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Biomedical subjects

H Iwamoto

Publications and source records attributed to H Iwamoto.

At least 91 records · Page 5Linked to original sources
PubMed1999

Abundant expression and cytoplasmic aggregations of [alpha]1A voltage-dependent calcium channel protein associated with neurodegeneration in spinocerebellar ataxia type 6.

Spinocerebellar ataxia type 6 (SCA6) is one of the eight neurodegenerative diseases caused by a tri-nucleotide (CAG) repeat expansion coding polyglutamine (CAG repeat/polyglutamine diseases) and is characterized by late onset autosomal dominant cerebellar ataxia and predominant loss of cerebellar Purkinje cells. Although the causative, small and stable CAG repeat expansion for this disease has been identified in the [alpha]1A voltage-dependent calcium channel gene (CACNA1A), the mechanism which leads to predominant Purkinje cell degeneration is totally unknown. In this study, we show that the calcium channel mRNA/protein containing the CAG repeat/polyglutamine tract is most intensely expressed in Purkinje cells of human brains. In SCA6 brains, numerous oval or rod-shaped aggregates were seen exclusively in the cytoplasm of Purkinje cells. These cytoplasmic inclusions were not ubiquitinated, which contrasts with the neuronal intra-nuclear inclusions of other CAG repeat/polyglutamine diseases. In cultured cells, formation of perinuclear aggregates of the channel protein and apoptotic cell death were seen when transfected with full-length CACNA1A coding an expanded polyglutamine tract. The present study indicates that the mechanism of neurodegeneration in SCA6 is associated with cytoplasmic aggregations of the [alpha]1A calcium channel protein caused by a small CAG repeat/polyglutamine expansion in CACNA1A.

Animals↗
PubMed1999

Calpain inhibitor-1 reduces infarct size and DNA fragmentation of myocardium in ischemic/reperfused rat heart.

Myocardial ischemia/reperfusion activates a calcium-dependent protease, calpain, in the ischemic myocytes. It is not known whether calpain is involved in the mechanism of ischemia/reperfusion injury in hearts. Thus the purpose of this study was to clarify the effect of a selective calpain inhibitor (CAI) on infarct size and the extent of DNA damage in ischemic/reperfused rat hearts. Rats were divided in four groups (n = 7 each). In saline group, 0.3 ml of saline was administered (i.v.) 10 min before 30-min coronary occlusion followed by 6-h reperfusion. In vehicle group, 0.3 ml of 10% dimethyl sulfoxide (DMSO) was administered 10 min before the 30-min ischemia. CAI (0.5 mg/kg) was administered 10 min before the 30-min ischemia (CAI-A group) and 10 min before the 6-h reperfusion period (CAI-B group). Infarct size was detected with triphenyl tetrazolium chloride, and DNA fragmentation was detected by agarose gel electrophoresis and by in situ nick end labeling (ISEL). Infarct size was significantly smaller in the CAI-A group compared with the vehicle group (13+/-9% vs. 48+/-12%; p < 0.01), and the incidence of ISEL-positive myocyte nuclei in the subendocardial region was significantly reduced in the CAI-A group compared with the vehicle group (26+/-3% vs. 59+/-6%; p < 0.01). However, the effects of CAI in CAI-B group were not significant. Activation of calpain is involved in the mechanism of ischemia/reperfusion injury, and the preischemic administration of CAI was effective in reducing myocardial infarct size and the DNA damage of the myocytes in ischemic/reperfused rat heart.

Animals↗
PubMed1999

Clinical, neuropathological, and molecular study in two families with spinocerebellar ataxia type 6 (SCA6).

To clarify the clinical, neuropathological, and molecular characteristics of spinocerebellar ataxia type 6 (SCA6), two unrelated Japanese families with SCA6 were studied. A clinical feature of the two families was late onset "pure" cerebellar ataxia. Pathologically, three SCA6 brains consistently showed Purkinje cell dominant cortical cerebellar degeneration. Morphometric analysis showed that loss of the cerebellar granule cells and inferior olivary neurons were very mild compared with the severity of Purkinje cell loss. There was no obvious ubiquitin immunoreactive nuclear inclusions. All affected patients had identical expanded alleles, and the expansion was also homogeneously distributed throughout the brain without mosaicism. The present study showed that SCA6 is characterised by Purkinje cell dominant cortical cerebellar degeneration, highly stable transmission of the CAG repeat expansion, and lack of ubiquitin immunoreactive nuclear inclusions.

Adult↗
PubMed1999

Ischemic preconditioning attenuates apoptosis through protein kinase C in rat hearts.

To investigate the role of protein kinase C (PKC) in the mechanism of ischemic preconditioning (IP), infarct size and the incidence of apoptosis caused by ischemia-reperfusion were tested in four groups of Sprague-Dawley rats. Dimethyl sulfoxide (vehicle) or calphostin C (0.1 mg/ml) was administered 5 min before the 30-min coronary occlusion followed by a 6-h reperfusion. Three cycles of 3 min of ischemia followed by 3 min of reperfusion was performed as IP before the 30-min ischemia followed by a 6-h reperfusion with or without calphostin C pretreatment. Infarct size defined by triphenyltetrazolium chloride staining was reduced from 60 +/- 2 to 26 +/- 2% by IP (P < 0.01), but the effect of IP was abolished by calphostin C (51 +/- 3%). Apoptosis defined by in situ terminal deoxynucleotidyl transferase end-labeling (TUNEL) was reduced by IP from 44 +/- 3 to 13 +/- 2% in the subendocardial region (P < 0.01). This effect of IP was abolished by calphostin C (42 +/- 8%). Thus the effect of IP on reducing the infarct size and the incidence of apoptosis are both mediated by PKC in rat hearts.

Animals↗
PubMed1999

Cyclosporin A protects lung function from hyperoxic damage.

Cyclosporin A (CsA), an inhibitor of protein phosphatase 2B (calcineurin), has been shown to play a role in exocytosis and neutrophil mobility. Hyperoxia (>95% oxygen for 72 h) causes lung injury and reduces lung compliance. This model is indicative of deficiencies in surfactant and elicits a vigorous immune response leading to further damage. We examined the effects of CsA on surfactant-secreting lung alveolar type II cells. CsA enhances ATP-stimulated increases in whole cell capacitance in the presence of 2 mM extracellular Ca2+. This measurement corresponds with increases in exocytosis. Because of its effect on the immune system and exocytosis from type II cells, CsA was examined for its protective effects against hyperoxia-induced lung damage in mice. We found that CsA (50 mg. kg-1. day-1) attenuated hyperoxia-induced reductions in lung compliance when administered before or during 72 h of >95% oxygen (P < 0.05). CsA (10 mg. kg-1. day-1) also had a protective effect against hyperoxia-induced changes in neutrophil infiltration, capillary congestion, edema, and hyaline membrane formation. Wet lung weight-to-dry lung weight ratios did not show any significant changes after hyperoxia or hyperoxia plus CsA (P < 0. 05). CsA may be useful to treat patients undergoing prolonged high-oxygen therapy and possibly other lung injuries.

Adenosine Triphosphate↗
PubMed1999

Prevalence of intracranial saccular aneurysms in a Japanese community based on a consecutive autopsy series during a 30-year observation period. The Hisayama study.

BACKGROUND AND PURPOSE: Subarachnoid hemorrhage is a life-threatening disease that occurs mostly because of the rupture of intracranial saccular aneurysms. However, little is known about the prevalence of ruptured and unruptured aneurysms in the general population. The aim of the present study was to examine the prevalence of intracranial aneurysms on the basis of a consecutive autopsy series over a 30-year observation period in a general Japanese population in Hisayama. METHODS: We evaluated 1230 consecutive autopsy cases with craniotomy among the total deaths of Hisayama residents during 1962 through 1991 (overall autopsy rate, 80.1%). RESULTS: A total of 73 intracranial saccular aneurysms were found in 57 cases (4.6%). The prevalence of aneurysms for women was 2.4 times higher than that for men (7.1% versus 2.9%). Among men, the prevalence of aneurysms remained unchanged across the range of age groups. In contrast, there were 2 peaks in the prevalence of aneurysms for women falling in the 40- to 49-year (14.3%) and 60- to 69-year age groups (14.5%). The most common site of the aneurysms was the middle cerebral artery (31.5%), followed by the anterior communicating artery (30.1%), anterior cerebral artery (15.1%), vertebrobasilar artery (12.3%), and internal carotid artery (11.0%). Among these 73 aneurysms, 29 (39.7%) were ruptured. Ruptured aneurysms were common in subjects <80 years of age, whereas unruptured aneurysms were prevalent in those >/=80 years of age. The frequency of ruptured aneurysms was highest in the vertebrobasilar system (66.7%) and lowest in the middle cerebral artery (13.0%). CONCLUSIONS: Our data suggest that intracranial aneurysms are more frequent in women in the general Japanese population. Aneurysms are more prevalent in the middle cerebral artery, but the risk of rupture is highest in the vertebrobasilar system.

Adolescent↗
PubMed1999

[An autopsy case of pulmonary embolism due to renal angiomyolipoma in an elderly woman].

An 86-year-old woman with a 13-year history of hypertension was admitted because of consciousness disturbance, hypotension, tachycardia, and cyanosis at her extremities. Enhanced computed tomography showed a thrombus in the truncus pulmonalis and right pulmonary artery, and also showed a left renal mass and a right renal cyst. Under a diagnosis of pulmonary embolism we started anticoagulant therapy, but the patient died five days after admission. At autopsy, a saddle-like thrombus was found in the truncus pulmonalis and bilateral trunks of pulmonary arteries. Microscopic examination showed smooth muscle cells in the thrombus. We could not find any other thrombus in the inferior vena cava, intrapelvic veins, nor in veins of lower extremities by milking. We also found tumors in both kidneys. Microscopically all tumors were diagnosed as angiomyolipoma. There were many fibrin thrombi in the sinuses of the tumors but there was no evidence of malignancy. We finally diagnosed pulmonary embolism due to renal angiomyolipoma because there was no other thrombus origin and microscopically the same smooth muscle cells were found both in the renal tumor and the pulmonary thrombus. There is only one case report concerning pulmonary embolism due to renal angiomyolipoma which happened during operative treatment. The treatment method of renal angiomyolipoma is determined by tumor size and symptoms, and usually intensive treatment is not performed in cases without symptoms. Our patient had no symptoms until the onset of severe complication of pulmonary embolism, suggesting that radical treatment is necessary for renal angiomyolipoma with a thrombus even when there are no symptoms.

Aged↗
PubMed1998

Importance of left anterior descending coronary artery curvature in determining cross-sectional plaque distribution assessed by intravascular ultrasound.

We assessed the relation between the circumferential distribution of coronary atherosclerotic plaques and the structure of the epicardial coronary arteries in patients with coronary artery disease using intravascular ultrasound in vivo. Coronary atherosclerosis preferentially formed at the inner arc of the curved coronary vessels, and greater vessel curvatures were associated with greater distributions of atherosclerotic lesions along the inner coronary artery wall.

Adult↗
PubMed1998

High-Resolution X-ray Diffraction of Muscle Using Undulator Radiation from the Tristan Main Ring at KEK.

High Energy Accelerator Research Organization (KEK), Tsukuba, Japan.High-resolution X-ray diffraction studies on striated muscle fibres were performed using a hard X-ray undulator installed in the Tristan main ring at KEK, Tsukuba, Japan. The performance of the undulator, along with an example experiment which exploited the unique characteristics of undulator radiation, are reported. The vertical divergence angle of the first harmonic of the undulator was approximately 10 micro rad under 8 GeV multi-bunch operating conditions and the peak photon flux density was estimated to be approximately 3 x 10(16) photons s(-1) mrad(-2) (0.1% bandwidth)(-1) (10 mA)(-1). The well collimated X-ray beam from the undulator made it possible to resolve clearly, with high angular resolution ( approximately 700 nm), the closely spaced diffraction peaks on the meridional axis in the X-ray patterns arising from the thick filaments of a striated muscle under static conditions. By fitting the meridional intensity pattern, a model for the molecular arrangement of the constituent proteins in the thick filaments is proposed. These studies of muscle demonstrate the promise of undulator radiation from third-generation sources for high-resolution diffraction studies.

Journal Article↗
PubMed1998

Enzymatic synthesis of a neoglycoconjugate by transglycosylation with Arthrobacter endo-beta-N-acetylglucosaminidase: a substrate for colorimetric detection of endo-beta-N-acetylglucosaminidase activity.

The transglycosylation activity of endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae was used for the enzymatic synthesis of a novel oligosaccharide, Man6GlcNAc-p-nitrophenyl-alpha-D-glucose (Man6GlcNAc-Glc-pNP). The reaction was efficiently induced in aqueous solution containing dimethyl sulfoxide. In the medium containing 20% (v/v) dimethyl sulfoxide with 0.1 M Glc-pNP as an acceptor, the transglycosylation attained yields of 75% by high-performance anion-exchange chromatography. The structure of Man6GlcNAc-Glc-pNP was confirmed by ion mass spectrometry and 400 MHz 1H NMR spectrometry. Various endo-beta-N-acetylglucosaminidases hydrolyzed this oligosaccharide and Man6GlcNAc and Glc-pNP were released from the oligosaccharide by endo-beta-N-acetylglucosaminidase digestion. We have established a new procedure for the colorimetric detection of endo-beta-N-acetylglucosaminidase activity using Man6Glc-NAc-Glc-pNP, which is simple as that for other exoglycosidase assays with pNP-glycosides as substrates.

Acetylglucosaminidase↗
PubMed1998

Evidence for the load-dependent mechanical efficiency of individual myosin heads in skeletal muscle fibers activated by laser flash photolysis of caged calcium in the presence of a limited amount of ATP.

Although a contracting muscle regulates its energy output depending on the load imposed on it ("Fenn effect"), the mechanism underlying the load-dependent energy output remains obscure. To explore the possibility that the mechanical efficiency, with which chemical energy derived from ATP hydrolysis is converted into mechanical work, of individual myosin heads changes in a load-dependent manner, we examined the auxotonic shortening of glycerinated rabbit psoas muscle fibers, containing ATP molecules almost equal in number to the myosin heads, after laser-flash photolysis of caged calcium. Immediately before laser-flash activation, almost all of the myosin heads in the fiber are in the state M.ADP.Pi, and can undergo only one ATP hydrolysis cycle after activation. When the fibers were activated to shorten under various auxotonic loads, the length, force, and power output changes were found to be scaled according to the auxotonic load. Both the power and energy outputs were maximal under a moderate auxotonic load. The amount of M.ADP.Pi utilized at a time after activation was estimated from the amount of isometric force developed after interruption of fiber shortening. This amount was minimal in the isometric condition and increased nearly in proportion to the distance of fiber shortening. These results are taken as evidence that the efficiency of chemomechanical energy conversion in individual myosin heads changes in a load-dependent manner.

Acetates↗
PubMed1998

Effect of a cardiotonic agent, MCI-154, on the contractile properties of skinned skeletal muscle fibers.

We have studied the effect of a cardiotonic agent, MCI-154 (6-[4-(4-pyridylamino)phenyl]-4,5-dihydro-3(2H)-pyridazinone hydrochloride trihydrate), on the contractile properties and adenosine triphosphatase (ATPase) activity of chemically skinned rabbit skeletal muscle fibers. As in cardiac muscle, MCI-154 potentiated isometric tension and improved isometric tension cost at full Ca2+ activation. It showed little Ca2+-sensitizing effect. In contrast to its effect on cardiac muscle, however, MCI-154 decreased all the kinetic parameters tested (shortening velocity, the rate of rise of tension, and actomyosin ATPase activity). All the results are explainable if MCI-154 acts directly on skeletal actomyosin and inhibits a reaction step(s) of the ATPase cycle later than the force-generating event. The qualitative difference between cardiac and skeletal muscles in the responsiveness to this class of cardiotonic agents (MCI-154 and EMD 53998, a thiadiazinone derivative) is most readily understood if the agents have two independent actions, one on troponin and the other on actomyosin itself, the latter being dominant in skeletal muscle.

Adenosine Triphosphatases↗
PubMed1998

Innervation of blood vessels in the rat incisor pulp: a scanning electron microscopic and immunoelectron microscopic study.

Although two types of nerve endings have been proposed to innervate blood vessels in the dental pulp, the precise innervation pattern is not well understood. This is mainly due to the lack of information regarding the positional relationships of nerve fibers with blood vessels at the electron microscopic level. The rat incisor pulp was investigated by scanning electron microscopy (SEM) after connective tissue digestion and by transmission electron microscopy after immunohistochemical localization with polyclonal anti-PGP 9.5 antibody. SEM specimens revealed that unmyelinated nerve fibers passed through the tunica adventitia of the blood vessel in the center of the pulp and that these fibers then entered the tunica media of the smaller arterioles. The nerve fibers divided into many collaterals, and these terminated on the surface of smooth muscle cells and endothelial cells. The fibers extended toward smaller vessels. The terminal fibers then reached the subodontoblastic capillary plexus and terminated on pericytes, while the odontoblastic capillary plexus had no direct innervation. In the pulpal venules, nerve fibers were located adjacent to the pericytes and endothelial cells, and they extended toward postcapillary venules. The same results were confirmed by immunoelectron microscopy. The present study demonstrated that in the rat incisor pulp the microvasculature as well as larger vessels were directly innervated by free nerve endings, suggesting that the local regulation of blood flow could take place not only at larger vessels but also at the level of the microvasculature in this tissue.

Animals↗
PubMed1998

Load-dependent mechanical efficiency of individual myosin heads in skeletal muscle fibers activated by laser flash photolysis of caged calcium in the presence of a limited amount of ATP.

Although a contracting muscle regulates its energy output depending on the load imposed on it ("Fenn effect"), the mechanism underlying the load-dependent energy output remains obscure. To explore the possibility that the mechanical efficiency, with which chemical energy derived from ATP hydrolysis is converted into mechanical work, of individual myosin heads changes in a load-dependent manner, we examined the auxotonic shortening of glycerinated rabbit psoas muscle fibers, containing ATP molecules almost equal in number to the myosin heads, following laser flash photolysis of caged calcium. Immediately before laser flash activation, almost all of the myosin heads in the fiber are in the state, M.ADP.Pi, and can undergo only one ATP hydrolysis cycle after activation. When the fibers were activated to shorten under various auxotonic loads, the length, force and power output changes were found to be scaled according to the auxotonic load. Both the power and energy outputs were maximal under a moderate auxotonic load. The amount of M.ADP.Pi utilized at a time after activation was estimated from the amount of isometric force developed after interruption of fiber shortening. This amount was minimal in the isometric condition, and increased nearly in proportion to the distance of fiber shortening. These results are taken as evidence that the efficiency of chemo-mechanical energy conversion in individual myosin heads changes in a load-dependent manner.

Adenosine Triphosphate↗
PubMed1998

Comparison of genetic susceptibility to experimental allergic/immune-mediated blepharoconjunctivitis between Lewis and Fischer rats.

BACKGROUND: Fischer rats were less susceptible to experimental autoimmune uveoretinitis (EAU) than Lewis rats, although both strains have the same MHC molecules. The purpose of this study was to compare the susceptibility of experimental allergic/immune-mediated blepharoconjunctivitis (EAC) between these two strains. METHODS: Male Lewis and Fischer rats were immunized with either ovalbumin (OVA) or OVA peptide (OVA323-339) in complete Freund's adjuvant (CFA). Three weeks later, they were challenged with OVA by eye drops. Twenty-four hours later, after clinical evaluation, they were killed and eyes, blood and lymph nodes were harvested for histology, antibody titer and proliferation assay, cytokine production or flow cytometric analysis, respectively. RESULTS: Fischer rats developed mild EAC compared with Lewis rats. Cellular proliferative responses, IFN-gamma production of culture supernatant and serum IgG specific for OVA were basically the same between the two strains. The same OVA peptides were selected as immunodominant. Flow cytometric analysis demonstrated the same cellular profile of lymph node cells in the two strains. CONCLUSION: EAC in Fischer rats was milder than that in Lewis rats, although no apparent differences in immunological parameters between these two strains were detected. These data suggest that factors unrelated to immunological parameters may depend on the susceptibility of EAC.

Allergens↗
PubMed1998

Thin filament cooperativity as a major determinant of shortening velocity in skeletal muscle fibers.

The mechanism underlying the calcium sensitivity of the velocity of shortening of skeletal muscle fibers was investigated using a multiple shortening protocol: within a single contraction, skinned rabbit psoas fibers were made to shorten repetitively under a light load by briefly stretching back to their initial length at regular intervals. At saturating [Ca2+], the initial fast shortening pattern was repeated reproducibly. At submaximal [Ca2+], the first shortening consisted of fast and slow phases, but only the slow phase was observed in later shortenings. When the fibers were held isometric after the first shortening, the velocity of the second shortening recovered with time. The recovery paralleled tension redevelopment, implying a close relationship between the velocity and the number of the preexisting force-producing cross-bridges. However, this parallelism was lost as [Ca2+] was increased. Thus, the velocity was modified in a manner consistent with the cooperative thin filament activation by strong binding cross-bridges and its modulation by calcium. The present results therefore provide evidence that the thin filament cooperativity is primarily responsible for the calcium sensitivity of velocity. The effect of inorganic phosphate to accelerate the slow phase of shortening is also explained in terms of the cooperative activation.

Animals↗
PubMed1998

Inhibition of integrin signaling with Arg-Gly-Asp motifs in rat hepatic stellate cells.

BACKGROUND/AIMS: Activation of hepatic stellate cells plays a key role in liver fibrogenesis. Disruption of normal hepatic stellate cell-matrix interactions may contribute to this process. However, little is known about the molecular events leading from integrin-extracellular matrix interaction to hepatic stellate cell function. Therefore, we investigated the role of integrin signaling in tyrosine phosphorylation of focal adhesion kinase and cytoskeletal assembly in rat hepatic stellate cells using soluble Arg-Gly-Asp containing peptides. METHODS: Hepatic stellate cells were isolated from normal rat livers. Integrin alpha5beta1 expression in hepatic stellate cells was analyzed by immunoprecipitation and immunocytochemistry. The cytoskeletal assembly and tyrosine phosphorylation of focal adhesion kinase were determined by immunocytochemistry and immunoblotting. We also analyzed the effect of Arg-Gly-Asp containing peptides on the expression of smooth muscle alpha actin by immunocytochemistry and immunoblotting. RESULTS: We identified integrin alpha5beta1 in rat hepatic stellate cells. Stress fiber formation and cell shape were different when hepatic stellate cells were plated on various extracellular matrix components. Treatment of hepatic stellate cells with soluble Arg-Gly-Asp peptides diminished the adhesion-induced tyrosine phosphorylation of focal adhesion kinase and inhibited the formation of stress fibers. The peptides also reduced the expression of smooth muscle alpha actin. CONCLUSIONS: Our results demonstrate that adhesion to extracellular matrix induces tyrosine phosphorylation of focal adhesion kinase and promotes actin stress fiber formation and focal adhesion assembly in rat hepatic stellate cells, and that these events are disturbed by soluble Arg-Gly-Asp peptides.

Animals↗