IntroductionCardiac shock-wave therapy (CSWT) is a non-invasive regenerative treatment method based on low-frequency ultrasound waves, which stimulate angiogenesis. Current data about the effects of revascularization procedures on angiogenesis biomarkers is limited. Recently, an association of catestatin and endocan with coronary collateral development was shown in several trials. In this study, we aimed to evaluate the impact of CSWT on the dynamics of catestatin and endocan levels and to assess their correlation with parameters of myocardial perfusion and function.MethodsProspective, randomized, triple-blind, sham procedure-controlled study enrolled 72 adult subjects who complied with defined inclusion criteria (NCT02339454). We measured biomarkers in 48 patients with stable angina (24 patients of CSWT group, 24 patients of sham-procedure group). Additionally, patients were divided into responders and non-responders according to improvement in myocardial perfusion and/or contractility assessed by myocardial scintigraphy and dobutamine echocardiography (30 and 13 patients, respectively). The blood samples were collected at baseline, after the last treatment procedure (9th treatment week) and at 6-month follow-up to evaluate biomarkers concentration and stored at –80° until analysis. Serum catestatin and endocan levels were determined by commercially available ELISA kits.ResultsSerum catestatin concentration significantly increased in all patients. While endocan levels significantly decreased in the responders sub-group. The increase in catestatin levels at 9th week and 6 months was positively associated with improvement in summed difference score (rho = 0.356, p = 0.028) and wall motion score, WMS (rho = 0.397, p = 0.009) at 6 months in the whole study population. Meanwhile, the decrease in endocan levels over 6 months was positively correlated with improvement in WMS at 3- and 6- months (r = 0.378, p = 0.015 and r = 0.311, p = 0.045, respectively). ROC analysis revealed that a change at 6 months in catestatin and endocan levels significantly predicted improvement in myocardial perfusion and contractile function with 68.9% sensitivity and 75.0% specificity (p = 0.039) and 51.7% sensitivity, and 91.7% specificity (p = 0.017), respectively. Baseline endocan concentration and its change at 6 months predicted response to CSWT with 68.8% sensitivity and 83.3% specificity (p = 0.039) and 81.3% sensitivity and 100% specificity (p < 0.0001), respectively.ConclusionThis study demonstrates the association of increase in catestatin and decrease in endocan levels with the improvement of myocardial perfusion and contractile function. The potential predictive value of catestatin and endocan dynamics for the response to regenerative therapy is shown.

Abstract Background: Cardiac shock-wave therapy (CSWT) is a non-invasive treatment based on low-frequency ultrasound waves that stimulate angiogenesis. Current data on the effects of revascularization procedures on angiogenesis biomarkers are limited. In this study, we aimed to characterize the serum protein profiles of patients with coronary artery disease after CSWT treatment in relation to changes in exercise and imaging parameters. Methods: The study population included 10 patients from a prospective, randomized, triple-blind, sham-procedure controlled study (NCT02339454), who received CSWT and had demonstrated improvement in myocardial perfusion and/or contractility at 6 months follow-up The blood samples were collected at baseline, after the last treatment procedure (9th treatment week), at 6-month follow-up and stored at −80°C until analysis. The cardiovascular-related proteins (n=92) were measured using the Olink Proseek Multiplex Cardiovascular III panel (Olink, Uppsala, Sweden). Results: The median age was 65.5 years, and 7 patients were males. A total of 20 biomarkers showed significant changes from baseline to 9th week and 6 months follow-up (p<0.05). We identified 28 proteins that showed clear association with an increase in exercise tolerance and improvement in myocardial contractility or perfusion after CSWT. Using a volcano plot and results from the paired Mann-Whitney U test at confidence level of 0.95, we identified 3 proteins (PON3, TR-AP, CD163) with elevated values corresponding to increase in exercise duration (p<0.05) and 2 proteins (CPA1, COL1A1) related to improved myocardial contractility (p<0.05). A pathway analysis including 28 proteins suggested that these biomarkers were related to immune and inflammatory response, cell adhesion, tissue remodeling, proteolysis and catabolic processes. Conclusions: This study demonstrates the association of an increase in protein levels with an improvement in exercise duration and contractile function. Proteomic analysis suggests that CSWT exerts biological effects including immune and inflammation response, cell adhesion and tissue remodeling, all of which may mediate angiogenesis. Trial registration: Clinicaltrials.gov (NCT02339454).

Background: The soluble form of the IL-33 receptor (sST2) and Galectin-3 (Gal-3) are fibrosis biomarkers with prognostic value in heart failure (HF). We investigated the prognostic capacity of sST2 when combined with Gal-3, and determined if the prognostic utility of sST2 is affected by mineralocorticoid receptor antagonist (MRA) therapy. Methods: sST-2 and Gal-3 were measured in 101 stable chronic HF (CHF) patients receiving MRA therapy and compared to 97 BNP and cardiovascular risk factor matched patients not treated with MRA. sST2 and Gal-3 levels were measured to determine the relationship with all-cause mortality at 6-year follow-up. Results: ROC curve cut-off points were defined as sST2 = 36.3 ng/mL, Gal-3 = 17.8 ng/mL, and BNP = 500 pg/mL, and had 6-year mortality hazard ratios (HR) of 7.3, 6.6 and 5.4, respectively. The combination of an elevated sST2 and Gal-3 had a HR = 4.4 [95% CI 1.9-8.9]. Combining sST2 and Gal-3 to a clinical model relevant for CHF prognosis allowed a significant reclassification of 1-year adverse outcome risk, even when BNP was included. Finally, prognostic prediction by sST2 was unaffected by MRA treatment. Conclusion: Simultaneous sST2 and Gal-3 elevation is associated with poorer prognosis compared to either alone, regardless of BNP levels, and the prognostic capacity of sST2 is independent of MRA therapy.

Background: Insulin-like Growth Factor Binding Protein 2 (IGFBP2) showed greater heart failure (HF) diagnostic accuracy than the "grey zone" B-type natriuretic peptides, and may have prognostic utility as well. Objectives: To determine if IGFBP2 provides independent information on cardiovascular mortality in HF. Methods: A retrospective study of 870 HF patients from 3 independent international cohorts. Presentation IGFBP2 plasma levels were measured by EUSA, and patients were followed from 1 year (Maastricht. Netherlands) to 6 years (Atlanta, GA, USA and Toulouse, France). Multivariate analysis, Net Reclassification Improvement (NRI) and Integrated Discrimination Improvement (IDI) were performed in the 3 cohorts. The primary outcome was cardiovascular mortality. Results: In multivariate Cox proportional hazards analysis, the highest quartile of IGFBP2 was associated with mortality in the Maastricht cohort (adjusted hazard ratio 1.69 (95% CI 1.18-2.41), p = 0.004) and in the combined Atlanta and Toulouse cohorts (adjusted hazard ratio 2.04 (95%CI, 13-3.3), p = 0.003). Adding IGFBP2 to a clinical model allowed a reclassification of adverse outcome risk in the Maastricht cohort (NRI - 18.7% p = 0.03; IDI - 3.9% p = 0.02) and with the Atlanta/Toulouse patients (NRI of 40.4% p = 0.01, 31,2% p = 0.04, 315% p = 0,02 and IDI of 2,9% p = 0,0005, 3.1% p = 0,0005 and 4,2%, p = 0.0005, fora follow-up of 1, 2 and 3 years, respectively). Conclusion: In 3 international cohorts, IGFBP2 level is a strong prognostic factor for cardiovascular mortality in HF, adding information to natriuretic monitoring and usual clinical markers, that should be further prospectively evaluated for patients optimized care. (C) 2019 Elsevier B.V. All rights reserved.

Diagnostic biomarkers for heart failure (HF) such as the natriuretic peptides (NPs) are widely used but have limitations. Innovative biomarkers could provide improved diagnostic performance. We launched a prospective case-control proteomic study and investigated for polypeptides specific to HF through a capillary electrophoresis-mass spectrometry (CE-MS) proteomic analysis. The putative biomarker was identified by Orbitrap liquid chromatography-MS, validated by western blot, then by ELISA using plasmas from multicentric international cohorts. A rat model of HF was tested for biomarker expression levels. We identified insulin like growth factor binding protein 2 (IGFBP2) as a new diagnostic biomarker for HF with a high sensitivity and specificity (AUC = 0.93; 95% CI, 0.89–0.96; P b 0.0001) in the local cohort and IGFBP2 levels provided an AUC of 0.943 (95% CI, 0.860–1.026) which gave a 87% sensitivity in AHF and 90% specificity at the cut off value previously determined in the discovery cohort, i.e. 556 ng/ml. ROC curve analysis of IGFBP2 and NTproBNP showed an AUC of 0.784 (95% CI, 0.744–0.820) for IGFBP2 and a significantly higher AUC of 0.927 (95% CI, 0.900–0.949) for NT-proBNP, P < 0.0001 in a Dutch cohort. In this cohort, the optimal cut off value for IGFBP2 gave a sensibility of 71% (95% CI, 66–76) and a specificity of 75% (95% CI, 65–83). IGFBP2 is a new biomarker to diagnose HF, which could be used to provide additional information to the NPs. Animals models, will help in the evaluation of the putative IGFBP2 regulated mechanisms in HF.

Metabolic syndrome (MetS) is a known risk factor for cardiovascular events. It is characterized by central obesity plus any two of dyslipidemia, insulin resistance, and hypertension. Obese and diabetic patients with MetS have low plasma insulin-like growth factor binding protein 2 (IGFBP2). Our first aim was to investigate IGFBP2 cardiac expression in MetS patients and in a mice model of MetS. The second aim was to assess if gene therapy with adeno-associated virus 9 carrying human IGFBP2 (AAV9-hIGFBP2) could reduce MetS associated left ventricular hypertrophy in mice. We measured plasma IGFBP2 by ELISA and cardiac mRNA IGFBP2 expression in MetS patients by RT-qPCR. Both plasma levels and heart expression of IGFBP2 were decreased in patients with MetS vs. control patients. Further, in a C57BL/6J mouse model of diet-induced MetS, found similar left ventricular mRNA IGFBP2 expression. Finally, we demonstrated for the first time that in MetS mice with decreased cardiac IGFBP2 mRNA levels, human IGFBP2 can be induced by a single AAV9-hIGFBP2 injection, and that the increased IGFBP2 prevents left ventricle wall thickening, hypertrophy and dysfunction. Human plasma and cardiac IGFBP2 are decreased in MetS patients. In mice, restoration of cardiac IGFBP2 expression level prevents MetS associated left ventricular dysfunction and hypertrophy. These clinical and animal data suggest that IGFBP2 is a new cardiac marker and therapeutic target in MetS to prevent heart remodeling consistent with heart failure.

About 77.9 million (1 in 4) American adults have high blood pressure. High blood pressure is the primary cause of left ventricular hypertrophy (LVH), which represents a strong predictor of future heart failure and cardiovascular mortality. Previous studies have shown an altered metabolic profile in hypertensive patients with LVH. The goal of this study was to identify blood metabolomic LVH biomarkers by 1H NMR to provide novel diagnostic tools for rapid LVH detection in populations of hypertensive individuals. This cross-sectional study included 48 hypertensive patients with LVH matched with 48 hypertensive patients with normal LV size, and 24 healthy controls. Two-dimensional targeted M-mode echocardiography was performed to measure left ventricular mass index. Partial least squares discriminant analysis was used for the multivariate analysis of the H-1 NMR spectral data. From the H-1 NMR-based metabolomic profiling, signals coming from methylene (-CH2-) and methyl (-CH3) moieties of aliphatic chains from plasma lipids were identified as discriminant variables. The -CH2-/-CH3 ratio, an indicator of the mean length of the aliphatic lipid chains, was significantly higher (P<0.001) in the LVH group than in the hypertensive group without LVH and controls. Receiver operating characteristic curve showed that a cutoff of 2.34 provided a 52.08% sensitivity and 85.42% specificity for discriminating LVH (AUC= 0.703, P-value<0.001). We propose the -CH2-/-CH3 ratio from plasma aliphatic lipid chains as a biomarker for the diagnosis of left ventricular remodeling in hypertension.

We investigated the impact of heart failure (HF) etiology on the outcome of cardiac rehabilitation (CR) assessed by functional and clinical parameters.Treatment of chronic HF requires multidisciplinary approaches with a recognized role for CR. INCARD is a French study aimed at evaluating the benefits of sustainable CR in coronary (C) and noncoronary patients (NC) treated and educated during a 24-month period of follow-up.Prospective, monocentric patients with HF underwent inpatient physical training followed by a home-based program. Evaluations were performed at inclusion, discharge, 3 months after discharge, and subsequently every 6 months over the 24 months of outpatient rehabilitation.A total of 147 HF patients with left ventricular ejection fraction (LVEF) <40 were admitted to the CR center, 63 accepted to join INCARD (29 C and 34 NC).Although the C participants C having both an echocardiographic LVEF and an initially lower peak VO2, inpatient rehabilitation improved all functional parameters. Only NC showed an improved LVEF during the first 3 months of outpatient-follow-up. The main outcome of the outpatient rehabilitation was a trend toward stabilization of clinical and laboratory parameters with no significant difference between C and NC.This study confirms the benefits of initial HF inpatient rehabilitation and encourages prolonged outpatient monitoring. The results on functional parameters suggest exercise training should be conducted regardless of the HF etiology.

Background: Diagnostic biomarkers for heart failure (HF) such as the natriuretic peptides (NPs) are widely used but have limitations. Innovative biomarkers could provide improved diagnostic performance.Methods: We launched a prospective case-control proteomic study and investigated for polypeptides specific to HF through a capillary electrophoresis-mass spectrometry (CE-MS) proteomic analysis. The putative biomarker was identified by Orbitrap liquid chromatography-MS, validated by western blot, then by ELISA using plasmas from multicentric international cohorts. A rat model of HF was tested for biomarker expression levels.Results: We identified insulin like growth factor binding protein 2 (IGFBP2) as a new diagnostic biomarker for HF with a high sensitivity and specificity (AUC = 0.93; 95% CI, 0.89-0.96; p < 0.0001) in the local cohort and IGFBP2 levels provided an AUC of 0.943 (95% CI, 0.860-1.026) which gave a 87 % sensitivity in AHF and 90 % specificity at the cut off value previously determined in the discovery cohort, i.e. 556 ng/ml. ROC curve analysis of IGFBP2 and NTproBNP showed an AUC of 0.784 (95% CI, 0.744-0.820) for IGFBP2 and a significantly higher AUC of 0.927 (95% CI, 0.900-0.949) for NT-proBNP, p < 0.0001 in a Dutch cohort. In this cohort, the optimal cut off value for IGFBP2 gave a sensibility of 71% (95% CI, 66-76) and a specificity of 75% (95% CI, 65-83).Conclusion: IGFBP2 is a new biomarker to diagnose HF which could be used to provide additional information to the NPs. Animals models will help in the evaluation of the putative IGFBP2 regulated mechanisms in HF. (C) 2014 The Authors. Published by Elsevier Ireland Ltd. This is an open access article under the CC BY-NC-SA license (http://creativecommons. org/licenses/by-nc-sa/3.0/).

Background: It has been suggested that the adipokine resistin links obesity and insulin resistance, although how resistin acts on muscle metabolism is controversial. We aimed to quantitatively analyse the effects of resistin on the glucose metabolic flux profile and on insulin response in L6E9 myotubes at the metabolic level using a tracer-based metabolomic approach and our in-house developed software, Isodyn.Results: Resistin significantly increased glucose uptake and glycolysis, altering pyruvate utilisation by the cell. In the presence of resistin, insulin only slightly increased glucose uptake and glycolysis, and did not alter the flux profile around pyruvate induced by resistin. Resistin prevented the increase in gene expression in pyruvate dehydrogenase-E1 and the sharp decrease in gene expression in cytosolic phosphoenolpyruvate carboxykinase-1 induced by insulin.Conclusions: These data suggest that resistin impairs the metabolic activation of insulin. This impairment cannot be explained by the activity of a single enzyme, but instead due to reorganisation of the whole metabolic flux distribution.

Objective Galectin-3 (Gal-3) is considered as a myocardial fibrosis biomarker with prognostic value in heart failure (HF). Since aldosterone is a neurohormone with established fibrotic properties, we aimed to investigate if mineralocorticoid receptor antagonists (MRAs) would modulate the prognostic value of Gal-3. Methods The IBLOMAVED cohort comprised 427 eligible chronic HF patients (CHF) with echocardiography and heart failure biomarkers assessments (BNP). After propensity score matching CHF patients for cardiovascular risk factors, to form balanced groups, Gal-3 levels were measured at baseline in plasma from patients treated with MRAs (MRA-Plus, n=101) or not (MRA-Neg, n=101). The primary end point was all-cause mortality with a follow-up of 3 years. Results Gal-3 in plasma from these patients were similar with median values of 14.0 ng/mL [IQR, 9.9–19.3] and 14.4 ng/mL [IQR, 12.3–19.8] (P = 0.132) in MRA-Neg and MRA-Plus, respectively. Patients with Gal-3 ≤17.8 ng/mL had an HR of 1 (reference group) and 1.5 [0.4–5.7] in MRA-Neg and MRA-Plus, respectively (p=0.509). Patients with Gal-3 ≥ 17.8 ng/mL had an HR of 7.4 [2.2–24.6] and 9.0 [2.9–27.8] in MRA-Plus and MRA-Neg, respectively (p=0.539) and a median survival time of 2.4 years [95%CI,1.8–2.4]. Multivariate Cox proportional hazard analysis confirmed that MRA and the interaction term between MRA treatment and Gal-3 >17.8 ng/mL were not factors associated with survival. Conclusions MRA treatment did not impair the prognostic value of Gal-3 assessed with a 17.8 ng/mL cut off. Gal-3 levels maintained its strong prognostic value in CHF also in patients treated with MRAs. The significance of the observed lack of an interaction between Gal-3 and treatment effect of MRAs remains to be elucidated.

Diabetic cardiomyopathy is a secondary complication of diabetes with an unclear etiology. Based on a functional genomic evaluation of obesity-associated cardiac gene expression, we previously identified and cloned the gene encoding apolipoprotein O (APOO), which is overexpressed in hearts from diabetic patients. Here, we generated APOO-Tg mice, transgenic mouse lines that expresses physiological levels of human APOO in heart tissue. APOO-Tg mice fed a high-fat diet exhibited depressed ventricular function with reduced fractional shortening and ejection fraction, and myocardial sections from APOO-Tg mice revealed mitochondrial degenerative changes. In vivo fluorescent labeling and subcellular fractionation revealed that APOO localizes with mitochondria. Furthermore, APOO enhanced mitochondrial uncoupling and respiration, both of which were reduced by deletion of the N-terminus and by targeted knockdown of APOO. Consequently, fatty acid metabolism and ROS production were enhanced, leading to increased AMPK phosphorylation and Ppara and Pgc1a expression. Finally, we demonstrated that the APOO-induced cascade of events generates a mitochondrial metabolic sink whereby accumulation of lipotoxic byproducts leads to lipoapoptosis, loss of cardiac cells, and cardiomyopathy, mimicking the diabetic heart-associated metabolic phenotypes. Our data suggest that APOO represents a link between impaired mitochondrial function and cardiomyopathy onset, and targeting APOO-dependent metabolic remodeling has potential as a strategy to adjust heart metabolism and protect the myocardium from impaired contractility.

Blood heart failure (HF) biomarkers such as the natriuretic peptides (NP) are widely used but the NP biomarker family has limitations and HF diagnosis remains too often complicated because of atypical presentations and the need of specialized care access. More specific and accurate biomarkers with detection method allowing for facilitated large scale HF screenings are required. Indeed, 30 % of the patients admitted to emergency care for acute dyspnea have a BNP concentration in a « grey zone » that do not allow for diagnosis. Thus, diagnosis of these patients is costly and time-consuming while a rapid medical care of the patient has a positive impact on his health and reduces the treatment cost. Therefore, we have launched a prospective multicentric case-control study and investigated for urinary polypetides specific to acute (AHF) or chronic heart failure (CHF), using a capillary electrophoresis-mass spectroscopy strategy (CE-MS). Based on a screen of the urinary proteome of patients (CHF, AHF, and healthy controls matched for cardiovascular risk factors), we revealed a panel of polypeptides specific to HF. One polypeptide (X64, patent pending) seemed very relevant because it could discriminate AHF and CHF with a high specificity and specificity (AUC = 0,99; p<0,0001). X64 plasma levels were assessed within Atlanta (n=340, follow-up 3 years) and Toulouse (n=187, follow-up 6 years) cohorts. The Atlanta cohort revealed that X64 level is associated with major clinical events: HR: 1.23 per 100 ng/mL (95% CI: 1.06 to 1.43); P=0.005 and with HF admissions: IRR: 1.20 per 100 ng/mL (95% CI: 1.00 to 1.44); P=0.05. Moreover, the Toulouse cohort revealed that high X64 levels were strong predictors of cardiac death (p<0.0001). Our work provides with new biomarkers allowing for HF diagnosis in the BNP « grey zone » or prognosis and therefore allow for a better care of the HF patients at the hospital but also in general care.

targeted and targeted gas chromatography-mass spectrometry (MS) and liquid chromatography-MS. In a single center identification phase (243 HF patients, 83 matched healthy controls), metabolites were nominated for multimarker panels and their performance was verified in a separate validation cohort from three clinical centers (195 DCM, 184 ICM and 210 HFpEF patients, 255 matched healthy controls). Classification of patients and controls based on metabolite- and NT-pro-BNP levels was carried out with four different statistical approaches (penalized logistic regression, random forest, elastic net or mutual information). Results: Clear metabolic signatures were observed to discriminate DCM, ICM and HFpEF patients fromcontrols for symptomatic (NYHAII-III)as wellas asymptomatic patients (NYHA I). Compared to controls, numerous pathways, includ- ing lipid and amino acid metabolism, were altered in HF patients. Two types of biomarkers were defined in the identification phase: type 1 biomarkers consisting of 10 metabolites, and type 2 biomarkers of 9 metabolites in combination with NT-pro-BNP. In the separate validation cohort, a similar diagnostic performance of type 1 biomarkers compared to NT-pro-BNP was observed for several HF sub-groups. Compared to NT-pro-BNPalone, type2 biomarkers showed a significantly superior diagnostic performance for DCM and ICM patients (AUC=0.96 vs. 0.9, p < 0.001; 86% vs. 75% sensitivity at fixed 90% specificity), which was more pro- nounced for asymptomatic DCM and ICMpatients (AUC=0.95 vs. 0.86, p < 0.001), but also for all asymptomatic HF patients (AUC=0.90 vs. 0.84, p= < 0.001; 74% vs. 64% sensitivity at fixed 90% specificity). Conclusions: Metabolomic biomarkers offer clear potential for improved diagno- sis especially in asymptomatic HF patients in early stages of disease. Combina-tions of a limited number of metabolites with NT-pro-BNP may help to overcome the limitations of single feature markers in complex diseases like HF. Blood heart failure (HF) biomarkers such as the natriuretic peptides (NP) are widelyused butthe NPbiomarker familyhas limitationsand HFdiagnosis remains too often complicated because of atypical presentations and the need of special- ized care access. More specific and accurate biomarkers with detection method allowing for facilitated large scale HF screenings are required. Indeed, 30% of the patients admitted to emergency care for acute dyspnea have a BNP concentration in a "grey zone" that do not allow for diagnosis. Thus, diagnosis of these patients is costly and time-consuming while a rapid medical care of the patient has a positive impact on his health and reduces the treatment cost. Therefore, we have launched a prospective monocentric case-control study and investigated for urinary polypetides specific to acute (AHF) or chronic heart failure (CHF), us- ing a capillary electrophoresis-mass spectroscopy strategy (CE-MS). Based on a screen of the urinary proteome of 80 patients (CHF, AHF, and healthy controls matched for cardiovascular risk factors), we revealed a panel of polypeptides specific with idiopathic DCM. All endomyocar-dialbiopsy and samples of blood fromthe ventricle and peripheral blood obtained. Then histopathological and immunohistochemical study of the biopsy samples was performed. In 40 (of 100) patients’ clinical evaluation and/or results from the basic histopathological and immunohistochemical investigation indicated myocarditis. From those patients specimens were investigated using real-time PCR in order to detect viral nucleic acids. Real-time PCR was performed with use of TaqMan and HybProbe probes. Results: In biopsy specimens, viral nucleic acids were detected in 15 (of 40) patients. We detected in 7 patients parvovirus B19 (PB19V), in 4 enterovirus, in 3 adenovirus and in 2 human herpes virus 6 (HHV-6). In 1 patient coinfection of adeno- and enterovirus was detected. In blood samples from the ventricle in 6 (of 40) patients viral nucleic acids were detected. We detected in 3 adenovirus, in 2 enterovirus and in 1 HHV-6. In peripheral blood samples in 7 (of 40) patients viral nucleic acids were detected. We detected in 4 adenovirus, in 3 enterovirus and in 1 HHV-6. We observed PB19V only in biopsy specimen, none PB19V was detected in blood samples from the ventricle or from the peripheral blood. In 1 patient we detected HHV-6 in many copies as well as in blood samples from the ventricle or from the peripheral blood. In other cases we observed no correlation. Conclusions: Our findings suggest frequency of PB19V, entero-, adenovirus and HHV-6 DNA in patients with myocarditis and DCM more commonly than previ- ously expected. Those viruses should be therefore recognized as a potential car-diotropic pathogen in patients of all ages and as a potential therapeutic target. Proper diagnosis of DCM and myocarditis should be therefore based on perform-ing endomyocardial biopsy. Purpose: Clinical and experimental studies suggest an association between in- fectious agents and idiopathic dilated cardiomyopathy (DCM), but other data question this relation. The aim of this study was to investigate the presence of infectious agents in endomyocardial biopsy (EMB) specimens of patients with idiopathic DCM and other specific etiologies, compared to a control group of heart donors. Methods and results: Between 2008 and 2011 were studied EMB specimens from hospitalized patients with idiopathic DCM in evaluation for heart transplan-tation, donors and explanted hearts from different etiologies. 2 groups were de- fined: donors (29 cases) and DCM (55 cases, including 32 with idiopathic DCM, 9 chagasic, 6 ischemic and 8 other etiologies). Were studied by immunohisto- chemistry: enterovirus, adenovirus, herpes simplex, Epstein-Barr virus (EBV), parvovirus B19 (PB19), HHV6, hepatitis B and C, mycoplasma, chlamydia and borrelia. Results were presented as median, interquartile range p25, p75 (per-centage of positive area), and comparison between donors and DCM were made by Mann Whitney U test. There were an increased expression of enterovirus antigens in donors compared to DCM [2 (0.56 to 5.7) x 0.61 (0.28 - 2.45), p = 0.0075] and increased expression of hepatitis C antigens [1.31 (0.5 to 3.6) x 0.6 (0.37 - 1.41), p = 0.02] in DCM compared to donors. By molecular biology, were investigated: adenovirus, EBV, cytomegalovirus (CMV), HHV6, PB19, my-coplasma, chlamydia and borrelia. The results were presented by percentage of genome positivity, and comparison between groups were made by qui-square and Fisher’s exact tests. There was a high genome positivity of microorganisms, including co-infections, with higher positivity in donors comparing to DCM for adenovirus (83.3% vs. 58.7%, p = 0.035) and HHV6 (86.4% vs. 49%, p = 0.0015). To our knowledge, this study is pioneer demonstrating the presence of virus genome in cardiac tissue of chagasic DCM (adenovirus 55%, EBV 40%, CMV 20%, HHV6 75%, PB19 57%). Conclusion: The presence of infectious agents in the myocardium of patients with idiopathic DCM is frequent, and similarly in donors and DCM from other etiologies, including ischemic and chagasic. Based on our results the causal re- lationship between the presence of infectious agents in cardiac tissue and the development of DCM is controversial. Additional studies are needed to determine the real role of infectious agents in the pathogenesis of DCM. HFPEF,and potential biomarkers subjects in each group). Thereafter, the distinctly expressed proteins were fur- therly measured in a larger population (eighteen HFPEF patients and seventeen healthy controls). Results: Interestingly, the results of protein chip detection showed that the serum concentrations of 17 proteins (e.g., activin A, activin B, angiogenin, artemin, CD80) differed considerably between HFPEF patients and both hypertensive pa- tients and controls (P < 0.05), while a protein expression pattern distinct from that in non-HFPEF subjects might be associated with HFPEF. Moreover, of the 17 distinctly expressed proteins, angiogenin was furtherly measured in 35 subjects, which verified the upregulation of angiongenin in HFPEF patients than that in the healthy subjects (P < 0.05). Meanwhile, angiogenin distinguished HFPEF from control with an area under the receiver operating characteristic (ROC) curve of 0.73 (P < 0.05) and a diagnostic cut-off point of 426 ng/ml. Additionally, we found that the serum angiogenin level in HFPEF patients was positively correlated with N-terminal pro-B-type natriuretic peptide (NT-proBNP) (P < 0.05). Conclusion: Our pilot study indicate that these 17 potential biomarkers, such as angiogenin, may hold great promise forboth diagnosis and prognosis assessment of HFPEF. Purpose: The contribution of regulatory T cells (Tregs) to the pathogenesis of Acute Coronary Syndrome (ACS) remains poorly understood. One core obstacle is the lack of Treg-specific markers that distinguish Tregs and activated conven- tional T cells. Ahighly conserved CpG enriched element 1 (FOXP3 i l) is unmethylated only in Tregs, and measuring the unmethylation of FOXP3 i l can be used to identify the role of Tregs in clinical diseases. This study investigated whether analyzing the demethylation status of FOXP3 i 1 is a more reliable means than using Treg-specific surface markers in ACS. Methods: We evaluated circulating Tregs percentages on different levels includ- ing cell frequencies (CD4+CD25highFOXP3+Tregs and CD4+CD25highCD45+ naïve Tregs) or FOXP3 mRNA, FOXP3 i 1 demethylation status and related cy- tokine secretion in 90 patients with ACS and 40 controls. Results: FOXP3 i1 demethylation assay showed that the amount of Tregs in ACS patients was significantly reduced than that in controls (p=0.0005). This test dis-played a sensitivity of 94.7% and specificity of 80% in distinguishing ACS patients from the controls (p < 0.001). However, flow cytometry analysis did not identify any reduction of CD4+CD25highFOXP3+Tregs in ACS patients. N

Objective: Mortality in heart failure (AHF) remains high, especially during the first days of hospitalization. New prognostic biomarkers may help to optimize treatment. The aim of the study was to determine metabolites that have a high prognostic value.Methods: We conducted a prospective study on a training cohort of AHF patients (n = 126) admitted in the cardiac intensive care unit and assessed survival at 30 days. Venous plasmas collected at admission were used for H-1 NMR-based metabonomics analysis. Differences between plasma metabolite profiles allow determination of discriminating metabolites. A cohort of AHF patients was subsequently constituted (n = 74) to validate the findings.Results: Lactate and cholesterol were the major discriminating metabolites predicting 30-day mortality. Mortality was increased in patients with high lactate and low total cholesterol concentrations at admission. Accuracies of lactate, cholesterol concentration and lactate to cholesterol (Lact/Chol) ratio to predict 30-day mortality were evaluated using ROC analysis. The Lact/Chol ratio provided the best accuracy with an AUC of 0.82 (P < 0.0001). The acute physiology and chronic health evaluation (APACHE) II scoring system provided an AUC of 0.76 for predicting 30-day mortality. APACHE II score, Cardiogenic shock (CS) state and Lact/Chol ratio >= 0.4 (cutoff value with 82% sensitivity and 64% specificity) were significant independent predictors of 30-day mortality with hazard ratios (HR) of 1.11, 4.77 and 3.59, respectively. In CS patients, the HR of 30-day mortality risk for plasma Lact/Chol ratio >= 0.4 was 3.26 compared to a Lact/Chol ratio of < 0.4 (P = 0.018). The predictive power of the Lact/Chol ratio for 30-day mortality outcome was confirmed with the independent validation cohort.Conclusion: This study identifies the plasma Lact/Chol ratio as a useful objective and simple parameter to evaluate short term prognostic and could be integrated into quantitative guidance for decision making in heart failure care.

Fatima Smih (1), Franck Desmoulin (2), Matthieu Berry (1), Annie Turkieh (2), Romain Harmancey (1), Jason Iacovoni (2), Charlotte Trouillet (1), Clement Delmas (1), Atul Pathak (2), Olivier Lairez (2), Francois Koukoui(1), Pierre Massabuau (1), Jean Ferrieres (3), Michel Galinier (1), Philippe Rouet [Orateur] (1) (1) INSERM U1048, Equipe 7 “Obesite et insuffisance cardiaque”, Toulouse, France – (2) I2MC Inserm UMR 1048, Toulouse, France – (3) Inserm U1027, CHU Toulouse, Cardiologie, Toulouse, France