bluuebunny/crossref_metadata_2025_split huggingface dataset:
P6263Results of percutaneous coronary intervention of unprotected left main coronary disease in octogenarian patients
Authors: Romero Reyes M J , Moya Martin A , Gonzalez Alemany N , Sanchez Burguillos F J , Perez Santigosa P , Rufian Andujar S , Molano Casimiro F J 
Published: October 2019

Abstract

Introduction
Conservative treatment unprotected left main coronary (uLMCA) disease has a high mortality rate (50% at 3 years). Since octogenarian patients are often dismissed for surgical treatment, they tend to adopt a more conservative attitude in this population.

Purpose
We report medium and long-term outcomes of percutaneous coronary intervention (PCI) for uLCMA stenosis in elderly patients.

Methods
Retrospective cohort study of consecutive patients ≥80 years with uLMCA stenosis, treated with PCI at a single center between June 2005 and February 2017.

Results
A total of 100 patients were included in the study. 58% were male, with a mean age of 83.8±3 years. There were 86% hypertensive, 63% diabetic and 68% dyslipidemic. 14% of the patients had an LVEF ≤35%. Unstable angina (45%) and acute coronary syndromes withouth ST-segment elevation (44%) were the most common presentation. In 9% of the cases, cardiogenic shock was the initial presentation form. The distal left main coronary was the most frequent localitation of the lesion (46%) followed by the ostium (33%). In 63% of the cases, a multivessel coronary disease was detected and in 47% the revascularization was incomplete.
The survival rate after a year follow up was 79% and after three years follow up was 65%. However, in most of the cases the cause of death was due to other comorbidities, with cardiac death being 10% per year and 13% at 3 years of follow-up. The rate of non-fatal acute myocardial infarction was 13% per year, increasing to 20% at 3 years of follow-up. There was a 9% stent restenosis implanted at 3 years. The presence of severe left ventricular systolic dysfunction was the main predictor of mortality in long-term follow-up (OR 1.39 [95% CI 1.10–1.752], p<0.001). Incomplete revascularization was not associated with a higher mortality rate.

Conclusion
PCI is a safety option for revascularization in uLMCA stenosis in elderly patientes with excellent short-term results, as well as acceptable long-term results. Age should not be a handicap to consider uLMCA revascularization in this population.
P6273Inhibitory phosphorylation of RIP1 at Ser320 induces nuclear translocation of TFEB, leading to suppression of necroptosis in cardiomyocytes
Authors: Abe K , Yano T , Sato T , Kouzu H , Kuno A , Tanno M , Miki T , Miura T 
Published: October 2019

Abstract

Background
Necroptosis, a form of programmed necrosis, has been shown to contribute to the pathogenesis of various diseases including ischemia/reperfusion injury and heart failure. We recently reported that necroptotic signals suppresses autophagy in cardiomyocytes and that rapamycin, an mTORC1 inhibitor, not only promotes autophagy but also protect the cells from necroptosis.

Purpose
We examined the mechanism by which rapamycin suppresses necroptosis of cardiomyocytes, focusing on regulation of RIP1 activity and autophagic flux.

Methods and results
In H9c2 cardiomyoblasts, necroptosis was induced by treatment with TNF and z-VAD-fmk (zVAD) for 24 h, and cell death was determined by LDH release (as % of total). The treatment with TNF/zVAD increased LDH release from 3.4±1.3% to 46.1±2.3%, and LDH release was suppressed by necrostatin-1 (5.9±0.9%), a RIP1 inhibitor, and by rapamycin (23.5±1.4%). The protective effect of rapamycin was mimicked by Ku-0063794, an mTORC1/2 inhibitor. TNF/zVAD induced RIP1-RIP3 complex formation, together with suppression of TNF-induced RIP1 cleavage, which was mitigated by rapamycin. In addition, rapamycin not only suppressed TNF/zVAD-induced phosphorylation of RIP1-Ser166, an index of RIP1 activation, but also increased phosphorylation of RIP1-Ser320, an inhibitory phosphorylation site. In cells transfected with RIP1-S320A, which lack Ser320 for inhibitory phosphorylation, rapamycin failed to suppress TNF/zVAD-induced RIP1-RIP3 binding and cell death. Immunoblot analyses showed that TNF/zVAD significantly increased level of LC3-II. The accumulation of LC3-II protein was not further increased by bafilomycin A1 (100 nM), an inhibitor of lysosomal protein degradation, indicating that accumulation of LC3-II by TNF/zVAD reflected suppression of autophagic flux. Inhibition of RIP1 by necrostatin-1 attenuated TNF/zVAD-induced accumulation of LC3 II. The restoration of autophagic flux in TNF/zVAD-treated cells by necrostatin-1 was confirmed by monitoring tandem RFP-GFP-LC3 transfected cells; necrostatin-1 increased a ratio of RFP-LC3-puncta (autolysosomes) to RFP-GFP-LC3-puncta (autophagosomes) in TNF/zVAD-treated cells. In addition, necrostatin-1 and rapamycin induced nuclear translocation of TFEB, a regulator of lysosome biogenesis, which was associated with upregulation of MCOLN1 mRNA, a downstream target of TFEB. Restoration of autophagic flux in TNF/zVAD-treated cells by necrostatin-1 was inhibited by siRNA-mediated knockdown of TFEB.

Conclusion
Activation of TFEB by inhibitiory phosphorylation of RIP1-Ser320 is a primary mechanism of cytoprotection afforded by mTORC1 inhibition against necroptosis.
P6276Impact of the cardiac specific deletion of AMPKalpha2 on the contractile and metabolic phenotype of the heart in male and female mice
Authors: Grimbert L , Sanz M N , Rucker-Martin C , Novotova M , Gressette M , Gomez S , Solgadi A , Dumont F , Ventura-Clapier R , Veksler V , Piquereau J , Garnier A 
Published: October 2019

Abstract

Introduction
Mitochondrial dysfunction plays a major role in the Heart Failure (HF) pathophysiology.The AMP activated protein kinase (AMPK) is activated by a high AMP-ADP/ATP ratio and regulates a number of metabolic pathways. Many studies have highlighted a protective role of AMPK in HF, but its relevance to cardiac tissue, its metabolic part and its sex specificity are not well established.

Purpose
Then, the aim of this study is to determine the role of AMPK in the healthy and failing heart in male and female mice.

Methods
We developed and validated a mouse strain with an adult-inducible cardiac-specific deletion of AMPKα2, the major cardiac isoform, using the Cre-Lox system (40mg/kg tamoxifen injection on two consecutive days at adult age). At four months after the deletion, cardiac contractility, morphology and metabolism were studied in control and KO mice from both sexes.

Results
We observed only in male KO mice a decrease of left ventricular ejection fraction (−10%), an increase of the total fibrosis (+64%) and defects in mitochondrial structures. Male KO mice also showed a reduced (−28%) mitochondrial respiration via complex I associated with a different cardiolipin species distribution.

Conclusion
Our results reveal in adult healthy hearts, a sex-specificity in the effects of AMPKα2 deletion, leading to impaired contractile function related to metabolic and non-metabolic alterations only in male mice.
P6281Over expression of HINT2 preserves the mitochondria function of cardiomyocytes via regulating the NAD+ homeostasis
Authors: Mengkang F , Qian Q J Y 
Published: October 2019

Abstract

Background
HINT2, a member of the histidine triad proteins family, has been proved to positively regulate mitochondrial function and energy metabolism.However, its effect in cardiomyocytes is largely unknown.

Purpose
To explore the effects of HINT2 on mitochondrial function and ascertain the underlying mechanism in cardiomyocytes

Methods
The neonatal mouse cardiomyocytes was isolated and cultured for subsequent research. Hypoxia as a adverse factor was used to stimulate the cardiomyocytes for 12 hours. The expression of HINT2 in neonatal mouse cardiomyocytes was determined by real-time quantitative polymerase chain reaction and immunoblotting. We used adenovirus vector to over express or knock down HINT2 in cardiomyocytes.Cell viability were assayed using CCK-8 kit. The mitochondria function was evaluated via detection of oxygen consumption rates and mitochondrial membrane potential. The mitochondrial membrane potential was observed using confocal microscopy after TMRM fluorescence probe labeling. The OCR were measured with a Seahorse Extracellular Flux (XF)-96 Bioanalyzer. The NAD+,as a hydrogen carrier for oxidation respiratory chain, was measured by colorimetric kit.The crystal structure data of HINT2 protein and NAD+ were subjected to rigid body molecular docking using AUTODOCK4.

Results
The expression of HINT2 was down-regulated in cardiomyocytes after hypoxic stimulus for 12 hours. Over expression of HINT2 protected the cardiomyocytes from hypoxia induced reduced cell viability. Moreover,the mitochondrial membrane potential was higher and the basal, ATP-linked and maximum OCR were increased in HINT2 overexpressing cells under hypoxic stimulus. Conversely,the mitochondrial membrane potential, the basal, ATP-linked and maximum OCR were decreased in cell with HINT2 under expression in normoxia condition. Furthermore,the NAD+ levels was increased in HINT2 overexpressing cells and decreased in cells with knock-down expression of HINT2. Besides,The beneficial effects of HINT2 overexpression can be blocked by nicotinamide phosphoribosyltransferase inhibitor. Moclecular docking results showed that HINT2, as a nucleotide binding protein could docking with NAD+, and the conformation with the strongest binding force has 2 hydrogen bonds which was at ARG99:HH21 and GLN102:HE22 in HINT2.

Conclusions
Our results suggest that overexpression of HINT2 preserve the mitochondrial function in cardiomyocytes via maintaining NAD+ homeostasis.

Acknowledgement/Funding
Natural Science Foundation of China
P6283Downregulation of Tim44 exacerbates oxidative stress-induced ROS production and cardiomyocytes death by reducing mitochondrial SOD2
Authors: Ikeda S , Matsushima S , Okabe K , Ishikita A , Tadokoro T , Enzan N , Yamamoto T , Sada M , Deguchi H , Ikeda M , Ide T , Tsutsui H 
Published: October 2019

Abstract

Background
Mitochondrial dysfunction has been highlighted as a critical driver of cardiac remodeling and failure. Mitochondria contains about 1500 proteins, 99% of which are encoded in the nuclear genome. Therefore, protein import into mitochondria is essential to maintain mitochondrial function. Previous reports suggest that nuclear-encoded mitochondrial precursor proteins import into mitochondria by multiple complex; translocase of outer membrane (TOM), translocase of inner membrane (TIM), and protein associated motor (PAM). However, the role of these protein import machineries of mitochondria in cardiac remodeling remains to be elucidated.

Objective
The purpose of this study was to elucidate the role of TOM, TIM, and PAM complex in cardiac remodeling and cardiomyocyte death.

Methods and results
C57BL/6J mice were subjected to myocardial infarction (MI) by permanent ligation of left anterior descending artery. Four weeks after operation, MI-mice demonstrated left ventricular (LV) dilation (LV end-diastolic dimension: 3.91 vs. 5.54 mm, n=8–11, p<0.05) and dysfunction (LV fractional shortening: 33.3 vs. 7.7%, n=8–11, p<0.05). Tim44 protein levels, a component of PAM complex, in mitochondrial fraction from non-infarcted left ventricle were significantly decreased compared with those in the heart from sham-operated mice by 39% (p<0.05), whereas other proteins related to TOM, TIM and PAM complex such as Tom20, Tom22, Tom40, Tom70, Tim22, Tim23 and mtHSP70 were not altered between MI-mice and sham-mice. In addition, blue-native polyacrylamide gel electrophoresis revealed that a protein complex associated to Tim44 was significantly decreased in non-infarcted LV by 40% (p<0.05). Superoxide dismutase 2 (SOD2), a mitochondrial matrix protein, was decreased in mitochondrial fraction from non-infarcted LV by 20% (p<0.05), accompanied by enhancing protein carbonylation, a marker of oxidative stress, by 40% (p<0.05). To assess the role of Tim44, it was downregulated by small interfering RNA in cultured neonatal rat ventricular myocytes (NRVMs). Knockdown of Tim44 significantly decreased SOD2 protein levels in mitochondrial fractionation (22%, p<0.05), with no significant changes in its mRNA levels. Furthermore, knockdown of Tim44 significantly increased protein carbonylation (20%, p<0.05) and cleaved caspase 3 (47%, p<0.05) and decreased cell viability (69%, p<0.05), assessed by cell titer assay, in H2O2-treatred NRVMs.

Conclusions
Downregulation of Tim44 exacerbates oxidative stress-induced ROS production and cardiomyocytes death, which is associated with a decrease in mitochondrial SOD2. Endogenous Tim44 might play a protective role in cardiac remodeling by attenuating oxidative stress and cardiomyocyte death via SOD2 import into mitochondria.
P6287Analysis of novel cardiac markers sST2 and IL-33 in chronic heart failure with reduced ejection fraction
Authors: Jirak P , Lichtenauer M , Wernly B , Paar V , Jung C , Yilmaz A , Hoppe U , Schulze P C , Kretzschmar D , Pistulli R 
Published: October 2019

Abstract

Background
Soluble (s) ST-2 has been recently evaluated as a monitoring parameter in heart failure (HF). Besides being a marker for cardiac strain and hemodynamic stress, studies also found an influence of ST2 on the immune system, above all mediated through its Janus-Face ligand IL-33, an alarmin released under stress conditions or by cellular death. In contrast to sST2, the role of IL-33 in HF is yet unknown.

Objective
In this project, we aimed for an analysis of the ST2/IL33 pathway in patients with heart failure with reduced ejection fraction (HFrEF).

Methods
In total, 200 patients were included in the study: 59 with ischemic (ICM), 65 with dilated (DCM) cardiomyopathy (mean LVEF 38%), as well as 76 control patients without coronary artery disease or signs of heart failure. Serum samples were analyzed by use of ELISA after informed consent.

Results
sST2 showed a significant elevation in all HF patients (p<0.0001) compared to the control group. No significant differences in levels of sST2 were observed between ICM and DCM patients. In contrast to sST2, no differences between HF patients and control group were observed for IL-33. Furthermore, sST2 showed a significant correlation with CRP (p<0.001, r=0.28), NT-pro-BNP (p<0.0001, r=0.40) and an inverse correlation with ejection fraction (p<0.0001, r=−0.40). Additionally, sST2 showed a significant elevation in patients in NYHA stages I-II (p=0.030) and NYHA stages III-IV (p<0.01). Again, no significant correlations were observed between IL-33 and parameters mentioned above.
Analysis of sST2 in heart failure

Conclusions
We observed a significant increase and correlation with disease severity of sST2 in chronic HFrEF patients of both ischemic and non-ischemic origin, but contrary to our expectations, no significant changes in serum levels of IL-33. Thus, a mechanism independent of ST2/IL33 axis could be responsible of sST2 secretion in HF. Further studies including acute decompensated patients could provide a better understanding of the IL-33 role in HF.
P6289Neuregulin-1 improves cardiac function through protecting the loss of endothelial barrier in septic rats via RhoA/ROCK pathway
Authors: Wang L O N G , Kang W E N , Cheng Y U E , Wang X I 
Published: October 2019

Abstract

Background
The main path of sepsis-induced cardiomyopathy is the loss of endothelial barrier function. Neuregulin-1 (NRG-1) has beneficial effects on endothelial function.

Purpose
To investigate the effect of NRG-1 treatment on the protection of cardiac endothelial cells and the changes of RhoA/ROCK signaling in sepsis.

Methods
Rats were randomly divided into three groups: sham, LPS, and NRG-1. After successful induction of sepsis by lipopolysaccharide (LPS, 10mg/kg), rats were administered either a vehicle or recombinant human neuregulin-1 (rhNRG-1, 10μg/kg/d) for one or two days. We recorded their survival rate at 48h after sepsis. Hemodynamic methods were performed to assess cardiac function. We used immunofluorescence assay to detect von Willebrand Factor (vWF). Intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial growth factor (VEGF)levels in serum were measured by enzyme-linked immunosorbent assay (ELISA), and serum nitric oxide (NO) was detected by reductase method. We used transmission electron microscopy to observe changes in myocardial ultrastructure and western blot to assess expression of RhoA and ROCK1 protein.

Results
Sepsis impaired endothelial function manifested by increased ICAM-1, NO and VEGF levels in serum, NRG-1 treatment could significantly alleviate these increase (P<0.05). Compared with the vehicle, NRG-1 significantly decreased cardiac vascular permeability through increasing the expression of vWF on endothelial lining (P<0.05). Moreover, NRG-1 alleviated damages of ultrastructure of myocardial cells and suppressed the expression of RhoA and ROCK1 protein (P<0.05). Ultimately, NRG-1 improved the survival,and prevented hemodynamic derangement.
Levels of endothelial biomarker in serum ICAM-1 (pg/ml) VEGF (pg/ml) NO (pg/ml) Sham 24h 1564.74±94.41 0.84±0.28 203.27±1.81 Sham 48h 322.92±7.92 1.78±0.61 6.57±0.38 LPS 24h 5139.85±284.15† 3.11±0.04† 679.05±78.59† LPS 48h 950.41±25.23†‡ 4.65±0.30† 180.40±30.08†‡ NRG 24h 2772.18±164.45§ 0.30±0.04§ 355.14±23.41§ NRG 48h 578.57±28.97§‡ 1.02±0.45§ 104.67±2.13§‡ †p<0.05 vs. the sham group, §p<0.05 vs. the LPS group and ‡p<0.05 vs. the 24h group.
Effect of NRG-1 on endothelial cells

Conclusions
NRG-1 could alleviate endothelial injury in sepsis by strengthening the barrier function of vascular, reducing the secretion of endothelial-related biomarkers and inhibiting oxidative stress, thus improving cardiac function and survival rate, these effects may base on RhoA/ROCK signaling pathway. These may contribute to reverse the impaired endothelial cells in sepsis in the future.

Acknowledgement/Funding
The national natural science foundation of China (81772044)
P6292Role of CXCL12gamma isoform and its interactions with heparan-sulfates in post-ischemic cardiac remodeling
Authors: Duval V , Zlatanova I , Sun Y , Alayrac P , Gomez I , Lemitre M , Vilar J , Silvestre J S , Levoye A 
Published: October 2019

Abstract

Introduction
Myocardial infarction (MI) is a severe ischemic disease precipitating long-term adverse remodeling and heart failure. The chemokine CXCL12/SDF-1 is essential for cardiovascular system development and plays a prominent role in physio-pathological processes such as inflammation, angiogenesis and tissue fibrosis. In addition to the binding to its cognate receptors CXCR4 and CXCR7, CXCL12 interacts with heparan-sulfates (HS) which coordinate its biological activity. We have previously highlighted the essential role of CXCL12/HS interactions in vascular growth and remodeling in the setting of critical limb ischemia. In addition, studies in experimental model of MI revealed a protective role for the CXCL12α isoform, through the regulation of cardiomyocyte survival and recruitment of inflammatory cells. However, in mice, three CXCL12 isoforms (α, β and γ) have been identified and, among them, the CXCL12γ isoform shows an unchallenged ability to cooperate with HS, suggesting a putative pivotal role in tissue repair.

Objectives
The aim of the study is to analyze the role of CXCL12γ isoform and the importance of CXCL12/HS interactions in post-ischemic cardiac remodeling in an acute model of MI.

Methods
MI was induced by permanent ligation of the left ascending coronary artery in mice carrying a Cxcl12 gene mutation that precludes interactions with HS (Cxcl12Gagtm) and in Cxcl12γ knock-in animals (Cxcl12γ-KI) harboring CXCL12γ deficiency. Alternatively, the impact of CXCL12γ overexpression and the importance of its interactions with HS was also evaluated in wild-type (WT) mice receiving transcutaneous echo-guided injections of adenovirus encoding WT Cxcl12γ or HS-binding-disabled Cxcl12γ in cardiac tissue. Cardiac function and remodeling have been assessed through echocardiography analysis, evaluation of infarct size, interstitial fibrosis, vascular growth (capillary and arteriole densities) and inflammatory cell infiltration into the cardiac tissue.

Results
After MI, Cxcl12Gagtm and Cxcl12γ-KI animals exhibit reduction in cardiac function and adverse left ventricular remodeling when compared to their respective WT littermates. Interestingly, overexpression of CXCL12γ in WT mice cardiac restored cardiac function by reducing the size of the infarcted area, interstitial fibrosis and promoting vascular growth. In sharp contrast, HS–binding disabled CXCL12gamma mutants failed to improve cardiac function and to abrogate adverse left ventricular remodeling.

Conclusion
We show that CXCL12γ isoform plays an important role in the regulation of post-ischemic cardiac function and remodeling and that its interactions with HS are essential for adequate cardiac repair in the setting of acute MI.
P6298Pericardial transplantation of adipose-derived stromal cells with self-assembling peptides scaffold prevents cardiac remodeling after myocardial infarction in mice
Authors: Kondo N , Nagai T , Aoki I , Kanda M , Takahashi T , Kobayashi Y 
Published: October 2019

Abstract

Introduction and purpose
Cardiac cell therapy to patients with heart failure after myocardial infarction (MI), has shown little improvement in cardiac function. The rate of engraftment after cell transplantation (TX) to the ischemic heart is low because of the leakage of transplanted cells without scaffold, the friction between the graft and thorax, and poor vascularization to the graft. We created a novel TX method (Figure 1) to transplant adipose-derived stromal cells (ASCs) with scaffold into the pericardial space where the pericardium protected the graft. We evaluated the therapeutic efficacy of pericardial TX of ASCs on the cardiac function and remodeling after MI.

Methods
We isolated ASCs from the brown adipose tissue of donor mice (C57BL6-Tg (CAG-EGFP), five weeks). ASCs increased and differentiated into spontaneously beating myocytes, endothelial cells, and pericytes three weeks after ex vivo culture. Cells were trypsinized and mixed with self-assembling peptides scaffold. Each graft (100μl gel scaffold) had 1×106 ASCs. To make recipient MI mice (C57BL6, 12 weeks), we ligated the left coronary artery through small chest incision without cutting rib bone to avoid postoperative adhesions. We confirmed the deterioration of cardiac function by echocardiography (n=21) and MRI (n=4) three weeks after MI. Then, recipient MI mice had TX (syngeneic, no immunosuppression). We opened the pericardium and transplanted the graft into the pericardial space. By suturing the pericardium, the graft was fixed on the MI scar area (Figure 1). We labeled donor cells with GFP and the scaffold with biotin. We evaluated cardiac function of TX group (n=10) and control group (MI with sham TX, n=11) by echocardiography.

Results
In the scaffold, donor cells increased three days to two weeks after TX, and slightly decreased four weeks after TX. The graft thickness was 0.9±0.2mm (two weeks after TX) and 0.7±0.2mm (four weeks after TX). There were many vWF positive vessels in the scaffold and some of which were GFP positive. Echocardiography showed that left ventricular diastolic dimension (LVDd) of TX group, did not increase four weeks after TX (ΔLVDd = −0.02mm, P=0.80). While, LVDd of control group significantly increased (ΔLVDd = +0.23mm, P=0.02) due to cardiac remodeling after MI. MRI confirmed the increase in left ventricular wall thickness in the MI scar area up to 4 weeks in TX group (n=2).

Conclusions
Pericardial transplantation of ASCs prevents cardiac remodeling. Its beneficial effect might be mediated by improved rate of engraftment, neovascularization, and increased ventricular wall thickness in the MI scar area.

Acknowledgement/Funding
Grants-in-Aid for Scientific Research