Phosphoproteomic of the acetylcholine pathway enables discovery of the PKC-ß-PIX-Rac1-PAK cascade as a stimulatory signal for aversive learning.

Acetylcholine is a neuromodulator critical for learning and memory. The cholinesterase inhibitor donepezil increases brain acetylcholine levels and improves Alzheimer's disease (AD)-associated learning disabilities. Acetylcholine activates striatal/nucleus accumbens dopamine receptor D2-expressing medium spiny neurons (D2R-MSNs), which regulate aversive learning through muscarinic receptor M1 (M1R). However, how acetylcholine stimulates learning beyond M1Rs remains unresolved. Here, we found that acetylcholine stimulated protein kinase C (PKC) in mouse striatal/nucleus accumbens. Our original kinase-oriented phosphoproteomic analysis revealed 116 PKC substrate candidates, including Rac1 activator ß-PIX. Acetylcholine induced ß-PIX phosphorylation and activation, thereby stimulating Rac1 effector p21-activated kinase (PAK). Aversive stimulus activated the M1R-PKC-PAK pathway in mouse D2R-MSNs. D2R-MSN-specific expression of PAK mutants by the Cre-Flex system regulated dendritic spine structural plasticity and aversive learning. Donepezil induced PAK activation in both accumbal D2R-MSNs and in the CA1 region of the hippocampus and enhanced D2R-MSN-mediated aversive learning. These findings demonstrate that acetylcholine stimulates M1R-PKC-ß-PIX-Rac1-PAK signaling in D2R-MSNs for aversive learning and imply the cascade's therapeutic potential for AD as aversive learning is used to preliminarily screen AD drugs.

Peak Work Rate during Exercise Could Detect Frailty Status in Elderly Patients with Stable Heart Failure.

The Kihon Checklist (KCL) is a reliable tool for determining frailty status in the elderly. However, there is no information in the literature about the relationship between frailty status and exercise capacity. Here, we examined the associations between cardiopulmonary exercise testing parameters and frailty status in elderly patients with stable heart failure (HF).Ninety-two elderly patients with stable HF were evaluated using cardiopulmonary exercise testing and the KCL. A KCL score of 0-3 was classified as robust, 4-7 as pre-frail, and ≥ 8 as frail.Mean age, peak VO2, and KCL score were 81.7 years, 13.2 mL/kg/minute, and 10.7, respectively. KCL score was significantly correlated with peak VO2 (r = -0.527, P < 0.001) and peak work rate (r = -0.632, P < 0.001). In patients with frailty (n = 63), the peak work rate (WR) was significantly lower than it was in patients without frailty (n = 29; 39.9 versus 69.5 W, respectively; P < 0.001). Multivariate analysis revealed that peak WR and peak systolic blood pressure were significant, independent predictors of frailty (ß = -0.108 and -0.045, respectively). In a diagnostic performance plot analysis, a cutoff value for peak WR of 51.9 W was the best predictor of frailty.Frailty status was significantly associated with peak WR and peak systolic blood pressure in elderly patients with stable HF. Therefore, cardiopulmonary exercise testing may be useful for assessing frailty status in this patient population.

Decreased Serum Albumin Predicts Bleeding Events in Patients on Antiplatelet Therapy After Percutaneous Coronary Intervention.

BACKGROUND: Antiplatelet therapy (APT) after percutaneous coronary intervention (PCI) prevents ischemic events with increased risk of bleeding. Little is known about the relationship between hypoalbuminemia and bleeding risk in patients receiving APT after PCI. This study investigated the association between serum albumin level and bleeding events in this population.Methods and Results:We enrolled 438 consecutive patients who were prescribed dual APT (DAPT; aspirin and thienopyridine) beyond 1 month after successful PCI without adverse events. The patients were divided into 3 groups according to serum albumin tertile: tertile 1, ≤3.7 g/dL; tertile 2, 3.8-4.1 g/dL; and tertile 3, ≥4.2 g/dL. Adverse bleeding events were defined as Bleeding Academic Research Consortium criteria types 2, 3, and 5. During the median follow-up of 29.5 months, a total of 30 adverse bleeding events were observed. Median duration of DAPT was 14 months. The tertile 1 group had the highest risk of adverse bleeding events (event-free rate, 83.1%, 94.3% and 95.8%, respectively; P<0.001). On Cox proportional hazards modeling, serum albumin independently predicted adverse bleeding events (HR, 0.10, 95% CI: 0.027-0.39, P=0.001, for tertile 3 vs. tertile 1). CONCLUSIONS: Decreased serum albumin predicted bleeding events in patients with APT after PCI.

Nutrition Status Predicts Severity of Vascular Calcification in Non-Dialyzed Chronic Kidney Disease.

BACKGROUND: Vascular calcification is a major complication in chronic kidney disease (CKD) that increases the risk of adverse clinical outcomes. Geriatric nutritional risk index (GNRI) is a simple nutritional assessment tool that predicts poor prognosis in elderly subjects. The purpose of the present study was to evaluate the correlation between GNRI and severity of vascular calcification in non-dialyzed CKD patients.Methods and Results:We enrolled 323 asymptomatic CKD patients. To evaluate abdominal aortic calcification (AAC), we used aortic calcification index (ACI) determined on non-contrast computed tomography. The patients were divided into three groups according to GNRI tertile. Median ACI significantly decreased with increasing GNRI tertile (15.5%, 13.6%, and 7.9%, respectively; P=0.001). On multivariate regression analysis GNRI was significantly correlated with ACI (ß=-0.15, P=0.009). We also investigated the combination of GNRI and C-reactive-protein (CRP) for predicting the severity of AAC. Low GNRI and high CRP were significantly associated with severe AAC, compared with high GNRI and low CRP (OR, 4.07; P=0.004). CONCLUSIONS: GNRI was significantly associated with AAC in non-dialyzed CKD patients.

Combinatorial screening of halide perovskite thin films and solar cells by mask-defined IR laser molecular beam epitaxy.

As an extension of combinatorial molecular layer epitaxy via ablation of perovskite oxides by a pulsed excimer laser, we have developed a laser molecular beam epitaxy (MBE) system for parallel integration of nano-scaled thin films of organic-inorganic hybrid materials. A pulsed infrared (IR) semiconductor laser was adopted for thermal evaporation of organic halide (A-site: CH3NH3I) and inorganic halide (B-site: PbI2) powder targets to deposit repeated A/B bilayer films where the thickness of each layer was controlled on molecular layer scale by programming the evaporation IR laser pulse number, length, or power. The layer thickness was monitored with an in situ quartz crystal microbalance and calibrated against ex situ stylus profilometer measurements. A computer-controlled movable mask system enabled the deposition of combinatorial thin film libraries, where each library contains a vertically homogeneous film with spatially programmable A- and B-layer thicknesses. On the composition gradient film, a hole transport Spiro-OMeTAD layer was spin-coated and dried followed by the vacuum evaporation of Ag electrodes to form the solar cell. The preliminary cell performance was evaluated by measuring I-V characteristics at seven different positions on the 12.5 mm × 12.5 mm combinatorial library sample with seven 2 mm × 4 mm slits under a solar simulator irradiation. The combinatorial solar cell library clearly demonstrated that the energy conversion efficiency sharply changes from nearly zero to 10.2% as a function of the illumination area in the library. The exploration of deposition parameters for obtaining optimum performance could thus be greatly accelerated. Since the thickness ratio of PbI2 and CH3NH3I can be freely chosen along the shadow mask movement, these experiments show the potential of this system for high-throughput screening of optimum chemical composition in the binary film library and application to halide perovskite solar cell.

Sirolimus-eluting stent vs. everolimus-eluting stent for coronary intervention in patients on chronic hemodialysis.

BACKGROUND: Even in the drug-eluting stent era, adverse cardiac events, including restenosis after percutaneous coronary intervention (PCI), have been more frequently seen in patients on hemodialysis (HD) than in non-HD patients. The objective of this study was to compare the sirolimus-eluting stent (SES) and everolimus-eluting stent (EES) for prevention of adverse cardiac events, including restenosis, in HD patients. METHODS AND RESULTS: A total of 100 consecutive patients on HD who underwent PCI were enrolled and randomly assigned to receive SES or EES. Although there was no difference between the 2 groups in baseline patient and lesion characteristics, the angiographic restenosis rate at 8-month follow-up was 21.2% in the SES group and 8.7% in the EES group (P = 0.041). Significant differences were also seen in % diameter stenosis (%DS), minimal lumen diameter, and late lumen loss at 8-month follow-up (P = 0.0024, P = 0.0040, and P = 0.033, respectively). During the 1-year follow-up, major adverse cardiac events occurred in 11 (22.0%) patients in the SES group and in 5 (10.0%) patients in the EES group (P = 0.10). CONCLUSIONS: The use of EES was as safe as that of SES. Moreover, EES significantly prevented restenosis in patients on maintenance HD compared with SES.

Dynamic Control of Intramolecular Rotation by Tuning the Surrounding Two-Dimensional Matrix Field.

The intramolecular rotation of 4-farnesyloxyphenyl-4,4-difluoro-4-bora-3a,4a-diaza- s-indacene (BODIPY-ISO) was controlled by tuning its local physical environment within a mixed self-assembled monolayer at an air-water interface. Intramolecular rotation was investigated by considering the twisted intramolecular charge transfer (TICT) fluorescence of BODIPY-ISO, which increases in intensity with increasing viscosity of the medium. In situ fluorescence spectroscopy was performed on mixed monolayers of BODIPY-ISO with several different lipids at the air-water interface during in-plane compression of the monolayers. Depending on the identity of the lipid used, the fluorescence of the mixed monolayers could be enhanced by mechanical compression, indicating that the rotation of BODIPY-ISO can be controlled dynamically in mixtures with lipids dispersed at the air-water interface. Taken together, our findings provide insight into strategies for controlling the dynamic behavior of molecular machines involving mechanical stimuli at interfaces.

Impact of Skeletal Muscle Mass on Long-Term Adverse Cardiovascular Outcomes in Patients With Chronic Kidney Disease.

Sarcopenia, defined as skeletal muscle loss and dysfunction, is attracting considerable attention as a novel risk factor for cardiovascular events. Although the loss of skeletal muscle is common in chronic kidney disease (CKD) patients, the relation between sarcopenia and cardiovascular events in CKD patients is not well defined. Therefore, we aimed to investigate the relation between skeletal muscle mass and major adverse cardiovascular events (MACE) in CKD patients. We enrolled 266 asymptomatic CKD patients (median estimated glomerular filtration rate: 36.7 ml/min/1.73 m2). To evaluate skeletal muscle mass, we used the psoas muscle mass index (PMI) calculated from noncontrast computed tomography. The patients were divided into 2 groups according to the cut-off value of PMI for MACE. There were significant differences in age and body mass index between the low and high PMI groups (median age: 73.5 vs 69.0 years, p = 0.002; median body mass index: 22.6 vs 24.2 kg/m2, p <0.001, respectively). During the follow-up period (median: 3.2 years), patients with low PMI had significantly higher risk of MACE than those with high PMI (31.7% and 11.2%, log-rank test, p <0.001). The Cox proportional hazard model showed that low PMI is an independent predictor of MACE in CKD patients (hazard ratio 3.98, 95% confidence interval 1.65 to 9.63, p = 0.0022). In conclusion, low skeletal muscle mass is an independent predictor of MACE in CKD patients. The assessment of skeletal muscle mass may be a valuable screening tool for predicting MACE in clinical practice.

Correlations between geriatric nutritional risk index and peripheral artery disease in elderly coronary artery disease patients.

AIM: Malnutrition is associated with the development of atherosclerosis and an increased risk of cardiovascular mortality in elderly patients. The present study aimed to investigate the association between the Geriatric Nutritional Risk Index (GNRI), a simple nutritional assessment tool, and the prevalence of peripheral artery disease (PAD) in elderly coronary artery disease patients. METHODS: We evaluated 228 elderly coronary artery disease patients (mean age 74.0 ± 5.7 years). Ankle-brachial index (ABI) measurements were routinely carried out to investigate the prevalence of lower extremity PAD. Patients showing ABI <0.9 were defined as having PAD. RESULTS: Based on our findings, 20.6% of the study patients had PAD. The median GNRI values were significantly lower in patients with PAD than those in patients without PAD (93.8 vs 100.0, P < 0.001). Even after multivariate adjustment, GNRI values were independently associated with PAD (odds ratio 0.94; 95% confidence interval 0.89-0.99; P = 0.024). Furthermore, patients with low GNRI and high C-reactive protein levels had a 5.5-fold higher risk of having PAD than those with high GNRI and low C-reactive protein levels. CONCLUSIONS: GNRI values showed a strong relationship with PAD in elderly coronary artery disease patients. These data reinforce the utility of GNRI as a screening tool in clinical practice. Geriatr Gerontol Int 2017; 17: 1057-1062.

Impact of nutritional assessment and body mass index on cardiovascular outcomes in patients with stable coronary artery disease.

BACKGROUND: An inverse association between obesity, as defined by body mass index (BMI) and prognosis has been reported in patients with cardiovascular disease ("obesity paradox"). The aim of this study was to investigate whether adding nutritional information to BMI provides better risk assessment in patients undergoing elective percutaneous coronary intervention (PCI). METHOD: This study comprised 1004 patients undergoing elective PCI. We calculated each patient's controlling nutritional status (CONUT) score for nutritional screening at baseline. Patients were divided into 4 groups based on CONUT score (low, 0-1 [<75th percentile]; or high, ≥2 [≥75th percentile]) and BMI (normal, 18.5-24.9kg/m2; or high, ≥25kg/m2). The endpoint was major adverse cardiac events (MACE) defined as cardiac death and/or myocardial infarction. RESULTS: Low CONUT score+normal BMI, low CONUT score+high BMI, high CONUT score+normal BMI, and high CONUT score+high BMI were determined in 374, 242, 275, and 113 patients, respectively. During a median follow-up of 1779 days, 73 events occurred. High CONUT score+normal BMI showed a 2.72-fold increase in the incidence of MACE (95% CI 1.46-5.08, p=0.002) compared with low CONUT score+normal BMI after adjusting for confounding factors. On the other hand, no significant difference in the incidence of MACE was observed in the other three groups. CONCLUSION: The combination of CONUT score and BMI was a useful predictor of MACE in this population. Using BMI to assess the cardiovascular risk may be misleading unless the nutritional information is considered.

Impact of Geriatric Nutritional Risk Index on cardiovascular outcomes in patients with stable coronary artery disease.

BACKGROUND: The association between malnutrition and cardiovascular prognosis in patients with stable coronary artery disease remains unclear. The aim of this study was to evaluate the association between Geriatric Nutritional Risk Index (GNRI), a simple tool to assess nutritional risk, and long-term outcomes after elective percutaneous coronary intervention (PCI). METHODS: This study consisted of 802 patients (age, 70±10 years, male, 69%) who underwent elective PCI. GNRI was calculated at baseline as follows: GNRI=[14.89×serum albumin (g/dl)+[41.7×(body weight/body weight at body mass index of 22)]]. Patients were then divided into three groups as previously reported: GNRI <92, 92 to ≤98, and >98. The endpoint of this study was the composite of cardiac death or non-fatal myocardial infarction. RESULTS: During a median follow-up period of 1568 days, 56 cardiac events occurred. Using Kaplan-Meier analysis, the 4-year event-free rates were found to be 79% for GNRI <92, 90% for GNRI 92 to ≤98, and 97% for GNRI >98 (log-rank test p<0.001). GNRI <92 and GNRI 92 to ≤98 showed 6.76-fold [95% confidence interval (CI) 3.13-14.56, p<0.001] and 3.03-fold (HR 3.03, 95%CI 1.36-6.78, p=0.007) increase in the incidences of cardiac death or non-fatal myocardial infarction compared with GNRI >98 after adjusting for confounding factors. CONCLUSION: GNRI significantly associated with cardiac events after elective PCI. Further studies should be performed to establish appropriate therapeutic strategies for this vulnerable patient group.

Impact of Paradoxical Decrease in High-density Lipoprotein Cholesterol Levels After Statin Therapy on Major Adverse Cardiovascular Events in Patients with Stable Angina Pectoris.

PURPOSE: Statin therapy usually increases HDL-C levels. However, a paradoxical decrease in HDL-C levels after statin therapy is often seen in clinical settings. The relationship between a paradoxical decrease in HDL-C levels after statin therapy and adverse cardiovascular events in patients with stable angina pectoris (SAP) is not well understood. The purpose of this study was to analyze the relationship between paradoxical HDL-C decreases after statin therapy and major adverse cardiovascular events (MACEs) in patients undergoing percutaneous coronary intervention (PCI) for SAP. METHODS: Between January 2006 and March 2015, 867 patients underwent PCI for SAP. Of them, we enrolled 209 patients who were newly started on statin therapy before PCI. We excluded patients who had started statin therapy earlier than 6 months before PCI, patients who had not started statin therapy after PCI, and patients who were diagnosed with acute coronary syndrome. They were divided into 2 groups according to the change in their HDL-C levels between baseline and 6 to 9 months after the index PCI: decreased HDL group after statin treatment (80 patients) and increased HDL group (129 patients). The primary end points were MACEs defined as a composite of all-cause death, nonfatal acute myocardial infarction, and target vessel revascularization (TVR). FINDINGS: Using Kaplan-Meier analysis, the 7-year event rate for composite MACEs in the decreased HDL group was found to be higher than that for the increased HDL group (38% versus 24%, log-rank P = 0.02). TVR occurred more frequently in the decreased HDL group than in the increased HDL group (32% versus 12%, log-rank P = 0.01). With the use of multivariate analysis, changes in HDL-C levels after statin therapy indicated a significant inverse association with the increased risk of MACEs, (hazard ratio [HR] = 0.94; 95% CI, 0.92-0.97; P < 0.01). The incidence of MACEs was more strongly associated with ΔHDL than with ΔLDL. Moreover, BMS usage also independently predicted MACEs (HR = 2.18; 95% CI, 1.14-4.17; P < 0.01). IMPLICATIONS: A paradoxical decrease in HDL-C levels after statin therapy might be a risk factor for MACEs, especially TVR, in patients with SAP.

Predictors of abdominal aortic calcification progression in patients with chronic kidney disease without hemodialysis.

BACKGROUND AND AIMS: Abdominal aortic calcification (AAC) is an important predictor of cardiovascular mortality in patients with chronic kidney disease (CKD). However, little is known regarding AAC progression in these patients. This study aimed to identify risk factors associated with AAC progression in patients with CKD without hemodialysis. METHODS: We recruited 141 asymptomatic patients with CKD without hemodialysis [median estimated glomerular filtration rate (eGFR), 40.3 mL/min/1.73 m2] and evaluated the progression of the abdominal aortic calcification index (ACI) over 3 years. To identify risk factors contributing to the rate of ACI progression, the associations between baseline clinical characteristics and annual change in ACI for each CKD category were analyzed. The annual change of ACI (ΔACI/year) was calculated as follows: (second ACI - first ACI)/duration between the two evaluations. RESULTS: Median ΔACI/year values significantly increased in advanced CKD stages (0.73%, 0.87%, and 2.24%/year for CKD stages G1-2, G3, and G4-5, respectively; p for trend = 0.041). The only independent risk factor for AAC progression in mild to moderate CKD (G1-3, eGFR ≥ 30 mL/min/1.73 m2) was pulse pressure level (ß = 0.258, p = 0.012). In contrast, parathyroid hormone (PTH) level was significantly correlated with ΔACI/year (ß = 0.426, p = 0.007) among patients with advanced CKD (G4-5, eGFR < 30 mL/min/1.73 m2). CONCLUSIONS: This study suggests that the AAC progression rate was significantly accelerated in patients with advanced CKD. In addition, measuring PTH is useful to evaluate both bone turnover and AAC progression in patients with advanced CKD.

Impact of adipose tissue composition on cardiovascular risk assessment in patients with stable coronary artery disease.

BACKGROUND AND AIMS: Visceral adipose tissue (VAT), unlike subcutaneous adipose tissue (SAT), is highly correlated with cardiovascular risk factors. This study aimed to evaluate the predictive value of adipose tissue composition, as measured by computed tomography, for cardiovascular events in patients with stable coronary artery disease. METHODS: 357 consecutive patients who underwent 64-slice computed tomography and elective percutaneous coronary intervention (PCI) were recruited. The ratio of visceral to subcutaneous adipose tissue (VAT/SAT) was calculated. Patients were divided into three groups in accordance with VAT/SAT (low VAT/SAT, <0.55 [<25th percentile]; moderate VAT/SAT, 0.55-1.03 [25th-75th percentile]; high VAT/SAT, ≥1.03 [≥75th percentile]). The investigated risk factors were hypertension, hyperglycaemia, and dyslipidaemia. We analysed the incidence of major adverse cardiovascular events (MACE), defined as the composite of cardiac death, myocardial infarction, and any revascularization. RESULTS: The rate of patients with two or more concomitant risk factors was significantly higher in the high VAT/SAT group (p = 0.006). During 1480 person-years, 109 events were documented. There was a significant association between the incidence of MACE and VAT/SAT, with the worst event-free survival rate in the high VAT/SAT group (log-rank, p = 0.01). In Cox analysis, the hazard ratio of high VAT/SAT for MACE was 2.72 (95% confidence interval 1.04-7.09, p = 0.04) compared with the low VAT/SAT after adjustment for confounding factors. CONCLUSIONS: Increased VAT/SAT is independently associated with the incidence of MACE, indicating that adipose tissue composition is a useful predictor of cardiovascular outcome, after elective PCI.

Sirolimus- vs. paclitaxel-eluting stent to coronary intervention in dialysis patients.

BACKGROUND: Patients on maintenance hemodialysis (HD) are at high risk for restenosis after percutaneous coronary intervention (PCI) even if treated with a sirolimus-eluting stent (SES). The aim of this study was to compare the effects of SES and paclitaxel-eluting stent (PES) in preventing restenosis in HD patients with coronary artery disease. METHODS: A total of 100 consecutive patients on HD who underwent PCI were enrolled into the study. They were randomly assigned to receive either SES or PES. We compared follow-up angiographic outcomes between the SES and PES groups at 8-month follow-up. RESULTS: The angiographical restenosis rate, defined as % diameter stenosis>50% at 8-month follow-up, was 19.7% in the SES group and 20.0% in the PES group (p=0.97). Late loss was also similar between the two groups (0.49±0.70 mm vs. 0.48±0.91 mm, P=0.94). There were no significant differences in the rates of all-cause death, non-fatal myocardial infarction, or TLR due to stent restenosis-induced ischemia between the two groups (2.0% vs. 4.0%, p=0.56, 2.0% vs. 4.0%, p=0.56, and 16.0% vs. 12.0%, p=0.57, respectively). CONCLUSIONS: There was no significant difference in angiographical outcome at 8-month follow-up between HD patients treated with SES and PES. Even if treated with DES including SES and PES, patients on HD are at high risk of restenosis after PCI.