Calmodulin (CaM) and Ca(2+)/CaM-dependent protein kinase II (CaMKII) play important roles in the development of heart failure. In this study, we evaluated the effects of CaM on mitochondrial membrane potential (DeltaPsi(m)), permeability transition pore (mPTP) and the production of reactive oxygen species (ROS) in permeabilized myocytes; our findings are as follows. (1) CaM depolarized DeltaPsi(m) dose-dependently, but this was prevented by an inhibitor of CaM (W-7) or CaMKII (autocamtide 2-related inhibitory peptide (AIP)). (2) CaM accelerated calcein leakage from mitochondria, indicating the opening of mPTP, however this was prevented by AIP. (3) Cyclosporin A (an inhibitor of the mPTP) inhibited both CaM-induced DeltaPsi(m) depolarization and calcein leakage. (4) CaM increased mitochondrial ROS, which was related to DeltaPsi(m) depolarization and the opening of mPTP. (5) Chelating of cytosolic Ca(2+) by BAPTA, the depletion of SR Ca(2+) by thapsigargin (an inhibitor of SERCA) and the inhibition of mitochondrial Ca(2+) uniporter by Ru360 attenuated the effects of CaM on mitochondrial function. (6) CaM accelerated Ca(2+) extrusion from mitochondria. We conclude that CaM/CaMKII depolarized DeltaPsi(m) and opened mPTP by increasing ROS production, and these effects were strictly regulated by the local increase in cytosolic Ca(2+) concentration, initiated by Ca(2+) releases from the SR. In addition, CaM was involved in the regulation of mitochondrial Ca(2+) homeostasis.

Although mitochondrial oxidative catabolism of fatty acid (FA) is a major energy source for the adult mammalian heart, cardiac lipotoxity resulting from elevated serum FA and enhanced FA use has been implicated in the pathogenesis of heart failure. To investigate the effects of intermediates of FA metabolism [palmitoyl-l-carnitine (Pal-car) and palmitoyl-CoA (Pal-CoA)] on mitochondrial function, we measured membrane potential (DeltaPsi(m)), opening of the mitochondrial permeability transition pore (mPTP), and the production of ROS in saponin-treated rat ventricular myocytes with a laser scanning confocal microscope. Our results revealed that 1) lower concentrations of Pal-car (1 and 5 muM) caused a slight hyperpolarization of DeltaPsi(m) [tetramethylrhodamine ethyl ester (TMRE) intensity increased to 115.5 +/- 5.4% and 110.7 +/- 1.6% of baseline, respectively, P < 0.05] but did not open the mPTP, 2) a higher concentration of Pal-car (10 microM) depolarized DeltaPsi(m) (TMRE intensity decreased to 61.9 +/- 12.2% of baseline, P < 0.01) and opened the mPTP (calcein intensity decreased to 70.7 +/- 2.8% of baseline, P < 0.01), 3) Pal-CoA depolarized DeltaPsi(m) without opening the mPTP, and 4) only the higher concentration of Pal-car (10 muM) increased ROS generation (2',7'-dichlorofluorescein diacetate intensity increased to 3.4 +/- 0.3-fold of baseline). We concluded that excessive exogenous intermediates of long-chain saturated FA may disturb mitochondrial function in different ways between Pal-car and Pal-CoA. The distinct mechanisms of the deteriorating effects of long-chain FA on mitochondrial function are important for our understanding of the development of cardiac diseases in systemic metabolic disorders.

DE-MRI is useful to diagnose the cardiac involvement of sarcoidosis and to evaluate cardiac function. It is likely that the distribution of DE in mid- to epi-myocardium is the characteristic of cardiac sarcoidosis, and the larger DE area may be correlated with poor LV function.

BackgroundRecently, time-resolved 3D phase contrast magnetic resonance imaging (4D-flow) allows flow dynamics in patients with pulmonary arterial hypertension to be measured. Abnormal flow dynamics, such as vortex blood flow pattern in the pulmonary artery (PA), may reflect progression of pulmonary arterial hypertension (PAH). Some reports suggested that abnormal blood flow parameters including wall shear stress (WSS) could be markers of PAH. However, it was not fully assessed clinical usefulness of these variables. We aimed to assess whether these flow dynamic parameters, such as vortex formation time (VFT) and WSS, were associated with right ventricular (RV) function.ResultsFifteen subjects, nine with PAH and six healthy volunteers, underwent 4D-flow. Differences of Blood flow patterns, blood flow velocities and WSS between PAH patients and healthy volunteers were evaluated. We also assessed the association between VFT, WSS and RV function in PAH patients. Both vortex blood flow patterns and early systolic retrograde flow in the main PA were observed in all patients with PAH. The PA flow velocities and WSS in patients with PAH were lower than those in healthy volunteers, but that blood flow volumes in the MPA, RPA and LPA and SV in the MPA were broadly comparable between the groups. The mean VFT was 35.0 ± 16.6 % of the cardiac cycle. The VFT significantly correlated with RV ejection fraction, RV end systolic volume, and RV end systolic volume index (RVEF = 75.1 + (−85.7)·VFT, p = 0.003, RVESV = 12.4 + 181.8·VFT, p = 0.037 and RVESVI = 10.6 + 114.8·VFT, p = 0.038, respectively) in PAH patients, whereas WSS did not correlate with RV function.ConclusionsWe confirmed that abnormal blood flow dynamics, including the vortex formation and the early onset of retrograde flow, low WSS in the PA were characteristics of PAH. The VFT may be associated with right ventricular dysfunction, whereas WSS was not. Our results suggest that 4D-flow is an effective means of detecting right heart failure as well as diagnosing PAH.Clinical trial registration URL: https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr.cgi. Unique identifier: UMIN000011128Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-016-2755-7) contains supplementary material, which is available to authorized users.

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Objective Many studies have revealed that white blood cell count (WBC) is related to insulin resistance which is a central mechanism of metabolic syndrome (MetS). However, few cohort studies have examined the role of WBC in the development of MetS. We hypothesized that WBC is associated with the future development of MetS, and investigated the longitudinal incidence of MetS in healthy workers. Methods WBC was measured in 5,073 workers (mean age 42.5 years) without MetS at baseline. The incidence of MetS was monitored over 7 years of follow-up, in relation to quartiles of WBC. During the followup, 925 participants were diagnosed as MetS. Results Incidence of MetS was increased in participants with higher WBC: the rates of incidence of MetS were 22.6, 32.9, 42.9, and 57.5 per 1,000 person-years of follow-up in the 1st, 2nd, 3rd, and 4th quartiles of WBC, respectively. After adjustments for confounding factors, the adjusted hazards ratio (95% confidence interval) for MetS was 1.00 (reference), 1.22 (0.98 to 1.51), 1.52 (1.24 to 1.87), and 1.66 (1.35 to 2.04) through the quartiles of WBC, respectively, (p <0.001). This relationship was consistent among current smokers and never smokers, and among male and female genders, respectively. Conclusion WBC is useful in predicting the future development of MetS which leads to atherosclerotic diseases.

ObjectiveObesity is a risk factor for chronic kidney disease (CKD) and cardiovascular disease. The association between waist to height ratio (WheiR) and CKD is unclear. This study evaluated the association between WheiR and CKD.Design and MethodsIn this longitudinal cohort study, 4841 Japanese workers (3686 males, 1155 females) 18 to 67 years of age in 2008 were followed up until 2011. CKD was defined as an estimated glomerular filtration rate of <60 mL/min/1.73 m2 (by the Modification of Diet in Renal Disease equation for Japanese) or dipstick proteinuria (≥1+). Cox proportional hazards models were used to examine the relationship between WheiR and development of CKD.ResultsA total of 384 (7.9%) participants (300 men and 84 women) were found to have new CKD. The incidence of CKD was 13.7, 24.2, 37.9 and 43.7 per 1000 person-years of follow-up in the lowest, second, third and highest quartiles of WheiR, respectively. After adjustment for potential confounders, the adjusted hazard ratios (95% confidence interval) for CKD were 1.00 (reference), 1.23 (0.85, 1.78), 1.59 (1.11, 2.26) and 1.62 (1.13, 2.32) through the quartiles of WheiR, respectively. WheiR had a significant predictive value for the incidence of both proteinuria and low estimated glomerular filtration rate. After subdivision according to gender, the relationship between WheiR and the incidence of CKD was statistically significant in the unadjusted model. However, after adjusting for potential confounders, WheiR was significantly associated with the incidence of CKD in females, whereas it was not significant in males.ConclusionsWheiR, which is commonly used as an index of central obesity, is associated with CKD. There was a significant gender difference in the relationship between CKD and WheiR.

Background: Previous studies demonstrated that calcium/calmodulin (Ca2+/CaM) activates nicotinamide adenine dinucleotide phosphate oxidases (NOX). In endothelial cells, the elevation of intracellular Ca2+ level consists of two components: Ca2+ mobilization from the endoplasmic reticulum (ER) and the subsequent store-operated Ca2+ entry. However, little is known about which component of Ca2+ increase is required to activate NOX in endothelial cells. Here, we investigated the mechanism that regulates NOX-derived reactive oxygen species (ROS) production via a Ca2+/CaM-dependent pathway. Methods: We measured ROS production using a fluorescent indicator in endothelial cells and performed phosphorylation assays. Results: Bradykinin (BK) increased NOX-derived cytosolic ROS. When cells were exposed to BK with either a nominal Ca2+-free or 1 mM of extracellular Ca2+ concentration modified Tyrode’s solution, no difference in BK-induced ROS production was observed; however, chelating of cytosolic Ca2+ by BAPTA/AM or the depletion of ER Ca2+ contents by thapsigargin eliminated BK-induced ROS production. BK-induced ROS production was inhibited by a CaM inhibitor; however, a Ca2+/CaM-dependent protein kinase II (CaMKII) inhibitor did not affect BK-induced ROS production. Furthermore, BK stimulation did not increase phosphorylation of NOX2, NOX4, and NOX5. Conclusions: BK-induced NOX-derived ROS production was mediated via a Ca2+/CaM-dependent pathway; however, it was independent from NOX phosphorylation. This was strictly regulated by ER Ca2+ contents.