2. Universidad Cardenal Herrera-CEU

Despite the reduction of cardiovascular events, including the risk of death, associated with sodium/glucose cotransporter 2 inhibitors (SGLT2i), their basic action remains unclear. Sodium/ hydrogen exchanger (NHE) has been proposed as the mechanism of action, but there are controversies related to its function and expression in heart failure (HF).We hypothesized that sodium transportedrelated molecules could be altered in HF and modulated through SGLT2i. Transcriptome alterations in genes involved in sodium transport in HF were investigated in human heart samples by RNAsequencing. NHE11 and NHE1 protein levels were determined by ELISA; the effect of empagliflozin on NHE11 and NHE1 mRNA levels in rats’ left ventricular tissues was studied through RT-qPCR.We highlighted the overexpression of SLC9C2 and SCL9A1 sodium transport genes and the increase of the proteins that encode them (NHE11 and NHE1). NHE11 levels were correlated with left ventricular diameters, so we studied the effect of SGLT2i on its expression, observing that NHE11 mRNA levels were reduced in treated rats. We showed alterations in several sodium transports and reinforced the importance of these channels in HF progression. We described upregulation in NHE11 and NHE1, but only NHE11 correlated with human cardiac dysfunction, and its levels were reduced after treatment with empagliflozin. These results propose NHE11 as a potential target of SGLT2i in cardiac tissue.

A controversial understanding of the state of the DNA methylation machinery exists in ischaemic cardiomyopathy (ICM). Moreover, its relationship to other epigenetic alterations is incomplete. Therefore, we carried out an in-depth study of the DNA methylation process in human cardiac tissue. We showed a dysregulation of the DNA methylation machinery accordingly with the genome-wide hypomethylation that we observed: specifically, an overexpression of main genes involved in the elimination of methyl groups (TET1, SMUG1), and underexpression of molecules implicated in the maintenance of methylation (MBD2, UHRF1). By contrast, we found DNMT3B upregulation, a key molecule in the addition of methyl residues in DNA, and an underexpression of miR-133a-3p, an inhibitor of DNMT3B transcription. However, we found many relevant alterations that would counteract the upregulation observed, such as the overexpression of TRAF6, responsible for Dnmt3b degradation. Furthermore, we showed that molecules regulating Dnmts activity were altered; specifically, SAM/SAH ratio reduction. All these results are in concordance with the Dnmts normal function that we show. Our analysis revealed genome-wide hypomethylation along with dysregulation in the mechanisms of addition, elimination and maintenance of methyl groups in the DNA of ICM. We describe relevant alterations in the DNMT3B system, which promote a normal Dnmt3b function despite its upregulation.

The use of cadavers is essential for veterinary anatomy learning. However, facing an animal corpse can be stressful for veterinary students because of their empathy toward animals. The objective of this study was to evaluate veterinary medicine students' emotions, feelings, and anxiety levels related to practicals with dog cadavers. Two questionnaires were administered to 1st year students (n = 168) at CEU Cardenal Herrera University in Valencia (Spain) before and after their first practical session with cadavers. The application of State–Trait Anxiety Inventory questionnaires showed that “state anxiety” decreased significantly (p < 0.05), from a score of 14.8 before the practical to 10.4 after, and that female students showed higher but not significantly different levels than males. Most (64%) of the students were not willing to donate the bodies of their pets, and those students were more stressed before the practical than their peers, although their anxiety levels significantly decreased by the end of the session. The majority of the students answered positively about emotions, such as feeling calm, safe, not nervous, relaxed and not worried before the practical, and this increased significantly to more than 80% by the end of the session. The visualization of educational videos prior to the session was evaluated positively by students. These results agree with those reported in other health science disciplines, showing that students face practical sessions with corpses in a similar way and suggesting that the use of videos can help decrease anxiety and enhance their learning experience.

BACKGROUND: Acute rejection is one of the most important direct contributors to mortality after heart transplantation. Advances in the development of novel non-invasive approaches for the early identification of allograft rejection are necessary. We conducted a non-targeted proteome characterization focused on identifying multiple plasmatic protein differences to evaluate their diagnostic accuracy for rejection episodes. METHODS: We included consecutive plasma samples from transplant recipients undergoing routine endomyocardial biopsies. A liquid chromatography−tandem mass spectrometry analysis using isobaric tags (tandem mass tag 10-plex) was performed and concentrations of CD5L were validated using a specific sandwich enzyme-linked immunosorbent assay. RESULTS: A total of 17 altered proteins were identified as potential markers for detecting heart transplant rejection, most involved in inflammation and immunity. CD5L, an apoptosis inhibitor expressed by macrophages, showed the best results in the proteomic analysis (n = 30). We confirm this finding in a larger patient cohort (n = 218), obtaining a great diagnostic capacity for clinically relevant rejection (≥Grade 2R: area under the curve = 0.892, p < 0.0001) and preserving the accuracy at mild rejection (Grade 1R: area under the curve = 0.774, p < 0.0001). CD5L was a strong independent predictor, with an odds ratio of 14.74 (p < 0.0001), for the presence of rejection. CONCLUSIONS: Episodes of acute cardiac allograft rejection are related to significant changes in a key inhibitor of apoptosis in macrophages, CD5L. Because of its precision to detect acute cellular rejection, even at mild grade, we propose CD5L as a potential candidate to be included in the studies of molecule combination panel assays. This finding could contribute to improving the diagnostic and preventive methods for the surveillance of cardiac transplanted patients.

Accumulating evidence has confrmed that the expression of sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) is downregulated in heart failure and cardiac allograft rejection. Although many SERCA2arelated genes and proteins involved in the regulation of myocardial Ca2+ fuxes have been explored, its related metabolites remain poorly studied. Our main objective was to identify circulating SERCA2arelated metabolites altered in cardiac allograft rejection and to determine whether these could serve as non-invasive biomarkers. Sixty plasma samples from adult heart transplant were included in a metabolomic analysis. Sphingosine-1 phosphate (S1P), metabolite closely related with SERCA, were increased in patients with cardiac rejection (p<0.0001). S1P discriminated between patients with and without rejection: normal grafts vs. all rejecting grafts (AUC=0.911, p<0.0001), normal grafts vs. Grade 1R (AUC=0.819, p<0.01), Grade 2R (AUC=0.911, p<0.0001), Grade 3R (AUC=0.996, p<0.0001). In addition, we found changes in key enzymes and receptors of S1P pathway analysed on explanted hearts from heart failure patients. This preliminary study reveals that circulating S1P determination could be a novel approach to detect cardiac rejection, showing a robust capability for detection that improves gradually with the severity of rejection. These alterations could be relevant to better understand the involvement of calcium regulation on the pathophysiology of rejection.

Transcriptomic signature ofXPO1was highly expressed and inversely related to left ventricular function in ischemic cardiomy-opathy patients. We hypothesized that treatment with AAV9-shXPO1 attenuates left ventricular dysfunction and remodeling in amyocardial infarction rat model. We induced myocardial infarction by coronary ligation in Sprague-Dawley rats (n= 10), whichreceived AAV9-shXPO1 (n= 5) or placebo AAV9-scramble (n= 5) treatment. Serial echocardiographic assessment was per-formed throughout the study. After myocardial infarction, AAV9-shXPO1-treated rats showed partial recovery of left ventricularfractional shortening (16.8 ± 2.8 vs 24.6 ± 4.1%,P< 0.05) and a maintained left ventricular dimension (6.17 ± 0.95 vs 4.70 ±0.93 mm,P< 0.05), which was not observed in non-treated rats. Furthermore, lower levels of EXP-1 (P< 0.05) and lowercollagen fibers and fibrosis in cardiac tissue were observed. However, no differences were found in the IL-6 or TNFR1 plasmalevels of the myocardium of AAV9-shXPO1 rats. AAV9-shXPO1 administration attenuates cardiac dysfunction and remodelingin rats after myocardial infarction, producing the gene silencing ofXPO1.