Biomedical subjects

Mitsuaki Isobe

Publications and source records attributed to Mitsuaki Isobe.

At least 55 records · Page 3Linked to original sources

PubMed2005

Identification of quantitative trait loci for cardiac hypertrophy in two different strains of the spontaneously hypertensive rat.

Cardiac hypertrophy and left ventricular hypertrophy are known to be substantially controlled by genetic factors. As an experimental model, we undertook genome-wide screens for cardiac mass in F2 populations bred from the stroke-prone spontaneously hypertensive rats (SHRSP) and normal spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) of a Japanese colony. Two F2 cohorts were independently produced: F2(SHRSP x WKY) (110 male and 110 female rats) and F2(SHR x WKY) (151 male rats). The ratio of heart weight to body weight (Hw/Bw) was evaluated at 12 months of age in F2(SHRSP x WKY) after salt-loading for 7 months, and at around 15 weeks of age in F2(SHR x WKY) who had been fed a normal rat chow diet. Subsequent to an initial screen with 251 markers in F2(SHRSP x WKY) male progeny, 170 and 161 markers were selected and characterized in F2(SHRSP x WKY) female progeny and F2(SHR x WKY) male progeny, respectively. Markers from four chromosomal regions showed suggestive or significant linkage to Hw/Bw. The strongest and the most consistent linkage was found in the vicinity of D3Mgh16 on rat chromosome (RNO) 3 (a maximal log of the odds score reached 4.0 to 6.6 across the F2 populations studied). In the other three regions on RNO6, RNO10 and RNO13, the degree of linkage was more prominent in either males or females. These data provide solid evidence for a "principal" RNO3 quantitative trait loci regulating Hw/Bw in SHRSP and SHR, and also suggest the possible presence of sexual dimorphism in regard to genetic susceptibility for cardiac hypertrophy.

Animals↗

PubMed2005

Dietary consumption of green tea catechins attenuate hyperlipidaemia-induced atherosclerosis and systemic organ damage in mice.

OBJECTIVE: The effects of consuming green tea catechins on the development of hyperlipidaemia-induced systemic organ damage have not been well studied; we investigated the effect using low density lipoprotein receptor knockout mice. METHODS AND RESULTS: Mice were treated with high cholesterol food containing 0.2 or 4% catechins and they were supplemented for 35 weeks. High plasma cholesterol levels, liver and renal dysfunctions were observed in no catechin fed mice, while chow containing catechin suppressed these levels and damages. Severe atherosclerosis of the aorta, fatty liver and renal injury were also shown in the control mice; inflammatory factors were enhanced in these lesions of nontreated mice. The lesions were attenuated with suppression of the inflammatory factors in the chow-contained catechin treatment group. CONCLUSION: Dietary consumption of tea catechins attenuated the development of the systemic organ damage; thus, this has a clinical effect against systemic inflammatory diseases.

Administration, Oral↗

PubMed2005

[Immunological tolerance].

New immunosuppressive agents have significantly improved early survival after organ transplantation for patients with end-stage organ failure. However, a major problem in transplantation is continued to be graft rejection. Chronic allograft rejection threatens the long-term survival of organ transplant recipients. Therefore, induction of donor specific tolerance is very important for improving long-term survival after transplantation. The costimulatory pathways are crucial in regulating T cell activation and donor specific tolerance. The costimulatory pathways are potential therapeutic targets for inducing donor specific tolerance and improving long-term survival after transplantation.

Animals↗

PubMed2004

Nitric oxide inhibits myocardial apoptosis by preventing caspase-3 activity via S-nitrosylation.

Two protein signaling systems, phosphorylation and S-nitrosylation, influence most aspects of cellular physiology. S-nitrosylation, which generates a nitrosothiol linkage on cysteine residues, is caused by nitric oxide (NO). NO is believed to act as an anti-apoptotic agent by inhibiting caspase activity in cardiomyocytes, but there is little direct evidence for this. We investigated whether apoptosis inhibition by NO involved S-nitrosylation of caspases in doxorubicin (DOX)-induced myocardial apoptosis. Cardiomyocytes were treated with 1 micromol/l of DOX to induce apoptosis. Pretreatment with an NO donor, S-nitroso-N-acetyl-penicillamine (SNAP) reduced the apoptosis. This effect was attenuated by treatment with 100 micromol/l of mercury dichloride (HgCl2), which is an agent of denitrosylation. After 24 h DOX-treatment, SNAP reduced the increased caspase-3 activity by 63%, and this effect was reversed by treatment with HgCl2. Immunoblot analysis showed that accumulation of the cleaved caspase-3 protein, an active form that induces apoptosis was inhibited significantly by SNAP. To elucidate nitrosothiol formation on caspase-3 by NO, we did several experiments. First, we prepared an immunoprecipitate of caspase-3 and measured the concentration of NO released from the precipitated complex by HgCl2. Second, S-nitrosylated proteins, purified by immunoprecipitation of caspase-3, were biotinylated and the biotin concentration was estimated by immunoblotting. Third, dual immunofluorescent staining was done with antibodies for S-nitrosocysteine and caspase-3. Results showed that formation of nitrosothiol in caspase-3 in DOX-treated cardiomyocytes with SNAP was increased significantly compared with untreated cardiomyocytes. We reported here that exogenous NO produces an anti-apoptotic effect by suppression of caspase activity via S-nitrosylation in cardiomyocytes.

Animals↗

PubMed2004

HMG-CoA reductase inhibitor attenuates experimental autoimmune myocarditis through inhibition of T cell activation.

OBJECTIVE: This study tested the hypothesis that 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitor affects T cell-mediated autoimmunity through inhibition of nuclear factor-kappaB (NFkappaB) and reduces the severity of experimental autoimmune myocarditis (EAM). METHODS: EAM was induced in Lewis rats by immunization with myosin. High-dose or low-dose fluvastatin or vehicle was administered orally for 3 weeks to rats with EAM. RESULTS: Fluvastatin reduced the pathophysiological severity of myocarditis. Fluvastatin inhibited expression of NFkappaB in the nuclei of myocardium in EAM. Fluvastatin reduced production of Th1-type cytokines, including interferon (IFN)-gamma and interleukin (IL)-2, and inhibited expression of inflammatory cytokine mRNAs in the myocardium. Infiltration of CD4-positive T cells into the myocardium and T cell proliferative responses were suppressed by fluvastatin. Plasma lipid levels did not differ between the groups. CONCLUSIONS: Fluvastatin ameliorates EAM by inhibiting T cell responses and suppressing Th1-type and inflammatory cytokines via inactivation of nuclear factor-kappaB, and this activity is independent of cholesterol reduction.

Animals↗

PubMed2004

E-selectin blockade decreases adventitial inflammation and attenuates intimal hyperplasia in rat carotid arteries after balloon injury.

OBJECTIVE: Inflammation is one of the initial repair processes after vascular injury. E-selectin facilitates adherence of leukocytes to vascular endothelium at the site of inflammation. Because the role of E-selectin in this process is not fully understood, we studied the role of E-selectin in vascular injury with a flow chamber model and a rat model of carotid artery injury. METHODS AND RESULTS: We established a rat aortic endothelial cell (RAEC) culture system from the aortas of adult male rats. When rat myelomonocytes were suspended in a flow chamber, rolling and adhesion to lipopolysaccharide (LPS)-stimulated RAECs were observed. Cell rolling and adhesion were greatly reduced by addition of anti-E-selectin monoclonal antibody (mAb). We then induced balloon injury in the left carotid arteries of rats. E-selectin expression was enhanced in endothelial cells at adventitial small vessels 7 days after injury. Rats with balloon injury were injected intraperitoneally with anti-E-selectin mAb for 8 days. Inflammatory cell infiltration was reduced by anti-E-selectin mAb treatment at the adventitia at 7 days after injury. This reduction was associated with attenuation of intimal hyperplasia in the rats treated with the mAb. CONCLUSIONS: These data suggest that E-selectin regulates adventitial inflammation through leukocyte adhesion and contributes to the process of intimal hyperplasia after balloon injury.

Animals↗

PubMed2004

Blockade of the interaction between PD-1 and PD-L1 accelerates graft arterial disease in cardiac allografts.

BACKGROUND: Programmed death-1 (PD-1), a member of the CD28 family, has been identified. PD-1 is involved in the negative regulation of some immune responses. We evaluated the role of PD-ligand 1 (PD-L1) in graft arterial disease (GAD) of cardiac allografts and in smooth muscle cells (SMCs). METHODS AND RESULTS: C57BL/6 murine hearts were transplanted into B6.C-H2 KhEg mice for examination of GAD. PD-L1 was expressed in SMCs of the thickened intima in the graft coronary arteries, and administration of anti-PD-L1 monoclonal antibody (mAb) enhanced the progression of GAD (luminal occlusion: 55+/-5.0% versus 9.8+/-4.3%, P<0.05). The expressions of interferon gamma (IFN-gamma) and tumor necrosis factor alpha of cardiac allografts were upregulated in response to anti-PD-L1 mAb treatment. In vitro, PD-L1 expression was induced in SMCs in response to IFN-gamma stimulation. Sensitized splenocytes increased SMC proliferation, and anti-PD-L1 mAb in combination with IFN-gamma stimulation increased this proliferation. CONCLUSIONS: The PD-L1 pathway regulates both the proliferation of SMCs and GAD. Thus, control of this interaction is a promising strategy for suppression of GAD.

Animals↗

PubMed2004

Suppression of acute and chronic rejection by hepatocyte growth factor in a murine model of cardiac transplantation: induction of tolerance and prevention of cardiac allograft vasculopathy.

BACKGROUND: Although treatment with immunosuppressive agents has contributed to overcoming acute rejection and improving the midterm survival of transplanted hearts, cardiac allograft vasculopathy (CAV) has remained the main cause of primary graft failure. Recent approaches have shown that hepatocyte growth factor (HGF) exhibits cardiotrophic functions. We therefore addressed whether HGF would regulate acute and chronic rejection in cardiac transplantation. METHODS AND RESULTS: We used a murine heterotopic cardiac transplantation model between fully incompatible strains and administered 500 microg x kg(-1) x d(-1) HGF during the initial 14 days after transplantation. The HGF-treated allografts showed significantly prolonged survival (42.3+/-4.1 days, P<0.001) compared with the controls (11.1+/-0.6 days), with tolerance induction in 47.4%. Histopathologically, the number of infiltrating cells was significantly decreased and myocardial necrosis was less prominent with a reduction of apoptosis in the allografts by HGF treatment during acute rejection. In the long-term surviving allografts, HGF significantly inhibited the development of CAV and interstitial fibrosis. With respect to intragraft cytokine mRNA expression, HGF treatment reduced the early expression of interferon-gamma and enhanced the expression of transforming growth factor-beta1 during the acute phase and of interleukin-10 continuously through the acute phase to the chronic phase. CONCLUSIONS: Our findings demonstrate that HGF can prolong the survival of allografts by its cardioprotective and immunomodulative potencies. Thus, HGF administration may constitute a new therapeutic approach to preventing cardiac graft failure that has not been overcome by conventional immunosuppressive agents.

Acute Disease↗

PubMed2004

Inhibition of IkappaB phosphorylation in cardiomyocytes attenuates myocardial ischemia/reperfusion injury.

OBJECTIVE: Reperfusion injury is related closely to inflammatory reactions such as activation of inflammatory cells and expression of cytotoxic cytokines. We investigated the efficacy of IkappaB phosphorylation blockade in a rat myocardial ischemia/reperfusion injury model. METHODS AND RESULTS: IMD-0354 inhibited phosphorylation of IkappaBalpha and nuclear translocation of nuclear factor-kappa B (NF-kappaB) induced by tumor necrosis factor-alpha (TNF-alpha) in cultured cardiomyocytes. TNF-alpha-induced production of interleukin-1beta and monocyte chemoattractant protein-1 from cultured cardiomyocytes was reduced significantly by IMD-0354. Transient left coronary artery occlusion (30 min) and reperfusion (24 h) were carried out in Sprague-Dawley rats. IMD-0354 (1, 5, 10 mg/kg) was injected intraperitoneally 5 min before the start of reperfusion. Treatment with IMD-0354 resulted in a significant dose-dependent reduction of the infarction area/area at risk ratio (vehicle, 47.0+/-3.4%; 10 mg/kg of IMD-0354, 19.4+/-4.0%; P<0.01) and the preservation of fractional shortening ratio (vehicle, 25.0+/-1.5%; 10 mg/kg of IMD-0354, 42.3+/-1.7%; P<0.01). Histological analysis showed that accumulation of polymorphonuclear neutrophils in the area at risk was decreased significantly. CONCLUSIONS: Inhibition of nuclear translocation of NF-kappaB by IkappaBalpha phosphorylation blockade could provide an effective approach to attenuation of ischemia/reperfusion injury. The cardioprotective effects of IMD-0354 include not only reduction of harmful neutrophil accumulation in myocardium but also inhibition of harmful cytokine and chemokine production by cardiomyocytes.

Animals↗

PubMed2004

Attenuation of graft arterial disease by manipulation of the LIGHT pathway.

OBJECTIVE: The tumor necrosis factor (TNF) superfamily member LIGHT, which binds herpes virus entry mediator (HVEM) and lymphotoxin beta receptor (LTbetaR), plays important roles in regulating the immune response. To clarify the mechanism underlying graft arterial disease (GAD), we investigated the role of the LIGHT pathway in the progression of GAD. METHODS AND RESULTS: Hearts from Bm12 mice were transplanted into C57BL/6 (B/6) mice (class II mismatch). Recipients were injected intraperitoneally with HVEMIg (100 microg per treatment) every 7 days for 8 weeks. Treatment with HVEMIg significantly attenuated GAD (luminal occlusion=16.5+/-7.7% versus control allograft=62.6+/-12.1%, P<0.05), and significantly decreased intragraft IL-4, IL-6, and interferon-gamma (IFN-gamma) mRNA expression compared with controls. LTbetaR was expressed in smooth muscle cells (SMCs) with or without cytokine stimulation, whereas HVEM was detected in SMCs stimulated by IFN-gamma. Coculture of SMCs with T cells after transplantation induced SMC proliferation, and addition of HVEMIg resulted in inhibition of SMC proliferation. CONCLUSIONS: These results indicate that the LIGHT pathway plays important roles in the regulation not only of T-cell activation but also of SMC proliferation. Blockade of the LIGHT pathway is a promising avenue for the prevention of GAD.

Animals↗

PubMed2004

Novel C-terminus frameshift mutation, 1122fs/147, of HERG in LQT2: additional amino acids generated by frameshift cause accelerated inactivation.

OBJECTIVE: The function of the C-terminus region of the human ether-a-go-go related gene (HERG) has not been well characterized except for its involvement in trafficking. To understand further the role of C-terminus region, we performed a functional analysis of a novel frameshift mutation (1122fs/147) identified in a Japanese long QT syndrome 2 (LQT2) patient who had recurrent episodes of syncope. METHODS: Wild type (WT) and mutant HERG plasmids were transfected into human embryonic kidney (HEK-293) cells, and whole-cell current was recorded by the patch-clamp technique. Confocal microscopy was performed to examine the membrane distribution of channel protein using a green fluorescent protein tagged to the N-terminus of HERG. RESULTS: The mutant 1122fs/147 alone could express current, but reduced density by 74% of control. No dominant negative effect was noted with co-expression of WT and 1122fs/147. Activation and deactivation time constants were not changed, while inactivation was accelerated in 1122fs/147 compared to WT, and V(1/2) of steady-state inactivation curve shifted by 11 mV in the negative direction. Current density of 1123stop mutant revealed 49% reduction compared to WT and showed no shift in steady-state inactivation. Confocal microscopy revealed reduced protein expression on the cell surface both in 1122fs/147 and 1123stop mutants compared to WT. CONCLUSION: Frameshift mutation at the C-terminus region with additional 147 amino acids evoked a loss of function of the HERG channel. A negative shift in steady-state inactivation induced by the additional 147 amino acids and trafficking defect contribute to a reduced current amplitude of 1122fs/147.

Adolescent↗

PubMed2004

Diagnostic criteria for penetrating atheromatous ulcer of the thoracic aorta.

An autopsy case of penetrating atheromatous ulcer of the ascending thoracic aorta is presented. The triplet imaging findings of mediastinal dilatation, interstitial pulmonary hemorrhage and pericardial effusion were diagnostic for penetrating atheromatous ulcer at the posterior wall of the ascending thoracic aorta. Immediate surgical treatment is indicated when these three findings emerge during examination.

Aged↗

PubMed2004

Clinical assessment of antiarrhythmic agents for paroxysmal atrial fibrillation guided by modification of electrophysiologic arrhythmogenicity.

INTRODUCTION: Rapid atrial pacing alters atrial electrophysiology, promoting initiation and maintenance of atrial fibrillation (AF). The aim of this study was to assess differences in the electrophysiologic properties of atrial tissue between patients with and without AF episodes and to determine whether electrophysiologic properties can predict the clinical efficacy of antiarrhythmic agents. METHODS AND RESULTS: Sixty patients were studied, 33 with documented episodes of paroxysmal atrial fibrillation (PAF) and 27 control patients. Atrial effective refractory period (AERP), atrial vulnerability, and intra-atrial conduction time were measured at baseline and after rapid constant atrial pacing for 5 minutes at rates of 130, 150, 170, and 190 beats/min. The clinical efficacy of antiarrhythmic agents for PAF prophylaxis was assessed over 14 months with an antiarrhythmic agent identical to that administered intravenously, and the antiarrhythmic agent effects on AERP, atrial vulnerability, and intra-atrial conduction time were assessed. AERP shortening and atrial vulnerability increase were significantly larger in the PAF group. Antiarrhythmic agents that were clinically effective in suppressing PAF significantly attenuated AERP shortening, but antiarrhythmic agents that were clinically ineffective did not. CONCLUSION: Changes in AERP and atrial vulnerability observed after rapid atrial pacing are considered indicative of the electrophysiologic substrate of PAF. Attenuation of AERP and atrial vulnerability by antiarrhythmic agents might be useful in predicting their clinical efficacy.

Anti-Arrhythmia Agents↗

PubMed2004

Clinical study of the efficacy of a cooled-tip catheter ablation system for common atrial flutter.

BACKGROUND: Radiofrequency catheter ablation (RF-CA) of common atrial flutter (AFL) requires the creation of a transmural incision to create a bidirectional conduction block in the cavotricuspid isthmus (ITH). METHODS AND RESULTS: RF-CA of the ITH using a cooled-tip system was carried out in 40 patients. In the 'conventional' mode (CONV) of the system, RF energy was applied for 2 min with the temperature set at 60 degrees C and power of up to 50 W, and in the failed cases the 'cooled-tip' mode (COOL) was utilized at 45 degrees C with up to 30 W (with a 15 ml/min saline flow rate). Of the 40 patients, 29 crossed over from the CONV to the COOL after a failed ablation of the AFL. As a result, in all 40 patients a complete linear incision could be created with either the COOL or the CONV, resulting in the successful abolition of the AFL. Complete bi-directional block was successfully created in all patients. No significant side effects occurred. CONCLUSIONS: The COOL was found to be more effective and just as safe as the CONV for AFL ablation, thus facilitating the rapid and complete elimination of the AFL.

Adult↗

PubMed2004

Giant organized thrombus in the left sinus of valsalva causing intermittent left coronary obstruction: an unusual case of acute myocardial infarction.

A 48-year-old Japanese man was admitted to hospital for acute myocardial infarction associated with a giant organized thrombus occupying the left sinus of Valsalva. Cardiac catheterization revealed no organic stenosis in either coronary artery, but left ventriculography and aortography showed a filling defect above the left coronary cusp. Transesophageal echocardiography was immediately performed and showed a round mass filling the left sinus of Valsalva. A solid, round mass approximately 2.5 cm in diameter was removed during emergency surgery and determined to be a thrombus on the basis of microscopic findings. This is the second report of a giant organized thrombus occupying the entire left sinus of Valsalva, obstructing the ostium of the left coronary artery intermittently, and leading to acute myocardial infarction.

Coronary Angiography↗

PubMed2004

Transvenous catheter cryoablation of the atrioventricular node and visual assessment of freezing of cardiac tissue using intracardiac echocardiography.

We investigated the use of a catheter-based cryoablation system on atrioventricular (AV) junction ablation in dogs. In five dogs, the cryoablation catheter was introduced to the AV junction area in order to create transient high degree or complete AV block. Cryo-freezing energy was applied by lowering the temperature to -75 degrees C for five minutes as a single cycle. This cycle was repeated until significant impairment of the AV conduction appeared. Transient high degree and complete AV block was obtained in all five dogs without any adverse effects. The iceball formation was identified by intracardiac echocardiography. Ablation of the AV junction is effective with several freeze-thaw cycles using a transvenous catheter cryoablation system.

Animals↗

PubMed2004

Ventricular unipolar potential in radiofrequency catheter ablation of idiopathic non-reentrant ventricular outflow tachycardia.

We conducted this study to verify the efficacy of ventricular unipolar potential (V-uni) for ablation of idiopathic non-reentrant ventricular tachycardia (idio-VT). The morphology of V-uni at the successful and unsuccessful sites was analyzed in 27 patients with idio-VT [20 with right ventricular outflow tachycardia (RVOVT) and 7 with left ventricular outflow tachycardia (LVOVT)]. The usefulness of V-uni was compared with a pacemapping method and the V-QRS interval. The incidence of QS-pattern V-uni at the successful and best unsuccessful sites were 100 versus 25% (P = 0.000005) in RVOVT and 86 versus 29% (P = 0.10) in LVOVT. The pacemapping scores at the successful and best unsuccessful sites were 11.5/12 versus 11.2/12; NS in RVOVT, and 11.2/12 versus 11.1/12; NS in LVOVT. The mean V-QRS interval at the successful and the best unsuccessful sites were 22.5 +/- 3.8 versus 21.6 +/- 3.4 msec; NS in RVOVT, 15.1 +/- 3.2 versus 12.5 +/- 3.3 msec; NS in LVOVT. The sensitivity (sen) and specificity (spe) of QS-pattern V-uni to determine the optimum target sites were 1.0 and 0.89 in RVOVT and 0.86 and 0.83 in LVOVT, respectively. In the ablation of idio-VT, QS-pattern V-uni is simply and visually identifiable, is very useful, and should be given a high priority when determining the optimum target site.

Adolescent↗

PubMed2004

Anti-vascular cell adhesion molecule-1 and anti-very late antigen-4 monoclonal antibodies inhibit neointimal hyperplasia in the murine model of arterial injury.

OBJECTIVE: Arterial restenosis after angioplasty limits long-term survival of patients. Murine arterial injury models develop neointimal hyperplasia similar to that observed in clinical coronary arterial restenosis after angioplasty. Adhesion of vascular cell adhesion molecule (VCAM)-1 and its ligand very late antigen (VLA)-4 plays an important role in neointimal formation after vascular injury. METHODS AND RESULTS: To evaluate the effectiveness of blocking VCAM-1 and VLA-4 adhesion to prevent neointimal formation, mice with induced abdominal aortic injury were treated with the combined anti-VCAM-1 and anti-VLA-4 mAbs (n = 8) or not treated (n = 6). Injured arteries were harvested at day 14. Immunohistochemistry and in situ reverse transcriptase polymerase chain reaction were performed for detection of VCAM-1, intercellular adhesion molecule (ICAM)-1 and platelet-derived growth factor (PDGF)-B mRNA expression in the arteries. Arteries without treatment showed significant neointimal formation with enhancement of ICAM-1, VCAM-1 and PDGF-B mRNA, while expression of these and intimal thickening were almost nonexistent in the recipients of mAbs to VCAM-1 and VLA-4. CONCLUSION: Anti-VCAM-1 and anti-VLA-4 mAbs prevent arterial neointimal formation after arterial injury.

Animals↗