Naoyuki Hasebe

This phase III, multicenter, randomized, double-blind, parallel-group study compared the efficacy and safety of nifedipine controlled-release (CR) 40 mg twice daily (b.i.d.) and once daily (q.d.) in 325 Japanese patients with essential hypertension uncontrolled with nifedipine CR 40 mg q.d. (ClinicalTrials.gov record: NCT01287260). The primary endpoint was the change from baseline in trough seated diastolic blood pressure (DBP) after 8 weeks. Nifedipine CR 40 mg b.i.d. showed significantly greater reductions in trough seated DBP (-7.7±0.6 mm Hg vs. -3.6±0.6 mm Hg) and trough seated systolic blood pressure (BP) (-11.1±0.9 mm Hg vs. -3.7±0.9 mm Hg) after 8 weeks of treatment compared with nifedipine CR 40 mg q.d. (both P<0.0001). At week 8, BP target achievement and responder rates were higher with nifedipine CR 40 mg b.i.d. (21.5% and 42.4% vs. 10.3% and 19.5%, respectively). Adverse events considered related to the study drug were reported in 9.0 and 9.7% of patients receiving nifedipine CR 40 mg b.i.d. and q.d., respectively. The frequency of drug-related adverse events commonly reported with nifedipine CR (headache, hot flush, palpitations, peripheral edema, hypotension, dizziness, tachycardia) was low and the results were similar between the treatment groups. In conclusion, a higher dose of nifedipine CR was associated with greater efficacy and a safety profile similar to that of the currently approved dose (40 mg q.d.) in Japanese patients with essential hypertension, and it may offer a valuable treatment choice for patients who do not achieve target BP levels with standard treatment.

The Japanese Society of Hypertension Guidelines for Self-monitoring of Blood Pressure at Home (Second Edition)

To assess the association between cerebral microbleeds (CMBs) and future spontaneous intracerebral hemorrhage (ICH) risk in ischemic stroke patients with nonvalvular atrial fibrillation (AF) taking oral anticoagulants.This was a meta-analysis of cohort studies with >50 patients with recent ischemic stroke and documented AF, brain MRI at baseline, long-term oral anticoagulation treatment, and ≥6 months of follow-up. Authors provided summary-level data on stroke outcomes stratified by CMB status. We estimated pooled annualized ICH and ischemic stroke rates from Poisson regression. We calculated odds ratios (ORs) of ICH by CMB presence/absence, ≥5 CMBs, and CMB topography (strictly lobar, mixed, and strictly deep) using random-effects models.We established an international collaboration and pooled data from 8 centers including 1,552 patients. The crude CMB prevalence was 30% and 7% for ≥5 CMBs. Baseline CMB presence (vs no CMB) was associated with ICH during follow-up (OR 2.68, 95% confidence interval [CI] 1.19-6.01, p = 0.017). Presence of ≥5 CMB was related to higher future ICH risk (OR 5.50, 95% CI 2.07-14.66, p = 0.001). The pooled annual ICH incidence increased from 0.30% (95% CI 0.04-0.55) among CMB-negative patients to 0.81% (95% CI 0.17-1.45) in CMB-positive patients (p = 0.01) and 2.48% (95% CI 1.2-6.2) in patients with ≥5 CMBs (p = 0.001). There was no association between CMBs and recurrent ischemic stroke.The presence of CMB on MRI and the dichotomized cutoff of ≥5 CMBs might identify subgroups of ischemic stroke patients with AF with high ICH risk and after further validation could help in risk stratification, in anticoagulation decisions, and in guiding randomized trials and ongoing large observational studies.

[Figure: see text].

Prostacyclin (PGI(2)) is one of the important vascular prostanoids, the effects of which counteract those of thromboxane (TXA(2)), and these 2 prostanoids provide an important balance in cardiovascular homeostasis. The clinical experience of COX-2 selective inhibitors having unexpected adverse effects in patients with cardiovascular risk has opened up a debate about the role of COX-2-derived prostanoids in vascular pathophysiology. PGI(2) is a major anti-atherogenic prostanoid produced by COX-2 in vascular cells, including endothelial and vascular smooth muscle cells. The balance between COX-2-derived PGI(2), COX-1-derived TXA(2), and other COX-2-mediated atherogenic prostanoids is a crucial factor in determining pathophysiological outcomes. Recent studies using stable PGI(2) analogs and genetically deficient mice have revealed novel effects of PGI(2) on its target cells, such as endothelial and endothelial progenitor cells. The role PGI(2) in the physiology and pathophysiology of vascular diseases is reviewed and the recent findings linking PGI(2), COX-2 and atherothrombosis are summarized.

Inflammatory responses in the kidney lead to tubulointerstitial fibrosis, a common feature of chronic kidney diseases. Here we examined the role of prostaglandin E2 (PGE2) in the development of tubulointerstitial fibrosis. In the kidneys of wild-type mice, unilateral ureteral obstruction leads to progressive tubulointerstitial fibrosis with macrophage infiltration and myofibroblast proliferation. This was accompanied by an upregulation of COX-2 and PGE2 receptor subtype EP4 mRNAs. In the kidneys of EP4 gene knockout mice, however, obstruction-induced histological alterations were significantly augmented. In contrast, an EP4-specific agonist significantly attenuated these alterations in the kidneys of wild-type mice. The mRNAs for macrophage chemokines and profibrotic growth factors were upregulated in the kidneys of wild-type mice after ureteral obstruction. This was significantly augmented in the kidneys of EP4-knockout mice and suppressed by the EP4 agonist but only in the kidneys of wild-type mice. Notably, COX-2 and MCP-1 proteins, as well as EP4 mRNA, were localized in renal tubular epithelial cells after ureteral obstruction. In cultured renal fibroblasts, another EP4-specific agonist significantly inhibited PDGF-induced proliferation and profibrotic connective tissue growth factor production. Hence, an endogenous PGE2–EP4 system in the tubular epithelium limits the development of tubulointerstitial fibrosis by suppressing inflammatory responses. Inflammatory responses in the kidney lead to tubulointerstitial fibrosis, a common feature of chronic kidney diseases. Here we examined the role of prostaglandin E2 (PGE2) in the development of tubulointerstitial fibrosis. In the kidneys of wild-type mice, unilateral ureteral obstruction leads to progressive tubulointerstitial fibrosis with macrophage infiltration and myofibroblast proliferation. This was accompanied by an upregulation of COX-2 and PGE2 receptor subtype EP4 mRNAs. In the kidneys of EP4 gene knockout mice, however, obstruction-induced histological alterations were significantly augmented. In contrast, an EP4-specific agonist significantly attenuated these alterations in the kidneys of wild-type mice. The mRNAs for macrophage chemokines and profibrotic growth factors were upregulated in the kidneys of wild-type mice after ureteral obstruction. This was significantly augmented in the kidneys of EP4-knockout mice and suppressed by the EP4 agonist but only in the kidneys of wild-type mice. Notably, COX-2 and MCP-1 proteins, as well as EP4 mRNA, were localized in renal tubular epithelial cells after ureteral obstruction. In cultured renal fibroblasts, another EP4-specific agonist significantly inhibited PDGF-induced proliferation and profibrotic connective tissue growth factor production. Hence, an endogenous PGE2–EP4 system in the tubular epithelium limits the development of tubulointerstitial fibrosis by suppressing inflammatory responses. The worldwide increase in the number of patients with chronic kidney disease and the resultant end-stage renal failure necessitating renal replacement therapy, such as transplantation and dialysis, is threatening to reach almost pandemic proportions.1.Meguid ElNahasA Bello A.K. Chronic kidney disease: the global challenge.Lancet. 2005; 365: 331-340Abstract Full Text Full Text PDF PubMed Scopus (895) Google Scholar Tubulointerstitial fibrosis, as well as glomerulosclerosis, is a major predictor of the progression to end-stage renal failure in various types of chronic kidney disease.2.Zeisberg M. Neilson E.G. Mechanisms of tubulointerstitial fibrosis.J Am Soc Nephrol. 2010; 21: 1819-1834Crossref PubMed Scopus (669) Google Scholar For example, interstitial fibrosis, along with glomerulosclerosis, is one of the most important factors determining the progression and prognosis of diabetic nephropathy.3.Gilbert R.E. Cooper M.E. The tubulointerstitium in progressive diabetic kidney disease: more than an aftermath of glomerular injury?.Kidney Int. 1999; 56: 1627-1637Abstract Full Text Full Text PDF PubMed Scopus (564) Google Scholar In immunoglobulin A (IgA) nephropathy, which is the most common type of chronic glomerulonephritis characterized by glomerular IgA1 deposition, tubulointerstitial fibrosis is more closely related to prognosis of the disease than to the glomerular damage itself.4.Donadio J.V. Grande J.P. IgA nephropathy.N Engl J Med. 2002; 347: 738-748Crossref PubMed Scopus (679) Google Scholar Therefore, clarification of the mechanism leading to tubulointerstitial fibrosis is an important issue for the management of chronic kidney disease. Tubular epithelial cells are thought to play a pivotal role in the process of tubulointerstitial fibrosis. These cells produce several types of chemokines in response to various stimuli and attract macrophages and lymphocytes into the renal interstitial space.5.Wynn T.A. Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases.J Clin Invest. 2007; 117: 524-529Crossref PubMed Scopus (1113) Google Scholar Among these chemokines, MCP-1 (monocyte chemoattractant protein-1) and RANTES (regulated on activation, T cell expressed, and secreted), which belong to the CC chemokine family, have emerged as important mediators of macrophage recruitment in various types of renal diseases.6.Anders H.J. Vielhauer V. Frink M. et al.A chemokine receptor CCR-1 antagonist reduces renal fibrosis after unilateral ureter ligation.J Clin Invest. 2002; 109: 251-259Crossref PubMed Scopus (209) Google Scholar, 7.Anders H.J. Vielhauer V. Schlondorff D. Chemokines and chemokine receptors are involved in the resolution or progression of renal disease.Kidney Int. 2003; 63: 401-415Abstract Full Text Full Text PDF PubMed Scopus (222) Google Scholar, 8.Bascands J.L. Schanstra J.P. Obstructive nephropathy: insights from genetically engineered animals.Kidney Int. 2005; 68: 925-937Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar The recruited macrophages induce proliferation of resident fibroblasts and their differentiation into myofibroblasts through the production of profibrotic growth factors, such as transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF).9.Eddy A.A. Molecular insights into renal interstitial fibrosis.J Am Soc Nephrol. 1996; 7: 2495-2508Crossref PubMed Google Scholar,10.Wolf G. Renal injury due to renin-angiotensin-aldosterone system activation of the transforming growth factor-β pathway.Kidney Int. 2006; 70: 1914-1919Abstract Full Text Full Text PDF PubMed Scopus (259) Google Scholar TGF-β1 is also known as the main player in the phenotypic conversion of tubular epithelial cells into mesenchymal fibroblasts, a process known as epithelial–mesenchymal transition (EMT).11.Kalluri R. Neilson E.G. Epithelial-mesenchymal transition and its implications for fibrosis.J Clin Invest. 2003; 112: 1776-1784Crossref PubMed Scopus (2106) Google Scholar Therefore, tubular epithelial cells, as well as resident fibroblasts and pericytes, are able to serve as a source of myofibroblasts.12.Kalluri R. Weinberg R.A. The basics of epithelial-mesenchymal transition.J Clin Invest. 2009; 119: 1420-1428Crossref PubMed Scopus (7315) Google Scholar,13.Iwano M. Plieth D. Danoff T.M. et al.Evidence that fibroblasts derive from epithelium during tissue fibrosis.J Clin Invest. 2002; 110: 341-350Crossref PubMed Scopus (1724) Google Scholar However, recently, Humphreys et al.14.Humphreys B.D. Lin S.L. Kobayashi A. et al.Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis.Am J Pathol. 2010; 176: 85-97Abstract Full Text Full Text PDF PubMed Scopus (1093) Google Scholar indicated no evidence of EMT in a unilateral ureteral obstruction (UUO) model using a novel fate-mapping technique, indicating some arguments about the role of EMT at least in tubulointerstitial fibrosis induced by UUO. Differentiated and activated myofibroblasts then produce interstitial matrices and collagen fiber, resulting in tubulointerstitial fibrosis. Although these sequential events take place in the development of tubulointerstitial fibrosis, the factors participating in the regulation of these events have not been fully characterized. UUO invariably induces tubulointerstitial fibrosis in the ureter-obstructed kidney (UOK), a well-established model used to investigate the mechanism leading to tubulointerstitial fibrosis.8.Bascands J.L. Schanstra J.P. Obstructive nephropathy: insights from genetically engineered animals.Kidney Int. 2005; 68: 925-937Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar, 15.Sato M. Muragaki Y. Saika S. et al.Targeted disruption of TGF-β1/Smad3 signaling protects against renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction.J Clin Invest. 2003; 112: 1486-1494Crossref PubMed Scopus (698) Google Scholar, 16.Chevalier R.L. Forbes M.S. Thornhill B.A. Ureteral obstruction as a model of renal interstitial fibrosis and obstructive nephropathy.Kidney Int. 2009; 75: 1145-1152Abstract Full Text Full Text PDF PubMed Scopus (712) Google Scholar It has been reported that prostaglandin E2 (PGE2) production increases significantly in the kidney during the development of tubulointerstitial fibrosis induced by UUO.17.Whinnery M.A. Shaw J.O. Beck N. Thromboxane B2 and prostaglandin E2 in the rat kidney with unilateral ureteral obstruction.Am J Physiol. 1982; 242: F220-F225PubMed Google Scholar,18.Okegawa T. Jonas P.E. Deschryver K. et al.Metabolic and cellular alterations underlying the exaggerated renal prostaglandin and thromboxane synthesis in ureter obstruction in rabbits. Inflammatory response involving fibroblasts and mononuclear cells.J Clin Invest. 1983; 71: 81-90Crossref PubMed Scopus (73) Google Scholar However, the role of PGE2 in the development of tubulointerstitial fibrosis remains to be determined. PGE2 exerts a variety of actions in the body by binding to its specific rhodopsin-type receptors.19.Narumiya S. Sugimoto Y. Ushikubi F. Prostanoid receptors: structures, properties, and functions.Physiol Rev. 1999; 79: 1193-1226Crossref PubMed Scopus (0) Google Scholar There are four subtypes of PGE2 receptors encoded by different genes: EP1, EP2, EP3, and EP4. EP1 couples to Gq and raises intracellular Ca2+ concentration. EP2 and EP4 couple to Gs and raise intracellular cAMP concentration ([cAMP]i). Notably, EP4 has another signaling pathway involving phosphatidylinositol 3-kinase, leading to the activation of Akt and extracellular signal–regulated kinases.20.Fujino H. West K.A. Regan J.W. Phosphorylation of glycogen synthase kinase-3 and stimulation of T-cell factor signaling following activation of EP2 and EP4 prostanoid receptors by prostaglandin E2.J Biol Chem. 2002; 277: 2614-2619Crossref PubMed Scopus (273) Google Scholar,21.Regan J.W. EP2 and EP4 prostanoid receptor signaling.Life Sci. 2003; 74: 143-153Crossref PubMed Scopus (382) Google Scholar In contrast, EP3 couples mainly to Gi and inhibits the increase in [cAMP]i. On the basis of the finding that the expression levels of mRNAs for cyclooxygenase (COX)-2, a rate-limiting enzyme of prostanoid synthesis, and EP4 were significantly upregulated in the kidney after UUO, we set out to clarify the EP4-mediated role of PGE2 in the development of tubulointerstitial fibrosis using mice that lack EP4 (EP4–/– mice) and ONO-4819, a specific EP4 agonist. We found that both endogenous PGE2 and ONO-4819 significantly suppress UUO-induced tubulointerstitial fibrosis via EP4 in vivo. In addition, a specific EP4 agonist ONO-AE1-329 significantly inhibited platelet-derived growth factor (PDGF)-BB-induced fibroblast proliferation and TGF-β1-induced EMT in vitro. In wild-type (WT) mice, hydronephrotic changes, including dilation of the renal pelvis and thinning of the renal parenchyma, occurred in UOK after UUO (Figure 1a). In addition, these changes were accompanied by marked collagen deposition in the renal interstitial area (Figure 1b), indicating the development of tubulointerstitial fibrosis in the present UUO model. We investigated whether prostanoids and their receptor system in the kidney respond to stress burdened by UUO in WT mice. First, we examined the expressions of mRNAs for COX-1 and COX-2. The expression level of mRNA for COX-2, an inducible isoform of COX, increased significantly in UOK on day 1 of UUO compared with that in the control kidney and remained to a similar extent on days 4 and 7 of UUO (Figure 1c), suggesting that COX-2-derived prostanoids play a role in the pathogenesis of tubulointerstitial fibrosis after UUO. Interestingly, a significant increase in the expression level of mRNA for COX-1, a constitutive isoform of COX, was also found in the UOK on days 4 and 7 of UUO (Figure 1c). Next, we examined the expression levels of mRNAs for EPs in UOK before and after UUO. Before UUO, mRNAs for all EPs were detected. After UUO, the expression level of EP4 mRNA increased significantly and reached a level of up to sevenfold on day 7 of UUO (Figure 1d), suggesting an EP4-mediated role of PGE2 in the development of tubulointerstitial fibrosis. Although the expression level of EP2 mRNA increased significantly to a level of up to twofold on day 7 of UUO, those of EP1 and EP3 mRNAs did not change or decreased significantly, respectively (Figure 1d). There were no significant differences in the expression levels of EP4 mRNA between the upper and lower parts of the kidney before and 7 days after UUO (data not shown). On the basis of these results, we set out to clarify the EP4-mediated role of PGE2 in tubulointerstitial fibrosis using the UUO model in the following examinations. To assess the degree of fibrotic response to UUO, interstitial collagen deposition was examined histologically in Sirius red–stained renal sections, and hydroxyproline content was measured for quantification of fibrosis. In the kidneys of WT mice (WT kidneys), tubulointerstitial fibrosis developed progressively after UUO, and renal hydroxyproline content increased twofold over that of control kidneys on day 7 of UUO (Figure 2a and e). In the kidneys of EP4–/– mice (EP4–/– kidneys), however, tubulointerstitial fibrosis was significantly augmented compared with that in WT kidneys on day 7 of UUO (Figure 2c and e), indicating that endogenous PGE2 has a suppressive role in tubulointerstitial fibrosis via EP4. Furthermore, when EP4 was stimulated by administration of ONO-4819, an EP4-specific agonist, the degree of tubulointerstitial fibrosis in WT kidneys was significantly reduced (Figure 2d and e). In addition, the effect of ONO-4819 was completely abolished in EP4–/– kidneys (Figure 2e), indicating that a potent suppressive effect of ONO-4819 on tubulointerstitial fibrosis was mediated by EP4. There was no apparent increase in interstitial collagen deposition in the contralateral kidney (CLK) on day 7 of UUO (Figure 2b). In tubulointerstitial fibrosis, macrophage infiltration is a characteristic feature of local inflammation, and infiltrated macrophages have a critical role in the succeeding myofibroblast proliferation and fibrosis.7.Anders H.J. Vielhauer V. Schlondorff D. Chemokines and chemokine receptors are involved in the resolution or progression of renal disease.Kidney Int. 2003; 63: 401-415Abstract Full Text Full Text PDF PubMed Scopus (222) Google Scholar, 8.Bascands J.L. Schanstra J.P. Obstructive nephropathy: insights from genetically engineered animals.Kidney Int. 2005; 68: 925-937Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar, 12.Kalluri R. Weinberg R.A. The basics of epithelial-mesenchymal transition.J Clin Invest. 2009; 119: 1420-1428Crossref PubMed Scopus (7315) Google Scholar Therefore, we next examined macrophage infiltration into the renal interstitial area in the UUO model (Figure 3). In WT kidneys, UUO induced progressive macrophage infiltration, which was noted exclusively in the peritubular area (Figure 3a and e). This infiltration of macrophages was detected as early as day 1 of UUO (data not shown). In EP4–/– kidneys, however, the degree of macrophage infiltration was significantly greater than that in WT kidneys on days 4 and 7 of UUO (Figure 3c and e), suggesting an EP4-mediated inhibitory effect of endogenous PGE2 on macrophage recruitment. In addition, ONO-4819 significantly suppressed the UUO-induced macrophage infiltration in WT kidneys, an effect absent in EP4–/– kidneys (Figure 3d and e). In control kidneys and CLKs, a small number of macrophages were detected in the interstitial area, but there was no significant difference in the macrophage number between WT and EP4–/– kidneys (Figure 3b and e). In accordance with macrophage infiltration, UUO induced a remarkable proliferation of interstitial myofibroblasts in WT kidneys after UUO (Figure 4a and e). In EP4–/– kidneys, however, myofibroblast proliferation was significantly augmented compared with that in WT kidneys on days 4 and 7 of UUO (Figure 4c and e). As anticipated, ONO-4819 significantly suppressed the UUO-induced myofibroblast proliferation in WT kidneys, an effect absent in EP4–/– kidneys (Figure 4d and e). In control kidneys and CLKs, myofibroblasts could not be found in the interstitial space (Figure 4b and e). These results indicate that the PGE2–EP4 system negatively regulates macrophage infiltration, thereby succeeding myofibroblast proliferation after UUO. The results therefore suggest the involvement of chemokine(s) for macrophages as the target molecule of the system. MCP-1 and RANTES are thought to be the major chemokines responsible for macrophage recruitment into the renal interstitium under inflammatory conditions.7.Anders H.J. Vielhauer V. Schlondorff D. Chemokines and chemokine receptors are involved in the resolution or progression of renal disease.Kidney Int. 2003; 63: 401-415Abstract Full Text Full Text PDF PubMed Scopus (222) Google Scholar,8.Bascands J.L. Schanstra J.P. Obstructive nephropathy: insights from genetically engineered animals.Kidney Int. 2005; 68: 925-937Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar Therefore, we examined the renal expression of MCP-1 and RANTES mRNAs after UUO. In WT kidneys, expression levels of MCP-1 and RANTES mRNAs increased remarkably after UUO (Figure 5). In EP4–/– kidneys, however, the expression levels of these mRNAs were significantly higher than those in WT kidneys on days 4 and 7 of UUO (Figure 5), indicating an EP4-mediated suppressive action of endogenous PGE2 on the UUO-induced chemokine induction. Accordingly, ONO-4819 significantly suppressed the UUO-induced renal expression of MCP-1 and RANTES mRNAs in WT mice, an effect absent in EP4–/– mice (Figure 5). In control kidneys and CLKs, there were no significant differences in the expression levels of these chemokine mRNAs between WT and EP4–/– mice. There were no significant differences in the expression levels of RANTES mRNA between the upper and lower parts of the kidney before and 7 days after UUO in both WT and EP4–/– mice (data not shown). These results indicate that the PGE2–EP4 system indeed suppresses chemokine induction and limits the progression of tubulointerstitial fibrosis. Recruited macrophages, as well as tubular epithelial cells, are the major source of TGF-β1 and CTGF, the representative profibrotic growth factors responsible for fibroblast proliferation and for the differentiation of fibroblasts into myofibroblasts.16.Chevalier R.L. Forbes M.S. Thornhill B.A. Ureteral obstruction as a model of renal interstitial fibrosis and obstructive nephropathy.Kidney Int. 2009; 75: 1145-1152Abstract Full Text Full Text PDF PubMed Scopus (712) Google Scholar,22.Bani-Hani A.H. Campbell M.T. Meldrum D.R. et al.Cytokines in epithelial-mesenchymal transition: a new insight into obstructive nephropathy.J Urol. 2008; 180: 461-468Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar Because macrophage recruitment was significantly augmented in EP4–/– kidneys, we next examined whether renal production of these macrophage-derived growth factors increases in EP4–/– mice after UUO. In WT kidneys, the expression levels of TGF-β1 and CTGF mRNAs increased remarkably after UUO (Figure 6). In EP4–/– kidneys, however, the expression level of TGF-β1 mRNA was significantly higher than that in WT kidneys on day 7 of UUO (Figure 6a). The expression level of CTGF mRNA in EP4–/– kidneys was also significantly higher than that in WT kidneys on days 4 and 7 of UUO (Figure 6b), confirming an EP4-mediated suppressive action of endogenous PGE2 on the UUO-induced induction of profibrotic growth factors. As anticipated, ONO-4819 significantly suppressed the UUO-induced expression of TGF-β1 and CTGF mRNAs in WT kidneys, an effect absent in EP4–/– kidneys (Figure 6). In control kidneys and CLKs, there were no significant differences in the expression levels of these growth factors between WT and EP4–/– mice. These results suggest that the PGE2–EP4 system negatively regulates the TGF-β1/CTGF axis in the development of tubulointerstitial fibrosis by reducing macrophage recruitment. We next examined by immunohistochemical analysis where the upregulation of COX-2 and MCP-1 occurs within WT kidneys after UUO. We found intense COX-2 immunoreactivities mainly in tubular epithelial cells on day 7 of UUO (Figure 7b). Intense immunoreactivities for MCP-1 were also found mainly in tubular epithelial cells on day 7 of UUO (Figure 7b). Importantly, COX-2 and MCP-1 immunoreactivities were colocalized in some tubular epithelial cells (Figure 7b). In control kidneys, we could detect only slight immunoreactivities for COX-2 and MCP-1 (Figure 7a). These results indicate that renal tubular epithelial cells are the major cells expressing both COX-2 and MCP-1 after UUO and suggest that COX-2-derived PGE2 is also produced in tubular epithelial cells. Next, we examined the expression pattern of EP4 mRNA within the kidney after UUO by in situ hybridization analysis. In WT control kidneys, EP4 mRNA expression was barely detectable (Figure 8a). However, we could detect slight signals for EP4 mRNA in the glomeruli and vascular walls of WT control kidneys in other in situ hybridization analyses using a radioisotope (data not shown). On day 7 of UUO, signals for EP4 mRNA increased remarkably throughout WT kidneys (Figure 8b). Notably, these signals were localized mainly to tubular epithelial cells and to a lesser extent to interstitial cells and glomeruli (Figure 8c). To further confirm tubular localization of EP4 mRNA, E-cadherin, a representative marker for tubular epithelium, was immunostained in a contiguous tissue section (Figure 8d). Signals for EP4 mRNAs were detected mainly in the structure that coincides with tubular epithelium expressing E-cadherin (Figure 8d). There were no signals for EP4 mRNA in EP4–/– kidneys on day 7 of UUO (data not shown). These results indicate that the renal tubular epithelium is the major site expressing EP4 after UUO, while recruited macrophages likely express EP4. In renal tubular epithelial cells, TGF-β1 is a representative inducer of EMT, which was characterized by changes in the expression of a multitude of proteins, such as α-smooth muscle actin (α-SMA), and transcription factors including Twist, as well as morphological changes. We first examined the effect of TGF-β1 on the expression level of EP4 mRNA in cultured renal tubular epithelial cells isolated from WT kidneys. TGF-β1 significantly increased the expression level of EP4 mRNA (Figure 9a), suggesting upregulation of EP4 in response to a proinflammatory stimulus. Next, to test whether EP4 activation affects EMT, we examined the effect of an EP4-selective agonist ONO-AE1-329 on the TGF-β1-induced expression of α-SMA and Twist mRNAs in cultured renal tubular epithelial cells. In both tubular epithelial cells isolated from WT and EP4–/– kidneys, TGF-β1 significantly increased the expression levels of α-SMA and Twist mRNAs (Figure 9b and c), indicating induction of EMT. In tubular epithelial cells from WT kidneys, ONO-AE1-329 significantly suppressed the TGF-β1-induced expressions of α-SMA and Twist mRNAs. In contrast, ONO-AE1-329 did not have these effects on tubular epithelial cells from EP4–/– kidneys (Figure 9b and c), suggesting a suppressive role of the PGE2–EP4 system in EMT. Next, we examined whether EP4 works also in renal fibroblasts as well as in tubular epithelial cells. In cultured renal fibroblasts isolated from WT kidneys, ONO-AE1-329 significantly suppressed PDGF-BB-induced proliferation in a concentration-dependent manner (Figure 10a). In cultured renal fibroblasts isolated from EP4–/– kidneys, however, the suppressive effect of ONO-AE1-329 disappeared (Figure 10a). In addition, ONO-AE1-329 significantly inhibited TGF-β1-induced CTGF production by fibroblasts isolated from WT kidneys but not that by fibroblasts isolated from EP4–/– kidneys (Figure 10b). These results indicate that the PGE2–EP4 system would play a suppressive role in the development of tubulointerstitial fibrosis after UUO acting on fibroblasts in addition to tubular epithelial cells. Finally, we examined whether an EP4 agonist could protect the kidney in another model of kidney disease as well, folic acid–induced renal injury. Blood urea nitrogen and serum creatinine concentrations increased significantly in folic acid–injected balb/c mice (Figure 11d and e), indicating induction of renal injury by folic acid. ONO-4819, however, significantly suppressed the increases in blood urea nitrogen and creatinine concentrations (Figure 11d and e), suggesting the protective effect of ONO-4819 in folic acid–induced renal injury. In addition, substantial renal interstitial fibrosis was observed after folic acid injection by histological examination with Sirius red staining (Figure 11a and b); this fibrosis was apparently suppressed by ONO-4819 (Figure 11c). Accordingly, renal hydroxyproline content increased significantly on day 7 of folic acid injection (Figure 11f). Notably, ONO-4819 significantly suppressed the increase in renal hydroxyproline content (Figure 11f), indicating that ONO-4819 has an antifibrotic effect also in this model. Suppression of the development of tubulointerstitial fibrosis has emerged as an important issue for the management of chronic kidney diseases because it determines prognosis of the diseases, and there is little antifibrotic therapy for preventing the progression of chronic kidney diseases to end-stage renal failure. Therefore, extensive research has been conducted to clarify the mechanism of tubulointerstitial fibrosis, and a number of signaling molecules have been proposed to participate in the pathogenesis of this condition. These signaling molecules, such as chemokines, inflammatory cytokines, and profibrotic growth factors, mediate the sequential events taking place during the development of tubulointerstitial fibrosis.6.Anders H.J. Vielhauer V. Frink M. et al.A chemokine receptor CCR-1 antagonist reduces renal fibrosis after unilateral ureter ligation.J Clin Invest. 2002; 109: 251-259Crossref PubMed Scopus (209) Google Scholar, 7.Anders H.J. Vielhauer V. Schlondorff D. Chemokines and chemokine receptors are involved in the resolution or progression of renal disease.Kidney Int. 2003; 63: 401-415Abstract Full Text Full Text PDF PubMed Scopus (222) Google Scholar, 23.Misseri R. Rink R.C. Meldrum D.R. et al.Inflammatory mediators and growth factors in obstructive renal injury.J Surg Res. 2004; 119: 149-159Abstract Full Text Full Text PDF PubMed Scopus (69) Google Scholar Among these mediators, TGF-β1 and CTGF constitute an established signaling axis in the process of tubulointerstitial fibrosis.15.Sato M. Muragaki Y. Saika S. et al.Targeted disruption of TGF-β1/Smad3 signaling protects against renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction.J Clin Invest. 2003; 112: 1486-1494Crossref PubMed Scopus (698) Google Scholar,22.Bani-Hani A.H. Campbell M.T. Meldrum D.R. et al.Cytokines in epithelial-mesenchymal transition: a new insight into obstructive nephropathy.J Urol. 2008; 180: 461-468Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar In addition, several factors, such as angiotensin II and bradykinin, have been reported as upstream key molecules controlling the events.8.Bascands J.L. Schanstra J.P. Obstructive nephropathy: insights from genetically engineered animals.Kidney Int. 2005; 68: 925-937Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar,10.Wolf G. Renal injury due to renin-angiotensin-aldosterone system activation of the transforming growth factor-β pathway.Kidney Int. 2006; 70: 1914-1919Abstract Full Text Full Text PDF PubMed Scopus (259) Google Scholar However, the role of PGE2 as such a key molecule remains to be determined. In EP4–/– kidneys, tubulointerstitial fibrosis was significantly augmented compared with that in WT kidneys after UUO, indicating that endogenous PGE2 plays a suppressive role in the progression of tubulointerstitial fibrosis via EP4. In addition, when EP4 was pharmacologically activated by an EP4-specific agonist, ONO-4819, UUO-induced tubulointerstitial fibrosis was significantly diminished in WT kidneys. Furthermore, this effect was completely absent in EP4–/– kidneys, indicating that the potent suppressive effect of ONO-4819 on tubulointerstitial fibrosis is mediated by EP4. To our knowledge, this is the first report demonstrating that endogenous PGE2 and an EP4 agonist exert a suppressive action via EP4 on the development of tubulointerstitial fibrosis in vivo. The renal expression level of COX-2 mRNA increased significantly after UUO, suggesting that COX-2-derived prostanoids including PGE2 participate in the pathogenesis of tubulointerstitial fibrosis. This result is in good agreement with reported results that state that production of PGE2 increa

NLRP3 inflammasomes recognize non-microbial danger signals and induce release of proinflammatory cytokine interleukin (IL)-1β, leading to sterile inflammation in cardiovascular disease. Because sterile inflammation is involved in doxorubicin (Dox)-induced cardiotoxicity, we investigated the role of NLRP3 inflammasomes in Dox-induced cardiotoxicity. Cardiac dysfunction and injury were induced by low-dose Dox (15 mg/kg) administration in NLRP3-deficient (NLRP3(-/-)) mice but not in wild-type (WT) and IL-1β(-/-) mice, indicating that NLRP3 deficiency enhanced the susceptibility to Dox-induced cardiotoxicity independent of IL-1β. Although the hearts of WT and NLRP3(-/-) mice showed no significant difference in inflammatory cell infiltration, macrophages were the predominant inflammatory cells in the hearts, and cardiac IL-10 production was decreased in Dox-treated NLRP3(-/-) mice. Bone marrow transplantation experiments showed that bone marrow-derived cells contributed to the exacerbation of Dox-induced cardiotoxicity in NLRP3(-/-) mice. In vitro experiments revealed that NLRP3 deficiency decreased IL-10 production in macrophages. Furthermore, adeno-associated virus-mediated IL-10 overexpression restored the exacerbation of cardiotoxicity in the NLRP3(-/-) mice. These results demonstrated that NLRP3 regulates macrophage IL-10 production and contributes to the pathophysiology of Dox-induced cardiotoxicity, which is independent of IL-1β. Our findings identify a novel role of NLRP3 and provided new insights into the mechanisms underlying Dox-induced cardiotoxicity.

Background Although the impact of malnutrition in patients with acute stroke has been reported, its significance after rehabilitation is not well understood. The geriatric nutritional risk index (GNRI) is a simple and well-established nutritional screening tool that predicts poor prognosis in elderly patients and in those with a high risk of cardiovascular events. We investigated the associations between GNRI and all-cause mortality, cardiovascular events, and infectious diseases in patients with stroke after rehabilitation. Methods This study included 138 patients aged 80 years or below who were discharged between 2010 and 2013 in a single center, and followed up for more than 1 year. Malnutrition was defined as a GNRI of 96 or lower. Results The mean age was 63.9 ± 11.0 years, the mean GNRI at discharge was 98.8 ± 6.5, and the mean total functional independence measure (FIM) score at discharge was 91.8 ± 25.8. Among the patients, 37 (27%) had malnutrition. During the follow-up period, all-cause mortality, cardiovascular events, and infectious diseases were recorded in 11 (8%), 21 (15%), and 20 (15%) patients, respectively. Kaplan–Meier curves showed a significantly higher incidence of each outcome in patients with a GNRI of 96 or lower. In the Cox proportional analysis, GNRI was an independent determinant of all-cause mortality (hazard ratio [HR], .71; 95% confidence interval [CI], .61-.83), cardiovascular events (HR, .87; 95% CI, .80-.95), and infectious diseases (HR, .80; 95% CI, .74-.87) after adjusting for age, gender, and total FIM score. Conclusions Malnutrition has a negative impact on prognosis in patients with stroke even after rehabilitation. Although the impact of malnutrition in patients with acute stroke has been reported, its significance after rehabilitation is not well understood. The geriatric nutritional risk index (GNRI) is a simple and well-established nutritional screening tool that predicts poor prognosis in elderly patients and in those with a high risk of cardiovascular events. We investigated the associations between GNRI and all-cause mortality, cardiovascular events, and infectious diseases in patients with stroke after rehabilitation. This study included 138 patients aged 80 years or below who were discharged between 2010 and 2013 in a single center, and followed up for more than 1 year. Malnutrition was defined as a GNRI of 96 or lower. The mean age was 63.9 ± 11.0 years, the mean GNRI at discharge was 98.8 ± 6.5, and the mean total functional independence measure (FIM) score at discharge was 91.8 ± 25.8. Among the patients, 37 (27%) had malnutrition. During the follow-up period, all-cause mortality, cardiovascular events, and infectious diseases were recorded in 11 (8%), 21 (15%), and 20 (15%) patients, respectively. Kaplan–Meier curves showed a significantly higher incidence of each outcome in patients with a GNRI of 96 or lower. In the Cox proportional analysis, GNRI was an independent determinant of all-cause mortality (hazard ratio [HR], .71; 95% confidence interval [CI], .61-.83), cardiovascular events (HR, .87; 95% CI, .80-.95), and infectious diseases (HR, .80; 95% CI, .74-.87) after adjusting for age, gender, and total FIM score. Malnutrition has a negative impact on prognosis in patients with stroke even after rehabilitation.

Chronic kidney disease (CKD) is one of the most significant risk factors for cardiovasculardisese. Malnutrition has been recognized as a significant risk factor for cardiovascular disease in patients with CKD, including those on chronic dialysis. Current studies showed higher all-cause and cardiovascular mortality rates in patients with CKD and malnutrition. Geriatric nutritional risk index (GNRI), a simple and validated nutritional screening measure for both elderly people and patients on dialysis, is based only on three objective parameters: body weight, height, and serum albumin level. Recently, we demonstrated that the cutoff GNRI for predicting all-cause and cardiovascular mortality was 96 in patients on hemodialysis. Moreover, together with left ventricular hypertrophy and low estimated glomerular filtration rate, the utility of GNRI as a significant determinant of cardiovascular events was demonstrated in non-dialysis-dependent patients with CKD. In the present review, we summarize available evidence regarding the relationship of GNRI with all-cause and cardiovascular mortality in patients with CKD including those on dialysis.

Persistent reduction of renal perfusion pressure induces renovascular hypertension by activating the renin-angiotensin-aldosterone system; however, the sensing mechanism remains elusive. Here we investigated the role of PGI2 in renovascular hypertension in vivo, employing mice lacking the PGI2 receptor (IP-/- mice). In WT mice with a two-kidney, one-clip model of renovascular hypertension, the BP was significantly elevated. The increase in BP in IP-/- mice, however, was significantly lower than that in WT mice. Similarly, the increases in plasma renin activity, renal renin mRNA, and plasma aldosterone in response to renal artery stenosis were all significantly lower in IP-/- mice than in WT mice. All these parameters were measured in mice lacking the four PGE2 receptor subtypes individually, and we found that these mice had similar responses to WT mice. PGI2 is produced by COX-2 and a selective inhibitor of this enzyme, SC-58125, also significantly reduced the increases in plasma renin activity and renin mRNA expression in WT mice with renal artery stenosis, but these effects were absent in IP-/- mice. When the renin-angiotensin-aldosterone system was activated by salt depletion, SC-58125 blunted the response in WT mice but not in IP-/- mice. These results indicate that PGI2 derived from COX-2 plays a critical role in regulating the release of renin and consequently renovascular hypertension in vivo.

The aim was to evaluate the potential of single-photon emission computed tomography (SPECT) to predict cardiac death in chronic hemodialysis patients using the iodinated fatty acid analogue iodine-123 123I-beta-methyl iodophenyl-pentadecanoic acid (BMIPP).We previously reported that BMIPP SPECT could detect asymptomatic coronary artery disease with high sensitivity in hemodialysis patients.We prospectively enrolled 375 asymptomatic hemodialysis patients who had undergone dual SPECT using 123I-BMIPP and 201thallium (Tl) chloride. Patients who had a clinical history of myocardial infarction and/or coronary revascularization were excluded from the study. Uptake on SPECT images was graded in 17 segments on a 5-point scale (0 normal, 4 absent) and assessed as summed BMIPP or Tl scores.During a 3.6 +/- 1.0-year follow-up, 57 patients who had undergone coronary revascularization within 60 days of SPECT were excluded from the analysis. Among the remaining 318 patients (male/female: 170/148; 64 +/- 12 years of age), 50 died of cardiac events (acute myocardial infarction 22, congestive heart failure 17, cardiac sudden death 11). Stepwise Cox hazard analysis associated cardiac death with age (> or =70 years) and with severely abnormal BMIPP SPECT images (BMIPP summed scores > or =12: hazard ratio 21.894; p < 0.0001). Kaplan-Meier analysis showed that the cardiac death-free survival rates at 3 years were 61% and 98% in patients with BMIPP summed scores of > or =12 and <12, respectively.Severely impaired myocardial fatty acid metabolism, which might mainly reflect repetitive myocardial ischemia, can identify a high-risk group of cardiac death among hemodialysis patients.

Endothelial progenitor cells (EPCs) play an important role in the self-healing of a vascular injury by participating in the reendothelialization that limits vascular remodeling. We evaluated whether prostaglandin I(2) plays a role in the regulation of the function of EPCs to limit vascular remodeling.EPCs (Lin(-)cKit(+)Flk-1(+) cells) were isolated from the bone marrow (BM) of wild-type (WT) mice or mice lacking the prostaglandin I(2) receptor IP (IP(-/-) mice). Reverse transcription-polymerase chain reaction analysis showed that EPCs among BM cells specifically express IP. The cellular properties of EPCs, adhesion, migration, and proliferation on fibronectin were significantly attenuated in IP-deficient EPCs compared with WT EPCs. In contrast, IP agonists facilitated these functions in WT EPCs, but not in IP-deficient EPCs. The specific deletion of IP in BM cells, which was performed by transplanting BM cells of IP(-/-) mice to WT mice, accelerated wire injury-mediated neointimal hyperplasia in the femoral artery. Notably, transfused WT EPCs, but not IP-deficient EPCs, were recruited to the injured vessels, participated in reendothelialization, and efficiently rescued the accelerated vascular remodeling.These findings clearly indicate that the prostaglandin I(2)-IP system is essential for EPCs to accomplish their function and plays a critical role in the regulation of vascular remodeling.

Atherosclerosis is considered an "inside-out" response, that begins with the dysfunction of intimal endothelial cells and leads to neointimal plaque formation. The adventitia of large blood vessels has been recognized as an active part of the vessel wall that is involved in the process of atherosclerosis. There are characteristic changes in the adventitial vasa vasorum that are associated with the development of atheromatous plaques. However, whether vasa vasorum plays a causative or merely reactive role in the atherosclerotic process is not completely clear. Recent studies report that the vascular wall contains a number of stem/progenitor cells that may contribute to vascular remodeling. Microvessels serve as the vascular niche that maintains the resident stem/progenitor cells of the tissue. Therefore, the vasa vasorum may contribute to vascular remodeling through not only its conventional function as a blood conducting tube, but also its new conceptual function as a stem cell reservoir. This brief review highlights the recent advances contributing to our understanding of the role of the adventitial vasa vasorum in the atherosclerosis and discusses new concept that involves vascular-resident factors, the vasa vasorum and its associated vascular-resident stem cells, in the atherosclerotic process.

Background Detecting myocardial ischemia in hemodialysis patients is crucial given the high incidence of silent ischemia and the high cardiovascular mortality rates. Abnormal myocardial fatty acid metabolism as determined by imaging with 123I-labeled BMIPP (β-methyl iodophenyl-pentadecanoic acid) might be associated with cardiac-derived death in hemodialysis patients. Study Design Prospective observational study. Setting & Participants Asymptomatic hemodialysis patients with one or more cardiovascular risk factors, but without known coronary artery disease, were followed up for 3 years at 48 Japanese hospitals (406 men, 271 women; mean age, 64 years). Predictor Baseline BMIPP summed scores semiquantified using a 17-segment 5-point system (normal, 0; absent, 4). Outcomes Cardiac-derived death, including cardiac and sudden death. Measurements HRs were estimated using a Cox model for associations between BMIPP summed scores and cardiac-derived death, adjusting for potential confounders of age, sex, body mass index, dialysis duration, and cardiovascular risk factors. Results Rates of all-cause mortality and cardiac-derived death were 18.5% and 6.8%, respectively. Cardiac-derived death (acute myocardial infarction [n = 10], congestive heart failure [n = 13], arrhythmia [n = 2], valvular heart disease [n = 1], and sudden death [n = 20]) accounted for 36.8% of all-cause deaths. Cardiac-derived death (n = 46) was associated with age, history of heart failure, and BMIPP summed scores of 4 or higher (HR, 2.9; P < 0.001). Three-year cardiac-derived death–free survival rates were 95.7%, 90.6%, and 78.8% when BMIPP summed scores were 3 or lower, 4-8, and 9 or higher, respectively. BMIPP summed score also was a predictor of all-cause death (HR, 1.6; P = 0.009). Limitations Sudden death of unknown cause was considered to have been cardiac derived, although a coronary origin was not confirmed. Conclusions Abnormal myocardial fatty acid metabolism is associated with cardiac-derived death in hemodialysis patients. BMIPP single-proton emission computed tomography appears clinically useful for predicting cardiac-derived death in this population. Detecting myocardial ischemia in hemodialysis patients is crucial given the high incidence of silent ischemia and the high cardiovascular mortality rates. Abnormal myocardial fatty acid metabolism as determined by imaging with 123I-labeled BMIPP (β-methyl iodophenyl-pentadecanoic acid) might be associated with cardiac-derived death in hemodialysis patients. Prospective observational study. Asymptomatic hemodialysis patients with one or more cardiovascular risk factors, but without known coronary artery disease, were followed up for 3 years at 48 Japanese hospitals (406 men, 271 women; mean age, 64 years). Baseline BMIPP summed scores semiquantified using a 17-segment 5-point system (normal, 0; absent, 4). Cardiac-derived death, including cardiac and sudden death. HRs were estimated using a Cox model for associations between BMIPP summed scores and cardiac-derived death, adjusting for potential confounders of age, sex, body mass index, dialysis duration, and cardiovascular risk factors. Rates of all-cause mortality and cardiac-derived death were 18.5% and 6.8%, respectively. Cardiac-derived death (acute myocardial infarction [n = 10], congestive heart failure [n = 13], arrhythmia [n = 2], valvular heart disease [n = 1], and sudden death [n = 20]) accounted for 36.8% of all-cause deaths. Cardiac-derived death (n = 46) was associated with age, history of heart failure, and BMIPP summed scores of 4 or higher (HR, 2.9; P < 0.001). Three-year cardiac-derived death–free survival rates were 95.7%, 90.6%, and 78.8% when BMIPP summed scores were 3 or lower, 4-8, and 9 or higher, respectively. BMIPP summed score also was a predictor of all-cause death (HR, 1.6; P = 0.009). Sudden death of unknown cause was considered to have been cardiac derived, although a coronary origin was not confirmed. Abnormal myocardial fatty acid metabolism is associated with cardiac-derived death in hemodialysis patients. BMIPP single-proton emission computed tomography appears clinically useful for predicting cardiac-derived death in this population.

Atrial fibrillation (AF) is a cardiac arrhythmia that frequently induces ischemic strokes. Nowadays, non-vitamin K antagonist oral anticoagulants (NOACs) have come into widespread use for cardiogenic embolism prevention in place of warfarin. Recently, cerebral microbleeds (CMBs) have been noticed for their potential implication in cerebral small vessel disease. We hypothesized that NOACs do not have an unfavorable influence over cerebral small vessels and investigated whether NOACs increase CMBs in AF patients in a prospective manner.We performed baseline magnetic resonance imaging (MRI) examinations on the 69 enrolled AF patients and re-examined second round of MRI 1 year later. The enrolled patients continued the same anticoagulation therapy during the meantime.CMBs did not develop in the 23 patients with NOACs for 1 year. Nine patients with antiplatelets also did not develop CMBs. On the other hand, 3 of 21 patients continued on warfarin and 3 of 9 with warfarin and antiplatelets had CMBs. When divided into 2 groups according to whether the CMBs developed, significant differences in the incidence of using NOACs were observed between the 2 groups (P = .02). A multivariate regression analysis showed that warfarin was independently related to the new development of CMBs (hazard ratio, 10.75; 95% confidence interval, 1.22-94.99; P = .03).This is the first report to clarify that NOACs do not increase CMBs in AF patients longitudinally in 1 year. Further consideration will be continued with a much longer follow-up in large samples.

Cytokines induce apoptosis in vascular disease lesions through enhancement of inducible nitric oxide (NO) synthase (iNOS) activation. The thiazolidinediones, novel insulin-sensitizing agents, have been demonstrated to modulate cytokine-induced NO production. We have investigated the role of pioglitazone in the apoptosis of vascular smooth muscle cells (VSMCs) in vitro and developed intimal hyperplasia in vivo.Pioglitazone (0.1 to 10 micromol/L) significantly enhanced cytokine-induced expression of iNOS and NO production in a dose-dependent manner in rat VSMCs, but 15-deoxy-Delta(12,14)-prostaglandin J2 (up to 10 micromol/L), a native peroxisome proliferator-activated receptor-gamma ligand, showed no effect. Pioglitazone also significantly enhanced reduction of cell viability, as evidenced by the increase in the number of TUNEL-positive cells. All of these effects of pioglitazone were blocked by treatment with N-monomethyl-L-arginine, an NO synthesis inhibitor. In an in vivo study with a balloon-injured rat carotid artery, neointimal thickness had reached maximum levels at 2 weeks after injury. Then, rats were fed with or without pioglitazone (3 mg. kg(-1). d(-1)) for an additional week. The ratio of intima to media area of carotid artery was significantly decreased by 30%, and the ratio of apoptotic cells in neointima was significantly increased in pioglitazone-treated rats compared with vehicle-treated control rats.Pioglitazone enhanced apoptosis in an NO-dependent manner in cytokine-activated VSMCs and induced significant regression of intimal hyperplasia in balloon-injured rat carotid artery. It appears that pioglitazone is a potent apoptosis inducer in vascular lesions, providing a novel pharmacological strategy to prevent restenosis after vascular intervention.

On the hypothesis that coronary sinus occlusion (CSO) may reduce myocardial ischemia, we examined the effects of CSO on coronary collateral blood flow and on the distribution of regional myocardial blood flow (RMBF) in dogs. Thirty-eight anesthetized dogs underwent occlusion of the left anterior descending coronary artery with or without CSO and intact vasomotor tone. We measured RMBF and intramyocardial pressure (IMP) in the subendocardium (Endo) and subepicardium (Epi) separately. With intact vasomotor tone, CSO during ischemia significantly increased RMBF in the ischemic region (IR), particularly in Endo from 0.17 +/- 0.03 to 0.33 +/- 0.05 ml x min(-1) x g(-1) (P < 0.05), and increased the Endo/Epi from 0.59 +/- 0.10 to 1.15 +/- 0.15 (P < 0.01). These effects of CSO were partially abolished by adenosine. However, the Endo/Epi was still increased from 0.90 +/- 0.13 to 2.09 +/- 0.30 (P < 0.01). The changes in RMBF in IR were significantly correlated with the peak CS pressure during CSO. The Endo/Epi of IMP in IR was significantly decreased during CSO. In conclusion, CSO potentially enhances coronary collateral flow, and preserves the ischemic myocardium, especially in Endo.

Impaired endothelial-dependent vascular responses after coronary artery occlusion (CAO) and reperfusion (CAR) have been investigated extensively. However, it is not known whether impaired endogenous endothelium-derived relaxing factor production affects postischemic myocardial dysfunction, ie, myocardial stunning.Eight dogs were instrumented with an intracoronary catheter and an hydraulic occluder on the left circumflex coronary artery. The effects of a 10-minute CAO randomized with and without intracoronary administration of NG-nitro-L-arginine (L-NA), a nitric oxide (NO) synthesis inhibitor, were compared in the same conscious dogs. Postischemic regional contractile dysfunction in subendocardial and subepicardial as well as transmural wall thickening was measured with ultrasonic dimension crystals, and myocardial blood flow was measured with radioactive microspheres. Intracoronary infusion of L-NA did not affect systemic hemodynamics, and transmural myocardial blood flow was reduced slightly (-8%), but significantly, only in the left circumflex territory. The recovery of wall thickening was significantly delayed in the presence of L-NA compared with the absence of L-NA, eg, at 30-minute CAR, not only in the subendocardium (-76 +/- 9% versus -49 +/- 9%) but also in the subepicardium (-52 +/- 8% versus -29 +/- 7%). During CAO, blood flow was decreased identically in both conditions, and during CAR, the differences in blood flow were minor (7%).Inhibition of NO synthesis enhanced myocardial stunning transmurally in conscious dogs, potentially independent of its effects on blood flow.

Objective To compare the clinical efficacy of low-dose controlled-release (CR) nifedipine (20 mg/day) plus candesartan (8 mg/day) combination therapy with that of up-titrated candesartan (12 mg/day) monotherapy. Design Randomized, double-blind study. Setting Outpatient study. Patients and participants Patients with essential hypertension, who did not achieve their target blood pressure with baseline treatment of candesartan 8 mg/day for 8 weeks. Main outcome measures Blood pressure, pulse pressure, urinary microalbumin excretion. Results Blood pressure was significantly reduced in both groups (P < 0.05), but the reduction was significantly greater in the combination therapy group (12.1 ± 1.4/8.7 ± 0.9 mmHg) than in the up-titrated monotherapy group (4.1 ± 1.4/4.6 ± 0.9 mmHg) (P < 0.0001). The reduction in pulse pressure was significantly greater in the combination therapy group (3.3 ± 1.2 mmHg) than in the up-titrated monotherapy group (0.7 ± 1.2 mmHg) (P = 0.0031). Urinary microalbumin excretion decreased significantly in the combination therapy group (from 61.9 to 40.5 mg/g creatinine; P < 0.05), but not in the up-titrated monotherapy group. Conclusions These findings suggest that the low-dose combination therapy of nifedipine CR and candesartan is superior to the up-titrated monotherapy of candesartan in terms of blood pressure control and renal protection in patients with essential hypertension.

Atrial fibrillation (AF) is a cardiac arrhythmia that does not infrequently induce ischemic strokes; however, little research has been reported on focal cerebral microangiopathic lesions in patients with AF. Recently cerebral microbleeds (CMBs) have been noticed for their potential implication in cerebral small vessel disease. Therefore, we had 2 goals in the present study: (1) to compare the prevalence of CMBs in patients with AF with that in patients without AF, and (2) to prove that CMBs could be a clinical predictive factor for the development of future cerebral microangiopathy in patients with AF without a history of symptomatic cerebral infarction in a prospective manner.We performed yearly brain magnetic resonance imaging (MRI) assessments for a maximum of 5 years in 131 patients with AF and 112 control patients. Seventy-seven patients with AF underwent more than 3 yearly MRI scans.The Kaplan-Meier curve showed that the development of an asymptomatic cerebral infarction (ACI) was associated with the baseline presence of a CMB (P=.004). A multivariate Cox regression analysis revealed that the CMBs at baseline were significantly associated with an increment in not only the occurrence of ACIs (hazard ratio [HR], 5.414; 95% confidence interval [CI], 1.03-28.43; P=.046) but also in the consecutive development of CMBs (HR, 6.274; 95% CI, 1.43-27.56; P=.015).Patients with AF had a significantly higher prevalence of CMBs. The presence of CMBs in the baseline MRI may predict the consequent onset of an ACI and increase in CMBs in patients with AF.

Conifer and broadleaf trees emit volatile organic compounds in the summer. The major components of these emissions are volatile monoterpenes. Using solid phase microextraction fiber as the adsorbant, monoterpenes were successfully detected and identified in forest air samples. Gas chromatography/mass chromatogram of monoterpenes in the atmosphere of a conifer forest and that of serum from subjects who were walking in a forest were found to be similar each other. The amounts of α-pinene in the subjects became several folds higher after forest walking. The results indicate that monoterpenes in the atmosphere of conifer forests are transferred to and accumulate in subjects by inhalation while they are exposed to this type of environment.

BackgroundAlthough anticoagulation effectively prevents stroke in patients with atrial fibrillation (AF), it has been underused in elderly AF patients for many reasons, mainly because of knowledge gaps regarding cardiovascular treatment of these populations with multiple comorbidities and poor prognosis. The objectives of the All Nippon AF In the Elderly (ANAFIE) Registry are to collect real-world information about the clinical status of patients with non-valvular AF (NVAF) aged ≥75 years, current status of anticoagulant therapy, and prognosis with/without anticoagulation to establish a database for this specific patient population that is increasing remarkably worldwide.Methods and designThe ANAFIE Registry is an observational, multicenter, prospective study of Japanese patients with NVAF aged ≥75 years that will include 30,000 patients and have the primary endpoint of composite of stroke and systemic embolism over a 2-year follow-up period. In parallel with the main study, seven sub-cohort studies will be conducted with assessments including coagulation-fibrinolysis markers, echocardiography, heart rate, hypertension, cognitive function, frailty, and medication adherence. Subgroup analyses will be performed, and stratified by renal function, HbA1c, and maximum number of drugs used. The study was started in October 2016, with a planned 2-year recruitment period. As of January 31, 2018, 33,213 patients were enrolled; the recruitment was therefore ended 8 months earlier than the original plan.ConclusionsThe ANAFIE Registry will provide a valuable database for the clinical status, management, and outcomes of mortality, stroke, systemic embolism, and hemorrhagic events with/without anticoagulation in the increasing population of elderly NVAF patients, and will identify risk factors associated with these clinical events.

Cardiac glycosides such as digoxin are Na+/K+-ATPase inhibitors that are widely used for the treatment of chronic heart failure and cardiac arrhythmias; however, recent epidemiological studies have suggested a relationship between digoxin treatment and increased mortality. We previously showed that nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasomes, which regulate caspase-1-dependent interleukin (IL)-1β release, mediate the sterile cardiovascular inflammation. Because the Na+/K+-ATPase is involved in inflammatory responses, we investigated the role of NLRP3 inflammasomes in the pathophysiology of cardiac glycoside-induced cardiac inflammation and dysfunction. The cardiac glycoside ouabain induced cardiac dysfunction and injury in wild-type mice primed with a low dose of lipopolysaccharide (LPS), although no cardiac dysfunction was observed in mice treated with either ouabain or LPS alone. Ouabain also induced cardiac inflammatory responses, such as macrophage infiltration and IL-1β release, when mice were primed with LPS. These cardiac manifestations were all significantly attenuated in mice deficient in IL-1β. Furthermore, deficiency of NLRP3 inflammasome components, NLRP3 and caspase-1, also attenuated ouabain-induced cardiac dysfunction and inflammation. In vitro experiments revealed that ouabain induced NLRP3 inflammasome activation as well as subsequent IL-1β release from macrophages, and this activation was mediated by K+ efflux. Our findings demonstrate that cardiac glycosides promote cardiac inflammation and dysfunction through NLRP3 inflammasomes and provide new insights into the mechanisms underlying the adverse effects of cardiac glycosides.

Chronic kidney disease (CKD) is associated with an increased risk of cardiovascular disease, and thus is a major worldwide public health problem. Recently, an estimated glomerular filtration rate (eGFR) using the Modification of Diet in Renal Disease equation for Japanese patients was proposed by the Japanese Society of Nephrology. However, the role of eGFR in the assessment of atherosclerosis is not well understood in Japanese patients. We analyzed the relationship between eGFR and severity of arterial stiffness using brachial-ankle pulse wave velocity (baPWV) in 647 adult Japanese patients. baPWV correlated significantly and positively with age, hypertension, diabetes, prior cardiovascular disease, blood pressure, pulse pressure and heart rate, and negatively with eGFR (r=-0.405, p<0.0001). A multiple regression analysis revealed that baPWV correlated independently with eGFR. Furthermore, there was a stepwise increase in baPWV, corresponding to advances in CKD through stages 1 to 5. When CKD stage 3 was divided at eGFR 45 mL/min/1.73 m2, the baPWV of stage 3b (eGFR 30 to 44) was significantly higher than that of stage 3a (eGFR 45 to 59) independent of traditional risk factors, suggesting that an eGFR of 45 mL/min/1.73 m2 may be a critical cut off value to predict arterial stiffness in CKD. In conclusion, the newly proposed eGFR is significantly associated with arterial stiffness, independent of traditional risk factors for cardiovascular disease.

Although malnutrition indicates an unfavorable prognosis in some clinical settings, the synergistic impact of nutritional state, renal dysfunction and left ventricular hypertrophy (LVH) on cardiovascular events is unknown. Among 338 patients aged 40-80 years who underwent echocardiographic evaluation between 2003 and 2005, 161 patients who were followed for >7 years were recruited. Malnutrition was defined as a geriatric nutritional risk index (GNRI) of ⩽96. The mean patient age was 63.5±9.2 years; the mean estimated glomerular filtration rate (eGFR) was 72.9±18.7 ml min(-1) per 1.73 m(2); the mean LV mass index was 114±33 g m(-)(2); and the mean GNRI was 100.4±6.0. Among the patients, 25% (n=40) had an eGFR of <60 ml min(-1) per 1.73 m(2), 29% (n=46) exhibited chronic kidney disease (CKD) and 37% (n=59) had LVH. During the follow-up period (median: 96 months), cardiovascular events were observed in 15 patients (9%). Kaplan-Meier curves showed a significantly higher incidence of cardiovascular events in patients with an eGFR of <60 ml min(-1) per 1.73 m(2) (log-rank P=0.007), a GNRI of ⩽96 (P=0.003) or LVH (P=0.010). In a Cox regression analysis, eGFR, LVH and GNRI were independent determinants of cardiovascular event incidence after adjusting for age, gender and the presence of hypertension and diabetes. Furthermore, the combination of LVH and lower GNRI was significantly associated with a higher rate of cardiovascular events not only in all patients but also in patients with CKD. In conclusion, malnutrition, low eGFR and LVH were independent determinants of cardiovascular event incidence; they synergistically increased rates of these events in the long term. The evaluation and management of LVH progression and the improvement of nutritional status are critical for preventing cardiovascular complications even in non-dialysis patients.

Abstract We conducted a survey to examine the gaps between Japanese physician and patient perspectives on hypertension management and to investigate important factors that may help solve the “hypertension paradox” in Japan. Web-based surveys of patients and physicians were conducted in Japan between October 19 and 31, 2017. The data collected included physician and patient perspectives on hypertension education, adherence to lifestyle modifications and antihypertensive medication, and reasons for treatment adherence/nonadherence. Factors relating to specific patient behaviors (e.g., monitoring their home blood pressure [BP] daily) were analyzed by multivariate logistic regression analysis. Of the 541 physicians and 881 patients included in the analyses, both groups recognized that the extent of lifestyle changes was insufficient. Approximately 80% of physicians reported that they fully or sufficiently provided education to patients about reasons for hypertension treatment and its associated risks, target BP levels, and lifestyle modifications. Only 40–50% of patients considered those topics having been fully or sufficiently discussed. Logistic regression analyses revealed that positive lifestyle modifications (daily home BP monitoring, salt intake &lt;6 g/day, and daily aerobic exercise for ≥30 min) were positively associated with receiving feedback from physicians about specific lifestyle modifications and patient motivation for maintaining their target BP. In conclusion, perception of the amount of education provided by physicians on hypertension management was lower in patients than in physicians. In addition to effective regular follow-up regarding lifestyle modifications, patient motivation by physicians is an important factor for improving lifestyle modifications and achieving effective hypertension management in Japan.

&lt;i&gt;Background:&lt;/i&gt; The production of reactive oxygen species (ROS) has been suggested to play an important role in the progression of chronic kidney disease (CKD). An oral adsorbent, AST-120, removes uremic toxins such as indoxyl sulfate (IS) and delays the progression of CKD, but the effect on ROS production is unknown. The present study aimed to determine whether AST-120 reduces oxidative stress in uremic rat kidneys using markers of ROS production such as acrolein and 8-hydroxy-2′-deoxyguanosine (8-OHdG). &lt;i&gt;Methods:&lt;/i&gt; Daily administration of AST-120 was started 6 weeks after 5/6 nephrectomy and continued for 18 weeks. The changes in metabolic data, serum and urine IS levels, urinary excretion of markers of oxidative stress, and renal histological findings were investigated in uremic rats with or without AST-120 treatment. &lt;i&gt;Results:&lt;/i&gt; In parallel with the increase in serum and urine IS, the serum creatinine, urinary protein and acrolein levels started to increase at 6 weeks, but urinary 8-OHdG remained unchanged and significantly increased at 18 weeks in uremic rats. AST-120 markedly and significantly attenuated increases in uremic toxins and oxidative stress levels as well as the histological changes in glomerular sclerosis, interstitial fibrosis, and the tubular staining of 8-OHdG. &lt;i&gt;Conclusion:&lt;/i&gt; AST-120 suppressed the progression of CKD, at least in part, via attenuation of oxidative stress induced by uremic toxin.

Background: Prostacyclin (PGI2) enhances angiogenesis, especially in cooperation with bone marrow (BM)-derived endothelial progenitor cells (EPCs). However, the mechanisms of PGI2 in EPC-mediated angiogenesis in vivo remain unclear. The purpose of this study was to clarify the role of PGI2 in EPC-mediated angiogenesis using BM-specific IP deletion mice. Methods and Results: Hind limb ischemia (HLI) was induced in wild-type (WT) mice transplanted with IP-deleted BM (WT/BM(IP–/–). Recovery of blood flow (RBF) in WT/BM(IP–/–) was impaired for 28 days after HLI, whereas RBF in IP–/–/BM(WT) was attenuated for up to 7 days compared with WT/BM(WT). The impaired RBF in WT/BM(IP–/–) was completely recovered by intramuscular injection of WT EPCs but not IP–/– EPCs. The impaired effects of IP–/– EPCs were in accordance with reduced formation of capillary and arterioles in ischemic muscle. An ex vivo aortic ring assay revealed that microvessel formation was enhanced by accumulation/adhesion of EPCs to perivascular sites as pericytes. IP–/–EPCs, in which expression of integrins was decreased, had impaired production of angiogenic cytokines, adhesion to neovessels and their angiogenic effects. The small-interfering RNA (siRNA)-mediated knockdown of integrin β1 in WT EPCs attenuated adhesion to microvessels and their in vivo and in vitro angiogenic effects. Conclusions: PGI2 may induce persistent angiogenic effects in HLI through adhesion of EPCs to perivascular sites of neovessels via integrins in addition to paracrine effects. (Circ J 2013; 77: 1053–1062)

Patients with chronic kidney disease (CKD) are at a high risk for cardiovascular diseases including stroke. However, the characteristics of the stroke subtypes in patients with CKD remain to be determined.We investigated the associations between stroke subtypes and estimated glomerular filtration rate (eGFR), and traditional risk factors in 451 (males, 239; mean age, 71.1 y) acute stroke patients at our hospital between 2006 and 2010.The stroke subtype was cardiogenic cerebral embolism in 129 (29%), cerebral hemorrhage in 104 (23%), unclassified cerebral infarction in 65 (14%), lacunar infarction in 65 (14%), subarachnoid hemorrhage in 41 (9%), atherothrombosis in 30 (7%), and transient ischemic attacks in 17 (4%). Among the 451 patients, 134 (30%) had CKD with eGFR <60 mL/min/1.73 m2. Compared with a group without CKD, mean age (75.9 vs. 69.0 years, p<0.05), the prevalence of atrial fibrillation (AF) (44% vs. 21%, p<0.01) and a history of cardiovascular disease (37% vs. 19%, p<0.01) were significantly higher in that with CKD. A comparison of stroke subtypes revealed a significantly higher incidence of cardiogenic cerebral embolism (36% vs. 26%, p<0.05) in the group with, than without CKD.We showed the prevalence of CKD in about 30% of acute stroke patients, and those patients were older, had a significantly higher prevalence of AF and a higher rate of cardiogenic cerebral embolism compared with patients without CKD. Thus, strict control of blood pressure and management of AF should be important to prevent stroke among patients with CKD.

Metabolic syndrome confers an increased risk of cardiovascular disease (CVD) in the general population. The relationship between adiponectins, and clinical outcomes in patients undergoing hemodialysis remains controversial. We investigated whether adiponectins, biomarkers of inflammation, nutrition status and clinical features predict the mortality of patients undergoing hemodialysis for 6 years. We measured baseline plasma total and high-molecular-weight (HMW) adiponectins, tumor necrosis factor (TNF)-α, serum high sensitivity C-reactive protein (hsCRP), and clinical characteristics including visceral fat area (VFA) and the Geriatric Nutritional Risk Index (GNRI) in 133 patients undergoing chronic hemodialysis. Forty-one of the 133 patients died during follow-up. The deceased patients were significantly older, had more prior CVD and diabetes, higher TNF-α and hsCRP levels but lower GNRI. VFA, and total and HMW adiponectin did not significantly differ between the two groups. TNF-α and hsCRP levels and GNRI score were significant for predicting all-cause and cardiovascular mortality in receiver operating curve analyses. When stratified by a GNRI score of 96, Cox proportional hazards analyses identified TNF-α as a significant predictor of all-cause mortality (hazard ratio [HR] 1.23; P = 0.038) and hsCRP as a significant predictor of all-cause and cardiovascular mortality (HR, 2.32, P = 0.003; HR 2.30, P = 0.012, respectively) after adjusting for age, sex, diabetes mellitus, and prior CVD, only in malnourished patients. These results demonstrate that malnutrition and the inflammatory markers TNF-α and hsCRP, but not metabolic markers, including VFA and adiponectins have a significant impact on 6-year all-cause and cardiovascular mortality in Japanese patients undergoing hemodialysis.

Background:Capillary pericytes (cPCs), the mural cells of microvessels, play an important role in the formation and maintenance of microvessels; however, little is known about the mechanisms of how cPCs regulate angiogenesis. To identify factors that modulate cPC function, genes whose levels were altered in cPCs during neovessel formation were identified through a microarray screen.Methods and Results:Ninjurin1 (nerve injury-induced protein, Ninj1) was selected as a candidate factor for angiogenesis regulation. Ninj1 was expressed in capillary cells including endothelial cells (cECs) and was expressed at a higher level in cPCs. Hypoxia induced the gene expression of Ninj1 in addition of vascular endothelial growth factor (VEGF) in cPCs. When cPCs were co-incubated with a thoracic aorta in a three-dimensional Matrigel system, the length of the EC-tubes sprouting from the aorta was increased. Small interfering RNA-mediated downregulation of Ninj1 in cPCs enhanced these cPCs-mediated angiogenic effects, whereas overexpression of Ninj1 attenuated their effects. The production of angiogenic growth factors, such as VEGF and angiopoietin 1, by cPCs was enhanced by the downregulation of Ninj1, and reduced by the overexpression of Ninj1.Conclusions:Ninj1 is a novel regulator for the angiogenic effect of PCs. Specifically, Ninj1 negatively regulates the formation of neovessels, that is, the EC-tube, by reducing the trophic effects of cPCs. (Circ J 2015; 79: 1363–1371)

Minimal change nephrotic syndrome (MCNS) is a common form of nephrotic syndrome (NS). We herein present the case of a 57-year-old woman with advanced lung adenocarcinoma treated with the tyrosine kinase inhibitor (TKI) gefitinib who developed NS. A renal biopsy revealed minor glomerular abnormalities, and the patient's symptoms improved exclusively with the discontinuation of gefitinib. Therefore, we diagnosed her with MCNS associated with gefitinib treatment. A few months later, however, she developed recurrent lung tumors. Following the challenging initiation of the TKI erlotinib, she achieved remission without proteinuria. We thus conclude that erlotinib is a potential treatment option in patients with NS associated with gefitinib therapy.

Adventitial microvessels, vasa vasorum in the vessel walls, have an active role in the vascular remodeling, although its mechanisms are still unclear. It has been reported that microvascular pericytes (PCs) possess mesenchymal plasticity. Therefore, microvessels would serve as a systemic reservoir of stem cells and contribute to the tissues remodeling. However, most aspects of the biology of multipotent PCs (mPCs), in particular of pathological microvessels are still obscure because of the lack of appropriate methods to detect and isolate these cells. In order to examine the characteristics of mPCs, we established immortalized cells residing in adventitial capillary growing at the injured vascular walls. We recently developed in vivo angiogenesis to observe adventitial microvessels using collagen-coated tube (CCT), which also can be used as an adventitial microvessel-rich tissue. By using the CCT, CD146- or NG2-positive cells were isolated from the adventitial microvessels in the injured arteries of mice harboring a temperature-sensitive SV40 T-antigen gene. Several capillary-derived endothelial cells (cECs) and PCs (cPCs) cell lines were established. cECs and cPCs maintain a number of key endothelial and PC features. Co-incubation of cPCs with cECs formed capillary-like structure in Matrigel. Three out of six cPC lines, termed capillary mPCs demonstrated both mesenchymal stem cell- and neuronal stem cell-like phenotypes, differentiating effectively into adipocytes, osteoblasts, as well as schwann cells. mPCs differentiated to ECs and PCs, and formed capillary-like structure on their own. Transplanted DsRed-expressing mPCs were resident in the capillary and muscle fibers and promoted angiogenesis and myogenesis in damaged skeletal muscle. Adventitial mPCs possess transdifferentiation potential with unique phenotypes, including the reconstitution of capillary-like structures. Their phenotype would contribute to the pathological angiogenesis associated with vascular remodeling. These cell lines also provide a reproducible cellular tool for high-throughput studies on angiogenesis, vascular remodeling, and regeneration as well.

Abstract Overcoming the insufficient survival of cell grafts is an essential objective in cell-based therapy. Apurinic/apyrimidinic endonuclease/redox factor 1 (APE1) promotes cell survival and may enhance the therapeutic effect of engrafted cells. The aim of this study is to determine whether APE1 overexpression in cardiac progenitor cells (CPCs) could ameliorate the efficiency of cell-based therapy. CPCs isolated from 8- to 10-week-old C57BL/6 mouse hearts were infected with retrovirus harboring APE1-DsRed (APE1-CPC) or a DsRed control (control-CPC). Oxidative stress-induced apoptosis was then assessed in APE1-CPCs, control-CPCs, and neonatal rat ventricular myocytes (NRVMs) cocultured with these CPCs. This analysis revealed that APE1 overexpression inhibited CPC apoptosis with activation of transforming growth factor β-activated kinase 1 (TAK1) and nuclear factor (NF)-κB. In the coculture model, NRVM apoptosis was inhibited to a greater extent in the presence of APE1-CPCs compared with control-CPCs. Moreover, the number of surviving DsRed-positive CPC grafts was significantly higher 7 days after the transplant of APE1-CPCs into a mouse myocardial infarction model, and the left ventricular ejection fraction showed greater improvement with attenuation of fibrosis 28 days after the transplant of APE1-CPCs compared with control-CPCs. Additionally, fewer inflammatory macrophages and a higher percentage of cardiac α-sarcomeric actinin-positive CPC-grafts were observed in mice injected with APE1-CPCs compared with control-CPCs after 7 days. In conclusion, antiapoptotic APE1-CPC graft, which increased TAK1-NF-κB pathway activation, survived effectively in the ischemic heart, restored cardiac function, and reduced cardiac inflammation and fibrosis. APE1 overexpression in CPCs may serve as a novel strategy to improve cardiac cell therapy. Significance Improving the survival of cell grafts is essential to maximize the efficacy of cell therapy. The authors investigated the role of APE1 in CPCs under ischemic conditions and evaluated the therapeutic efficacy of transplanted APE1-overexpressing CPCs in a mouse model of myocardial infarction. APE1 hindered apoptosis in CPC grafts subjected to oxidative stress caused in part by increased TAK1-NF-κB pathway activation. Furthermore, APE1-CPC grafts that effectively survived in the ischemic heart restored cardiac function and attenuated fibrosis through pleiotropic mechanisms that remain to be characterized. These findings suggest that APE1 overexpression in CPCs may be a novel strategy to reinforce cardiac cell therapy.

Kidney damage is a common consequence of arterial hypertension, but is also a cause of atherogenesis. Dysfunction and/or harm of the endothelium in glomeruli and tubular interstitium damage the function of these structures and translates into dynamic changes of filtration fraction, with progressive reduction in glomerular filtration rate, expansion of extracellular fluid volume, abnormal ion balance, and hypoxia, ultimately leading to chronic kidney disease. Considering the key role played by endothelial dysfunction in chronic kidney disease, the Working Group on Endothelin and Endothelial Factors of the European Society of Hypertension and the Japanese Society of Hypertension have critically reviewed available knowledge on the mechanisms underlying endothelial cell injury. This resulted into two articles: in the first, we herein examine the mechanisms by which endothelial factors induce vascular remodeling and the role of different players, including endothelin-1, the renin–angiotensin–aldosterone system and their interactions, and of oxidative stress; in the second, we discuss the role of endothelial dysfunction in the major disease conditions that affect the kidney.

Background Both endocardial trigger elimination and epicardial substrate modification are effective in treating ventricular fibrillation (VF) in Brugada syndrome. However, the primary approach and the characteristics of patients who respond to endocardial ablation remain unknown. Methods Among 123 symptomatic Brugada syndrome patients (VF, 63%; syncope, 37%), ablation was performed in 21 VF/electrical storm patients, the majority of whom were resistant to antiarrhythmic drugs. Results Careful endocardial mapping revealed that 81% of the patients had no specific findings, whereas 19% of the patients, who experienced the most frequent VF episodes with notching of the QRS in lead V1, had delayed low-voltage fractionated endocardial electrograms. Ablation of VF triggers followed by endocardial substrate modification was performed in the right ventricular outflow tract in 85% of the cases and in the right ventricle in 15%. VF triggers could not be completely eliminated in 1 patient and VF became noninducible in 14 (88%) patients among 16 patients who underwent VF induction with normalization of Brugada-type ECG in 3. During follow-up (56.14±36.95 months), VF recurrence was observed in 7 patients. Importantly, all patients who had nothing of QRS in lead V1 did not respond to endocardial ablation despite presence of VF-triggering ectopic beats during ablation. Conclusions With careful documentation of VF-triggering ectopic beats and detailed endocardial mapping, endocardial VF trigger elimination followed by endocardial substrate modification has an excellent long-term outcome, whereas presence of QRS notching in lead V1 was associated with high VF recurrence suggesting epicardial substrate ablation as effective initial approach.

Appropriate regimens of antithrombotic therapy for patients with atrial fibrillation (AF) and coronary artery disease (CAD) have not yet been established.To compare the total number of thrombotic and/or bleeding events between rivaroxaban monotherapy and combined rivaroxaban and antiplatelet therapy in such patients.This was a post hoc secondary analysis of the Atrial Fibrillation and Ischemic Events With Rivaroxaban in Patients With Stable Coronary Artery Disease (AFIRE) open-label, randomized clinical trial. This multicenter analysis was conducted from February 23, 2015, to July 31, 2018. Patients with AF and stable CAD who had undergone percutaneous coronary intervention or coronary artery bypass grafting 1 or more years earlier or who had angiographically confirmed CAD not requiring revascularization were enrolled. Data were analyzed from September 1, 2020, to March 26, 2021.Rivaroxaban monotherapy or combined rivaroxaban and antiplatelet therapy.The total incidence of thrombotic, bleeding, and fatal events was compared between the groups. Cox regression analyses were used to estimate the risk of subsequent events in the 2 groups, with the status of thrombotic or bleeding events that had occurred by the time of death used as a time-dependent variable.A total of 2215 patients (mean [SD] age, 74 [8.2] years; 1751 men [79.1%]) were included in the modified intention-to-treat analysis. The total event rates for the rivaroxaban monotherapy group (1107 [50.0%]) and the combination-therapy group (1108 [50.0%]) were 12.2% (135 of 1107) and 19.2% (213 of 1108), respectively, during a median follow-up of 24.1 (IQR, 17.3-31.5) months. The mortality rate was 3.7% (41 of 1107) in the monotherapy group and 6.6% (73 of 1108) in the combination-therapy group. Rivaroxaban monotherapy was associated with a lower risk of total events compared with combination therapy (hazard ratio, 0.62; 95% CI, 0.48-0.80; P < .001). Monotherapy was an independent factor associated with a lower risk of subsequent events compared with combination therapy. The mortality risk after a bleeding event (monotherapy, 75% [6 of 8]; combination therapy, 62.1% [18 of 29]) was higher than that after a thrombotic event (monotherapy, 25% [2 of 8]; combination therapy, 37.9% [11 of 29]).Rivaroxaban monotherapy was associated with lower risks of total thrombotic and/or bleeding events than combination therapy in patients with AF and stable CAD. Tapered antithrombotic therapy with a sole anticoagulant should be considered in these patients.ClinicalTrials.gov Identifier: NCT02642419.

In response to oxidative stress, the pathogenesis of a number of cardiovascular events and several genes are stimulated by extracellular signal-regulated kinases (ERK1/2). Biphasic (early, 10 min; and delayed, 120 min) ERK1/2 activation by H2O2, a reactive oxygen species, was observed in cultured neonatal rat cardiomyocytes. We investigated the hypothesis that the delayed activation of ERK1/2 depends on a factor secreted by oxidative stress (FSO). The delayed activation was inhibited by calphostin C, a protein kinase C inhibitor. Conditioned medium (CM) obtained from cells stimulated with H2O2 induced rapid and monophasic ERK1/2 activation, which was not inhibited by calphostin C. In contrast, calphostin C-pretreated CM did not activate ERK1/2. Macrophage migration inhibitory factor (MIF) was one of the candidate FSOs activating ERK1/2. The existence of MIF in CM, the recombinant MIF-stimulated ERK1/2 rapid activation, and anti-MIF neutralizing antibody-induced inhibition of the delayed activation implied that MIF could be the FSO. Pretreatment of cardiomyocytes with a mitogen-activated protein kinase/ERK kinase (MEK) inhibitor did not suppress the MIF secretion, although it prevented the ERK1/2 activation by H2O2. These results indicate that MIF is secreted from cardiomyocytes as a result of oxidative stress and activates ERK1/2 through a MEK1/2-dependent mechanism, although the secretion is not regulated by ERK1/2 but by protein kinase C. In response to oxidative stress, the pathogenesis of a number of cardiovascular events and several genes are stimulated by extracellular signal-regulated kinases (ERK1/2). Biphasic (early, 10 min; and delayed, 120 min) ERK1/2 activation by H2O2, a reactive oxygen species, was observed in cultured neonatal rat cardiomyocytes. We investigated the hypothesis that the delayed activation of ERK1/2 depends on a factor secreted by oxidative stress (FSO). The delayed activation was inhibited by calphostin C, a protein kinase C inhibitor. Conditioned medium (CM) obtained from cells stimulated with H2O2 induced rapid and monophasic ERK1/2 activation, which was not inhibited by calphostin C. In contrast, calphostin C-pretreated CM did not activate ERK1/2. Macrophage migration inhibitory factor (MIF) was one of the candidate FSOs activating ERK1/2. The existence of MIF in CM, the recombinant MIF-stimulated ERK1/2 rapid activation, and anti-MIF neutralizing antibody-induced inhibition of the delayed activation implied that MIF could be the FSO. Pretreatment of cardiomyocytes with a mitogen-activated protein kinase/ERK kinase (MEK) inhibitor did not suppress the MIF secretion, although it prevented the ERK1/2 activation by H2O2. These results indicate that MIF is secreted from cardiomyocytes as a result of oxidative stress and activates ERK1/2 through a MEK1/2-dependent mechanism, although the secretion is not regulated by ERK1/2 but by protein kinase C. reactive oxygen species mitogen-activated protein kinase extracellular signal-regulated kinase MAPK/ERK kinase factor secreted by oxidative stress conditioned medium macrophage migration inhibitory factor protein kinase C N-[2-hydroxy-1,1-bis- (hydroxymethyl)ethyl]glycine Oxidative stress is important in the pathogenesis of ischemic/reperfusion injury (1Turners J.A. Bovanis A. Biochem. J. 1980; 156: 434-444Google Scholar, 2Xia Y. Katchikian G. Zweier J.L. J. Biol. Chem. 1996; 271: 10096-10102Abstract Full Text Full Text PDF PubMed Scopus (107) Google Scholar), apoptosis (3Gottlieb R.A. Burleson K.O. Kloner R.A. Bobior B.M. Engler R.L. J. Clin. Invest. 1994; 94: 1621-1628Crossref PubMed Scopus (1357) Google Scholar, 4Tanaka M. Ito H Adachi S. Akimoto H. 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It also acts as a proinflammatory cytokine produced by macrophages in response to a variety of inflammatory stimuli and as a factor stimulating cell growth in many cell types (18Calandra T. Bernhagen J. Metz C.N. Spiegel L.A. Bacher M. Donnelly T. Cerami A. Bucala R. Nature. 1995; 377: 68-71Crossref PubMed Scopus (1050) Google Scholar, 21Bernhagen J. Calandra T. Mitchell R.A. Martin S.B. Tracey K.J. Voelter W. Manogue K.R. Cerami A. Bucala R. Nature. 1993; 365: 756-759Crossref PubMed Scopus (931) Google Scholar). The responses of cardiovascular systems to MIF have not been reported except for mRNA and protein expression of MIF in the atherosclerotic region in a hypercholesterolemic model of rabbit vessels (22Lin S.G., Yu, X.Y. Chen Y.X. Huang X.R. Metz C. Bucala R. Lau C.P. Lan H.Y. Circ. Res. 2000; 87: 1202-1208Crossref PubMed Scopus (119) Google Scholar). Recently, increased serum concentrations of MIF have been reported in patients with acute myocardial infarction in which reperfusion injury may be involved (23Yu C. Lau C. Lai K.W. Huang X. Chen W. Lan H.Y. Am. J. Cardiol. 2001; 88: 774-777Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar). We hypothesized that cardiomyocytes could secrete factors increasing ERK1/2 activity in response to ROS and that MIF could be an important secreted ROS-induced factor in cardiomyocytes. To explore this hypothesis, we used cultured cardiomyocytes from neonatal rat to examine whether H2O2 induced MIF secretion in cardiomyocytes and to investigate the mechanism and the relationship between MIF and H2O2-induced ERK1/2 activation. Reagents for tissue culture were obtained from Invitrogen. Polyclonal antibodies against phosphospecific and total ERK1/2 and phosphospecific and total MEK1 or 1/2 were from Santa Cruz Biotechnology (Santa Cruz, CA). PD98059, geldanamycin, and calphostin C were purchased from Calbiochem. Rat recombinant MIF was expressed in Escherichia coli and purified to homogeneity as described previously (24Nishihira J. Kuriyama T. Sakai M. Nishi S. Ohki S. Hikichi K. Biochim. Biophys. Acta. 1995; 1247: 159-162Crossref PubMed Scopus (74) Google Scholar). A polyclonal anti-rat MIF antibody was generated by immunizing New Zealand White rabbits with recombinant rat MIF as reported previously (25Nishio Y. Mianami A. Kato H. Kaneda K. Nishihira J. Biochim. Biophys. Acta. 1999; 1453: 74-82Crossref PubMed Scopus (46) Google Scholar). Cardiomyocytes were isolated from 1–2-day-old Sprague-Dawley rat ventricles by an enzymatic method as described previously (26Booz G.W. Dostal D.E. Singer H.A. Baker K.M. Am. J. Physiol. 1994; 267: C1308-C1318Crossref PubMed Google Scholar, 27Hunt R.A. Bhat G.J. Baker K.M. Hypertension. 1999; 34: 603-608Crossref PubMed Scopus (24) Google Scholar, 28Fukuzawa J. Booz G.W. Hunt R.A. Shimizu N. Karoor V. Baker K.M. Dostal D.E. Hypertension. 2000; 35: 1191-1196Crossref PubMed Scopus (84) Google Scholar). Twenty-four hours after isolation, serum-containing medium was changed to Dulbecco's modified Eagle's medium/F-12 with a serum substitute as described previously (29Booz G.W. Dostal D.E. Baker K.M. Hypertension. 1996; 28: 635-640Crossref PubMed Scopus (202) Google Scholar). Twenty-four hours before experiments, cells were given the same medium without the serum substitute. Using these methods, cultures that contained 90–95% myocytes were obtained (26Booz G.W. Dostal D.E. Singer H.A. Baker K.M. Am. J. Physiol. 1994; 267: C1308-C1318Crossref PubMed Google Scholar). Cardiomyocytes were lysed after each stimulation with a buffer as described previously (30Bhat G.J. Thekkumkara T.J. Thomas W.G. Conrad K.M. Baker K.M. J. Biol. Chem. 1994; 269: 31443-31449Abstract Full Text PDF PubMed Google Scholar). The lysed samples were subjected to Tris-glycine-SDS-PAGE (8–10%). After separation by electrophoresis, samples were transferred to nitrocellulose membranes (Amersham Biosciences) and subjected to immunoblot analysis using each indicated antibody (30Bhat G.J. Thekkumkara T.J. Thomas W.G. Conrad K.M. Baker K.M. J. Biol. Chem. 1994; 269: 31443-31449Abstract Full Text PDF PubMed Google Scholar). Signals were visualized with enhanced chemiluminescence (PerkinElmer Life Sciences). For detection of MIF, the samples were subjected to Tris-Tricine SDS-PAGE as described previously (31Schagger H. von Jagow G. Anal. Biochem. 1987; 166: 368-379Crossref PubMed Scopus (10480) Google Scholar). In each experiment, three independent analyses were performed to confirm the reproducibility. Conditioned medium was collected and concentrated according to a method described elsewhere (14Jin Z.G. Melaragno M.G Liao D.F. Yan C. Haendeler J. Suh Y.A. Lambeth J.D. Berk B.C. Circ. Res. 2000; 87: 789-796Crossref PubMed Scopus (334) Google Scholar). Briefly, cells were washed three times with Hanks' balanced salt solution (NaCl 130 mm, KCl 5 mm, CaCl2 1.5 mm, MgCl2 1 mm, HEPES 20 mm, pH 7.4) and were equilibrated for 24 h. H2O2 (1 mm) was added to the Hanks' balanced salt solution pretreated with/or without calphostin C (1 μm) or PD98059 (50 μm) or control medium from the cells unstimulated with H2O2, were collected and centrifuged to remove the debris at 800 × g at 4 °C for 10 min. The conditioned medium pretreated with or without calphostin C or PD98059 and the control medium were concentrated 100-fold by using a CentriprepTM centrifugal filter device (Millipore, Bedford, MA). The concentration (degradation) of the exogenously added H2O2 in the medium with or without cells was measured with a Bioxytech H2O2-560TM (Oxis International Inc., Portland, OR) according to the manufacturer's protocol. Exposure of cardiomyocytes to H2O2 (1 mm) stimulated ERK1/2 phosphorylation (activation) with a peak at 10 min. The ERK1/2 activity returned to the base-line level at 45–60 min. After returning to the base line, a second peak of ERK1/2 activation appeared at 120 min (Fig.1 A). The concentration of H2O2 that was added exogenously to the medium decreased exponentially and returned to the basal (zero) level within 90 min after the exogenous administration (Fig. 1 B). Concentration dependences of H2O2 in the early and late phases of ERK1/2 activation occurred in different manners (Fig. 1 C). To evaluate whether H2O2-induced ERK1/2 activation was mediated via an ROS-dependent or -independent mechanism, cells were treated with the antioxidant reagent, catalase (3000 units/ml) before exposure to H2O2 (1 mm). Catalase inhibited both the early and late phases of ERK1/2 activation in response to H2O2 (Fig. 1 D). To assess the upstream mechanism of H2O2-induced ERK1/2 activation, cardiomyocytes were pretreated with reagents that inhibit MAPK/ERK kinase (MEK), Raf-1, or protein kinase C (PKC) before exposure to H2O2. The addition of PD98059 (50 μm), an inhibitor of MEK, decreased both the early and late phases of ERK1/2 activation in response to H2O2 (Fig.2 A). The activity of MEK1/2 was also up-regulated biphasically by H2O2administration into cultured medium as was that of ERK1/2 (Fig.2 B). Geldanamycin (2 μm), which can bind to HSP-90 and disrupts the Raf-1-HSP90 multimolecular complex leading to destabilization of Raf-1 (32Schulte T.W. Blagosklonny M.V. Romanova L. Mushinski J.F. Monia B.P. Johnston J.F. Nguyen R. Trepei J. Neckers L.M. Mol. Cell. Biol. 1996; 16: 5839-5845Crossref PubMed Scopus (256) Google Scholar), also inhibited both peaks (Fig.2 C). However, calphostin C (1 μm), a specific inhibitor of protein kinase C, diminished only the delayed activation of ERK1/2 by H2O2 (Fig. 2 D). Conditioned medium obtained from the cells exposed to 1 mmH2O2 (CM+) stimulated ERK1/2 activation rapidly and monophasically (Fig.3 A), whereas H2O2-unstimulated control medium (CM−) did not stimulate ERK1/2 activation (Fig.3 B). The conditioned medium was fractionated on a molecular weight basis using a commercially available filter device. Adding each fraction to the medium stimulated ERK1/2 activation in cardiomyocytes. CM10–30, CM50, and CM30–50, to a lesser extent, could stimulate ERK1/2 activation (Fig. 3 C). In contrast to the H2O2-stimulated cells, catalase did not inhibit ERK1/2 activation in cells stimulated with CM+ (Fig.3 D). Calphostin C-pretreated conditioned medium obtained from the cells stimulated with H2O2 (1 mm) showed suppressed ERK1/2 activation (Fig.4 A). In contrast, the conditioned medium-stimulated rapid and monophasic activation of ERK1/2 was not inhibited by pretreatment with calphostin C (1 μm) (Fig. 4 B).Figure 4Secretion of H2O2-induced factor by a calphostin C-dependent mechanism and regulation of the secreted factor-induced ERK1/2 activation by a calphostin C-independent mechanism. A, medium derived from the cultured cardiomyocytes preincubated with calphostin C (1 μm) for 30 min followed by H2O2 (1 mm) for 120 min was collected and concentrated (CMcalphostin) as described under “Materials and Methods.” The concentrated medium was applied to the cells for the indicated times. The cells were subjected to immunoblot analysis using anti-phosphospecific ERK1/2 (upper panel) and anti-ERK (lower panel) antibodies. B, 30 min after calphostin C incubation, neonatal cultured cardiomyocytes were exposed to the conditioned medium derived from cells stimulated with H2O2 (1 mm) for 120 min (CM +), and the ERK1/2 activation was assessed.View Large Image Figure ViewerDownload Hi-res image Download (PPT) The MIF in the conditioned medium was detected by Tris-Tricine SDS-PAGE and immunoblotting, whereas no MIF was detectable in the control medium or mock-stimulated medium (Fig.5 A). To confirm the effect of ERK1/2 on MIF secretion induced by H2O2, PD98059 (50 μm)-pretreated conditioned medium was analyzed. PD98059 did not inhibit H2O2-induced MIF secretion from cardiomyocytes. However, protein kinase C inhibition by calphostin C (1 μm) inhibited H2O2-induced MIF secretion. MIF concentration in the conditioned medium increased from 90 min after H2O2 stimulation (Fig.5 B). By contrast, MIF was not secreted in the calphostin C-pretreated conditioned medium. To determine whether the H2O2-induced MIF secretion was controlled via a regulatory or constitutive mechanism (33Sadoshima J. Izumo S. Annu. Rev. Physiol. 1997; 59: 551-571Crossref PubMed Scopus (721) Google Scholar), the changes in the MIF concentration in cells stimulated with H2O2 (1 mm) were determined by using whole cell lysate. Intracellular MIF contents were decreased at 90 and 120 min after stimulation (Fig. 5 C). Cells stimulated with a high concentration of recombinant MIF showed rapid and monophasic activation of ERK1/2-like conditioned medium-stimulated cells (Fig.6 A). We also determined the effect of an anti-MIF antibody on H2O2-induced ERK activation. Anti-MIF antibody (1:1000) pretreatment inhibited the late phase activation of ERK1/2, whereas pretreatment of the cells with preimmune serum did not decrease the H2O2-induced late phase activation (Fig.6 B). In each ERK1/2 activation stimulated with the conditioned medium fractionated by the centrifugal filtration system, CM30–50-induced ERK1/2 activation was completely inhibited by the anti-MIF antibody, and the one stimulated with CM10–30 was partially inhibited (Fig. 3 C). The MAPK signaling pathway plays a pivotal role in the mediation of cellular responses to a variety of signaling molecules. ERK1/2 regulates cell growth, embryonic development, cell survival (9Xia Z. Dickens M. Raingeaud J. Davis R.J. Greenberg M.E. Science. 1995; 270: 1326-1331Crossref PubMed Scopus (5036) Google Scholar), and cell differentiation in different cell types through the stimulation of specific gene expression. The signaling pathway for ERK1/2 activation includes intracellular calcium, heterotrimeric G proteins, protein kinase C, JAK/STAT (Janus kinase/signal transducers and activators of transcription) pathways, and phosphatidylinositol 3-kinase. in cardiomyocytes the Src/Ras/Raf-1 cascade has also been reported as a pathway for the redox-sensitive regulation of ERK1/2 in the early phase (15Aikawa R. Komuro I. Yamazaki T. Zou Y. Kudoh S. Tanaka M. Shiojima I. Hiroi Y. Yazaki Y. J. Clin. Invest. 1997; 100: 1813-1821Crossref PubMed Scopus (630) Google Scholar). In the present study, we observed that exogenously supplied H2O2 (1 mm) caused biphasic activation of ERK1/2, which was inhibited by catalase, an antioxidant reagent, in cultured cardiomyocytes from neonatal rats. The mechanism of this phenomenon was inferred to be ROS-dependent. Pharmacological blockade of the Raf-1/MEK cascade activation suppressed the activation in both phases. On the other hand, inhibition of the PKC pathway by calphostin C blocked only the late-phase activation of ERK1/2 in response to H2O2. It was obvious that stimulation by regeneration of H2O2 could be excluded as a mechanism for the late phase activation of ERK1/2 by H2O2. The concentration of the exogenously applied H2O2in the medium decreased exponentially in the presence of the cells (Fig. 1 B). The phenomenon of biphasic ERK1/2 activation in various cells has already been reported. Like H2O2, basic fibroblast growth factor, nerve growth factor, and ROS generators such as LY83583 were reported to stimulate the biphasic ERK1/2 activation (12Meloche S. Seuwen K. Pages G. Pouyssegur J. Mol. Endocrinol. 1992; 6: 845-854Crossref PubMed Google Scholar, 13York R.D. Yao H. Dillon T. Ellig C.L. Eckert S.P. McCleskey E.W. Stork P.J. Nature. 1998; 392: 622-626Crossref PubMed Scopus (757) Google Scholar, 14Jin Z.G. Melaragno M.G Liao D.F. Yan C. Haendeler J. Suh Y.A. Lambeth J.D. Berk B.C. Circ. Res. 2000; 87: 789-796Crossref PubMed Scopus (334) Google Scholar). The Ras/Raf/MEK cascade is known as an ERK1/2 activation system. In the present study, Raf-1 and MEK were shown to be upper signaling cascades for both phases of the ERK1/2 activation. Because geldanamycin, which was used as an inhibitor of Raf-1 in the present experiments, could function as an inhibitor of HSP-90 (32Schulte T.W. Blagosklonny M.V. Romanova L. Mushinski J.F. Monia B.P. Johnston J.F. Nguyen R. Trepei J. Neckers L.M. Mol. Cell. Biol. 1996; 16: 5839-5845Crossref PubMed Scopus (256) Google Scholar), biphasic activation of ERK1/2 with H2O2 could also be explained by an HSP-90-related mechanism. Further examination will be needed to confirm whether the phenomenon is HSP-90-dependent. We have demonstrated that the late phase activation of ERK1/2 is PKC-dependent. Although PKC is also known as a Raf-1 stimulator followed by ERK1/2 activation, the early phase activation of ERK1/2 by H2O2 was not mediated through this cascade in the present experiments. This indicated that the mechanisms of H2O2-induced ERK1/2 activation were different in the early and late phases. This finding was supported by the differences in H2O2 concentration dependence of ERK1/2 activation between the early and late phases. PKC plays important roles in many cellular responses in various types of cells. These responses include contraction, migration, hypertrophy, proliferation, apoptosis, and secretion (34Dempsey E.C. Newton A.C. Mochly-Rosen D. Fields A.P. Reyland M.E. Insel P.A. Messing R.O. Am. J. Physiol. 2000; 279: L429-L438Crossref PubMed Google Scholar). In addition, lung surfactant phospholipid from alveolar type II cells is secreted via a conventional type PKC (PKC-α and -β)-dependent mechanism (35Linke M, J. Burton F.M. Fiedeldey D.T. Rice W.R. Am. J. Physiol. 1997; 272: L171-L177PubMed Google Scholar). Considering these reports, we hypothesized that the mechanism of the PKC-dependent late phase activation of ERK1/2 by H2O2 could be explained by a factor secreted by oxidative stress (FSO). To examine this hypothesis, we examined the effect of conditioned medium derived from the cells stimulated with H2O2 (1 mm) for 120 min on ERK1/2 activation in cardiomyocytes. As demonstrated in Fig. 3, concentrated control medium obtained from the cells unstimulated with H2O2 did not activate ERK1/2 in cardiomyocytes. In contrast, the concentrated conditioned medium rapidly and monophasically activated the ERK1/2. These facts confirmed the hypothesis that a secreted factor plays a key role in the H2O2-induced late phase activation of ERK1/2 in cardiomyocytes. Catalase, which abrogated H2O2-induced ERK activation, did not inhibit the rapid and monophasic activation of ERK1/2 induced by conditioned medium. This suggested that the mechanism of the conditioned medium-induced ERK1/2 activation was independent of ROS. The decay of H2O2 shown in Fig. 1 indicated that the mechanism of the residual H2O2 in the medium was excluded for the rapid and monophasic activation of ERK1/2 by conditioned medium. Conditioned medium pretreated with calphostin C did not activate ERK1/2 in cardiomyocytes, as demonstrated in Fig. 4. This, along with the result that the late phase activation of ERK1/2 by H2O2 was PKC-dependent, indicated that the mechanism of the oxidative stress-induced secretion was PKC-dependent. In the present study, we demonstrated that the ERK1/2 activation in conditioned medium could be fractionated into various molecular weight ranges and that each fraction induced ERK1/2 activation to some extent. This indicated that some secreted factors were responsible for the late phase activation. By means of a literature search (36Koong A.C. Denko N.C. Hudson K.M. Schindler C. Swiersz L. Koch C. Evans S. Ibrahim H., Le, Q.T. Terris D.J. Giaccia A.J. Cancer Res. 2000; 60: 883-887PubMed Google Scholar), we examined which growth factors or cytokines were activated by oxidative stress, including ischemia and hypoxia, and stimulated ERK1/2 activation. Considering the molecular weight, MIF could be one of the candidate molecules that activate ERK1/2. The molecular weight of MIF is 13,000; it forms oligomers, especially trimers, as confirmed by crystal structure analysis (37Sugimoto H. Suzuki M. Nakagawa A. Tanaka I. Nishihira J. FEBS Lett. 1996; 389: 145-148Crossref PubMed Scopus (64) Google Scholar, 38Sun H. Bernhagen J. Bucala R. Lolis E. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: 5191-5196Crossref PubMed Scopus (294) Google Scholar, 39Suzuki M. Sugimoto H. Nakagawa A. Tanaka I. Nishihira J. Sakai M. Nat. Struct. Biol. 1996; 3: 259-266Crossref PubMed Scopus (188) Google Scholar) and cross-linking studies (40Bendrat K., Al- Abed Y. Callaway D.J.I. Peng T. Calandra T. Metz C.N. Bucala R. Biochemistry. 1997; 36: 15363-15370Crossref PubMed Scopus (143) Google Scholar,41Mischke R. Kleemann R. Brunner H. Bernhagen J. FEBS Lett. 1998; 427: 85-90Crossref PubMed Scopus (52) Google Scholar). It has been reported to stimulate ERK1/2 in fibroblasts via a protein kinase A-dependent pathway (42Mitchel R.A. Metz C.M. Peng T. Bucala R. J. Biol. Chem. 1999; 274: 18100-18106Abstract Full Text Full Text PDF PubMed Scopus (417) Google Scholar). MIF was originally described as a T-cell-derived cytokine that inhibits macrophage migration in vitro and promotes macrophage accumulation in the delayed-type hypersensitivity reaction (16David J.R. Proc. Natl. Acad. Sci. U. S. A. 1966; 56: 72-77Crossref PubMed Scopus (1092) Google Scholar, 17Bloom B.R. Bennett B. Science. 1966; 153: 80-82Crossref PubMed Scopus (1274) Google Scholar). As shown in Fig.6, we observed that neutralizing antibodies against MIF partially inhibited the late phase activation of ERK1/2 by H2O2 and that recombinant MIF induced ERK1/2 activation in a manner similar to the ERK1/2 activation induced by conditioned medium. The medium fraction of CM30–50 (containing major parts of the MIF trimer)-induced ERK1/2 activation was completely inhibited and that of CM10–30 (containing a possible MIF monomer or dimer) was partially inhibited by the anti-MIF antibody. These results suggested that MIF can act as a monomer or oligomer and that some other factors, induced by H2O2 and increased ERK1/2 activation, existed in the CM10–30. These results indicated that MIF, either alone or with one or more other factors, could be the FSO. Factors secreted in response to oxidative stress mediate production of the oxidative stress-induced growth factor in several cell types (43Sano M. Fukuda K. Sato T. Kawaguchi H. Suematsu M. Matsuda S. Koyasu S. Matsui H. Yamauchi-Takihara K. Harada M. Saito Y. Ogawa S. Circ. Res. 2001; 89: 661-669Crossref PubMed Scopus (262) Google Scholar). Although different growth factors are involved depending on the type of tissue as well as the type of oxidative stress, it is obvious that responses in an autocrine/paracrine manner could be a common mechanism utilized in oxidative stress-induced signal transduction. MIF mRNA expression in cardiomyocytes was up-regulated 6 h after H2O2 stimulation (44Takahashi M. Nishihira J. Shimpo M. Mizue Y. Ikeda U. Shimada K. Cardiovasc. Res. 2001; 52: 438-445Crossref PubMed Scopus (92) Google Scholar), and the concentration of MIF in the cardiomyocytes decreased after H2O2stimulation. It is likely that myocytes utilize the regulated type (preformed MIF within cells) but not the constitutive type of secretion for oxidative stress-induced MIF secretion, like atrial natriuretic factor secretion in response to endothelin (45Irons C.E. Sei C.A. Glembotski C.C. Am. J. Physiol. 1993; 264: H282-H285Crossref PubMed Google Scholar). This suggests that oxidative stress causes secretion of preformed MIF, although an oxidative stress-induced increase in MIF production is likely to occur at a later stage. Several possibilities could be considered for the mechanism of MIF secretion induced by oxidative stress. First, oxidative stress activates signaling pathways such as that of PKC, Ca2+, and small G protein (Rab), which may in turn stimulate growth factor secretion. The present work suggests this PKC-dependent explanation. Second, oxidative stress causes an alteration in myocyte sarcolemmal permeability, which indicates release of intracellular growth factors. Pacing-induced basic fibroblast growth factor release from adult rat ventricular cardiomyocytes accounts for this mechanism (46Kaye D. Pemental D. Prasad S. Maki T. Berger H.J. McNeil P.L. Smith T.W. Kelly R.A. J. Clin. Invest. 1996; 97: 281-291Crossref PubMed Scopus (118) Google Scholar). Although the plasma MIF level increases in patients with acute myocardial infarction, it does not increase in patients with other ischemic conditions such as unstable angina pectoris (23Yu C. Lau C. Lai K.W. Huang X. Chen W. Lan H.Y. Am. J. Cardiol. 2001; 88: 774-777Abstract Full Text Full Text PDF PubMed Scopus (35) Google Scholar). These data suggest that MIF is secreted from necrotic cardiomyocytes. Thus, these possibilities will have to be examined. Next, the question arises as to whether linkage between the early phase and late phase activation of ERK1/2 exists. PKC is known to stimulate Raf-1 followed by ERK1/2 in some types of cells (47Barrie A.P. Clohessy A.M. Buensuceso C.S. Rogers M.V. Allen J.M. J. Biol. Chem. 1997; 272: 19666-19671Abstract Full Text Full Text PDF PubMed Scopus (122) Google Scholar), indicating that PKC signaling could be the upper signaling cascade for each phase of ERK1/2 activation. As shown in Fig. 4, a PKC inhibitor, calphostin C, inhibited neither conditioned medium-induced (that is, FSO-induced) ERK1/2 activation nor H2O2-induced early phase activation of ERK1/2, indicating that PKC was not the upper signaling cascade for either early or late phase ERK activation evoked by H2O2. A similar result for the early phase activation of ERK1/2 was reported previously (15Aikawa R. Komuro I. Yamazaki T. Zou Y. Kudoh S. Tanaka M. Shiojima I. Hiroi Y. Yazaki Y. J. Clin. Invest. 1997; 100: 1813-1821Crossref PubMed Scopus (630) Google Scholar). Because we used calphostin C, known as a pan-specific PKC inhibitor, in the present study as well as in a previous one (15Aikawa R. Komuro I. Yamazaki T. Zou Y. Kudoh S. Tanaka M. Shiojima I. Hiroi Y. Yazaki Y. J. Clin. Invest. 1997; 100: 1813-1821Crossref PubMed Scopus (630) Google Scholar), additional experiments using isotype-selective PKC inhibitor such as Go-6983 will be needed to determine which PKC isozymes contribute to the H2O2-induced MIF secretion. PD98059, an inhibitor of MEK, and geldanamycin, an inhibitor of Raf-1, had inhibitory effects on H2O2-induced ERK1/2 activation in both phases as demonstrated in Fig. 2, indicating that the Raf-1/MEK pathway was the upper signaling cascade for ERK1/2 activation in both phases. On the other hand, ERK1/2 did not play a major role in MIF secretion, which had been suggested by the finding that H2O2-stimulated conditioned medium pretreated with PD98059 did not affect the secretion of MIF from cardiomyocytes, as shown in Fig. 5. These data indicate that the early phase activation of ERK1/2 by oxidative stress is not required for the late phase activation of ERK1/2. For the secretion of MIF, PKC plays a key role. The present study indicated a novel role for MIF in the mediation of the oxidative stress-induced signaling cascade up-regulation in cardiomyocytes. In summary, we have demonstrated that oxidative stress activates ERK1/2 biphasically in cardiomyocytes. A PKC-dependent mechanism contributes to the late phase activation of ERK1/2 by oxidative stress. Secretion of MIF from cardiomyocytes themselves, induced by oxidative stress, plays a key role in late phase activation of ERK1/2 through a MEK-dependent mechanism, although its secretion is not regulated by the MEK-ERK1/2 signaling cascade but by PKC. These phenomena provide important insights into the cellular response to oxidative stress. We thank M. Yashima for excellent technical assistance and the members of the First Department of Medicine, Asahikawa Medical College, for helpful discussions.

Calcineurin and calcium/calmodulin-dependent protein kinase (CaMK) II have been suggested to be the signaling molecules in cardiac hypertrophy. It was not known, however, whether these mechanisms are involved in cardiac hypertrophy induced by pressure overload without the influences of blood-derived humoral factors, such as angiotensin II. To elucidate the roles of calcineurin and CaMK II in this situation, we examined the effects of calcineurin and CaMK II inhibitors on pressure overload-induced expression of c-fos, an immediate-early gene, and protein synthesis using heart perfusion model. The hearts isolated from Sprague-Dawley rats were perfused according to the Langendorff technique, and then subjected to the acute pressure overload by raising the perfusion pressure. The activation of calcineurin was evaluated by its complex formation with calmodulin and by its R-II phosphopeptide dephosphorylation. CaMK II activation was evaluated by its autophosphorylation. Expression of c-fos mRNA and rates of protein synthesis were measured by northern blot analysis and by 14C-phenylalanine incorporation, respectively. Acute pressure overload significantly increased calcineurin activity, CaMK II activity, c-fos expression and protein synthesis. Cyclosporin A and FK506, the calcineurin inhibitors, significantly inhibited the increases in both c-fos expression and protein synthesis. KN62, a CaMK II inhibitor, also significantly prevented the increase in protein synthesis, whereas it failed to affect the expression of c-fos. These results suggest that both calcineurin and CaMK II pathways are critical in the pressure overload-induced acceleration of protein synthesis, and that transcription of c-fos gene is regulated by calcineurin pathway but not by CaMK II pathway.

This case report describes a short-coupled variant of Torsades de Pointes with a characteristic ECG pattern consisting of a prominent J wave in leads V3-V6, in which an electrical storm was evoked with autonomic receptor stimulation and a blockade test. The patient's frequent VF attacks were triggered by short-coupled premature ventricular contractions with a right bundle branch block morphology and left-axis deviation, and were suppressed by deep sedation followed by a combination therapy using verapamil and mexiletine. Interestingly, with the use of those drugs, the prominent J wave diminished. The mechanism underlying this syndrome is discussed.

Prevalent atrial fibrillation (AF) in patients with hypertrophic cardiomyopathy (HCM) represents an important issue with regard to stroke events caused by embolization and is associated with high mortality. Increased epicardial adipose tissue (EAT), which shows high metabolic activity, can locally influence the activity of the autonomic ganglia, enhancing autonomic dysregulation and increasing the likelihood of AF. We tested the hypothesis that EAT is associated with prevalent AF in HCM patients. Sixty-two patients with idiopathic HCM diagnosed on the basis of ultrasound cardiography findings and histopathological evaluation of myocardium obtained by right ventricular biopsy underwent cardiac magnetic resonance imaging to estimate the extent of EAT. EAT area was significantly higher in the group with AF episodes than in the group without. An increased incidence of AF was found to be significantly related to an increase in EAT, and this association persisted after adjustment for body mass index, sex, and age. Time domain measures of heart rate variability measured by Holter electrocardiography, standard deviation of normal to normal, and standard deviation of the average of normal to normal were negatively related to EAT area. EAT was positively correlated with intraventricular septal thickness and cystatin C level and negatively correlated with the 24-hour creatinine clearance rate. Increased EAT area in HCM patients is significantly related to the presence of AF, which is associated with changes in baseline autonomic nervous tone, left ventricular mass, and chronic kidney disease.

J Wave Syndromes . Introduction: Recently, great attention has been paid to the risk stratification of asymptomatic patients with an electrocardiographic early repolarization (ER) pattern. We investigated several repolarization parameters including the Tpeak‐Tend interval and Tpeak‐Tend/QT ratio in healthy individuals and patients with J wave syndrome who were aborted from sudden cardiac death. Methods and Results: Ninety‐two subjects were enrolled: 12 patients with ventricular fibrillation associated with J waves, 40 healthy subjects with an uneventful ER pattern and 40 healthy control subjects (C) without any evident J waves. Using ambulatory electrocardiogram recordings, the average QT interval, corrected QT interval (QTc), Tpeak‐Tend (Tp‐e) interval, which is the interval from the peak to the end of the T wave, and Tp‐e/QT ratio were calculated. Using ANOVA and post hoc analysis, there was no significant difference in the average QT and QTc in all 3 groups (QT; 396 ± 27 vs 405 ± 27 vs 403 ± 27 m, QTc; 420 ± 26 vs 421 ± 21 vs 403 ± 19 milliseconds in the C, ER pattern and J groups, respectively). The Tp‐e interval and Tp‐e/QT ratio were significantly more increased in the J wave group than the ER Pattern group (Tp‐e: 86.7 ± 14 milliseconds vs 68 ± 13.2 milliseconds, P &lt; 0.001, Tp‐e/QT; 0.209 ± 0.04 vs 0.171 ± 0.03, P &lt; 0.001), but they did not significantly differ between the C and ER pattern groups (Tp‐e: 68.6 ± 7.5 vs 68 ± 13.2, P = 0.97, Tp‐e/QT 0.174 ± 0.02 vs 0.171 ± 0.03, P = 0.4). Conclusion: As novel markers of heterogeneity of ventricular repolarization, Tpeak‐Tend interval and Tp‐Te/QT ratio are significantly increased in patients with J wave syndromes compared to age and sex‐matched uneventful ER. (J Cardiovasc Electrophysiol, Vol. 23 pp. 1109‐1114, October 2012)

Background The presence of a myocardial scar detected by late gadolinium enhancement (LGE) on cardiac magnetic resonance (CMR) has been described as a predictor of all-cause mortality in hypertrophic cardiomyopathy (HCM). However, the detailed spatial relationship between LGE site and electrical abnormality is unclear in high-risk HCM with malignant arrhythmia. Objective The purpose of this study was to elucidate the detailed relationship between the site on CMR imaging and the electrically damaged site, a potential origin of ventricular arrhythmias in patients with HCM. Methods Fifty consecutive HCM patients underwent contrast-enhanced CMR. Of those patients, 18 patients with ventricular tachycardia underwent electrophysiology study including endocardial mapping of the left ventricle (LV). The LGE area was calculated at 12 different LV sites: anterior, lateral, posterior, and septal segments of the basal, middle, and apical portions. At each LV site, the bipolar electrogram, effective refractory period (ERP), and monophasic action potential were recorded. Results LGE-positive segments demonstrated a significantly lower amplitude (4.0 ± 2.8 mV vs 7.3 ± 3.6 mV; P < .001), longer duration (54.7 ± 17.8 vs 40.6 ± 7.8 ms; P < .001), longer ERP (320 ± 42 ms vs 284 ± 37 ms; P = .001), and longer monophasic action potential duration measured at 90% repolarization (321 ± 19 ms vs 283 ± 25 ms; P < .001) than did LGE-negative segments. The LGE area negatively correlated with the amplitude (r = −0.59; P < .001) and positively correlated with the duration (r = 0.64; P < .001), ERP (r = 0.44; P < .001), and action potential duration measured at 90% repolarization (r = 0.63; P < .001). All the observed VTs originated from LGE-positive segments. Conclusion The spatial distribution of LGE significantly correlates with depolarizing and repolarizing electrical damage in high-risk HCM with malignant ventricular arrhythmia. The presence of a myocardial scar detected by late gadolinium enhancement (LGE) on cardiac magnetic resonance (CMR) has been described as a predictor of all-cause mortality in hypertrophic cardiomyopathy (HCM). However, the detailed spatial relationship between LGE site and electrical abnormality is unclear in high-risk HCM with malignant arrhythmia. The purpose of this study was to elucidate the detailed relationship between the site on CMR imaging and the electrically damaged site, a potential origin of ventricular arrhythmias in patients with HCM. Fifty consecutive HCM patients underwent contrast-enhanced CMR. Of those patients, 18 patients with ventricular tachycardia underwent electrophysiology study including endocardial mapping of the left ventricle (LV). The LGE area was calculated at 12 different LV sites: anterior, lateral, posterior, and septal segments of the basal, middle, and apical portions. At each LV site, the bipolar electrogram, effective refractory period (ERP), and monophasic action potential were recorded. LGE-positive segments demonstrated a significantly lower amplitude (4.0 ± 2.8 mV vs 7.3 ± 3.6 mV; P < .001), longer duration (54.7 ± 17.8 vs 40.6 ± 7.8 ms; P < .001), longer ERP (320 ± 42 ms vs 284 ± 37 ms; P = .001), and longer monophasic action potential duration measured at 90% repolarization (321 ± 19 ms vs 283 ± 25 ms; P < .001) than did LGE-negative segments. The LGE area negatively correlated with the amplitude (r = −0.59; P < .001) and positively correlated with the duration (r = 0.64; P < .001), ERP (r = 0.44; P < .001), and action potential duration measured at 90% repolarization (r = 0.63; P < .001). All the observed VTs originated from LGE-positive segments. The spatial distribution of LGE significantly correlates with depolarizing and repolarizing electrical damage in high-risk HCM with malignant ventricular arrhythmia.

An immature vasa vasorum in the adventitia of arteries has been implicated in induction of the formation of unstable atherosclerotic plaques. Normalization/maturation of the vasa vasorum may be an attractive therapeutic approach for arteriosclerotic diseases. Nerve growth factor (NGF) is a pleotropic molecule with angiogenic activity in addition to neural growth effects. However, whether NGF affects the formation of microvessels in addition to innervation during pathological angiogenesis is unclear. In the present study, we show a new role for NGF in neovessels around injured arterial walls using a novel in vivo angiogenesis assay. The vasa vasorum around arterial walls was induced to grow using wire-mediated mouse femoral arterial injury. When collagen-coated tube (CCT) was placed beside the injured artery for 7-14 days, microvessels grew two-dimensionally in a thin layer on the CCT (CCT-membrane) in accordance with the development of the vasa vasorum. The perivascular nerve was found at not only arterioles but also capillaries in the CCT-membrane. Biodegradable hydrogels containing VEGF and NGF were applied around the injured artery/CCT. VEGF significantly increased the total length and instability of microvessels within the CCT-membrane. In contrast, NGF induced regeneration of the peripheral nerve around the microvessels and induced the maturation and stabilization of microvessels. In an ex vivo nerve-free angiogenesis assay, although NGF potentially stimulated vascular sprouting from aorta tissues, no effects of NGF on vascular maturation were observed. These data demonstrated that NGF had potent angiogenic effects on the microvessels around the injured artery, and especially induced the maturation/stabilization of microvessels in accordance with the regeneration of perivascular nerves.

A 75‐year‐old woman was admitted to our hospital with rapidly deteriorating consciousness disturbance. She had a 7‐year history of rheumatoid arthritis (RA), which had been treated with methotrexate (MTX) and prednisolone. Brain T2‐weighted MRI showed diffuse high‐intensity lesions in the cerebral subcortical and deep white matter, bilateral basal ganglia and thalamus. A cerebrospinal fluid examination revealed elevated protein levels and positive Epstein–Barr virus (EBV) DNA. Human immunodeficiency virus was negative. Brain biopsy showed perivascular lymphocytic infiltration in the parenchyma and meninx with EBV‐encoded small RNA (EBER). Since this case did not fulfill the criteria for chronic active EBV infection (CAEBV), she was diagnosed with Epstein–Barr virus (EBV)‐associated vasculitis of the central nervous system. High‐dose methylprednisolone, acyclovir, ganciclovir and foscarnet were not effective. Although EBV is a causative agent of infectious mononucleosis (IM), lymphomas and nasopharyngeal carcinomas, vasculitic pathology of the central nervous system with EBV reactivation in the elderly is rare. Immunosuppressive drugs such as steroids and MTX are widely used to treat autoimmune disorders, but may exacerbate the reactivation of EBV. This is the first case of biopsy‐proven EBV‐positive/HIV‐negative vasculitis during the treatment of RA with MTX and steroids. This case indicates that EBV‐associated vasculitis needs to be considered as a differential diagnosis of CNS vasculitis.

Objective- Angiogenesis, entire step from endothelial cells (ECs) sprouts to vascular maturation, is a critical response to ischemia. To form functional mature vessels, interactions between ECs and pericytes are essential. Ninj1 (ninjurin1) is an adhesion molecule that contributes to the pathogenesis of neuroinflammation. We recently demonstrated that Ninj1 is expressed in pericytes during angiogenesis. However, the role of Ninj1 in angiogenesis under pathophysiological ischemic conditions has not yet been elucidated. Approach and Results- Ninj1 was detected in microvessels, and its expression was enhanced in ischemic tissues after mouse hindlimb ischemia. Knockdown of Ninj1 was performed by injection of biodegradable microspheres releasing Ninj1-small interfering RNA into muscle tissues. Alternatively, pericyte-specific Ninj1 knockout was induced by tamoxifen treatment of NG2-CreERT/Ninj1-flox mice. Ninj1 knockdown/knockout reduced the formation of blood-circulating functional vessels among total CD31+ microvessels within ischemic tissues and subsequently attenuated color Doppler-assessed blood flow recovery. Ninj1 overexpression enhanced expression of Anpt (angiopoietin) 1, whereas Ninj1 knockdown enhanced the endogenous Anpt1 antagonist, Anpt2 expression in pericytes and inhibited the association of pericytes with ECs and subsequent formation of capillary-like structure, that is, EC tube surrounded with pericytes in 3-dimensional gel culture. Conclusions- Our data demonstrate that Ninj1 is involved in the formation of functional matured vessels through the association between pericytes and ECs, resulting in blood flow recovery from ischemia. These findings further the current our understanding of vascular maturation and may support the development of therapeutics for ischemic diseases.

Ninjurin 1 (Ninj1) is identified as a peripheral nerve injury-induced protein. However, the role of Ninj1 in nerve regeneration is unclear. Schwann cells (SCs) and microvasculature are critical for peripheral nerve regeneration. SCs precursors and microvascular pericytes (PCs), which are nerve/glial antigen 2 (NG2)-positive cells are observed in peripheral nervous system. In this study, we investigated the role of Ninj1 in peripheral nerve regeneration using NG2+cell-specific inducible deletion of Ninj1 mouse model. The number of NG2+cells, which were associated with and without microvessels was increased after sciatic nerve crush injury. There was a significant increase in the expression of Ninj1 and EphA7 in the injured nerve tissue. This increase was mostly observed in NG2+cells. Genetic tracing of NG2+cells was performed using tamoxifen (Tam) treatment on NG2CreERT:R26R-tdTomato mice. The sciatic nerve was injured following the Tam-treatment, then tdTomato-expressing SCs were mostly observed in regenerated SCs at 21 days after nerve injury. Ninj1 gene knockout (Ninj1 KO) in NG2+cells was induced using NG2CreERT:Ninj1loxp mice. Tam-treated-NG2CreERT or Tam-nontreated NG2CreERT:Ninj1loxp mice were used as controls. Following Tam-treatment, the sciatic nerve in each group was injured. Ninj1KO significantly attenuated the expression of the myelin binding protein (MBP) as well as the number of myelinated axons. The expression of MBP in cultured SCs was significantly reduced by SiRNA-mediated Ninj1 knockdown (KD). Ninj1KD also attenuated the differentiation of SCs by isolated EphA7+multipotent PCs. The current data indicate that Ninj1 plays a vital role in peripheral nerve regeneration. This is observed particularly in the myelination process of NG2+cells including SCs precursors and multipotent PCs.

Although abnormalities of adventitial vasa vasorum are associated with vascular remodeling such as atherosclerosis, the mechanisms of vasa vasorum malformation and its role in vascular remodeling have not been fully clarified. The present study provides a line of novel evidence that ninjurin-1 contributes to adventitial microvascular maturation during vascular injury and regulates vascular remodeling.

Blood pressure (BP) fluctuates significantly in patients with atrial fibrillation (AF); office BP measurements seem insufficient to assess AF patient risk accurately. We hypothesized that home BP could better predict the risk of stroke/systemic embolic events (SEE) and major bleeding in patients with AF than office BP.In this prespecified subcohort study of the ANAFIE (All Nippon AF in the Elderly) Registry, we evaluated the impact of home BP on the risk of stroke/SEE, major bleeding, intracranial hemorrhage, all-cause death, and net cardiovascular outcome (a composite of stroke/SEE and major bleeding). At enrollment, home BP was measured twice in the morning and evening for 7 days.In total, 4933 elderly patients (aged ≥75 years) with nonvalvular AF participated. Incidences of net cardiovascular outcome, stroke/SEE, major bleeding, and intracranial hemorrhage increased significantly with increasing home systolic BP (H-SBP). Compared with H-SBP <125 mm Hg, ≥145 mm Hg was associated with increased risk of these events. The association between H-SBP and the events was observed only in patients with ≥20 H-SBP measurements.In elderly patients with nonvalvular AF, high H-SBP (≥145 mm Hg) was a significant predictor of stroke/SEE, major bleeding, and intracranial hemorrhage risk. Strict BP control guided by the increasing number of home BP measurements may provide an accurate clinical outcome risk assessment.URL: https://www.umin.ac.jp/ctr; Unique identifier: UMIN000024006.

The beneficial effects of thermal therapy have been reported in several cardiovascular diseases. However, it is unknown whether the thermal treatment has some beneficial roles against the development of atherosclerosis.The inflammatory arterial lesion was introduced by placement of a polyethylene cuff on femoral arteries of male Sprague-Dawley rats for 4 weeks. Thermal-treated group underwent daily bathing in 41 degrees C hot water for 15 minutes. Neointimal thickening along with immunohistochemical expression of heat-shock proteins (HSPs), monocyte chemoattractant protein-1 (MCP-1), and NADPH oxidase were compared with those of a thermally untreated (Control) group. Morphometric analysis demonstrated a significant suppression of neointimal thickening in thermal-treated group compared with the Control group (intimal/medial area ratios, 0.01+/-0.01 versus 0.31+/-0.04, P<0.01). Expression of MCP-1 and infiltration of ED-positive cells were enhanced in the adventitial layer of Control. More importantly, expression of HSP72 in media was enhanced by thermal treatment. Expression of p22-phox, the major membrane subunit of NADPH oxidase, and MCP-1 was augmented in cuff-injured adventitia of the Control but not the thermal-treated groups.Thermal treatment significantly attenuated infiltration of inflammatory cells in adventitia and suppressed neointimal thickening in cuff-injured arteries with the enhancement of HSP72 expression and suppression of oxidative stress.

Reactive oxygen species (ROS) enhance myocardial ischemia-reperfusion (I/R) injury. Ebselen, a seleno-organic glutathione peroxidase (GPx) mimetic, has a protective effect against tissue injury induced by ROS. However, the cardio-protective effect of orally administered ebselen has never been investigated in cardiac I/R injury. We investigated the effects and mechanisms of orally administered ebselen on experimental myocardial infarction. Isolated perfused rabbit hearts underwent 30 min of global ischemia and 60 min of reperfusion, with or without oral administration of ebselen 24 h before I/R, with or without enhanced oxidative stress by H2O2 infusion for the first 1 min of reperfusion. The recovery of left ventricular developed pressure (LVDP) was significantly improved, and the myocardial infarct size was significantly reduced by ebselen. The recovery of LVDP and the myocardial infarct size were markedly aggravated by H2O2 infusion. These enhancements by H2O2 were dose-dependently suppressed by ebselen, along with a reduction in myocardial 8-hydroxydeoxyguanosine levels, a marker for oxidative DNA damage. The myocardial reduced glutathione (GSH) level was preserved by ebselen. Ebselen markedly enhanced myocardial heat shock protein (HSP) 72 expression. The cardioprotective effect of ebselen-induced HSP72 was confirmed by MTT assay in isolated cardiomyocytes using KNK437, a novel HSP inhibitor. In conclusion, an oral administration of ebselen 24 h before I/R provided excellent cardioprotective effects, at least in part through HSP72 induction and GSH preservation.

We describe a case of early repolarization syndrome in which augmented J waves were documented during an electrical storm associated with hypokalemia. The patient was referred to our hospital for therapy to treat recurrent ventricular fibrillation (VF). The 12‐lead electrocardiogram showed giant J waves associated with hypokalemia during multiple episodes of VF. Although antiarrhythmic agents or deep sedation were not effective for the VF, an intravenous supplementation of potassium completely suppressed the VF with a reduction in the J‐wave amplitude. Our report discusses the possible relationship between hypokalemia and VF in early repolarization syndrome. (PACE 2012; 35:e234–e238)

Combination antihypertensive therapy with an angiotensin receptor blocker (ARB) and a calcium channel blocker (CCB) or diuretics is common. This subanalysis investigated blood pressure (BP) variability in patients receiving ARB-based combination therapy.In a prospective, randomized, open-label trial, hypertensive outpatients (≥65 years) who did not achieve their target BP with ARB monotherapy switched to losartan 50 mg/hydrochlorothiazide 12.5 mg (ARB + D) or ARB plus amlodipine 5 mg (ARB + C) for 12 months. Clinic BP and heart rate (HR), measured every 3 months, visit-to-visit variability and seasonal variation were evaluated.No significant between-group differences in average, maximum, or minimum systolic or diastolic BP, or HR, were found. Visit-to-visit BP variability (systolic) was significantly higher in the ARB + D group than in the ARB + C group. When each group was subdivided into two seasonal groups (summer and winter), no significant between-group differences in BP were found. Multivariate regression analyses showed a tendency toward negative correlation between outdoor temperature and urinary albumin:creatinine ratio and estimated glomerular filtration rate at 12 months in the ARB + D group.Combination therapy with an ARB plus a CCB may be preferable to that with an ARB plus diuretics for decreasing BP variability. As for seasonal variability, both treatments can be used safely regardless of season.

The incidence and age distribution of membranoproliferative glomerulonephritis (MPGN) vary throughout the world by race and ethnicity. We sought to evaluate the clinical features, pathogenesis, and age distribution of MPGN among a large nationwide data from the Japan Renal Biopsy Registry (J-RBR).A cross-sectional survey of 593 patients with MPGN (types I and III) registered in the J-RBR between 2007 and 2015 was conducted. Clinical parameters, and laboratory findings at diagnosis were compared between children (< 20 years), adults (20-64 years), and elderly patients (≥ 65 years).The median age of the patients was 59.0 years and mean proteinuria was 3.7 g/day. The rate of nephrotic syndrome was significantly higher in adults (40.4%) and elderly patients (54.0%) than in children (14.9%), whereas the rate of chronic glomerulonephritis was significantly higher in children (66.2%) than in adults (34.4%) and elderly patients (31.2%). According to the CGA risk classification, high-risk (red zone) cases accounted for 3.4% of children, 52.5% of adults and 84.1% of elderly patients with MPGN. As for pathogenesis, primary MPGN was most frequent (56.0%). Lupus nephritis was the most common disease among adult patients with secondary MPGN, whereas infectious disease was more common in elderly patients. Multiple regression analysis revealed that high systolic blood pressure and high proteinuria were independent factors associated with decreased estimated glomerular filtration rate (eGFR) in adults and elderly patients with MPGN.In Japan, adults and elderly patients with MPGN had a lower eGFR and severer proteinuria than children.

Abstract The presence of pericytes (PCs) with multipotency and broad distribution along capillary suggests that microvasculature plays a role not only as a duct for blood fluid transport but also as a stem cell niche that contributes to tissue maintenance and regeneration. The lack of an appropriate marker for multipotent PCs still limits our understanding of their pathophysiological roles. We identified the novel marker EphA7 to detect multipotent PCs using microarray analysis of an immortalized PC library. PCs were isolated from microvessels of mouse subcutaneous adipose tissues, then EphA7+ PCs called capillary stem cells (CapSCs) were separated from EphA7− control PCs (ctPCs) using fluorescence-activated cell sorting system. CapSCs had highly multipotency that enabled them to differentiate into mesenchymal and neuronal lineages compared with ctPCs. CapSCs also differentiated into endothelial cells and PCs to form capillary-like structures by themselves. Transplantation of CapSCs into ischemic tissues significantly improved blood flow recovery in hind limb ischemia mouse model due to vascular formation compared with that of ctPCs and adipose stromal cells. These data demonstrate that EphA7 identifies a subpopulation of multipotent PCs that have high angiogenesis and regenerative potency and are an attractive target for regenerative therapies. Significance statement The present study characterizes the multipotency of pericyte populations isolated from mammalian capillaries using a novel genetic marker, EphA7. It is shown that this marker can be used to isolate living cells and that EphA7+ pericytes, termed capillary stem cells (CapSCs), have capillary formation by themselves and cross-germ layer plasticity to differentiate into mesenchymal and neuronal lineages, indicating its potential use in both disease models and regenerative therapies.

Background: Although we and others have reported cases of patients with Anderson-Fabry disease (AFD) complicated by coronary spastic angina (CSA), the prevalence of CSA in these patients remains unknown. Methods and Results:We performed the acetylcholine-induced provocation test, according to the Japanese guidelines for the diagnosis and treatment of patients with CSA, in 9 consecutive patients having 5 independent AFD pedigrees.Coronary spasms were provoked in conjunction with symptoms and ECG ischemic changes in 8 of 9 (89%) patients with AFD. Conclusions:We found an unexpectedly high prevalence of CSA in patients with AFD.

The β 3 -adrenergic receptor (β 3 AR) is related to myocardial fatty acid metabolism and its expression has been implicated in heart failure. In this study, we investigated the role of β 3 AR in sepsis-related myocardial dysfunction using lipopolysaccharide (LPS)-induced endotoxemia as a model of cardiac dysfunction. We placed mice into three treatment groups and treated each with intraperitoneal injections of the β 3 AR agonist CL316243 (CL group), the β 3 AR antagonist SR59230A (SR group), or normal saline (NS group). Survival rates were significantly improved in the SR group compared with the other treatment groups. Echocardiography analyses revealed cardiac dysfunction within 6–12 h of LPS injections, but the outcome was significantly better for the SR group. Myocardial ATP was preserved in the SR group but was decreased in the CL-treated mice. Additionally, quantitative PCR analysis revealed that expression levels of genes associated with fatty acid oxidation and glucose metabolism were significantly higher in the SR group. Furthermore, the expression levels of mitochondrial membrane protein complexes were preserved in the SR group. Electron microscope studies showed significant accumulation of lipid droplets in the CL group. Moreover, inducible nitric oxide synthase (iNOS) protein expression and nitric oxide were significantly reduced in the SR group. The in vitro study demonstrated that β 3 AR has an independent iNOS pathway that does not go through the nuclear factor-κB pathway. These results suggest that blockading β 3 AR improves impaired energy metabolism in myocardial tissues by suppressing iNOS expression and recovers cardiac function in animals with endotoxin-induced heart failure. NEW &amp; NOTEWORTHY Nitric oxide production through stimulation of β 3 -adrenergic receptor (β 3 AR) may improve cardiac function in cases of chronic heart failure. We demonstrated that the blockade of β 3 AR improved mortality and cardiac function in endotoxin-induced heart failure. We also determined that LPS-induced inducible nitric oxide synthase has a pathway that is independent of nuclear factor-κB, which worsened cardiac metabolism and mortality in the acute phase of sepsis. Treatment with the β 3 AR antagonist had a favorable effect. Thus, the blockade of β 3 AR could offer a novel treatment for sepsis-related heart failure.

This study examined the usefulness of cerebrospinal fluid (CSF) neuron-specific enolase (NSE) levels as a candidate biomarker of neurodegeneration in Alzheimer's disease (AD), Parkinson's disease (PD), PD with dementia (PDD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA).We performed a systematic search of PubMed, the Cochrane Library, Scopus, and Google Scholar to find studies that measured CSF NSE levels in AD, PD, DLB, and/or MSA. For each disease, we pooled all available data and performed a meta-analysis, and meta-regression analyses of age and sex were conducted if the main analysis found a significant association.Twenty studies were included (13 for AD, 8 for PD/PDD/DLB, and 4 for MSA). Significantly elevated CSF NSE levels were detected in AD (Hedges' g = 0.822, 95% confidence interval [95% CI] 0.332 to 1.311, p = 0.0010), but the data exhibited high heterogeneity (I2 = 88.43%, p < 0.001). The meta-regression analysis of AD showed that age (p < 0.001), but not sex, had a significant effect on CSF NSE levels. A meta-analysis of the pooled data for PD/PDD/DLB did not show any significant changes in the CSF NSE level, but a sub-group analysis of PDD/DLB revealed significantly elevated CSF NSE levels (Hedges' g = 0.507, 95% CI 0.020 to 0.993, p = 0.0412). No significant changes in CSF NSE levels were detected in MSA.The CSF NSE level may be a useful biomarker of neurodegeneration in AD and PDD/DLB. Age was found to affect the CSF NSE levels of AD patients.

Abstract The prevalence of hypertension is high among patients undergoing dialysis. We extracted data of patients undergoing dialysis between 2012 and 2020 with recorded pre‐dialysis systolic blood pressure (SBP) using a web‐based national database in Japan. Following the 2019 Japanese Society of Hypertension guidelines, we classified SBP and assessed its trends over time based on sex, age, diabetes status, and the anti‐hypertensive medication use. Using the 2020 database, we examined 336,759 Japanese patients undergoing dialysis (114,249 female; 222,510 male). The mean age was 69.4 ± 12.5 years, and the mean SBP was 152.3 ± 24.7 mm Hg. The prevalence rate of pre‐dialysis hypertension was 70.2%, with 32.5%, 24.5%, and 13.2% of patients having grade I, grade II, and grade III hypertension, respectively. From 2014 to 2020, prevalence rate of pre‐dialysis hypertension and absolute values of pre‐dialysis SBP were higher in dialysis patients with diabetes than in those without diabetes across all age groups and sexes. Younger patients with diabetes or those on anti‐hypertensive medication exhibited an SBP of approximately 160 mm Hg. Cerebrovascular death in patients with diabetes was associated with a higher rate of pre‐dialysis hypertension than that in those without diabetes, and there was a significant difference in the prevalence of grade III hypertension between the two groups. In conclusion, the mean pre‐dialysis SBP among patients undergoing dialysis remained high, and younger patients with diabetes or those receiving anti‐hypertensive medications had poor blood pressure control. Optimal blood pressure management may be necessary to reduce the risk of cardiovascular mortality.

The effectiveness of ischemic postconditioning (iPoC) in patients with ST-elevation myocardial infarction (STEMI) without ischemic preconditioning has not been determined. Therefore, we investigated the impact of iPoC and its potential mechanism related to heat shock protein 72 (HSP72) induction on myocardial salvage in patients with STEMI without prodromal angina (PA).

Abstract MicroRNAs (miRNAs) are sequence-specific inhibitors of post-transcriptional gene expression. However, the physiological functions of these non-coding RNAs in renal interstitial mesenchymal cells remain unclear. To conclusively evaluate the role of miRNAs, we generated conditional knockout (cKO) mice with platelet-derived growth factor receptor-β (PDGFR-β)-specific inactivation of the key miRNA pathway gene Dicer. The cKO mice were subjected to unilateral ureteral ligation, and renal interstitial fibrosis was quantitatively evaluated using real-time polymerase chain reaction and immunofluorescence staining. Compared with control mice, cKO mice had exacerbated interstitial fibrosis exhibited by immunofluorescence staining and mRNA expression of PDGFR-β. A microarray analysis showed decreased expressions of miR-9-5p, miR-344g-3p, and miR-7074-3p in cKO mice compared with those in control mice, suggesting an association with the increased expression of PDGFR-β. An analysis of the signaling pathways showed that the major transcriptional changes in cKO mice were related to smooth muscle cell differentiation, regulation of DNA metabolic processes and the actin cytoskeleton, positive regulation of fibroblast proliferation and Ras protein signal transduction, and focal adhesion-PI3K/Akt/mTOR signaling pathways. Depletion of Dicer in mesenchymal cells may downregulate the signaling pathway related to miR-9-5p, miR-344g-3p, and miR-7074-3p, which can lead to the progression of chronic kidney disease. These findings highlight the possibility for future diagnostic or therapeutic developments for renal fibrosis using miR-9-5p, miR-344g-3p, and miR-7074-3p.

Although heparin-binding epidermal growth factor-like growth factor (HB-EGF) is thought to produce hypertrophy in isolated cardiomyocytes via an autocrine mechanism, the pathophysiological role of HB-EGF, in myocardial hypertrophy in vivo, is not yet known. To investigate the involvement of HB-EGF in cardiac remodeling associated with hypertension in vivo, we assayed the expression of HB-EGF mRNA and protein in the left ventricle (LV) during the development of left ventricular hypertrophy in spontaneously hypertensive rats (SHR).Prior to sacrifice and assay of HB-EGF and EGF-receptor (EGF-R) mRNA, morphologic and hemodynamic variables were measured in SHR and in age-matched Wistar Kyoto rats (WKY). At 5, 9 and 12 weeks of age, rats were killed, their hearts were removed, and the expression of HB-EGF and EGF-R mRNA and protein were measured. In addition, SHR and WKY were treated with enalapril, atenolol, or both for 4 weeks.In untreated SHR, double products (i.e. systolic blood pressure (sBP) multiplied by heart rate (HR), an index of mechanical load, peaked at 9 weeks. Expression of HB-EGF mRNA was also observed to peak in these animals at 9 weeks, while expression of EGF-R mRNA increased from 5 to 9 weeks, but remained constant thereafter. In untreated WKY, double products and EGF-R mRNA expression did not change over time, whereas the level of HB-EGF message increased gradually. Antibody to HB-EGF reacted primarily with myocyte membranes in SHR, whereas antibody to EGF-R reacted mainly with interstitial cells in these animals. The angiotensin-converting enzyme inhibitor, enalapril, markedly decreased sBP in SHR, whereas the beta 1-adrenoreceptor antagonist, atenolol, significantly decreased HR. While neither alone affected the expression of HB-EGF mRNA, their combination significantly reduced the expression of HB-EGF mRNA, as well as double products, in these rats, but had no effect on expression of EGF-R mRNA.The enhanced expression of HB-EGF mRNA and protein in LV of SHR suggest that this growth factor may play an important role during the early development of LV hypertrophy and cardiac fibrosis in SHR. The association between double products and HB-EGF expression suggest that the latter may be induced by increased mechanical load and may contribute, in turn, to cardiac remodeling.

The effects of exercise on regional myocardial blood flow and function were examined in the presence and absence of beta-adrenergic receptor blockade in 10 adult conscious dogs with severe left ventricular (LV) hypertrophy induced by aortic banding in puppies, which increased the LV weight/body weight ratio by 87%. Exercise at the most intense level studied increased LV systolic (+87 +/- 8 mm Hg) and end-diastolic (+28 +/- 5 mm Hg) pressures, systolic (+85 +/- 12 g/cm2) and diastolic (+49 +/- 11 g/cm2) wall stresses, and subepicardial wall thickening (+0.18 +/- 0.05 mm) but reduced subendocardial wall thickening (-0.45 +/- 0.12 mm) and full wall thickening (-0.42 +/- 0.13 mm). This was associated with a fall in the subendocardial/subepicardial (endo/epi) blood flow ratio to 0.87 +/- 0.06 from 1.24 +/- 0.08. Subendocardial dysfunction persisted during recovery, at a time when transmural blood flow distribution returned to baseline, suggesting myocardial stunning. At the least intense level of exercise studied, the endo/epi blood flow ratio did not fall (1.27 +/- 0.14), but increases in heart rate (+73 +/- 8 beats per minute) and LV systolic (+35 +/- 8 g/cm2) and diastolic (+27 +/- 3 g/cm2) wall stresses were observed, and subendocardial wall thickening fell significantly (-0.21 +/- 0.08 mm, p less than 0.05). With anticipation of exercise, subendocardial wall thickening was not changed. However, subendocardial dysfunction was even evident after 10 beats, i.e., the first 3 seconds of exercise, at a time when LV pressures and stresses had not increased. After beta-adrenergic receptor blockade with propranolol, the most intense level of exercise was associated with lesser increases in systolic and diastolic LV wall stresses, heart rate, and LV dP/dt, and the endo/epi blood flow ratio was no longer reduced below unity (1.17 +/- 0.09). In addition, there were no decreases in subendocardial or full wall thickening, and myocardial stunning was no longer observed. Thus, the subendocardial hypoperfusion and depression in subendocardial wall thickening observed during exercise in dogs with LV hypertrophy was prevented by pretreatment with beta-adrenergic receptor blockade. Furthermore, the subendocardial dysfunction occurred rapidly, before alterations in LV systolic or diastolic wall stress or an alteration in the endo/epi blood flow ratio.(ABSTRACT TRUNCATED AT 400 WORDS)

Clinical impact of magnesium (Mg) therapy remains controversial in acute myocardial infarction. We investigated the infarct size limiting effects of Mg and its mechanism in rabbits.Anesthetized rabbits underwent 30 min coronary occlusion and 3 h reperfusion in ten groups: (1) Control, (2) Low Mg, (3) Mg, (4) High Mg, (5) calcium (Ca), (6) Mg+Ca, (7) 8-phenyltheophylline (8PT), an adenosine receptor blockade, (8) 8PT+Mg, (9) alpha, beta-methylene-adenosine diphosphate (AOPCP), a selective inhibitor of ecto-5'-nucleotidase, and (10) AOPCP+Mg groups. Infract size (IS) to area at risk (AR) was measured by triphenyltetrazorium chloride method.The IS/AR ratio was significantly smaller in Mg, 27+/-3% (P<0.05) and High Mg, 24+/-2% (P<0.05) compared to Control, 50+/-3% and Low Mg, 42+/-4%. The IS limiting effects of Mg were abolished in 8PT+Mg, AOPCP+Mg and Mg+Ca. The IS/AR ratio correlated with neither rate-pressure products nor incidence of arrhythmia.Magnesium administration has an infarct size limiting effect independent of its effects on myocardial oxygen consumption and incidence of arrhythmia in rabbits. The infarct size limiting effect of magnesium is attributable, at least in part, to augmentation of adenosine mechanism.

&lt;i&gt;Purpose:&lt;/i&gt; To investigate the prevalence of cervical and intracranial atherosclerosis and silent brain infarction in patients with coronary artery disease (CAD).&lt;i&gt; Methods:&lt;/i&gt; Cervical and intracranial atherosclerotic lesions on magnetic resonance angiography (MRA) and silent brain infarctions on magnetic resonance imaging (MRI) were investigated in comparison with the findings of coronary angiography in 133 consecutive patients with CAD. &lt;i&gt;Results:&lt;/i&gt; The mean severity scores of cervical and intracranial MRA lesions were significantly higher in the three-vessel CAD (0.40 and 0.53, respectively) than in the zero-vessel CAD group (0.04 and 0.11). The mean scores of the maximal size and multiplicity of MRI lesion were also significantly greater in the two-vessel (1.00 and 1.44) and three-vessel CAD (0.94 and 1.26) than in the zero-vessel CAD group (0.27 and 0.50). The incidence of MRA lesion was markedly higher in patients with brain MRI lesion than in those without (51.1 vs. 6.5%). &lt;i&gt;Conclusions:&lt;/i&gt; Serious coronary artery lesions were commonly accompanied by latent atherosclerotic lesions in the cervical and intracranial arteries besides silent brain infarction in patients with CAD.

Reactive oxygen species (ROS) enhance myocardial ischemia-reperfusion (I/R) injury. Ischemic preconditioning (PC) provides potent cardioprotective effects in I/R. However, it has not been elucidated whether PC diminishes ROS stress in I/R and whether PC protects the myocardium from ROS stress transmurally and homogeneously. Isolated rabbit hearts perfused with Krebs-Henseleit buffer underwent 30 min of ischemia and 60 min of reperfusion. Hemodynamic changes and myocardial damage extent were analyzed in four groups. The control group underwent I/R alone. The H2O2 group underwent I/R with H2O2 infusion (50 microM) in the first minute of reperfusion to enhance oxidative stress. The PC and H2O2+PC groups underwent 5 min of PC before control and H2O2 protocols, respectively. Extracted myocardial DNA was analyzed for 8-hydroxydeoxyguanosine (8-OHdG), an indicator of oxidative DNA damage, with the use of the HPLC-electrochemical detection method. Glutathione peroxidase (GPX) activity and the reduced form of GSH were measured by spectrophotometric assays. The myocardial infarct size was significantly reduced in the PC group (19 +/- 2%) compared with the control group (37 +/- 4%; P < 0.05), particularly in the subendocardium. H2O2 transmurally increased the infarct size by 59 +/- 4% (P < 0.05), which was significantly diminished in the H2O2+PC group (31 +/- 4%; P < 0.01). The GSH levels, but not GPX activity, were well preserved transmurally in protocols with PC. The 8-OHdG levels were significantly decreased in PC and were significantly enhanced in H2O2 (P < 0.01). These changes in oxidative DNA damage were effectively diminished by PC. In conclusion, PC enhanced the scavenging activity of GSH against ROS transmurally, reduced myocardial damage, particularly in the subendocardium, and diminished the transmural difference in myocardial infarct size.

Sympathetic nervous activation is a crucial compensatory mechanism in heart failure. However, excess catecholamine may induce cardiac dysfunction and β-adrenergic desensitization. Although magnesium is known to be a cardioprotective agent, its beneficial effects on acute cardiac dysfunction remain to be elucidated. We examined the effects of magnesium on left ventricular (LV) dysfunction induced by a large dose of isoproterenol in dogs. Sixteen anesthetized dogs underwent a continuous infusion of isoproterenol (1 μg·kg −1 ·min −1 ) with or without a magnesium infusion (1 mg·kg −1 ·min −1 ). The dose response to small doses of isoproterenol (0.025–0.2 μg·kg −1 ·min −1 ) was tested hourly. A large dose of isoproterenol decreased LV systolic function, increased the time constant of LV isovolumic relaxation, and suppressed the dose response to small doses of isoproterenol in a time-dependent manner. Magnesium significantly attenuated isoproterenol-induced LV systolic and diastolic dysfunction and preserved the dose response to isoproterenol. Serum-ionized calcium significantly decreased with a large dose of isoproterenol but was fully maintained at baseline level with magnesium. A large dose of isoproterenol increased serum lipid peroxide levels and serological markers of myocardial damage, which were significantly suppressed by magnesium. In conclusion, magnesium significantly attenuated excess isoproterenol-induced acute cardiac dysfunction and β-adrenergic desensitization.

Inflammation and pulse wave velocity (PWV) are a potential risk factor and marker, respectively, for atherosclerosis in the primary prevention setting. Atherosclerosis is now generally accepted to be an inflammatory disorder of the arterial wall, and the high-sensitivity C-reactive protein (hs-CRP) level has been reported to be a strong predictor of cardiovascular events. High-sensitivity-CRP is associated with two factors related to inflammation: (1) the local production of CRP by atheromatous tissue or coronary artery smooth muscle cells and (2) adipose tissue as a potent source of inflammatory cytokines. Based on studies in North America and Europe, hs-CRP has been established as a cardiovascular risk factor and a cut-off value has been recommended. However, Japanese have lower hs-CRP values than their Western counterparts, partly because Japanese have a lower body mass index (BMI), which correlates positively to hs-CRP, and partly because lifestyle and genetic factors can affect hs-CRP values. Therefore, a cut-off value needs to be established by cohort studies for the Japanese population. Carotid-femoral PWV is most commonly measured by applanation tonometry, particularly in Europe, but this method is critically dependent upon the accurate placing of transducers over the arteries and is both time-consuming and complex. A novel device has been recently developed in Japan that measures brachial-ankle PWV (baPWV) using a volume-rendering method. Brachian-ankle PWV is a suitable screening method because of its technical simplicity and shorter measurement time. It is associated not only with conventional cardiovascular risk factors but also with new risk factors, such as inflammation, gamma-glutamyltransferase, chronic kidney disease, and psychosocial factors. However, a suitable cut-off value has yet to be established.

Recent evidence suggests an association between vasospastic angina and Brugada syndrome. Here we present two cases of coronary artery disease who presented with ECG abnormalities which might have been provoked or enhanced by ischemia of the conus branch of the right coronary artery. The 12-lead ECGs demonstrated normal sinus rhythm in these two cases. Interestingly, a saddle back or coved type ST segment elevation in leads V1-V3 was documented either in the percutaneous transluminal angioplasty procedure of the proximal right coronary artery or with an intracoronary acetylcholine (Ach) administration into the right coronary artery. These Brugada type ECG changes were restored to the baseline ECG waveform after improvement in the ischemia. In the second case, vasospasms of the conus branch of the right coronary artery were associated with a coved type ST segment elevation in leads V1 to V2. We discuss the possible interaction between ischemia caused by conus branch lesions and Brugada type electrocardiographic changes.

Marked reduction of RNA editing at the glutamine (Q)/arginine (R) site of the glutamate receptor subunit type 2 (GluR2) in motor neurons may be a contributory cause of neuronal death specifically in sporadic ALS. It has been shown that deregulation of RNA editing of several mRNAs plays a causative role in diseases of the central nervous system such as depression. We analyzed the effects of eight antidepressants on GluR2 Q/R site-RNA editing in a modified HeLa cell line that stably expresses half-edited GluR2 pre-mRNA. We also measured changes in RNA expression levels of adenosine deaminase acting on RNA type 2 (ADAR2), the specific RNA editing enzyme of the GluR2 Q/R site, and GluR2, in order to assess the molecular mechanism causing alteration of this site-editing. The editing efficiency at the GluR2 Q/R site was significantly increased after treatment with seven out of eight antidepressants at a concentration of no more than 10 microM for 24h. The relative abundance of ADAR2 mRNA to GluR2 pre-mRNA or to beta-actin mRNA was increased after treatment with six of the effective antidepressants, whereas it was unchanged after treatment with milnacipran. Our results suggest that antidepressants have the potency to enhance GluR2 Q/R site-editing by either upregulating the ADAR2 mRNA expression level or other unidentified mechanisms. It may be worth investigating the in vivo efficacy of antidepressants with a specific therapeutic strategy for sporadic ALS in view.

Background: Several animal experiments on acute myocardial infarction (AMI) have shown that the cardioprotective effects of ischemic preconditioning are more significant in hypertensive subjects. However, because there are no clinical data on the impact of hypertension on ischemic preconditioning in patients with AMI, whether clinical ischemic preconditioning of prodromal angina was beneficial in AMI patients with hypertension was investigated in the present study. Methods and Results: 125 patients with a first anterior AMI who had undergone successful reperfusion therapy were divided into 2 groups, with or without hypertension, and into 2 further subgroups based on the presence or absence of prodromal angina. Dual-isotope (thallium-201(TL)/Tc-99m pyrophosphate) single-photon emission computed tomography (SPECT) was performed within 1 week of reperfusion therapy. Left ventricular (LV) function and LV mass index (LVMI) were measured by left ventriculography and echocardiography, respectively. In patients without hypertension, prodromal angina resulted in significantly less myocardial damage on TL-SPECT, better LV ejection fraction and a greater myocardial blush grade compared to patients without prodromal angina. However, these cardioprotective effects of prodromal angina were significantly diminished in hypertensive patients. Importantly, the myocardial salvage effects of prodromal angina showed a significant negative correlation with LVMI, which was significantly greater in hypertensive patients. Conclusions: The cardioprotective effects of prodromal angina were attenuated in patients with hypertension. Hypertensive LV hypertrophy may crucially limit the effects of ischemic preconditioning in AMI. (Circ J 2011; 75: 1192-1199)

p53, a pivotal protein in the apoptotic pathway, has been identified as a mediator of transcriptional responses to ischemia-reperfusion (IR) injury. The characteristics and functional significance of the p53 response in vivo are largely unknown in IR-induced kidney injury. Therapeutic opportunities of delivering small interfering RNA (siRNA) via venous injection have gained recognition; however, systemic adverse effects of siRNA therapy should be considered. To prevent IR-induced kidney injury, we tested the efficacy of transarterial administration of siRNA targeting p53 (p53 siRNA). Female C57BL/6 mice underwent unilateral renal artery ischemia for 30 min, followed by reperfusion. siRNA experiments utilized short hairpin (sh) RNA plasmid-based approaches. Transfection of shRNA was performed using cationic polymer transfection reagent. Injection of synthetic p53 shRNA into the left renal artery just after ischemia improved tubular injury, apoptosis, and the swelling of mitochondria in cells of the thick ascending limb of Henle (mTALH) at the outer medullary regions. Staining of upregulated p53 was colocalized with the inducible expression of glycogen synthase kinase-3β (GSK-3β) at mTALH after IR injury. p53 shRNA inhibited GSK-3β expression and restored β-catenin expression at mTALH. For IR-induced kidney injury, transarterial delivery of p53 siRNA is an effective pharmacological intervention. Targeting siRNA to p53 leads to an attenuation of apoptosis and mitochondrial damage through the downregulation of GSK-3β expression and upregulation of β-catenin. Local delivery of vectors such as p53 siRNA through a transaortic catheter is clinically useful in reducing the adverse effect of siRNA-related therapy.

Prostaglandin I2 (PGI2) agonist has been reported to reduce tumor metastasis by modifying tumor angiogenesis; however, the mechanisms of how PGI2 affects the endothelial cells or pericytes in tumor vessel maturation are still unclear. The purpose of this study was to clarify the effects of PGI2 on tumor metastasis in a mouse lung metastasis model using Lewis lung carcinoma (LLC) cells. The mice were treated continuously with beraprost sodium (BPS), a PGI2 analog, for 3 weeks and then examined for lung metastases. The number and size of lung metastases were decreased significantly by BPS treatment. In addition, scanning electron microscopy and immunohistochemistry revealed that BPS increased the number of tumor‑associated pericytes and improved intratumor hypoxia. Collectively, this study suggests that BPS attenuated vascular functional maturation in metastatic tumors.

A 35-year-old male who had not previously suffered any major illnesses was admitted to our hospital because of general fatigue, fever, headache, vomiting, consciousness disturbance, and seizures. A neurological examination showed that he was in a semi-comatose state and exhibited neck stiffness. Brain magnetic resonance imaging detected high-intensity areas in the bilateral hippocampi and periventricular white matter. A cerebrospinal fluid examination revealed mononuclear pleocytosis, an elevated protein level, and positivity for human herpesvirus-7 (HHV-7) DNA. The patient's condition improved after the administration of methylprednisolone, intravenous immunoglobulins, and acyclovir. This is the first known case of limbic encephalitis associated with HHV-7 in an immunocompetent Japanese adult.

After examining in Part I the general mechanisms of endothelial cell injury in the kidney, the Working Group on Endothelin and Endothelial Factors of the European Society of Hypertension and the Japanese Society of Hypertension will herein review current knowledge on the role of endothelial dysfunction in multiple disease conditions that affect the kidney, including diabetes mellitus, preeclampsia, solid organ transplantation, hyperhomocysteinemia and antiangiogenic therapy in cancer. The few available randomized controlled clinical trials specifically designed to evaluate strategies for correcting endothelial dysfunction in patients with hypertension and/or chronic kidney disease are also discussed alongside their cardiovascular and renal outcomes.

Histone H3K4 trimethylation (H3K4 me3) is found in active euchromatic regions and plays an important role in podocyte function in which actin filaments are abundant in the foot processes. The pathogenesis of membranous nephropathy (MN), the most prevalent cause of primary nephrotic syndrome in the middle-aged and elderly, is podocyte dysfunction. We investigated the role of H3K4 me3 in podocyte dysfunction in nephrotic syndrome using cultured podocytes and a mouse proteinuria model induced by LPS. We examined renal biopsy specimens from six patients with nephrotic syndrome caused by Phospholipase-A2-Receptor-positive primary MN. H3K4 me3 exhibited a pattern of nuclear expression in podocytes of the kidneys from patients with MN. The overlapping expression of H3K4 me3 and cathepsin L (a potent endoprotease causing the breakdown of actin-associated protein within lysosomal compartments in kidney podocytes) were higher in patients with MN compared with the controls. Histone H3K4 me3 in kidney podocytes was negatively correlated with synaptopodin, an actin-associated protein in podocytes, and the expression was positively correlated with the proteinuria levels in patients with MN. Histone H3K4 me3 levels were elevated in podocytes of LPS-treated mice, combined with an increase in podocyte swelling, an elevation of serum creatinine and urine albumin, increased cathepsin L, and decreased synaptopodin expression. Histone H3K4 me3 levels at the cathepsin L promoter were elevated in LPS-exposed mouse kidneys. The administration of shRNA against MLL3 (an H3K4 methyltransferase) to LPS-treated mice and cultured podocytes co-cultured with LPS-stimulated macrophages ameliorated podocyte swelling, an elevation in the serum creatinine and urine albumin levels and an increased expression of histone H3K4 me3 and cathepsin L, and a decreased expression of synaptopodin and increase in histone H3K4 me3 levels at the cathepsin L promoter. Histone H3K4 me3 upregulation may be involved in podocyte dysfunction and the pathophysiology of MN. Targeting this epigenetic signature of histone H3K4 me3 followed by modulating the actin dynamics may be an effective strategy to ameliorate the consequences of MN.

Effects of the prostanoids on the growth of cultured aortic vascular smooth muscle cells (VSMCs) were examined using mice lacking prostanoid receptors. Proliferation of VSMCs was assessed by measuring [(3)H]-thymidine incorporation and the cell number, and their hypertrophy by [(14)C]-leucine incorporation and protein content. In VSMCs from wild-type mice, expressions of mRNAs for the EP(4) and TP were most abundant, followed by those for the IP, EP(3) and FP, when examined by competitive reverse transcriptase-PCR. Those for the EP(1), EP(2) and DP, however, could not be detected. AE1-329, an EP(4) agonist, and cicaprost, an IP agonist, inhibited platelet derived growth factor (PDGF)-induced proliferation of VSMCs from wild-type mice; these inhibitory effects disappeared completely in VSMCs from EP(4)(-/-) and IP(-/-) mice, respectively. In accordance with these effects, AE1-329 and cicaprost stimulated cAMP production in VSMCs from wild-type mice, which were absent in VSMCs from EP(4)(-/-) and IP(-/-) mice, respectively. Effects of PGE(2) on cell proliferation and adenylate cyclase were almost similar with those of AE1-329 in VSMCs from wild-type mice, which disappeared in VSMCs from EP(4)(-/-) mice. PGD(2) inhibited PDGF-induced proliferation of VSMCs from both wild-type and DP(-/-) mice to a similar extent. This action of PGD(2) was also observed in VSMCs from EP4(-/-) and IP(-/-) mice. In VSMCs from wild-type mice, I-BOP, a TP agonist, showed potentiation of PDGF-induced hypertrophy. I-BOP failed to show this action in VSMCs from TP(-/-) mice. The specific agonists for the EP(1), EP(2) or EP(3), and PGF(2)alpha showed little effect on the growth of VSMCs. These results show that PGE(2), PGI(2) and TXA(2) modulate PDGF-induced proliferation or hypertrophy of VSMCs via the EP(4), IP and TP, respectively, and that the inhibitory effect of PGD(2) on PDGF-induced proliferation is not mediated by the DP, EP(4) or IP.

Probucol administration of 4 weeks produced torsades de pointes associated with exacerbated QT interval prolongation in a 36-year-old woman with Romano-Ward syndrome. With discontinuance of probucol, the QT interval corrected for rate shortened from 620 msec to 500 msec and ventricular ectopic beats disappeared completely. Although probucol is known to prolong the QT interval, associated ventricular tachyarrhythmia has not been reported in humans as yet. This case suggests that one should be very careful in the administration of probucol to patients with long baseline QT intervals.

Treatment with a free radical scavenger could be a new option for ischemic brain attack, however, little is known about the alteration of oxidative stress markers induced by edaravone, a novel free radical scavenger, in human ischemic brain attack.We investigated the effects of edaravone on the oxidative stress markers in patients with ischemic brain attack. Twenty-one patients with ischemic brain attack and 19 controls were enrolled in this study. Blood samples were obtained just before and soon after the first administration of edaravone (30 mg) or ozagrel (40 mg). Intracellular reactive oxygen species of neutrophils were measured using 6-carboxy-2', 7'-dichlorodihydrofluorescin diacetate and a fluorescence-activated cell sorter. Superoxide from neutrophils, induced by phorbol myristate acetate (PMA), was determined by luminol-amplified chemiluminescence assay.Treatment with 30 mg of edaravone significantly decreased the intracellular reactive oxygen species (ROS) of neutrophils (Wilcoxon test, p=0.0001) and PMA-induced superoxide produced by neutrophils (Wilcoxon test, p=0.001). Ozagrel did not alter the intracellular ROS or superoxide production of neutrophils.Reduction of intracellular ROS and suppression of superoxide production in neutrophils provide a potential explanation for the clinical efficacy of edaravone in patients with ischemic brain attack.

We reported impaired QT-rate dependence in early repolarization syndrome (ERS); however, contemporary data have shown peak incidence of sudden cardiac death (SCD) in ERS and Brugada syndrome (BrS) at mid-night and early morning. Taken together, we analyzed the nocturnal QT-rate dependence in both syndromes.A total of 172 subjects were enrolled: 11 ERS, 11 BrS patients, 50 subjects with an uneventful ER pattern (ERP), and 100 non-J-wave control subjects. Ambulatory ECG-derived parameters (QT, QTc, and QT/RR slope) and day-night QT difference were analyzed and compared. Among the groups, there was no significant difference in the average QT or QTc; however, the 24-hour QT/RR slope was significantly smaller in ERS and BrS patients (0.103 ± 0.01 and 0.106 ± 0.01, respectively) than in the control group (0.156 ± 0.03, P < 0.001). Detailed analysis showed a lower day-night QT difference in ERS and BrS patients (19 ±18.7 and 24 ±14 milliseconds, respectively) than in the controls (40 ± 22 milliseconds, P = 0.007) with the lowest QT/RR slopes seen in the ERS and BrS groups from 0 to 3:00 am (QT/RR; 0.076 ± 0.02 vs. 0.092 ± 0.04 vs. 0.117 ± 0.04, for the ERS, BrS, and controls, respectively, P = 0.004) and from 3 to 6 am (QT/RR 0.074 ± 0.03 vs. 0.079 ± 0.02 vs. 0.118 ± 0.04, P < 0.001).In a large population of age- and gender-matched groups, both ERS and BrS patients showed attenuated QT-rate dependence and impaired QT day-night modulation that may provide a baseline reentrant substrate. Importantly, QT/RR maladaptation was most evident at mid-night and early morning, which may explain the propensity of such patients to develop SCD during this critical period.

A 69-year-old Japanese woman was admitted to our hospital with progressive muscle weakness and dysphagia.She was taking pitavastatin for dyslipidemia.Her serum creatine kinase was 6,300 U/L.Pitavastatin was stopped, but her symptoms deteriorated, and cardiac congestion appeared.A muscle biopsy showed necrotizing myopathy (NM), and anti-signal recognition particle (SRP) antibody was positive. 18Ffluorodeoxyglucose-positron emission tomography showed an abnormal uptake, and magnetic resonance imaging showed abnormal gadolinium enhancement in the left ventricular wall.An endomyocardial biopsy revealed inflammatory cardiomyopathy.Steroid, tacrolimus, and intravenous immunoglobulins were effective against the symptoms.This is the first case of biopsy-proven secondary cardiomyopathy due to anti-SRPpositive NM.

Sarcopenia is a pathophysiological malfunction induced by skeletal muscle atrophy. Several studies reported an association between sarcopenia-induced cardiac cachexia and poor prognosis in heart disease. However, due to lack of an established animal models, the underlying mechanism of disturbed cardiac repair accompanied with sarcopenia remains poorly understood. Here, we developed a novel sarcopenia-induced cardiac repair disturbance mouse model induced by tail suspension (TS) after cardiac ischemia and reperfusion (I/R). Importantly, we identified a specific exosomal-microRNA marker, miR-16-5p, in the circulating exosomes of I/R-TS mice. Of note, sarcopenia after I/R disturbed cardiac repair and raised the level of circulating-exosomal-miR-16-5p secreting from both the atrophic limbs and heart of TS mice. Likewise, miR-16-5p mimic plasmid disturbed cardiac repair in I/R mice directly. Additionally, in neonatal rat ventricular myocytes (NRVMs) cultured in vitro under hypoxic conditions in the presence of a miR-16-5p mimic, we observed increased apoptosis through p53 and Caspase3 upregulation, and also clarified that autophagosomes were decreased in NRVMs via SESN1 transcript interference-mediated mTOR activation. In conclusion, we show the pro-apoptotic effect of sarcopenia-derived miR-16-5p, which may be behind the exacerbation of myocardial infarction. Therefore, miR-16-5p can be a novel therapeutic target in the context of cardiac repair disturbances in sarcopenia-cachexia.

The effect of peripheral sympathetic block on the autonomic nerve function of the heart was studied using the head-up tilt test (HUTT) and right stellate ganglion block (RSGB). Blood pressure (BP), heart rate (HR) and the parameters of power spectral analysis of HR variability recorded during the HUTT were measured in 8 patients with chronic pain syndrome before and after RSGB. In the control state, the mean HR and the LF/HF component recorded during HUTT significantly increased whereas the HF component markedly decreased. Conversely, the mean HR and LF/HF and HF components during HUTT did not significantly alter after the RSGB procedure. There were no significant differences between the BP values before and after RSGB. These results suggest that RSGB suppresses cardiac sympathetic function without significantly affecting BP and thus may be a safe and effective therapy for the chronic pain syndrome.

We describe a case of a 67-year-old woman with a history of cerebral infarction and pulmonary embolism that presented with chest pain. Subsequent evaluation resulted in a diagnosis of acute myocardial infarction and occult DVT, and imaging revealed a rare congenital absence of the infra-renal portion of the inferior vena cava, with lower extremity venous drainage diverted via an ascending lumbar vein. Associations between congenital absence of the inferior vena cava and thrombosis are discussed.

The purpose of this study was to investigate whether the ideal control of atrial fibrillation (AF) associated with hypertensive patients depends on the usage of renin-angiotensin system (RAS) inhibitors or whether it occurs regardless of the kind of antihypertensive agents used. The control of AF was compared in 112 outpatients between 1) those with or without the administration of RAS inhibitors, and 2) those with an ideal or poor control of the blood pressure (BP) regardless of the kind of antihypertensive therapy used. The therapies with or without RAS inhibitors did not yield any significant difference in the AF control states, even though RAS inhibitors had been administered to the patient group with a high proportion of organic heart disease. The ideal BP control group exhibited a significantly better AF control in comparison to the poor BP control group. The former group had a significantly smaller left atrial diameter determined by ultrasonic echocardiography. BP control itself may essentially be important for preventing AF in the general patient population. Poor BP control seemed to have an affect on worsening AF possibly via left ventricular diastolic dysfunction, followed by left atrial overload.

Although metabolic syndrome confers an increased risk of cardiovascular disease in the general population, little is known about the alteration of abdominal adiposity and its association with adipocytokines in hemodialysis patients. We investigated the plasma high-molecular-weight (HMW) adiponectin level and its relationship to visceral fat area (VFA) and various markers of atherosclerosis in hemodialysis patients. In a cross-sectional study, conventional cardiovascular risk factors, plasma total and HMW adiponectin, the number of components of the metabolic syndrome and, using computed tomography, the distribution of abdominal adiposity were assessed in 144 hemodialysis patients (90 men and 54 women; mean age, 60.7 years) and 30 age- and sex-matched patients with chronic kidney disease (CKD). Plasma HMW adiponectin levels in hemodialysis patients were significantly higher than those in patients with CKD, negatively associated with VFA and serum triglycerides and positively associated with plasma total adiponectin, as well as the HMW-to-total adiponectin ratio in men and women (all P < 0.05) in a simple regression analysis. In a multiple regression analysis, VFA was a significant determinant of HMW adiponectin in hemodialysis patients. Furthermore, after adjustment for classical risk factors, HMW adiponectin levels were significantly higher in patients undergoing treatment with renin-angiotensin system inhibitors or calcium channel blockers compared with patients not undergoing such treatment. This study shows that plasma HMW adiponectin levels were negatively associated with VFA and positively associated with treatment with blockade of the renin-angiotensin system and of the calcium channel. Therefore, these drugs might be effective for improving adipocytokine-related metabolic abnormalities in hemodialysis patients.

Fetuin-A and osteoprotegerin (OPG) are arterial calcification regulators, which are related to cardiovascular survival in hemodialysis patients. We hypothesized that a balance of these calcification regulators might mediate the progression of left ventricular (LV) diastolic dysfunction in hemodialysis patients. We recruited 63 hemodialysis patients and measured their serum fetuin-A, OPG, arterial stiffness, aortic calcification and echocardiographic parameters, including the transmitral early diastolic velocity/tissue Doppler mitral annular early diastolic velocity ratio (E/E'), and analyzed the relationships between these variables. Fetuin-A levels were significantly and negatively correlated with the ankle-brachial pulse wave velocity (baPWV), aortic calcification score (AOCS), left atrial volume index (LAVI), LV mass index (LVMI) and E/E'. OPG levels and the ratio of OPG to fetuin-A levels were significantly and positively correlated with the baPWV, AOCS, LAVI and E/E'. A stepwise multiple regression analysis revealed that E/E' was independently correlated with fetuin-A levels (β=-0.334, P=0.02), OPG levels (β=0.367, P=0.01) and the ratio of OPG to fetuin-A (β=0.295, P=0.04). Categorizing the patients according to their serum fetuin-A and OPG levels revealed that patients with low fetuin-A and high OPG levels had the highest LAVI, LVMI and E/E' values after adjusting for potential confounders. Serum fetuin-A levels negatively reflected, whereas OPG levels and the ratio of OPG to fetuin-A positively reflected an increase in vascular and ventricular stiffness, leading to the aggravation of diastolic dysfunction. Therefore, based on our results, the balance of the tissue calcification regulators fetuin-A and OPG could mediate the progression of LV diastolic dysfunction in hemodialysis patients.

Systemic capillary leak syndrome (SCLS) is a life-threatening disorder which presents with periodic episodes of hypovolemic shock, due to plasma leakage to the extra-vascular space reflected by accompanying hypoalbuminemia, hemoconcentration and edema often with associated monoclonal gammopathy. We describe a 28-year-old woman with SCLS who required aggressive fluid resuscitation and was successfully treated with corticosteroid, terbutaline, and theophylline. At exacerbation, the levels of serum granulocyte colony-stimulating factor (G-CSF) were increased. Thus, G-CSF might play an important role and can be a useful biomarker for the severity of attacks in SCLS.

BackgroundProteinuria and reduced estimated glomerular filtration rate (eGFR) are associated with an increased risk of mortality from acute myocardial infarction (AMI). However, it is unknown whether there is a difference in prognostic value for all-cause mortality between proteinuria and eGFR during post-AMI.MethodsA consecutive series of 101 patients admitted with AMI who received angioplasty were enrolled. Dipstick proteinuria and eGFR were assessed on admission: (i) the patients were divided into 2 groups according to the presence of proteinuria (proteinuria, n = 25), or not (negative, n = 76), (ii) the patients were divided into 2 groups according to lower eGFR (GFR < 60 mL/min/1.73 m2, n = 31) or higher (GFR > 60 mL/min/1.73 m2, n = 70). Clinical characteristics and 3-year all-cause mortality estimated by Kaplan–Meier method were evaluated in each group. Additionally, a multivariate Cox proportional hazards model was applied to evaluate which factor was associated with all-cause mortality.ResultsMean follow-up period was 914 days. Higher brain natriuretic peptide (BNP) levels were shown in the proteinuria and lower eGFR groups, respectively (proteinuria, 301 ± 324 pg/mL; negative, 146 ± 159 pg/mL; p = 0.02; lower eGFR, 294 ± 305 pg/mL; higher eGFR, 142 ± 161 pg/mL; p = 0.02). Three-year all-cause mortality was higher in the proteinuria group than in the normal group (p < 0.001) and in the lower eGFR group than in the higher group (p = 0.006). In a Cox proportional hazards model, the presence of proteinuria [hazard ratio (95% confidence interval), 4.51 (1.07–18.96); p = 0.04] was selected as one of the predictors for all-cause mortality.ConclusionsDipstick proteinuria and lower eGFR in the early phase of AMI follow-up were related to increased plasma BNP level during the sub-acute phase and long-term adverse outcome. Dipstick proteinuria may be a prognostic marker for long-term all-cause mortality.

Abstract In the ANAFIE Registry home blood pressure subcohort, we evaluated 5204 patients aged ≥75 years with non‐valvular atrial fibrillation (NVAF) to assess blood pressure (BP) control, prevalence of masked hypertension, and anticoagulant use. Mean clinic (C) and home (H) systolic/diastolic BP(SBP/DBP) was 128.5/71.3 and 127.7/72.6 mm Hg, respectively. Overall, 77.5% of patients had hypertension; of these, 27.7%, 13.4%, 23.4%, and 35.6% had well‐controlled, white coat, masked, and sustained hypertension, respectively. Masked hypertension prevalence increased with diabetes, decreased renal function, age ≥80 years, current smoker status, and chronic obstructive pulmonary disease. By morning/evening average, 59.0% of patients had mean H‐SBP ≥ 125 mm Hg; 48.9% had mean C‐SBP ≥ 130 mm Hg. Early morning hypertension (morning H‐SBP ≥ 125 mm Hg) was found in 65.9% of patients. Although 51.1% of patients had well‐controlled C‐SBP, 52.5% of these had uncontrolled morning H‐SBP. In elderly NVAF patients, morning H‐BP was poorly controlled, and masked uncontrolled morning hypertension remains significant.