永井 良三

We found that cigarette smoking increased white blood cell count, and individuals which increased white blood cell count more likely to have metabolic syndrome in Japanese men. We investigated whether similar relationship can be observed also in women. We analyzed the data from 16,383 Japanese women who underwent general health screening. Age-adjusted logistic regression analysis showed that current smoking was positively associated with a highest white blood cell count quartile with an odds ratio of 2.40 (95% CI: 2.16-2.68, P &lt 0.0001). The white blood cell count showed a graded association with metabolic syndrome. On the other hand, the association between current smoking and metabolic syndrome was no longer significant after subdividing the individuals into groups according to the white blood cell quartile. These data collectively suggested that the association between current smoking and metabolic syndrome is heavily confounded by certain factors that increase the circulating white blood cell count in Japanese women, as in men. © 2007 Elsevier Ireland Ltd. All rights reserved.

Background: Despite male predominance in the prevalence of hypertrophic cardiomyopathy (HCM), repeated diagnosis at our institute indicates a possible higher prevalence of deep Q waves with HCM in women. Objective: The current study examined gender similarities and differences in the prevalence of deep Q waves in HCM and in the morphologic and electrocardiographic features of HCM with deep Q waves. Methods: Patients with HCM underwent cardiac magnetic resonance (CMR) imaging to identify the prevalence of deep Q waves in electrocardiographic limb leads, and to analyze the relationship between distribution patterns of deep Q waves and those of the localization of maximum amplitude of left ventricular (LV) hypertrophy. Contiguous LV short-axis images were obtained from the base toward the apex. Results: Of the 200 consecutive patients (172 males, aged 20-78 years 28 females, aged 16-79 years) with HCM who underwent CMR imaging, 10 male and 8 female patients had deep Q waves. Deep Q waves were more prevalent in females with HCM than in their male counterparts (28.6% vs 5.8%, respectively P &lt 0.001). Of the 18 patients with deep Q waves, maximum wall thickness was localized at either the basal anterior wall or the midventricular septum in 9 (90%) of the 10 male patients and 6 (75%) of the 8 female patients. In both sexes, the Q wave distribution pattern of I and aVL and of II and aVF indicated localization of maximum hypertrophy at the midventricular septum in 6 (75%) of the 8 patients with the former pattern, and at the basal anterior wall in 9 (90%) of the 10 patients with the latter pattern. Conclusions: Diagnostic deep Q waves were detected more frequently in female patients with HCM than in their male counterparts. In HCM with deep Q waves in limb leads, morphologic and electrocardiographic analysis showed similar features in both sexes. (Gend Med. Keywords: deep Q wave, hypertrophic cardiomyopathy, gender, cardiac magnetic resonance. © 2007 Excerpta Medica, Inc.

Objective: Cigarette smoking increases the circulating white blood cell (WBC) count and the prevalence of metabolic syndrome. We investigated the association between cigarette smoking, WBC count, and metabolic syndrome as defined by the Japanese criteria. Method: Cross-sectional data from 3,687 men undergoing general health screening between 2005 and 2006 were analyzed. Results: After adjustment for age and total cholesterol, former and current smoking were associated with the highest WBC quartile (≥6.3 × 103 cells/μL) with an odds ratio of 1.35 (95% CI 1.09-1.66, P=0.0055) and 4.45 (95% CI 3.69-5.37, P&lt 0.0001), respectively. It was found that increased WBC count was a risk factor for metabolic syndrome on the other hand, the current smoking was not found to be a predictor for metabolic syndrome, when each WBC count quartile was separately analyzed. Conclusions: Our data suggest that the risk for MetS, defined by Japanese criteria, might be estimated by the WBC count in Japanese men irrespective of their smoking status, although it should also be noted that the cigarette smoking increases the number of circulating WBC count. © 2007 The Japanese Society of Internal Medicine.

Serum albumin is a maker of nutritional status and possesses antioxidative properties. Here, we have sought to investigate the mode of association between serum albumin levels, metabolic syndrome, and carotid atherosclerosis by analyzing the data of the cross-sectional data from 8143 individuals who underwent general health screening test. After adjusting for age, total cholesterol, and smoking status, the highest quartile of serum albumin (>= 4.7 g/dL) was associated with increased prevalence of metabolic syndrome with an odds ratio of 1.80 (95% CI 1.41-2.23, P < 0.0001) in women, and 1.60 (95% CI 1.44-1.78, P < 0.0001) in men, when compared to the lowest serum albumin quartile (< 4.3 g/dL). By contrast, when compared with the lowest quartile, the highest quartile of serum albumin was associated with reduced prevalence of carotid plaque with an odds ratio of 0.62 (95% CI 0.42-0.9 1, P < 0.001) in women, and 0.76 (95% CI 0.62-0.93, P < 0.0 1) in men, and for carotid intima-media thickening with an odds ratio of 0.57 (95% CI 0.35-0.94, P < 0.05) in women, and 0.71 (95% CI 0.55-0.92, P < 0.0 1) in men. Our data showed that higher serum albumin was inversely associated with the prevalence of early carotid atherosclerosis, although it was positively associated with the prevalence of metabolic syndrome. Whether these observations are in part explained by the antioxidative properties of albumin requires further investigation. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

Objective: This study was designed to investigate the roles of programmed death-1 (PD-1) and PD-1ligands (PD-L) in the development of murine acute myocarditis caused by Coxsackievirus B3. PD-1/PD-L belong to the CD28/B7 superfamily, and the PD-1/PD-L pathway is known to transduce a negative immunoregulatory signal that antagonizes the T-cell receptor-CD28 signal and inhibits T-cell activation. Methods: We first analyzed the expression of PD-L1/PD-L2 on cardiac myocytes in vivo and in vitro. Second, we examined the effects of in vivo treatment with an anti-PD-1, PD-L1, or PD-L2 monoclonal antibodies on the development of myocardial inflammation in C3H/He mice infected with Coxsackievirus B3. Third, to investigate the effects of anti-PD-1 monoclonal antibody treatment on the activation of the infiltrating cells, we examined the expression of interleukin (IL)-2, interferon (IFN)-γ, CD40 ligand (CD40L), Fas ligand (FasL), and perforin as activation markers in mouse hearts by a semiquantitative PCR method. Results: PD-L1 was markedly induced on cardiac myocytes with acute myocarditis. In vivo anti-PD-1 or -PD-L1 blocking monoclonal antibody treatment increased the myocardial inflammation whereas anti-PD-1 stimulating monoclonal antibody treatment decreased the myocardial inflammation, and anti-PD-L2 monoclonal antibody treatment had no effect. Anti-PD-1 monoclonal antibody treatment significantly increased the expression of IFN-γ, FasL, CD40L, perforin, and Coxsackievirus B3 genomes in myocardial tissue. Conclusion: Our findings strongly suggest that the PD-1/PD-L1 pathway played a pivotal role in suppressing myocardial inflammation and raise the possibility of immunotherapy by stimulating the PD-1/PD-L1 pathway to prevent myocardial damage in viral myocarditis. © 2007 European Society of Cardiology.

OBJECTIVES: Acyclic retinoid (ACR) is a synthetic retinoid with a high safety profile that has been pursued with high expectations for therapeutic use in prevention (recurrence) and treatment of malignancies. With the objective of addressing the therapeutic potential in the cardiovasculature, namely neointima formation, effects of ACR on neointima formation and the involved mechanisms were investigated. METHODS AND RESULTS: ACR was administered to cuff-injured mice which showed inhibition of neointima formation. Investigation of involved mechanisms at the cellular and molecular levels showed that ACR induces apoptosis of neointimal cells and this to be mediated by selective induction of retinoic-acid receptor beta (RARbeta) which shows growth inhibitory and proapoptotic effects on smooth muscle cells. CONCLUSION: We show that ACR inhibits neointima formation by inducing RARbeta which in turn inhibits cell growth and induces apoptosis. The retinoid, ACR, may be potentially exploitable for treatment and prevention of neointima formation.

Adiponectin has been shown to stimulate fatty acid oxidation and enhance insulin sensitivity through the activation of AMP-activated protein kinase (AMPK) in the peripheral tissues. The effects of adiponectin in the central nervous system, however, are still poorly understood. Here, we show that adiponectin enhances AMPK activity in the arcuate hypothalamus (ARH) via its receptor AdipoR1 to stimulate food intake; this stimulation of food intake by adiponectin was attenuated by dominant-negative AMPK expression in the ARH. Moreover, adiponectin also decreased energy expenditure. Adiponectin-deficient mice showed decreased AMPK phosphorylation in the ARH, decreased food intake, and increased energy expenditure, exhibiting resistance to high-fat-diet-induced obesity. Serum and cerebrospinal fluid levels of adiponectin and expression of AdipoR1 in the ARH were increased during fasting and decreased after refeeding. We conclude that adiponectin stimulates food intake and decreases energy expenditure during fasting through its effects in the central nervous system.

Sterol regulatory element-binding protein (SREBP)-1c is now well established as a key transcription factor for the regulation of lipogenic enzyme genes such as FAS in hepatocytes. Meanwhile, the mechanisms of lipogenic gene regulation in adipocytes remain unclear. Here, we demonstrate that those in adipocytes are independent of SREBP-1c. In adipocytes, unlike in hepatocytes, the stimulation of SREBP-1c expression by liver X receptor agonist does not accompany lipogenic gene upregulation, although nuclear SREBP-1c protein is concomitantly increased, indicating that the activation process of SREBP-1c by the cleavage system is intact in adipocytes. Supportively, transcriptional activity of the mature form of SREBP-1c for the FAS promoter was negligible when measured by reporter analysis. As an underlying mechanism, accessibility of SREBP-1c to the functional elements was involved, because chromatin immunoprecipitation assays revealed that SREBP-1c does not bind to the functional SRE/E-box site on the FAS promoter in adipocytes. Moreover, genetic disruption of SREBP-1 did not cause any changes in lipogenic gene expression in adipose tissue. In summary, in adipocytes, unlike in hepatocytes, increments in nuclear SREBP-1c are not accompanied by transactivation of lipogenic genes; thus, SREBP-1c is not committed to the regulation of lipogenesis.

It has been shown previously that administration of angiogenic growth factors as genes or proteins can augment collateral growth in ischaemic tissues. In the present study, we have investigated the effect of ONO-1301, a synthetic prostacyclin agonist with thromboxane-synthase-inhibitory activity, on expression of endogenous growth factors and angiogenesis. ONO-1301 induced secretion of HGF (hepatocyte growth factor) and VEGF (vascular endothelial growth factor) from cultured normal human dermal fibroblasts in a dose-dependent manner. Dibutyryl cAMP, an analogue of cAMP, and forskolin, an adenylate cyclase activator, mimicked the effect of ONO-1301. Conversely, Rp-cAMP (adenosine 3',5'-cyclic monophosphorothioate), an inhibitor of cAMP, partially inhibited the effect of ONO-1301, suggesting that cAMP mediated the effect of ONO-1301 in up-regulating the expression of HGF and VEGF, at least in part. ONO-1301 promoted tube-like formation by HUVECs (human umbilical vein endothelial cells) when co-cultured with fibroblasts, and the angiogenic effect of ONO-1301 was abrogated by administration of a neutralizing antibody against HGF or VEGF. To generate a slow-releasing form of ONO-1301, ONO-1301 was mixed with poly(DL-lactic-co-glycolic acid). The slow-releasing form of ONO-1301 was injected directly into the ischaemic myocardium of mice immediately after ligation of the left anterior descending artery. The slow-releasing form of ONO-1301 up-regulated HGF and VEGF expression and increased capillary density in the border zone (342.7+/-29.7 capillaries/mm(2) in controls compared with 557.2+/-26.7 capillaries/mm(2) in treated animals; P<0.01) at 7 days. The slow-releasing form of ONO-1301 ameliorated left ventricular enlargement after 28 days and improved survival rate. In conclusion, our results indicate that ONO-1301 up-regulated endogenous growth factors and promoted angiogenesis in response to acute ischaemia. Therefore ONO-1301 might have a therapeutic potential in treating ischaemic diseases.

OBJECTIVE: The expansion of adipose tissue mass seen in obesity involves both hyperplasia and hypertrophy of adipocytes. However, little is known about how adipocytes, adipocyte precursors, blood vessels, and stromal cells interact with one another to achieve adipogenesis. RESEARCH DESIGN AND METHODS: We have developed a confocal microscopy-based method of three-dimensional visualization of intact living adipose tissue that enabled us to simultaneously evaluate angiogenesis and adipogenesis in db/db mice. RESULTS: We found that adipocyte differentiation takes place within cell clusters (which we designated adipogenic/angiogenic cell clusters) that contain multiple cell types, including endothelial cells and stromal cells that express CD34 and CD68 and bind lectin. There were close spatial and temporal interrelationships between blood vessel formation and adipogenesis, and the sprouting of new blood vessels from preexisting vasculature was coupled to adipocyte differentiation. CD34(+) CD68(+) lectin-binding cells could clearly be distinguished from CD34(-) CD68(+) macrophages, which were scattered in the stroma and did not bind lectin. Adipogenic/angiogenic cell clusters can morphologically and immunohistochemically be distinguished from crown-like structures frequently seen in the late stages of adipose tissue obesity. Administration of anti-vascular endothelial growth factor (VEGF) antibodies inhibited not only angiogenesis but also the formation of adipogenic/angiogenic cell clusters, indicating that the coupling of adipogenesis and angiogenesis is essential for differentiation of adipocytes in obesity and that VEGF is a key mediator of that process. CONCLUSIONS: Living tissue imaging techniques provide novel evidence of the dynamic interactions between differentiating adipocytes, stromal cells, and angiogenesis in living obese adipose tissue.

Hyperuricemia is postulated to be a risk factor for atherosclerotic diseases, although whether it is independent of classical atherogenic risk factors is controversial. The automatic computer-assisted measurement of brachial-ankle pulse wave velocity (baPWV) is a valid and reproducible method by which to assess arterial stiffness, a potential surrogate marker of early atherosclerosis. By analyzing cross-sectional data from 982 individuals who underwent health screening, we have investigated whether serum uric acid is associated with high baPWV, which was determined as the highest quartile of baPWV values, in a sex-specific manner. Multivariate analysis showed that the odds ratios (95% CI) of the highest baPWV quartile across the sex-specific quartiles of serum uric acid were 1.0, 2.80 (0.93-8.40), 2.13 (0.74-6.19), and 2.76 (1.01-7.55) in women, and 1.0, 1.10 (0.55-2.20), 1.97 (1.04-3.75), and 2.24 (1.10-4.56) in men after adjusting for age, total and HDL-cholesterol, BMI, systolic blood pressure, triglycerides, fasting glucose and smoking status. The association between uric acid and high baPWV was observed in both subjects with metabolic syndrome and those without. These data suggest that in both genders, serum uric acid level is associated with increased baPWV, a marker of arterial stiffness, and is in part independent of other conventional risk factors for atherosclerosis and metabolic syndrome. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

OPC-28326,4-(N-methyl-2-phenylethylamino)-1-(3,5-dimethyl-4-propionyl-aminobenzoyl) piperidine hydrochloride monohydrate, is a newly developed selective peripheral vasodilator and increases blood flow to lower extremities with alpha 2-adrenergic antagonist property. Here, we investigated the effect of OPC-28326 on ischemia-induced angiogenesis. OPC-28326 enhanced tube formation by human aortic endothelial cells (HAECs). Moreover, OPC-28326 enhanced the number of microvessels sprouting from aortic rings embedded in collagen gel. OPC-28326 markedly induced phosphorylation of endothelial nitric oxide synthase (eNOS) in HAECs via phosphatidylinositol-3 kinase PI3K/Akt (PI3K/ Akt) pathway. Next, the angiogenic effect of OPC-28326 was evaluated in a mouse hindlimb ischemia model. Blood flow recovery to the ischemic leg was significantly enhanced by OPC-28326. Furthermore, anti-CD31 immunostaining revealed that OPC-28326 increased capillary density in the ischemic muscle. However, OPC-28326 failed to promote blood flow recovery in ischemic hindlimb in eNOS-deficient mice. These results suggest that OPC-28326 promotes angiogenesis, which was associated with activation of eNOS via PI3K/Akt pathway. OPC-28326 might be promising to treat patients with ischemic vascular diseases. (C) 2006 Published by Elsevier Masson SAS.

Patients with neonatal lupus erythematosus (NLE) often have congenital heart block with or without heart failure and are born to mothers who have anti-SS-A and/or anti-SS-B antibodies. NLE has been considered to result from the placental transmission of maternal autoantibodies into the fetal circulation causing myocardial damage. We report a case of NLE with congenital heart block who had undergone pacemaker implantation at the age of 17, and then developed dilated cardiomyopathy (DCM) at the age of 19, which is much later than in most other cases. The patient's mother was positive for anti-SS-A and anti-SS-B antibodies, whereas the patient was negative for both anti-SS-A and anti-SS-B antibodies. There were some autoantibodies against cell surface antigens of cardiac myocytes in the serum from the patient, and annexin A6 was identified as one of the autoantigens. This is the first report demonstrating that annexin A6 is involved in the myocardial injury in patients with NLE. The results indicate that inhibition of annexin A6 function may prevent autoantibody-mediated myocardial injury in at least some cases of DCM.

INTRODUCTION: The prognostic value of identifying the retinal status of diabetic patients undergoing coronary implantation of drug-eluting stents is unknown. METHODS: We evaluated the outcomes of 318 consecutive patients undergoing implantation of sirolimus-eluting stents for coronary artery disease. Patients were divided into 5 groups according to the diabetic and retinal status: diabetic patients without retinopathy (43 patients); diabetic patients with nonproliferative retinopathy (34); diabetic patients with proliferative retinopathy (37); diabetic patients with unknown retinal status (30); and nondiabetic patients (174). RESULTS: During a mean follow-up of 385 days, 64 patients had target-vessel failure (defined as a composite of death from cardiac causes, myocardial infarction, and target-vessel revascularization). At 1 year, Kaplan-Meier estimates of the rate of target-vessel failure were 15.3% for diabetic patients without retinopathy, 56.6% for those with nonproliferative retinopathy, 17.3% for those with proliferative retinopathy, 19.0% for those with unknown retinal status, and 16.0% for nondiabetic patients. After adjustment for the potential confounders and differences between groups, the relation of nonproliferative retinopathy to target-vessel failure remained significant. In an analysis in which diabetic patients without retinopathy were used as the reference group, the hazard ratios for target-vessel failure were 3.9 for those with nonproliferative retinopathy, 1.3 for those with proliferative retinopathy, 1.1 for those with unknown retinal status, and 1.4 for nondiabetic patients (P for trend = 0.015). CONCLUSIONS: As compared with diabetic patients without retinopathy, those with nonproliferative retinopathy have an increased risk for target-vessel failure after coronary implantation of sirolimus-eluting stents.

AIMS/HYPOTHESIS: It has been suggested that transcription factor 7-like 2 protein (TCF7L2) plays an important role in glucose metabolism by regulating the production level of glucagon-like peptide-1, a hormone which modifies glucose-dependent insulin secretion. Recently, variants of TCF7L2 gene were reported to confer an increased risk of type 2 diabetes in three different samples from European and European-origin populations. We studied whether the single nucleotide polymorphisms (SNPs) in TCF7L2 were associated with type 2 diabetes in samples from a Japanese population. METHODS: Five SNPs were genotyped in three different sample sets. Association with type 2 diabetes was investigated in each, as well as in combined sample sets. RESULTS: The SNP rs7903146 was nominally associated with type 2 diabetes in the initial (p = 0.08) and two replication sample sets (p = 0.05 and 0.06). For the combined sample set, in which we successfully genotyped 1,174 type 2 diabetes patients and 823 control subjects, rs7903146 showed a significant association with type 2 diabetes (odds ratio = 1.69 [95% CI 1.21-2.36], p = 0.002) with the same direction as the previous reports in samples from European and European-origin populations. SNPs rs7903146 and rs7901695 were in complete linkage disequilibrium. The rest of the five SNPs (rs7895340, rs11196205 and rs12255372) did not show any significant associations with type 2 diabetes. CONCLUSIONS/INTERPRETATION: The consistent association between rs7903146 in TCF7L2 and type 2 diabetes in different ethnic groups, including the Japanese population, suggests that TCF7L2 is a common susceptibility gene for type 2 diabetes.

Aim: We have investigated whether metabolic syndrome is a risk factor for carotid atherosclerosis also in normotensive or prehypertensive individuals. Methods: We analyzed the data from 851 subjects who had a blood pressure of less than 140/90 mmHg and were not taking antihypertensive medication. Metabolic syndrome was defined according to three different criteria: Japan criteria (Japan-MetS); those of the National Cholesterol Education Program (NCEP)-Adult Treatment Panel M (ATP III) (NCEP-MetS); and modified NCEP-ATP III criteria in which body mass index was used as a surrogate for waist circumference (modified NCEP-MetS). Results: Japan-MetS, NCEP-MetS, and modified NCEP-MetS were found, respectively, in 1%, 4%, and 4%, of women, and in 10%, 5%, and 9%, of men. After the adjustment for gender and age, the association between MetS and carotid atherosclerosis did not reach statistical significance. Conclusion: Although the number of enrolled subjects was relatively small, these data may further support the importance of controlling blood pressure within the optimal range for the purpose of preventing atherosclerosis in individuals with metabolic syndrome.

Background - Plasma high-density lipoprotein (HDL) levels have an inverse correlation with incidence of ischemic heart disease as well as other atherosclerosis-related ischemic conditions. However, the molecular mechanism by which HDL prevents ischemic disease is not fully understood. Here, we investigated the effect of HDL on differentiation of endothelial progenitor cells and angiogenesis in murine ischemic hindlimb model. Methods and Results - Intravenous injection of reconstituted HDL (rHDL) significantly augmented blood flow recovery and increased capillary density in the ischemic leg. rHDL increased the number of bone marrow - derived cells incorporated into the newly formed capillaries in ischemic muscle. rHDL induced phosphorylation of Akt in human peripheral mononuclear cells. rHDL (50 to 100 mu g apolipoprotein A-I/mL) promoted differentiation of peripheral mononuclear cells to endothelial progenitor cells in a dose-dependent manner. The effect of rHDL on endothelial progenitor cells differentiation was abrogated by coadministration of LY294002, an inhibitor of phosphatidylinositol 3-kinase. rHDL failed to promote angiogenesis in endothelial NO - deficient mice. Conclusions - rHDL directly stimulates endothelial progenitor cell differentiation via phosphatidylinositol 3-kinase/Akt pathway and enhances ischemia-induced angiogenesis. rHDL may be useful in the treatment of patients with ischemic cardiovascular diseases.

Krüppel-like factor 5 (KLF5) is a transcription factor important in regulation of the cardiovascular response to external stress. KLF5 regulates pathological cell growth, and its acetylation is important for this effect. Its mechanisms of action, however, are still unclear. Analysis in KLF5-deficient mice showed that KLF5 confers apoptotic resistance in vascular lesions. Mechanistic analysis further showed that it specifically interacts with poly(ADP-ribose) polymerase-1 (PARP-1), a nuclear enzyme important in DNA repair and apoptosis. KLF5 interacted with a proteolytic fragment of PARP-1, and acetylation of KLF5 under apoptotic conditions increased their affinity. Moreover, KLF5 wild-type (but not a non-acetylatable point mutant) inhibited apoptosis as induced by the PARP-1 fragment. Collectively, we have found that KLF5 regulates apoptosis and targets PARP-1, and further, for acetylation to regulate these effects. Our findings thus implicate functional interaction between the transcription factor KLF5 and PARP-1 in cardiovascular apoptosis.

Histone chaperones assemble and disassemble nucleosomes in an ATP-independent manner and thus regulate the most fundamental step in the alteration of chromatin structure. The molecular mechanisms underlying histone chaperone activity remain unclear. To gain insights into these mechanisms, we solved the crystal structure of the functional domain of SET/TAF-Ibeta/INHAT at a resolution of 2.3 A. We found that SET/TAF-Ibeta/INHAT formed a dimer that assumed a "headphone"-like structure. Each subunit of the SET/TAF-Ibeta/INHAT dimer consisted of an N terminus, a backbone helix, and an "earmuff" domain. It resembles the structure of the related protein NAP-1. Comparison of the crystal structures of SET/TAF-Ibeta/INHAT and NAP-1 revealed that the two proteins were folded similarly except for an inserted helix. However, their backbone helices were shaped differently, and the relative dispositions of the backbone helix and the earmuff domain between the two proteins differed by approximately 40 degrees . Our biochemical analyses of mutants revealed that the region of SET/TAF-Ibeta/INHAT that is engaged in histone chaperone activity is the bottom surface of the earmuff domain, because this surface bound both core histones and double-stranded DNA. This overlap or closeness of the activity surface and the binding surfaces suggests that the specific association among SET/TAF-Ibeta/INHAT, core histones, and double-stranded DNA is requisite for histone chaperone activity. These findings provide insights into the possible mechanisms by which histone chaperones assemble and disassemble nucleosome structures.

Adiponectin plays a central role as an antidiabetic and antiatherogenic adipokine. AdipoR1 and AdipoR2 serve as receptors for adiponectin in vitro, and their reduction in obesity seems to be correlated with reduced adiponectin sensitivity. Here we show that adenovirus-mediated expression of AdipoR1 and R2 in the liver of Lepr(-/-) mice increased AMP-activated protein kinase (AMPK) activation and peroxisome proliferator-activated receptor (PPAR)-alpha signaling pathways, respectively. Activation of AMPK reduced gluconeogenesis, whereas expression of the receptors in both cases increased fatty acid oxidation and lead to an amelioration of diabetes. Alternatively, targeted disruption of AdipoR1 resulted in the abrogation of adiponectin-induced AMPK activation, whereas that of AdipoR2 resulted in decreased activity of PPAR-alpha signaling pathways. Simultaneous disruption of both AdipoR1 and R2 abolished adiponectin binding and actions, resulting in increased tissue triglyceride content, inflammation and oxidative stress, and thus leading to insulin resistance and marked glucose intolerance. Therefore, AdipoR1 and R2 serve as the predominant receptors for adiponectin in vivo and play important roles in the regulation of glucose and lipid metabolism, inflammation and oxidative stress in vivo.

Adipocyte-derived adiponectin has an antiatherosclerotic effect that acts independently of its antidiabetic effect. Plasma adiponectin levels are generally low in subjects with coronary artery disease. In this study, the relationship between the plasma adiponectin level and the severity of coronary artery disease, as assessed using the Gensini score, an index for the severity of coronary artery stenosis, was investigated. The subjects of the study were 104 patients (72 men and 32 women; BMI, 23.5 +/- 3.3 kg/m(2); age, 63.6 +/- 10.1 years) admitted to Tokyo University Hospital for coronary angiography. Plasma adiponectin levels were inversely correlated with the insulin resistance index HOMA-IR (P = 0.0127). The plasma adiponectin level was significantly associated with the Gensini score (P = 0.0332). After adjustment for conventional risk factors for cardiovascular diseases, the plasma adiponectin level tended to be inversely correlated with the Gensini score (P = 0.087). The measurement of plasma adiponectin levels may be useful for predicting the severity of coronary artery stenosis.

Purpose: Coronary artery vascular edge recognition on computed tomography (CT) angiograms is influenced by window parameters. A noninvasive method for vascular edge recognition independent of window setting with use of multi-detector row CT was contrived and its feasibility and accuracy were estimated by intravascular ultrasound (IVUS). Methods: Multi-detector row CT was performed to obtain 29 CT spatial profile curves by setting a line cursor across short-axis coronary angiograms processed by multi-planar reconstruction. IVUS was also performed to determine the reference coronary diameter. IVUS diameter was fitted horizontally between two points on the upward and downward slopes of the profile curves and Hounsfield number was measured at the fitted level to test seven candidate indexes for definition of intravascular coronary diameter. The best index from the curves should show the best agreement with IVUS diameter. Results: Of the seven candidates the agreement was the best (agreement: 16 ± 11%) when the two ratios of Hounsfield number at the level of IVUS diameter over that at the peak on the profile curves were used with water and with fat as the background tissue. These edge definitions were achieved by cutting the horizontal distance by the curves at the level defined by the ratio of 0.41 for water background and 0.57 for fat background. Conclusions: Vascular edge recognition of the coronary artery with CT spatial profile curves was feasible and the contrived method could define the coronary diameter with reasonable agreement. © 2005 The College of Radiographers.

Due to recent discoveries of novel genes involved in iron metabolism, our understanding of the molecular mechanisms underlying iron metabolism has dramatically increased. We have previously shown that the administration of angiotensin II alters iron homeostasis in the rat kidney, which may in turn aggravate angiotensin II-induced renal damage. Here we have investigated the effect of angiotensin II administration on the localization and expression of transferrin receptor (TfR), divalent metal transporter 1 (DMT1), ferroportin 1 (FPN), and hepcidin mRNA in the rat kidney. Weak expression of TfR, DMT1, FPN, and hepcidin mRNA was observed in the kidneys of control rats. In contrast, after 7 days of angiotensin II infusion by osmotic minipump, the expression of these mRNAs was more widely distributed. Staining of serial sections revealed that some, but not all, of the renal tubular cells positive for these genes contained iron deposits in the kidney of angiotensin II-infused animals. Real-time polymerase chain reaction (PCR) showed that the mRNA expression of TfR, iron-responsive element-negative DMT1, FPN, and hepcidin mRNA increased similar to 1.9-fold, similar to 1.7-fold, similar to 2.3-fold, and similar to 4.7-fold, respectively, after angiotensin II infusion as compared with that of untreated controls, and that these increases could be suppressed by the concomitant administration of losartan. Our data demonstrate that these genes were unequivocally expressed in the kidney and could be regulated by angiotensin II infusion. The relative contribution, if any, of these genes to renal and/or whole-body iron homeostasis in various disorders in which the renin angiotensin system is activated should be investigated in future studies.

Accelerated coronary arteriosclerosis remains a major problem for the long-term survival of cardiac transplant recipients. However, the pathogenesis of transplant vasculopathy is poorly understood and there is no effective therapy. HMG-CoA reductase inhibitors, or statins, are widely prescribed to lower plasma cholesterol level. Accumulating evidence indicates that statins have various effects on vascular cells which are independent of their lipid-lowering effect. We investigated whether orally administered atorvastatin, one of the most potent statins, inhibits the development of intima hyperplasia in a mouse model of cardiac transplantation. Cardiac allografts from DBA mice were transplanted heterotopically into B10.D2 mice. Mice were administered either vehicle or atorvastatin everyday by gavage. Morphometrical analysis revealed that atorvastatin significantly reduced the development of coronary arteriosclerosis on the cardiac allografts harvested at one month. Immunohistochemical analysis revealed that atorvastatin attenuated infiltration of inflammatory cells with reduced expression of TGF-beta and adhesion molecules. These results suggest that atorvastatin may be effective in preventing transplant-associated arteriosclerosis along with other immunosuppressive agents. (c) 2007 Elsevier Masson SAS. All rights reserved.

We have been examining the role of heat shock factor 1 (HSF1) in the pleiotropic effects of statins. In parallel studies, we found that statin induces the nuclear translocation of HSF1 and that a decoy oligonucleotide encoding the heat shock element inhibits the statin-induced expression of heat shock protein 70, endothelial nitric oxide synthase (eNOS) and thrombomodulin. Also, in vascular endothelial cells, increases in the expression of human HSF1 corresponded with elevated steady-state levels of eNOS and thrombomodulin and reduced levels of endothelin-1 and plasminogen activator inhibitor-1. We also found that heat shock proteins induced eNOS and thrombomodulin expression and reduced PAI-1 and ET-1 expression. In particular, a combination of HSP70 and HSP90 strongly induced eNOS expression and reduced PAI-1 expression. In the current studies, we generated a constitutively active form of HSF1 and found that it is more effective than the wild-type HSF at inducing thrombomodulin and eNOS expression and decreasing endothelin-1 and plasminogen activator inhibitor-1 expression. These results show that the wild-type and constitutively active forms of HSF1 induce anticoagulation and relaxation factors in vascular endothelial cells and could therefore be used to treat cardiovascular disease.

The cardiovascular diseases are closely related to circadian rhythm, which is under the control of the biological clock. Clock genes show circadian oscillation not only in the suprachiasmatic nucleus but also in peripheral tissues, suggesting the existence of the peripheral clock. We previously demonstrated that plasminogen activator inhibitor-1 (PAI-1) might be an output gene of the peripheral clock. To further elucidate the functional relevance of the peripheral clock in the cardiovascular system, we screened target genes of the peripheral clock by cDNA microarray analysis. A total of 29 genes including transcription factor, growth factors, and membrane receptors were upregulated by CLOCK/BMAL and showed circadian oscillation. These results suggest that cardiovascular systems have their own peripheral clocks, and at least in part, they may regulate the circadian oscillation of cardiovascular function directly. These results potentially provide a molecular basis for the circadian variation of cardiovascular function and novel strategies for the prevention and treatment of cardiovascular diseases.<br>

BACKGROUND: Recent evidence suggests that bone marrow (BM)-derived cells may differentiate into vascular cells that participate in arterial repair and/or lesion formation. However, it remains uncertain whether BM-derived cells also can participate in vascular remodeling associated with pulmonary arterial hypertension. METHODS AND RESULTS: The BM of Sprague-Dawley rats was reconstituted with that of green fluorescent protein-transgenic rats. The BM-chimeric rats were injected intraperitoneally with 60 mg/kg monocrotaline after unilateral subpneumonectomy, and they concurrently underwent wire-mediated endovascular injury in femoral artery. After 28 days, they had elevated right ventricular systolic pressure (58.8+/-5.4 versus 20.4+/-2.4 mm Hg in sham-control; P<0.01). The pulmonary arterioles were markedly thickened, with an infiltration of green fluorescent protein-positive macrophages into the perivascular areas. The endothelium of pulmonary arterioles contained only a few green fluorescent protein-positive cells, and green fluorescent protein-positive cells were seldom detected as smooth muscle cells in the lesions of thickened pulmonary arterioles. In contrast, BM-derived smooth muscle-like cells could be readily detected in the thickened neointima and media of the wire-injured femoral artery. Moreover, intravenous injection of 1x10(8) BM cells from young rats had no beneficial effects on pulmonary hypertension, pulmonary arterial remodeling, or survival in the aged rats treated with monocrotaline plus unilateral subpneumonectomy. No injected BM cell was identified as an endothelial cell or a smooth muscle cell. CONCLUSIONS: These results suggest that BM-derived cells can participate in arterial neointimal formation after mechanical injury, whereas they do not contribute substantially to pulmonary arterial remodeling associated with monocrotaline-induced pulmonary arterial hypertension in the pneumonectomized rats.

Mediastinal bronchogenic cysts are frequently detected incidentally in adults. Here, we present a unique case of atypical clinical course of bronchogenic cyst beneath the carina, which ruptured into the pericardium. We could not diagnose definitely at initial admission, even though the massive pericardial fluid was exudative with high carbohydrate antigen (CA) 19-9 production. The fluid cytology was negative for malignancy. Five years later, the regrown and unruptured bronchogenic cyst caused atrial fibrillation by impinging on the heart with a high level of serum CA19-9. After resection of the CA19-9-enriched cyst by thoracoscopic surgery, he was discharged without recurrence of atrial fibrillation or any chest symptoms, and the serum CA19-9 level decreased to within the normal range. Differential diagnosis of cytology negative pericardial effusion with high CA19-9 production may be difficult, however, intrapericardial rupture of bronchogenic cyst should be considered in some patients. © 2007 Elsevier Ltd. All rights reserved.

Objective: Iodine-123 MIBG imaging has been used to study cardiac sympathetic function in various cardiac diseases. Central sleep apnea syndrome (CSAS) occurs frequently in patients with chronic heart failure (CHF) and is reported to be associated with a poor prognosis. One of the mechanisms of its poor prognosis may be related to impaired cardiac sympathetic activity. However, the relationship between chemosensitivity to carbon dioxide, which is reported to correlate with the severity of CSAS, and cardiac sympathetic activity has not been investigated. Therefore, this study was undertaken to assess cardiac sympathetic function and chemosensitivity to carbon dioxide in CHF patients. Methods: The oxygen desaturation index (ODI) was evaluated in 21 patients with dilated cardiomyopathy (male/female: 19/2, LVEF &lt; 45%, 65 +/- 12 yr). Patients with an ODI &gt; 5 times/h underwent polysomnography. Patients with an apnea hypopnea index &gt; 15/h but without evidence of obstructive apnea were defined as having CSAS. Early (15 min) and delayed (4 hr) planar MIBG images were obtained from these patients. The mean counts in the whole heart and the mediastinum were obtained. The heart-to-mediastinum count ratio of the delayed image (HIM) and the corrected myocardial washout rate (WR) were also calculated. The central chemoreflex was assessed with the rebreathing method using a hypercapnic gas mixture (7% CO2 and 93% O-2). Results: Ten of the 21 patients had CSAS. The H/M ratio was similar in patients both with and without CSAS (1.57 +/- 0.18 vs. 1.59 +/- 0.14, p = 0.82). However, the WR was higher in patients with CSAS than in patients without CSAS (40 +/- 8% vs. 30 +/- 12%, p &lt; 0.05). ODI significantly correlated with central chemosensitivity to carbon dioxide. Moreover, there was a highly significant correlation between WR and central chemosensitivity (r = 0.65, p &lt; 0.05). However, there was no correlation between ODI and the WR (r = 0.36, p = 0.11). Conclusions: Cardiac sympathetic nerve activity in patients with CHF and CSAS is impaired. However, central sleep apnea might not directly increase cardiac sympathetic nerve activity. We suggest that central chemosensitivity, which is considered to be one of the mechanisms of CSAS, is correlated with cardiac sympathetic nerve activity.

Therapeutic angiogenesis has emerged as a promising therapy to treat patients with ischemic diseases. Transplantation of bone marrow cells (BMCs) is reported to augment collateral development in ischemic organs either by differentiating into vascular cells or by secreting angiogenic cytokines. Recent evidence suggests that adipose tissues secrete a number of humoral factors and contain pluripotent stem cells. Here, we evaluated the therapeutic potential of adipose tissue-derived cells to promote angiogenesis in a mouse model of hind limb ischemia. Stromal vascular fraction cells (SVFs) were isolated from inguinal adipose tissue. Endothelial-like cells or smooth muscle-like cells could be obtained from the culture of SVFs in the presence of growth factors. Freshly isolated BMCs, SVFs, or mature adipocytes were transplanted into the ischemic hind limb of mice. SVFs significantly augmented collateral development as determined by the restoration of blood perfusion and capillary density of the ischemic muscle. Angiogenic effects of SVFs were as potent as those of BMCs. Mature adipocytes showed no proangiogenic effects. The ischemic muscle contained endothelial cells or smooth muscle cells that derived from the transplanted SVFs and BMCs. These results suggest that SVFs might be used to promote angiogenesis in ischemic tissues. (c) 2006 Elsevier Inc. All rights reserved.

Glucokinase (Gck) functions as a glucose sensor for insulin secretion, and in mice fed standard chow, haploinsufficiency of beta cell-specific Gck (Gck(+/-)) causes impaired insulin secretion to glucose, although the animals have a normal beta cell mass. When fed a high-fat (HF) diet, wild-type mice showed marked beta cell hyperplasia, whereas Gck(+/-) mice demonstrated decreased beta cell replication and insufficient beta cell hyperplasia despite showing a similar degree of insulin resistance. DNA chip analysis revealed decreased insulin receptor substrate 2 (Irs2) expression in HF diet-fed Gck(+/-) mouse islets compared with wild-type islets. Western blot analyses confirmed upregulated Irs2 expression in the islets of HF diet-fed wild-type mice compared with those fed standard chow and reduced expression in HF diet-fed Gck(+/-) mice compared with those of HF diet-fed wild-type mice. HF diet-fed Irs2(+/-) mice failed to show a sufficient increase in beta cell mass, and overexpression of Irs2 in beta cells of HF diet-fed Gck(+/-) mice partially prevented diabetes by increasing beta cell mass. These results suggest that Gck and Irs2 are critical requirements for beta cell hyperplasia to occur in response to HF diet-induced insulin resistance.

Protein profiling would aid in better understanding the pathophysiology of metabolic disease. Here, we report on differential proteomic analysis using an animal model of diabetes mellitus and associated metabolic disorders (Otsuka Long-Evans Tokushima Fatty rat). Serum was analyzed by a new two-dimensional liquid chromatography system which separated proteins by chromatofocusing and subsequent reversed-phase chromatography. This is the first application of this approach to differential serum proteomics. Differentially expressed proteins, identified with MALDI-TOF mass spectrometry, included apolipoproteins and alpha2-HS-glycoprotein. These findings add to our understanding of the underlying pathophysiology. This new proteomic analysis is a promising tool to elucidate disease mechanisms.

We studied the possibility of using high-intensity focused ultrasound (HIFU) together with a microbubble agent to treat hepatocellular carcinoma. Development of liver tumors in rats was induced by administration of Dimethylnitrosamin (100 ppm). Rats with liver tumors were anesthetized, underwent laparotomy, and were given the microbubble agent Levovist or saline intravenously. After the injection, the liver was exposed to HIFU for 30 s (2.18 MHz, 600 W/cm2, 40 mm in diameter). Immediately after HIFU exposure, ultrasound images of the HIFU area were evaluated. Then the liver was excised and the volume of coagulated tissue was measured. The mean volumes of hyperechoic areas after HIFU were as follows (mm3, Levovist versus saline: 355.3 ± 180.7 versus 47.4 ± 35.6, P &lt 0.001, n = 13). The volumes of liver tissue coagulated by HIFU were as follows (mm3, Levovist versus saline: 275.3 ± 120.0 versus 60.1 ± 23.6, P &lt 0.001, n = 13). On microscopic examination of areas exposed to HIFU, implosion cysts were seen, and many cancer cells were found to have been destroyed completely (loss of cell membranes or nuclei). In conclusion, the microbubble agent Levovist can increase the volume of tissue coagulated by HIFU. © 2006 Elsevier Ireland Ltd. All rights reserved.

OBJECTIVE: To elucidate the interdependence between the mechanical state of the myocardium and its electrical activity, previous studies have been performed at the cellular level. However, the information to date has been limited by the technical difficulties associated with stretching single myocytes. METHODS: We solved this problem by combining two techniques, namely a carbon fiber technique for stretching rat myocytes with wide ranges of amplitude and speed, and ratiometric measurement of a fluorescent indicator (di8-ANEPPS) for evaluating the membrane potential in the non-contact mode. RESULTS: During systole, stretching caused depolarization that prolonged the action potential duration without affecting the peak amplitude, but the effect was only significant in the late phase. Application of a stretch to quiescent myocytes depolarized the membrane potential in amplitude- and speed-dependent manners, but the response was suppressed by cytochalasin D treatment, suggesting participation of the cytoskeleton in the mechanotransduction mechanism. Finally, ion replacement experiments revealed that although Na+ was the dominant charge carrier for large amplitude stretches, Ca2+ permeation was involved in small amplitude stretches, suggesting amplitude-dependent ion selectivity. CONCLUSIONS: Application of axial stretching to rat ventricular myocytes changed the membrane potential in phase-, amplitude- and speed-dependent manners. Amplitude may also modulate the ion selectivity of stretch-activated channels.

Lipid accumulation in the kidney is a marker of tissue damage and may play a role in the development of renal injury. We have previously shown that long-term administration of angiotensin 11 in rats causes increased expression of transforming growth factor-beta 1, coupled with an accumulation of lipids in the tubular and vascular wall cells in the kidney. In this study, we examine the regulation of expression of platelet-derived growth factor (PDGF) and its receptor system and their co-locallization with lipid deposits in the kidneys of angiotensin II-infused rats. Real-time RT-PCR showed that expression of PDGF-B, PDGF-D, and PDGIF receptor-beta (PDGFR-beta) mRNA was increased by angiotensin 11 infusion, and in situ hybridization showed the co-localization of these mRNAs. Tubular cells that had increased PDGF-B mRNA expression were positive for lipid deposition and also for cellular proliferation, which was indicated by the presence of proliferating cell nuclear antigen. By contrast, in the kidneys of angiotensin II-infused rats, apoptosis occurred in tubular cells that contained deposits of iron but not lipids. The deposition of lipids and upregulation of PDGF-B, PDGF-D, and PDGFR-beta induced by administration of angiotensin 11 were all suppressed by the selective angiotensin 11 type 1 (AT,) receptor antagonist losartan, but not by the nonspecific vasodilator hydralazine. The findings that lipid accumulation, upregulation of PDGF-B, PDGF-D, and PDGFR-beta, and cellular proliferation were topologically associated and regulated in an AT, receptor-dependent manner in the kidney of angiotensin II-infused rats suggests that these phenomena are related.

Adrenomedullin exerts not only vasodilatory effects, but also angiogenic effects. In the present study, we investigated the effects of adrenomedullin on collateral formation and circulating bone-marrow-derived cells after acute tissue ischaemia. Bone marrow of 8-10-week-old female C57BL/6J mice was replaced with that from GFP (green fluorescent protein) transgenic mice (GFP mice). At 8 weeks after transplantation, hindlimb ischaemia was induced by resecting the right femoral artery and a plasmid expressing human adrenomedullin (50 mug) was injected into the ischaemic muscle, followed by in vivo electroporation on a weekly basis. Overexpression of adrenomedullin significantly enhanced the blood flow recovery compared with controls (blood flow ratio, 1.0+/-0.2 compared with 0.6+/-0.3 respectively, at week 4; P<0.05) and increased capillary density in the ischaemic leg as determined by anti-CD31 immunostaining of the ischaemic muscle (567+/-40 compared with 338+/-65 capillaries/mm(2) respectively, at week 5; P<0.05). There were more GFP-positive cells in the thigh muscle of the mice injected with adrenomedullin than in that of the control mice (29.6+/-4.5 compared with 16.5+/-3.3 capillaries/mm(2) respectively, at week 5; P<0.05). We repeated the same experiments using LacZ-knock-in mice instead of GFP mice, and obtained similar results. These findings suggest that adrenomedullin may augment ischaemia-induced collateral formation with some effects on circulating bone-marrow-derived cells.