bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammatory mediators to predict left adverse ventricular remodelling in revascularized STEMI patients
Authors: Bevilacqua E , Galeano-Otero I , Guerrero-Marquez FJ , Falcon- Boyano D , Baron-Esquivias G , Smani T , Ordonez-Fernandez A , Del Toro R 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Institute of Carlos III the Spanish Ministry of Economy and Competitiveness

Background
Primary percutaneous coronary intervention (PPCI) in patients with ST-segment elevation myocardial infarction (STEMI) improves the survival of patients. Nevertheless, some patients develop left ventricular adverse remodeling (LVAR). The main objective of this study is to find reliable prognostic biomarkers for patients developing LVAR, focusing in the role of pro-inflammatory mediators, including cell populations and secreted molecules.

Methods
We assessed the level of pro-inflammatory cell subsets, before and after the revascularization, 1 and 6 months after the PPCI, using flow cytometry. We also performed miRNAs microarray in isolated peripheral blood mononuclear cells (PBMCs), and examined the levels of 27 cytokines in patients’ serum by multiplex ELISA.

Results
We observed that levels of pro-inflammatory monocytes were increased soon after PPCI, in patients who afterward developed LVAR. Furthermore, we detected significant increase in the concentration of secreted cytokines such as IL-1b, IL-17, IFN-g and VEGF in LVAR patients post-PPCI. Moreover, specific miRNAs levels regulating these cytokines secretion, also correlated with LVAR. Multivariate regression analysis and ROC curves indicated that these inflammatory mediators were good biomarkers to correlate with the appearance of LVAR in patients. In addition, we found that the combined analysis of these biomarkers increased the specificity and sensibility to identify patients with LVAR.

Conclusion
Our data suggest that the combined analysis of these inflammatory axes and related molecules, could reliably predict LVAR in STEMI patients.
E2F7/8 is involved in cardiomyocyte polyploidy but does not affect myocardial reperfusion injury recovery
Authors: Fang J , Van Der Geest JSA , Yao B , Yang Q , Chirico N , Brans MA , Roefs MT , Vader P , De Jager SCA , De Bruin A , Vink A , Van Mil A , Schiffelers RM , Lei Z , Sluijter JPG 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Other. Main funding source(s): CSC fellowship

Background
Polyploidy cells consist of more than two complete sets of homologous chromosomes. Although a characteristic feature of cardiomyocytes and observed in all mammalian species, its molecular mechanism and biological functions are still unknown. Cardiomyocytes polyploidy in rodents occurs mainly through incomplete cytokinesis and increases with age. Studies have demonstrated that E2F7/8 transcription factors are key regulators of polyploidy in the liver and pancreas, however, it remains unclear if E2F7/8 control the generation of polyploidy cardiomyocytes and what the functional consequence is post-myocardial infarction (MI).

Methods
By using a tamoxifen inducible Cre/LoxP approach in new-born mice, we deleted E2F7/8 transcription factors ubiquitously and evaluated the biological significance of postnatal E2F7/8 loss. Mice underwent myocardial ischemia reperfusion injury (IRI) and heart function was assessed by 4D-echocardiography. Cardiomyocyte nucleus polyploidy was measured by FACS and microscope.

Results
Deficiency of E2F7/8 significantly suppress cardiomyocyte mononucleated and multinucleated polyploidy, as well as dramatically decreased hepatocytes polyploidy. E2F7/8 defect also led to a decrease in cardiac stress related marker lever such as ANP, BNP, MMP2, β-MHC/α-MHC and an increase in CD31 expression level. Surprisingly, E2F7/8 deletion did not have impact on cardiac function and dimensions post-IRI.

Conclusion
In summary, we identified that E2F7/8 activity is involved in the cellular polyploidy in the heart but did not affect myocardial function after myocardial injury.
Implication of caveolae in the pathophysiology of human acute myocardial infarction: a histological study
Authors: Rios Navarro C , Cantos-Amores G , Ortega M , Gavara J , Marcos-Garces V , De Dios E , Perez-Sole N , Chorro FJ , Bodi V , Ruiz-Sauri A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This work has been funded by “Instituto de Salud Carlos III” and “Fondos Europeos de Desarrollo Regional FEDER” (Exp. PI20/00637) as well as by “Generalitat Valenciana” (Exp. PROMETEO/2021/008).

Introduction
Caveolae are lipid invaginations present in the membrane of most mammalian cells. They are mainly made up of three proteins: caveolin (Cav)-1, 2, and 3 and are involved in signal transduction and ion channels. Since caveolae regulate different signalling pathways to promote cardiac protection, the aim of this study is to compare the expression of Cav-1, 2, and 3 in the human myocardium of control subjects and in patients with myocardial infarction (MI).

Materials
Myocardial samples from human autopsies of 4 controls and 4 MI patients with more than 6 months of evolution were isolated. The infarction and control area were identified by haematoxylin-eosin and Masson’s trichromic stainings. The presence of Cav-1, 2 and 3 was detected by immunochemistry. Afterwards, 5 photographs were taken for each antibody and sample and the presence of each protein was morphometrically quantified using the image analyzer Image-Pro Plus. These results were also corroborated by immunofluorescence.

Results
A constitutive presence of Cav-1, 2, and 3 was observed in the myocardium of control patients, being Cav-1 and 3 higher expressed than Cav-2. Cav-2 and 3 were mainly found in cardiomyocytes, while Cav-1 was detected not only in cardiac muscle cells but also in endothelial cells. Comparing caveolae expression between the peri-infarct region of MI patients and controls, a significant reduction in the expression of the three proteins was observed by immunochemistry. Indeed, the decrease in Cav-2 and 3 could be also detected by immunofluorescence. Lastly, Cav-1 was more expressed in cardiomyocytes than in endothelial cells, but unlike muscle cells, its expression was not diminished after MI.

Conclusions
The presence of caveolin decreases in the myocardial tissue after MI. Further studies are needed to confirm the cardioprotective role of caveolae post-AMI and their use as potential therapeutic target.
Identification of cardioprotective miRNAs (protectomiRs) in a porcine model of acute myocardial infarction and cardioprotection by ischemic conditioning
Authors: Nagy RN , Makkos A , Baranyai T , Giricz Z , Kiss B , Puskas LG , Farago N , Lukovic D , Gyongyosi M , Gorbe A , Ferdinandy P 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Research, Development and Innovation Office of Hungary (NKFIA; NVKP-16-1-2016-0017 National Heart Program and OTKA-FK 134751)
MTA-SE System Pharmacology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, H-1089 Budapest, Hungary

Introduction
Changes in the expression profile of microRNAs contribute to cardioprotective signaling. We have previously identified cardioprotective microRNAs, termed protectomiRs, by a systematic analysis of microRNA expression pattern in myocardial infarction and cardioprotection induced by ischemic conditioning in rats. ProtectomiRs may be important therapeutics for cardioprotection.

Purpose
We aimed to identify protectomiRs in a translational porcine model of reperfused acute myocardial infarction (AMI) and cardioprotection, and validate their cardiocytoprotective effect.

Methods
We used cardiac tissue samples from our previous study in closed-chest AMI model in domestic pigs. Pigs were subjected to sham operation (Sham), ischemia/reperfusion to induce AMI (AMI) or preconditioning (IPreC), postconditioning (IPostC), and remote perconditioning (RIPerC). Tissue samples were collected from the infarcted region of the left ventricles. MiRNA expression pattern was detected by high-throughput qRT-PCR. Potential protectomiRs were selected by systematic comparison of significant expression changes due to different conditioning stimuli vs. AMI. To validate the cardiocytoprotective effect of potential protectomiRs, isolated rat cardiomyocytes were transfected with specific miRNA mimics or inhibitors (antagomiRs) of the selected protectomiRs, and the survival of cells was measured after simulated ischemia/reperfusion injury.

Results
Expression of 221 miRNAs was assessed. Expression of 57 microRNAs were changed by IPreC, 54 by IPostC and 68 by RIPerC as compared to AMI (min. 1,5×log2 fold-change, -log10p&gt;1,31 vs. AMI). Expression of 14 microRNAs changed significantly due to all three conditionings vs. AMI (10 miRNAs were downregulated and 4 upregulated). Rat homologs of these 14 protectomiR candidates were identified and 13 showed 100% sequence homology with the original pig miRNAs. The selected miRNAs (9 antagomiRs and 4 miRNA mimics) were transfected in isolated rat cardiomyocytes. Modulation of 2 mimics of these miRNAs significantly improved the survival of cells after ischemia/reperfusion injury (due to intellectual property protection, we do not disclose the protectomiRs here).

Conclusion
Here we identified 2 miRNAs in a clinically relevant porcine model of AMI that can be potential therapeutic protectomiRs.
Cardioprotection by limb remote ischemic preconditioning in rats: discrepancy between meta-analysis and a three-center in vivo study
Authors: Sayour N , Brenner GB , Makkos A , Kovacshazi CS , Gergely TG , Tian H , Zenkl V , Bencsik P , Heinen A , Schulz R , Baxter GF , Zuurbier CJ , Voko Z , Ferdinandy P , Giricz Z 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Heart Program funded by the National Research, Development and Innovation Office of Hungary
Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary, within the framework of the Therapeutic development thematic programme of the Semmelweis University

Purpose and aims
Despite promising preclinical results on cardioprotection by remote ischemic preconditioning (RIPC), large-scale clinical trials show no reduction in infarct size (IS). This translational gap might be due to unknown confounding factors and lack of sufficient high-quality in vivo preclinical proof-of-concept studies. Therefore, here we performed randomized, blinded in vivo studies in three study centers using the most often reported methodological settings, as well as a systematic review and meta-analysis of RIPC studies.

Methods and Results
Male Wistar rats were subjected to 20 to 45 min cardiac ischemia followed by 120 min reperfusion with or without preceding RIPC by 3 or 4×5-5 min occlusion/reperfusion of one or two femoral vessels by clamping, tourniquet, or pressure cuff. RIPC did not reduce IS, microvascular obstruction, or arrhythmias at any study centers. Systematic review and meta-analysis focusing on in vivo rat models of myocardial ischemia/reperfusion injury with limb RIPC showed that RIPC reduces IS by 21% on average; however, a tendency for publication bias towards positive results was found. In addition, our systematic review showed methodological heterogeneity and insufficient reporting quality in a high proportion of studies.

Conclusion
We report for the first time the lack of cardioprotection by RIPC in rats, assessed in individually randomized, blinded in vivo studies, involving multiple study centers. Although unanimously showing cardioprotection by RIPC, the majority of reviewed studies were inadequately reported, and under-publication of neutral studies is plausible. Methodological confounding factors leading to the discrepancy between meta-analysis and the present studies cannot be identified; insufficient reporting quality, heterogeneity of the applied settings, and selection bias against studies with neutral outcome may contribute to this discrepancy. More preclinical studies with adequate quality control, together with publication of neutral studies, are required to support development of clinically translatable cardioprotective interventions.
Ischemic postconditioning activates cardiac MMP-2 and decreases biglycan level as well as miR-34a-5p and miR-195-5p expression in early reperfusion in a porcine infarction model
Authors: Szabados T , Gomori K , Ferdinandy P , Gyongyosi M , Gorbe A , Bencsik P 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Kooperatív Doktori Program Doktori Hallgatói Ösztöndíj (KDP-2020)
Hungarian Scientific Research Fund-OTKA-138223

Background
Matrix metalloproteinase-2 (MMP-2) has been shown to be activated during myocardial ischemia-reperfusion (IR) injury leading to degradation of cardiac contractile proteins. In previous investigation, ischemic postconditioning (IPoC) failed to decrease myocardial necrosis in porcine model of acute myocardial infarction (AMI). However, IPoC significantly reduced the severity of myocardial microvascular obstruction (MVO) compared to ischemic control. Therefore, our aim was to reveal if two major extracellular players of cardiac pathophysiology, MMP-2 and biglycan (BGN) are involved in the MVO reducing effect of IPoC in a porcine AMI model.

Methods
AMI was induced in female domestic pigs (25-35 kg) with balloon catheter occlusion of the left descending coronary artery for 90 min in ischemic control group. IPoC group underwent 6×30 s I/R after 90-min occlusion. At the end of the 3-hour or 3-day reperfusion, infarcted left ventricular tissue samples were taken to determine MMP-2 activity and BGN levels as well as cardiac microRNA (miRNA) expression.

Results
Cardiac MMP-2 activity showed significant increase in IPoC group as compared to ischemic control after 3h reperfusion but after 3d reperfusion no significant change was observed between the two groups. Cardiac BGN level in IPoC was significantly decreased as compared to ischemic control after 3h reperfusion, which difference was also abolished after 3d reperfusion due to the decrease of BGN level of ischemic control group. Ischemic postconditioning altered the expression of three miRNAs: miR-34a-5p, and miR-195-5p were downregulated, and miR-15b-5p was upregulated as compared to ischemic control.

Conclusions
IPoC decreased MVO and BGN level and increased MMP-2 activity in the infarcted left ventricle during early reperfusion, and altered the expression on 3 cardiac miRNAs. Further increase was found in MMP activities after 3 days in both groups, which affected BGN level only in ischemic control group. Altered expression of miRNAs and early decrease in cardiac BGN level may contribute to the beneficial effect of IPoC on MVO.
Blockers of Hyperpolarization-activated Cyclic Nucleotide-gated channels exhibit antimuscarinic properties in human atrial cardiomyocytes
Authors: Gencarelli M , Laurino A , Balducci V , Spinelli V , Cameli M , Pecori R , Vistoli G , Matucci R , Cerbai E , Sartiani L 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): M.I.U.R. PRIN 2017 to E.C and ECRF to L.S.

Introduction
Ivabradine (Iva), Zatebradine (Zat) and ZD7288 (ZD) are blockers of Hyperpolarization-activated Cyclic Nucleotide-gated (HCN) channels, which play definite electrophysiological functions in cardiac pacemakers, working cardiomyocytes, peripheral and central neurons [1]. Among them, Iva is in clinical use as bradycardic agent for uncontrolled angina and heart failure with reduced ejection fraction. Accumulating evidence suggests Iva may have a more complex pharmacological profile, including targets different from HCN channels [2, 3]. Reports on experimental atrial fibrillation, detrusor contraction and bladder overactivity [4] led us to hypothesize that muscarinic receptors (mAChRs) may represent still unexplored targets of Iva and analogues. Present study was designed to identify the effect of Iva on acetylcholine-gated inward-rectifier K+-current (IKACh), the main effector of atrial mAChRs, and to assess whether Iva and analogues interact directly with human atrial and recombinant mAChRs.

Methods
Patch-clamp recordings were performed on human atrial myocytes (hAMs) isolated from atrial appendages of patients in sinus rhythm. Competition and kinetic radioligand binding assays were performed on hAM preparations and on mAChR(1,2,3) expressed in heterologous systems.

Results
Patch-clamp recordings on hAMs demonstrated Iva is able to suppress IKAch similarly to atropine, thus suggesting a functional antagonism of Iva on atrial mAChRs. Competition binding assays on hAM membranes revealed Iva, Zat and ZD bind to mAChRs with affinity values close to those of muscarinic ligands carbachol (Cch) and gallamine (Gal). Competition binding assays on recombinant mAChR(1,2,3), which are functionally relevant in hAMs, showed all blockers are ligands for each single mAChR type with affinity values close to those of CCh and Gal. Finally, using dissociation binding assays on mAChR(2) lacking the allosteric binding site, we showed Iva and Zat behave as allosteric ligands with affinity values similar to Gal. Doking studies confirmed Iva and Zat, but not ZD, have the structural requirements necessary to interact with the allosteric and the orthosteric site of mAChR(2).

Conclusions
This study demonstrates Iva directly interacts with mAChR expressed in hAMs, showing antimuscarinic activity on IKAch. This property is likely dependent on a double interaction of Iva with the orthosteric and the allosteric site of mAChR(2). Zat, but not ZD, shows similar binding properties compared to Iva. Our results identify a novel pharmacological target for Iva and analogues in human atria, which may have important consequences for the function of the healthy and diseased atrium.
ProANP31-67 ameliorates adverse cardiac remodeling and improves systolic and diastolic functions in a preclinical model of cardiorenal syndrome
Authors: Silva GJJ , Parvan R , Shen X , Frisk M , Altara R , Strand ME , Rypdal KB , Lunde IG , Louch WE , Aronsen JM , Stenslokken K-O , Stokke MK , Cataliotti A 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): South-Eastern Norway Regional Health Authority (HSØ-RHF, Project No. 25674)

Background
The cardiac hormone proANP31-67, a linear fragment of the N-terminal Atrial Natriuretic Peptide, has known enhancing renal effects. More recently, we described the cardio protective effects of this hormone in a model of chronic hypertension. More specifically, independently of the blood pressure level, proANP31-67 improved diastolic function, attenuated cardiac fibrosis, and reduced hypertrophy.

Purpose
The current study was designed to assess the cardiorenal effects of proANP31-67 in a rodent model of hampered renal function, followed by cardiac injury produced by ischemia/reperfusion (I/R).

Methods
Right uninephrectomy (UNX) was performed in Wistar rats (n=28). Sixteen weeks after UNX, rats underwent cardiac I/R injury and randomly assigned to proANP31-67 (50 ng/kg/day s.c., n=15) or Vehicle (n=13) for four weeks post I/R. Echocardiographic examinations were performed at baseline (before UNX), 16 weeks after UNX, and four weeks after I/R. At the end of the study, cardiomyocytes were isolated and tissue samples were collected.

Results
Chronic UNX resulted in diastolic impairment (E/A: 1.47±0.08 at baseline vs 0.98±0.14 at 16 wks post UNX, p=0.0010). I/R further accentuated the development of the cardiorenal syndrome, and induced a mild systolic dysfunction in the placebo treated animals. However, four weeks of treatment with proANP31-67 preserved systolic function (EF: 62±3% placebo vs 74±2% proANP31-67, p&lt;0.0001), and reverted the diastolic dysfunction (E/A: 0.72±0.15 placebo vs 1.24±0.11 proANP31-67, p=0.0134). ProANP31-67 ameliorated the adverse cardiac remodeling (i.e., reduction in the cardiomyocyte cross-sectional area and interstitial fibrosis), enhanced Ca2+ handling, and improved cardiomyocyte t-tubules´ structural changes compared to vehicle. At the cellular level, in vitro experiments demonstrated the direct effect of proANP31-67 on cardiomyocyte hypertrophy (assessed by [3H]-leucine incorporation) induced by endothelin 1 and angiotensin II.

Conclusion
ProANP31-67 has a direct cardiomyocyte protective effect, leading to an improvement in Ca2+ homeostasis and t-tubules´ structures and, prevents the development of systolic and diastolic dysfunction in a pre-clinical model of cardiorenal syndrome.
Towards advanced cardiac miRNA therapies through DNA-based nanostructures
Authors: Hernandez-Bellido N , Postigo A , Orrite C , Garrido-Huescar E , Garcia-Mendivil L , Pueyo E , Hernandez-Ainsa S , Ordovas L 
Published: June 2022

Abstract

Funding Acknowledgements
Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Proyecto MINECO-Ministerio de Ciencia e Innovación (Spain)- PID2019-105674RB-I00
Proyecto DGA- Diputación General de Aragón (Spain)- LMP128_21

Introduction
Cardiac microRNAs (miRNAs) have been found to be dysregulated in cardiac disease and aging both in animal models and humans. Thanks to their pleiotropic effects, miRNAs are spotlighted as promising therapeutic targets to treat cardiac conditions. However, their low stability and potential off-target effects in vivo difficult the development of effective and safe miRNA therapies. Nanotechnology enables the fabrication of biocompatible carriers to perform targeted delivery of both conventional and advanced therapies. In particular, nanostructures built with DNA (DNS) have been successfully developed for miRNA therapies in cancer, but they have not been applied in cardiac therapy yet [1].

Purpose
The main goal of this study is to demonstrate the capacity of DNS to deliver miRNA therapies in human cardiomyocytes.

Methods
Our group has identified MIR24-2 upregulated with age in the human left ventricle and miR24-2-5p to interact with genes related with the cardiac function, such as SERCA2 [2]. Different versions of DNS were designed and generated by self-assembly of DNA strands containing anti-miR24-2-5p capture segments. Physicochemical characterization of DNS was then conducted to determine their auto-assembling capacity, size, morphology and thermal stability. The characterization of the biological effect of the DNS was also conducted in vitro and in cell culture of HEK293 and human induced pluripotent stem cells (iPSC)-derived cardiomyocytes (iCM) to assess their cytotoxicity and internalization capacity.

Results
DNS were assembled correctly and reproducibly into carriers of around 30-50 nm. In vitro, the DNS showed thermal stability at physiological temperature, proper stability in serum and they disassembled specifically in the presence of the target miR24-2-5p sequence. In HEK293 and iCM cultures, the DNS showed lack of cytotoxicity demonstrating in vitro biocompatibility. DNS were efficiently internalized by HEK293, but the uptake capacity of DNS by iCM was remarkably lower.

Conclusions
We created and characterized biocompatible anti-miR-loaded DNS that effectively captured the target miR-24-2-5p in vitro. DNS internalized in iCM, but with lower efficiency than in HEK293 cells. Our results indicate that DNS are suitable candidates to carry miRNA therapies to human cardiac cells in vitro, but the observed cell-type specific differences suggest that future efforts need to be directed towards developing DNS functionalization capable of promoting efficient and specific carrier uptake in vivo by primary cardiac cells in vivo.