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Biomedical subjects

H Abe

Publications and source records attributed to H Abe.

At least 289 records · Page 16Linked to original sources

Characterization of K+ currents underlying pacemaker potentials of fish gonadotropin-releasing hormone cells.

Endogenous pacemaker activities are important for the putative neuromodulator functions of the gonadotropin-releasing hormone (GnRH)-immunoreactive terminal nerve (TN) cells. We analyzed several types of voltage-dependent K+ currents to investigate the ionic mechanisms underlying the repolarizing phase of pacemaker potentials of TN-GnRH cells by using the whole brain in vitro preparation of fish (dwarf gourami, Colisa lalia). TN-GnRH cells have at least four types of voltage-dependent K+ currents: 1) 4-aminopyridine (4AP)-sensitive K+ current, 2) tetraethylammonium (TEA)-sensitive K+ current, and 3) and 4) two types of TEA- and 4AP-resistant K+ currents. A transient, low-threshold K+ current, which was 4AP sensitive and showed significant steady-state inactivation in the physiological membrane potential range (-40 to -60 mV), was evoked from a holding potential of -100 mV. This current thus cannot contribute to the repolarizing phase of pacemaker potentials. TEA-sensitive K+ current evoked from a holding potential of -100 mV was slowly activating, long lasting, and showed comparatively low threshold of activation. This current was only partially inactivated at steady state of -60 to -40 mV, which is equivalent to the resting membrane potential. TEA- and 4AP-resistant sustained K+ currents were evoked from a holding potential of -100 mV and were suggested to consist of two types, based on the analysis of activation curves. From the inactivation and activation curves, it was suggested that one of them with low threshold of activation may be partly involved in the repolarizing phase of pacemaker potentials. Bath application of TEA together with tetrodotoxin reversibly blocked the pacemaker potentials in current-clamp recordings. We conclude that the TEA-sensitive K+ current is the most likely candidate that contributes to the repolarizing phase of the pacemaker potentials of TN-GnRH cells.

4-Aminopyridine↗

New method of bladder biopsy using combined aspiration and snare.

A method of multiple random biopsy combined with bladder wall aspiration and snare in bladder cancer is presented. Our method was used on 23 patients with clinically superficial bladder cancer and was found to have advantages in obtaining an adequate amount of tissue with minimal thermal degeneration.

Biopsy, Needle↗

Cardiomyocytes can be generated from marrow stromal cells in vitro.

We have isolated a cardiomyogenic cell line (CMG) from murine bone marrow stromal cells. Stromal cells were immortalized, treated with 5-azacytidine, and spontaneously beating cells were repeatedly screened. The cells showed a fibroblast-like morphology, but the morphology changed after 5-azacytidine treatment in approximately 30% of the cells; they connected with adjoining cells after one week, formed myotube-like structures, began spontaneously beating after two weeks, and beat synchronously after three weeks. They expressed atrial natriuretic peptide and brain natriuretic peptide and were stained with anti-myosin, anti-desmin, and anti-actinin antibodies. Electron microscopy revealed a cardiomyocyte-like ultrastructure, including typical sarcomeres, a centrally positioned nucleus, and atrial granules. These cells had several types of action potentials, such as sinus node-like and ventricular cell-like action potentials. All cells had a long action potential duration or plateau, a relatively shallow resting membrane potential, and a pacemaker-like late diastolic slow depolarization. Analysis of the isoform of contractile protein genes, such as myosin heavy chain, myosin light chain, and alpha-actin, indicated that their muscle phenotype was similar to that of fetal ventricular cardiomyocytes. These cells expressed Nkx2.5/Csx, GATA4, TEF-1, and MEF-2C mRNA before 5-azacytidine treatment and expressed MEF-2A and MEF-2D after treatment. This new cell line provides a powerful model for the study of cardiomyocyte differentiation.

Action Potentials↗

The C422F mutation of the growth hormone receptor gene is not responsible for short stature.

A missense mutation, C422F, was identified in the intracellular domain of GH receptor (GHR) in a Japanese short boy. Although this mutation was previously reported in a patient with GH insensitivity syndrome (GHIS), it has not been clear whether this mutation causes GH insensitivity. To clarify the effect of this mutation on GH signal transduction, mutant GHR was expressed in CHO cells, and its functional properties were investigated. There were no significant differences in GH-induced tyrosine phosphorylation of STAT5b (signal transducer and activator of transcription) between wild-type GHR (GHR-wt)- and mutant GHR (GHR-C422F)-expressing cells. Moreover, STAT5-mediated transcriptional activation of GHR-C422F-expressing cells was comparable to that of GHR-wt-expressing cells. These findings indicated that the C422F mutation of GHR affected neither GH-induced tyrosine phosphorylation nor the transcriptional activation of STAT5. In addition, the analysis of genotypes and phenotypes of his family revealed that body heights of family members with heterozygous or homozygous C422F mutations were all within normal ranges, with the single exception of the proband. These in vitro and in vivo results indicate that the C422F missense mutation of GHR is a polymorphism that does not result in GHIS.

Animals↗

Functional characterization of truncated growth hormone (GH) receptor-(1-277) causing partial GH insensitivity syndrome with high GH-binding protein.

We have previously reported a novel heterozygous donor splice site mutation in intron 9 of the GH receptor (GHR) gene in Japanese siblings who showed partial GH insensitivity and high serum GH-binding protein (GHBP) levels. This mutation caused the splicing abnormality and produced the truncated GHR consisting of 277 amino acids (GHR-277), which lacked most of the intracellular domain of GHR, including both boxes 1 and 2. In this study, we have characterized the function of GHR-277 expression in COS-7 and CHO cells in vitro. Scatchard analysis revealed that GHR-277 possessed approximately 1.5 times higher affinity to GH and twice the number of binding sites compared to wild-type full-length GHR (GHR-fl). The GHBP level in culture medium of GHR-277-expressing cells was approximately 3 times higher than that in GHR-fl-expressing cells. Interestingly, the ligand-induced internalization of GHR-277 was significantly reduced compared with that of GHR-fl. Moreover, in GH-induced tyrosine phosphorylation of signal transducer and activator of transcription-5 (STAT5), GHR-277 exerted a dominant negative effect when GHR-277 and GHR-fl were cotransfected. These in vitro data would well explain the clinical characteristics in our patients showing high serum GHBP levels and development of short stature despite a heterozygous mutation of the GHR gene.

Adolescent↗

XAIP1: a Xenopus homologue of yeast actin interacting protein 1 (AIP1), which induces disassembly of actin filaments cooperatively with ADF/cofilin family proteins.

We carried out affinity column chromatography using Xenopus ADF/cofilin (XAC), identified several polypeptides in oocytes specifically bound to this column with actin, and isolated a full-length cDNA clone for a 65 kDa protein in this fraction. The predicted amino acid sequence revealed that the 65 kDa protein has seven obvious WD repeats and exhibits striking homology with yeast actin interacting protein 1 (AIP1). Thus, we designated this protein Xenopus AIP1 (XAIP1). We purified XAIP1 from Xenopus oocytes, and its interaction with actin was characterized by a pelleting assay, photometrical analysis and electron microscopy. Although XAIP1 itself cosedimented with F-actin and increased unsedimented actin to some extent, it induced a rapid, drastic disassembly of actin filaments associated with XAC. Electron microscopic observation revealed that XAIP1 severs actin filaments in the presence of XAC. To elucidate the in vivo effects of XAIP1, the purified protein was injected into blastomeres at the two-cell stage. Although the localization of XAIP1 was similar to that of XAC, at the cortical cytoskeleton and diffusely in the cytoplasm, injection of a large amount of XAIP1 arrested development and abolished the strong cortical staining of both actin and XAC. From these results, we concluded that XAIP1 regulates the dynamics of the cortical actin cytoskeleton cooperatively with XAC in eggs.

Actin Depolymerizing Factors↗

Carnosine stimulates vimentin expression in cultured rat fibroblasts.

Two-dimensional electrophoretic gel profiles were compared between rat 3Y1 fibroblasts cultured in the presence and absence of 30 mM L-carnosine (beta-alanyl-L-histidine) for one week without any replenishment of medium. While a number of cellular proteins changed their expression levels by the addition of carnosine, we identified one of the most prominently varied proteins as vimentin. Immunoblot analysis with anti-vimentin antibody demonstrated that the vimentin levels increased about 2-fold after one-week culture in the presence of carnosine. We also confirmed that the increase of vimentin expression was dependent on the concentration of carnosine added to the medium. Moreover, when cultured cells were stained with anti-vimentin antibody and observed by light microscopy, most cells grown in the presence of carnosine were found to have markedly developed vimentin filaments. The increase of vimentin expression was also observed by adding with carnosine related dipeptides, N-acetylcarnosine and anserine.

Amino Acid Sequence↗

Photoinactivation of vesicular stomatitis virus with fullerene conjugated with methoxy polyethylene glycol amine.

Fullerene is known to effectively produce mainly singlet oxygen by absorbing light energy, and therefore may be useful as a virucidal photosensitizer. This study was designed to investigate the virucidal activity of a water-soluble fullerene derivative, which is conjugated with methoxy polyethylene glycol amine to enhance its water solubility. Vesicular stomatitis virus (VSV) was inactivated upon illumination with this water-soluble fullerene, in a concentration- or dose-dependent manner. The titer of VSV was reduced by >5log 10 in the presence of 10 mg/ml (400 microM) fullerene derivative with 120 J/cm2 white light irradiation. VSV inactivation was inhibited by oxygen removal or by the addition of sodium azide, a known singlet oxygen scavenger. The substitution of H2O by D2O, which is known to prolong the lifetime of singlet oxygen, promoted the virucidal activity. These results indicate that singlet oxygen may play a major role in VSV photoinactivation by the water-soluble fullerene derivative. The concentration needed for virus inactivation is higher than that of other sensitizers such as methylene blue.

Antiviral Agents↗

Identification of metabolites of NK-104, an HMG-CoA reductase inhibitor, in rat, rabbit and dog bile.

NK-104 is an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase with a very potent lipid-lowering effect. Biotransformation profiles of NK-104 in bile from rat, rabbit and dog given an intravenous infusion of NK-104 were investigated. Structural assignment was made by liquid chromatography (LC)-mass spectrometry (MS)-MS and proton NMR analyses. The predominant component was intact NK-104 in all the animals. At least eight other metabolites were present in rat, four in rabbit, and 10 in dog. These bile metabolites were purified and isolated by preparative HPLC. Biotransformation pathways elucidated for NK-104 were as follows: (a) lactonization ; (b) beta-oxidation of the side-chain; (c) hydroxylation of the quinoline ring; (d) conjugation with Beta-glucuronic acid and taurine. Beta-oxidative degradation of the side-chain in the case of other HMG-CoA reductase inhibitors is necessary for epimerization of the hydroxy group which has an R-configuration. However, M-16, glucuronide of the ketolactone derivative, was obtained as a key metabolite suggesting another beta-oxidation pathway for the side-chain.

Animals↗

Complete nucleotide sequence of the prophage VT2-Sakai carrying the verotoxin 2 genes of the enterohemorrhagic Escherichia coli O157:H7 derived from the Sakai outbreak.

The enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain RIMD 0509952, derived from an outbreak in Sakai city, Japan, in 1996, produces two kinds of verotoxins, VT1 and VT2, encoded by the stx1 and stx2 genes. In the EHEC strains, as well as in other VT-producing E. coli strains, the toxins are encoded by lysogenic bacteriophages. The EHEC O157:H7 strain RIMD 0509952 did not produce plaque-forming phage particles upon inducing treatments. We have determined the complete nucleotide sequence of a prophage, VT2-Sakai, carrying the stx2A and stx2B genes on the chromosome, and presumed the putative functions of the encoded proteins and the cis-acting DNA elements based on sequence homology data. To our surprise, the sequences in the regions of VT2-Sakai corresponding to the early gene regulators and replication proteins, and the DNA sequences recognized by the regulators share very limited homology to those of the VT2-encoding 933W phage carried by the EHEC O157:H7 strain EDL933 reported by Plunkett et al. (J. Bacteriol., p1767-1778, 181, 1999), although the sequences corresponding to the structural components are almost identical. These data suggest that these two phages were derived from a common ancestral phage and that either or both of them underwent multiple genetic rearrangements. An IS629 insertion was found downstream of the stx2B gene and upstream of the lysis gene S, and this might be responsible for the absence of plaque-forming activity in the lysate obtained after inducing treatments.

Bacterial Toxins↗

Functional prognosis after treatment of spinal dural arteriovenous fistulas.

Functional prognosis after treatment for spinal dural arteriovenous fistulas (SDAVFs) was retrospectively analyzed in 13 consecutive patients aged 38 to 73 years (mean 57 years) treated during the last 5 years. The duration of symptoms before diagnosis ranged from 3 to 72 months (mean 23 months). Neurological symptoms were examined before and 6 months after the treatment. Seven patients underwent embolization as the initial treatment. In four of six patients, N-butyl 2-cyanoacrylate (NBCA) embolization achieved complete obliteration of SDAVF. The other two patients with incomplete embolization and one embolized with polyvinyl alcohol particles underwent subsequent surgical treatment. Six patients were treated by direct surgery. Complete disappearance of SDAVF was confirmed in all nine patients treated surgically. Improvement of gait and micturition disturbance after the treatment was noted in six of 10 and three of six patients, respectively. Long duration of symptoms and high grade of neurological symptoms were associated with a poor functional outcome. NBCA embolization and surgery are curative treatments for SDAVF, but the functional prognosis is not always satisfactory. Embolization is the first choice of treatment for SDAVF because it is less invasive and relatively safe. However, when complete obliteration is not achieved, prompt surgery is recommended because a long duration of symptoms will result in a poor functional prognosis.

Adult↗

Galea-cranium suture method for prevention of the subcutaneous fluid collection--technical note.

A simple method is described for preventing cerebrospinal fluid (CSF) from collecting in the subcutaneous space following a supratentorial osteoplastic craniotomy. A few pairs of oblique holes through which suture materials may pass are made by drilling in the surface cortex of the bone flap. The galea aponeurotica is sutured to the bone flap by means of these holes. These galea-cranium sutures reduce the subgaleal free space in which pooling of CSF or hematoma may occur.

Cerebrospinal Fluid↗

Aggravation of brainstem symptoms caused by a large superior cerebellar artery aneurysm after embolization by Guglielmi detachable coils--case report.

An 81-year-old male presented with right oculomotor nerve paresis and left hemiparesis caused by a mass effect of a large superior cerebellar artery aneurysm. Endovascular treatment was performed using Guglielmi detachable coils. The patient subsequently suffered aggravation of the mass effect 3 weeks after the embolization. Bilateral vertebral artery occlusion was performed, which decreased the cerebral edema surrounding the aneurysm, but his neurological symptoms did not improve. Parent artery occlusion is recommended as the first choice of treatment for an unclippable large or giant aneurysm causing a mass effect on the brainstem.

Aged↗

Telemedicine in neurosurgery using international digital telephone services between Japan and Malaysia--technical note.

A new image transmission and teleconference system using international digital telephone services was established between Japan and Malaysia. This new system consists of an ordinary personal computer, image scanner, and terminal adapter for digital telephone lines. The quality of images transferred using this system was high enough for diagnosis and discussion except for images such as radiographs requiring huge data transfer. Transmission of one image took approximately 20 seconds. The cost performance was almost equal to the conventional mailing system. The most remarkable advantage of this new system is the high quality of transferred images, the cost and time performance, and security of the medical information. New communication systems using international digital networks including the internet may allow re-distribution of medical resources between advanced countries and developing countries in neurosurgery.

Academic Medical Centers↗

Anterior approach for dumbbell type cervical neurinoma.

A one-stage anterior approach was performed in four patients for total removal of dumbbell type neurinoma at the cervical level. In each case, the neurinoma compressed the spinal cord in the cervical canal, developed anteriorly through the intervertebral foramen, and compressed the vertebral artery. A conventional cervical anterior approach at the tumor site was performed, followed by confirmation of the tumor located outside the spinal canal. After identification of the vertebral artery, corpectomy was carried out and the extradural component of the tumor was resected. In cases with a portion of the tumor located also within the dura mater, the dura mater was opened for removal of the intradural tumor. We found the anterior approach to be effective for the total removal of some kinds of cervical dumbbell type neurinomas.

Adult↗

[Compliance concerning external protectors for hip fractures among the institutionalized elderly in Japan].

An external protector (EHP) which was developed in Europe to protect the femoral neck from direct impact on falling is available but has not been examined sufficiently in Japan. In order to explore the compliance of use of the EHP among the institutionalized elderly with in case of hip fracture who are at high risk of falling, we conducted the four-week intervention study using two types of a EHP. The subjects of a study consisted of 10 elderly people (2 men, 8 women) with a mean age 85.7 living in nursing home in a village. Informed consent was obtained from all participants. At the end of the study, rate of subjects who wore EHP was relatively high for the Finnish EHP (Safety Pants) group compared to the Danish EHP (Hip Protector) group. There were no significant differences between variables in age, sex, fall experience during the previous year, history of diseases etc. The reasons for dropout were first: difficult in wearing EHP and accompanying delay in toilet, secondly: taking much time to wear, thirdly: a sense of incongruity, too small or too tight'. If the EHP is redesigned to suit Japanese elderly, the compliance might increase. Thorough explanation to institutionalized elderly who may have cognitive impairment, physical problems, or both, is required.

Aged↗

Long-term patency of radial artery graft bypass for reconstruction of the internal carotid artery. Technical note.

Reconstruction of the carotid artery by using a radial artery graft is a useful option that can produce reliable long-term patency for the surgical treatment of giant and/or large aneurysms of the cavernous and paraclinoid internal carotid artery (ICA). During the past 10 years, 43 patients with intracavernous and paraclinoid giant aneurysms of the ICA have been treated by reconstruction of the ICA with radial artery grafts after ligation of the cervical ICA. The long-term patency of the grafted radial artery was evaluated over more than a 5-year period (mean 7.2 years) in 20 of these patients by using magnetic resonance angiography or conventional angiography. There was no late occlusion of the graft in any of these cases. Stenotic graft changes were observed in two cases.

Adult↗

Neuronal damage in gerbils caused by intermittent forebrain ischemia.

OBJECT: The purpose of this study was to investigate the possibility of preventing cumulative neuronal damage after repetitive severe ischemia. METHODS: The authors monitored ischemic depolarization in the gerbil hippocampus, which has recently been shown to be a good experimental model of the effects of brief ischemia on the brain, and evaluated neuronal damage in the CA1 subregion 7 days after the ischemic insult. In a single-ischemia paradigm, the results indicate that induction of ischemia-induced neuronal damage depended on the duration of ischemic depolarization. Neuronal damage can be detected in the CA1 subregion after a period of depolarization lasting 210 seconds. Using a double-ischemia paradigm in which the animals were subjected to two periods of ischemia, there was apparently no accumulation of neuronal damage from the first ischemic episode to the second, provided the duration of the first period of ischemic depolarization did not exceed 90 seconds. Neuronal damage accumulated when the duration of the first ischemia episode exceeded 90 seconds, regardless of the duration of the reperfusion interval between the two ischemic insults. Finally, when the ischemic insult was spread over four separate episodes, each lasting 90 seconds (with a reperfusion interval of 5 minutes), neuronal damage was not found when the total depolarization period was less than 420 seconds. CONCLUSIONS: The authors conclude that cumulative neuronal damage may be avoided by adopting an intermittent ischemia approach. The implications of these results for human surgery requiring temporary occlusion of the cerebral arteries are discussed.

Action Potentials↗