebellar membranes.

The kappa-opioid receptor agonists including U-50,488H and dynorphin A (1-17) in ranges of 0.1-100 nM inhibited the hydrolysis of GTP to GDP (P(i) release) inherent in GTP-binding proteins (G proteins) in guinea pig cerebellar membranes. U-50,488H inhibited only high-affinity GTPase activity, not low-affinity activity. The action of this agonist was found to be biphasic, and there was no inhibition at concentrations > 1 microM. The inhibition was abolished by pretreatment with preactivated pertussis toxin (PTX) at concentrations > 1 micrograms/ml but not with preactivated cholera toxin (30 micrograms/ml). Similar blockade of kappa-receptor-mediated inhibition was also observed when membranes were pretreated with a low concentration (8 microM) of N-ethylmaleimide (NEM) at low temperature (4 degrees C), which alkylates the cysteine residue to be ADP-ribosylated by PTX; but this treatment caused no significant change in kappa-agonist binding. When purified Gi1, but not G(o), was reconstituted into membranes pretreated with NEM, the kappa-receptor-mediated inhibition was recovered. These findings suggest that a subtype of kappa-opioid receptor is coupled to inhibition of intrinsic activity of Gi1.

Animals↗

PubMed1996

Exposure of astrocytes to hypoxia/reoxygenation enhances expression of glucose-regulated protein 78 facilitating astrocyte release of the neuroprotective cytokine interleukin 6.

Astrocytes exposed to hypoxia (H) or hypoxia/ reoxygenation (H/R) maintain cell viability and display changes in protein biosynthesis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of metabolically labeled astrocytes exposed to H showed induction of an approximately 78-kDa polypeptide that demonstrated sequence identity with glucose-regulated protein (GRP) 78. Cell lysates from H/R astrocytes displayed induction of neuroprotective interleukin (IL) 6, which was present in a high-molecular-weight complex also containing GRP78, suggesting that GRP78 might be functioning as a chaperone during cellular stress consequent on H/R. Introduction of antisense oligonucleotide to GRP78 into astrocytes prevented expression of the protein and suppressed H/R-induced astrocyte release of IL-6 by approximately 50%. These data indicate that modulation of astrocyte properties during oxygen deprivation results, in part, from intracellular glucose depletion and subsequent expression of GRP78, which sustains generation of neuroprotective IL-6 under the stress of H/R.

Animals↗

PubMed1996

Truncation of Vargula luciferase still results in retention of luminescence.

Significant amino acid sequence homology in two regions of Vargula hilgendorfii to one in apoaequorin was reported. The intra-amino acid homology in Vargula luciferase between residues 81-312 and 321-540 was 19.3%, and each of this intra-homologous region contained the region homologous to apoaequorin. In order to prove the possibility that only one of the homologous regions is sufficient for luminescence, we have produced a chimeric protein comprising of only the N-terminal homologous region of Vargula luciferase fused to protein A. Comparison of the luminescence of this truncate luciferase indicated that there was 38.5% retention in the bioluminescence of luciferase when compared to that of the mature form of luciferase. This fact may have interesting implications for further study of engineering luciferase.

Animals↗

PubMed1996

Magnetic resonance imaging of neurogenic tumors of the thoracic inlet: determination of the parent nerve.

To investigate the value of magnetic resonance imaging (MRI) in determining the parent nerves of neurogenic tumors in the thoracic inlet, analysis of MR images was performed in nine patients with surgically resected neurogenic tumors in the thoracic inlet (two neurofibromas and one schwannoma of the vagus nerve, three schwannomas of the brachial plexus, and two schwannomas and one ganglioneuroma of the sympathetic nerves). These MR images were compared with surgical and pathologic findings. The multidirectional capability and excellent tissue contrast of MRI facilitated recognition of the location, shape, and extent of the tumors. MRI, which permitted an easy understanding of the spatial relation between the tumors and the subclavian vessels, scalenus muscles, and brachial plexus, was useful in determining the nerves of origin. Two neurofibromas, four of six schwannomas, and one ganglioneuroma were recognized to extend along the axes of the parent nerves on MR images. MRI is useful in determining the parent nerve of neurogenic tumors in the thoracic inlet and is helpful in planning surgical treatment of these tumors.

Brachial Plexus↗

PubMed1996

Establishment of an undifferentiated leukemia cell line (Kasumi-3) with t(3;7)(q27;q22) and activation of the EVI1 gene.

A novel human leukemia cell line (Kasumi-3) was established from the blast cells of a 57-year-old man suffering from myeloperoxidase-negative acute leukemia. The cell line had five distinctive features, as follows. 1) Flow cytometric analyses showed cell surface expression of CD7, CD4, CD13, CD33, CD34, HLA-DR and c-Kit. This phenotype is compatible with that of acute myelocytic leukemia cells with the M0 subtype in the French-American-British classification. 2) Kasumi-3 cells carried chromosomal abnormalities of t(3;7)(q27:q22), del(5)(q15), del(9)(q32), and add(12)(p11). The breakpoint of 3q27 was located near the EVI1 gene, and a high level of expression of the EVI1 gene was observed. 4) Kasumi-3 cells treated with TPA showed maturation to monocytic lineage. 5) Treatment with either interleukin (IL)-2, IL-3, IL-4, granulocyte-macrophage colony-stimulating or stem cell factor induced the proliferation of Kasumi-3 cells. Thus, the Kasumi-3 cell line shows the characteristic features of undifferentiated leukemia. It should, therefore, be useful both for studying the biological characteristics of acute myelogenous leukemia M0 subtype and for investigating the role of the EVI1 gene in leukemogenesis.

Acute Disease↗

PubMed1996

Regulation of the EDG84A gene by FTZ-F1 during metamorphosis in Drosophila melanogaster.

The transcription factor FTZ-F1 is a member of the nuclear hormone receptor superfamily and is transiently expressed during the mid- and late prepupal periods in Drosophila melanogaster. A putative pupal cuticle gene, EDG84A, is expressed slightly following FTZ-F1 expression during the prepupal period and carries a strong FTZ-F1 binding site between bases 100 and 92 upstream of its transcription start site. In this study, EDG84A mRNA was found to be prematurely expressed upon heat induction of FTZ-F1 in prepupae carrying the heat shock promoter-FTZ-F1 cDNA fusion gene construct. Transgenic fly lines having the 0.8-kb region of the EDG84A promoter fused to lacZ expressed the reporter gene in a tissue- and stage-specific manner. Base substitutions in the FTZ-F1 binding site within the 0.8-kb promoter abolished expression of lacZ. These results strongly suggest that the EDG84A gene is a direct target of FTZ-F1. Deletion studies of the cis-regulatory region of the EDG84A gene revealed that space-specific expression in imaginal disc-derived epidermis is controlled by the region between bp -408 and -104 from the transcription start site. The region between bp -408 and -194 is necessary to repress expression in a posterior part of the body, while the region between bp -193 and -104 carries a positive element for activation in an anterior part of the body. These results suggest that FTZ-F1 governs expression of the EDG84A gene in conjunction with putative tissue-specific regulators.

Animals↗

PubMed1996

Proteoglycans in articular cartilage revealed with a quick freezing and deep etching method.

OBJECTIVES: To clarify the three dimensional ultrastructure of proteoglycans, and their relationship with other matrix components in articular cartilage. METHODS: Specimens from rat femoral heads were examined using three techniques: (1) Histochemical staining with cationic polyethyleneimine (PEI), using a pre-embedding or a postembedding method. Some tissues were pretreated with chondroitinase ABC or hyaluronidase. (2) Quick freezing and deep etching (QF-DE). Some specimens were fixed with paraformaldehyde and washed in buffer solution before quick freezing; others were frozen directly. (3) Ultrathin sections were studied after conventional preparation. RESULTS: Proteoglycans were observed as aggregated clumps with PEI staining by the pre-embedding method, but as fine filaments by the postembedding method. They were lost with enzyme digestion; this was also demonstrated by the QF-DE method. The ultrastructure was well preserved by the QF-DE method when fixation and washing procedures were included, but not without these procedures. A fine mesh-like structure was connected to the cell membrane in the pericellular matrix. Filamentous structures suggestive of aggrecans were observed among collagen fibrils. They had side chains, approximately 50 nm in length, which branched from the central filaments at intervals of 10-20 nm, and were occasionally linked to other structures. Many thin filaments were also attached to the collagen fibrils. CONCLUSIONS: The QF-DE method incorporating paraformaldehyde fixation and buffer washing procedures revealed three dimensional, extended structures suggestive of proteoglycans.

Animals↗

PubMed1996

Delayed neuronal death in ischemic hippocampus involves stimulation of protein tyrosine phosphorylation.

Glutamate triggers neuronal degeneration after ischemia-reperfusion in the brain. However, the details of intracellular signal transduction that propagates cell death remain unknown. The present work investigated whether protein tyrosine phosphorylation mediates neuronal death in the ischemic brain. Transient forebrain ischemia for 5-10 min in Mongolian gerbils or intoxication with the glutamate analogue kainic acid (12 mg/kg) in Sprague-Dawley rats caused neuronal death selectively in the hippocampus 2-4 days or 1 day later, respectively. Under these conditions, 160-, 115-, 105-, 92-, and 85-kDa proteins showed a significant increase in tyrosyl residue phosphorylation selectively in the hippocampus 3-12 h after ischemia or 4-8 h after kainic acid-induced seizures. Tyrosine kinases, including pp60c-src, were activated without a change of tyrosine phosphatases. Administration of radicicol, a selective inhibitor of tyrosine kinases, attenuated stimulation of tyrosine phosphorylation and hippocampal degeneration after ischemia or kainic acid injection. The results suggest that protein tyrosine phosphorylation might propagate delayed neuronal death in the mature hippocampus through glutamate overload after ischemia-reperfusion.

Animals↗

PubMed1996

Eustachian tube function and middle ear barotrauma associated with extremes in atmospheric pressure.

Eustachian tube (ET) function was studied by means of sonotubometry and tubotympano-aerodynamography (TTAG) prior to and following exposure to hypobaric or hyperbaric conditions. Forty normal adults were subjected to hypobaric pressure. Fifty adults who underwent hyperbaric oxygen (HBO) therapy also were studied. Following hypobaric exposure, 14 of 80 ears (17.5%) exhibited middle ear barotrauma. Following hyperbaric exposure, 34 of 100 ears (34%) exhibited middle ear barotrauma. Dysfunction of the ET, characterized by altered active and passive opening capacity, was more prevalent following exposure to extremes in atmospheric pressure compared to baseline. The ET function, which was impaired after the first HBO treatment, improved gradually over the next 2 hours. Overall, however, ET function was worse after the seventh treatment. The patients who developed barotrauma exhibited worse ET function prior to hypobaric or hyperbaric exposure. Thus, abnormal ET function can be used to predict middle ear barotrauma prior to exposure to hypobaric or hyperbaric atmospheric pressure.

Adult↗

PubMed1996

Synergistic response of cord blood myeloid progenitor cells to the combined administration of human granulocyte colony-stimulating factor and human stem cell factor in vitro.

We compared the in vitro response of myeloid progenitor cells [colony-forming units of culture (CFU-C)] that were prepared from human umbilical cord blood to the administration of the combination of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) versus that of CFU-C obtained from normal human bone marrow. Progenitors were purified according to CD34 expression; the number and size of colonies were evaluated by culture in agar or methylcellulose, respectively. In the presence of G-CSF alone, the mean number of colonies was significantly greater in the bone marrow culture versus that of cord blood. SCF alone bad little effect on colony formation, but in the presence of optimal or suboptimal concentrations of G-CSF, SCF significantly increased colony formation from both cell sources. Its effect on cord blood significantly exceeded that on bone marrow. SCF used in combination with G-CSF significantly increased the size of the colonies with cord blood CFU-C; this effect was less marked with bone marrow CFU-C. The percentage of cells that expressed c-Kit, the SCF receptor, did not appear to differ between the two sources of CFU-C. Results indicate that cord blood CFU-C showed a greater response to SCF in combination with G-CSF than did bone marrow CFU-C.

Cells, Cultured↗