Glycoprotein and Lipoprotein Profiles Assessed by 1H-NMR and Its Relation to Ascending Aortic Dilatation in Bicuspid Aortic Valve Disease

Bicuspid aortic valve; Glycoprotein; Lipoprotein metabolismVàlvula aòrtica bicúspide; Glicoproteïna; Metabolisme de les lipoproteïnesVálvula aórtica bicúspide; Glicoproteína; Metabolismo de las lipoproteínasIntroduction: The bicuspid aortic valve (BAV) confers a high risk of ascending aorta dilatation (AAoD), although its progression seems highly variable. Furthermore, the implication of lipoprotein metabolism and inflammation in the mechanisms that underlie AAoD is not fully established. The aim of this study consisted of evaluating the impact of the lipoprotein and glycoprotein profiles in AAOD as well as its progression in BAV aortopathy. Methods: Using 1H-nuclear magnetic resonance (1H-NMR), we analyzed and compared the lipoprotein and glycoprotein profiles of plasma samples from 152 BAV patients with dilated and nondilated ascending aorta. Additionally, these profiles were also compared for 119 of these patients who were prospectively followed-up clinically and by echocardiography in the long-term (5 years). Ascending aorta dilation velocity (mm/year) was calculated for this analysis. Results: Several parameters related to the lipoprotein profile including remnant cholesterol, small LDL and IDL-cholesterol were found to be significantly increased in the dilated group compared to those in the nondilated group. The glycoprotein A-nuclear magnetic resonance (NMR) signal, a novel inflammation biomarker, was also observed to be increased in the dilated group. After performing multivariate analysis, remnant cholesterol remained an independent variable related to AAoD. In the long-term follow-up, proatherogenic lipoprotein parameters were related to ascending aorta dilatation velocity ascending. After a lineal regression analysis, non-HDL particles remained as an independent predictor of ascending aorta dilation velocity. Conclusions: Patients with BAV and AAoD presented a more pro-atherogenic profile assessed by 1H-NMR, especially related to triglyceride-rich lipoproteins. This pro-atherogenic profile seems to contribute to the higher growth rate of ascending aorta diameter.This research was funded by “Proyectos de Investigación Clínica” from the Spanish Society of Cardiology, grant Bayer 2019. BA-G received a research scholarship within the Martí-Franquès Research Fellowship Program from the University of Rovira i Virgili

Different profiles of lipoprotein particles associate various degrees of cardiac involvement in adolescents with morbid obesity

Dyslipidemia secondary to obesity is a risk factor related to cardiovascular disease events, however a pathological conventional lipid profile (CLP) is infrequently found in obese children. The objective is to evaluate the advanced lipoprotein testing (ALT) and its relationship with cardiac changes, metabolic syndrome (MS) and inflammatory markers in a population of morbidly obese adolescents with normal CLP and without type 2 diabetes mellitus, the most common scenario in obese adolescents. Prospective case-control research of 42 morbidly obese adolescents and 25 normal-weight adolescents, whose left ventricle (LV) morphology and function had been assessed. The ALT was obtained by proton nuclear magnetic resonance spectroscopy, and the results were compared according to the degree of cardiac involvement - normal heart, mild LV changes, and severe LV changes (specifically LV remodeling and systolic dysfunction) - and related to inflammation markers [highly-sensitive C-reactive protein and glycoprotein A (GlycA)] and insulin-resistance [homeostatic model assessment for insulin-resistance (HOMA-IR)]. A second analysis was performed to compare our results with the predominant ALT when only body mass index and metabolic syndrome criteria were considered. The three cardiac involvement groups showed significant increases in HOMA-IR, inflammatory markers and ALT ratio LDL-P/HDL-P (40.0 vs. 43.9 vs. 47.1, p 0.012). When only cardiac change groups were considered, differences in small LDL-P (565.0 vs. 625.1 nmol/L, p 0.070), VLDL size and GlycA demonstrated better utility than just traditional risk factors to predict which subjects could present severe LV changes [AUC: 0.79 (95% CI: 0.54-1)]. In the second analysis, an atherosclerotic ALT was detected in morbidly obese subjects, characterized by a significant increase in large VLDL-P, small LDL-P, ratio LDL-P/HDL-P and ratio HDL-TG/HDL-C. Subjects with criteria for MS presented overall worse ALT (specially in triglyceride-enriched particles) and remnant cholesterol values. ALT parameters and GlycA appear to be more reliable indicators of cardiac change severity than traditional CV risk factors. Particularly, the overage of LDL-P compared to HDL-P and the increase in small LDL-P with cholesterol-depleted LDL particles appear to be the key ALT's parameters involved in LV changes. Morbidly obese adolescents show an atherosclerotic ALT and those with MS present worse ALT values

Free triiodothyronine levels and age influences the metabolic profile and COVID-19 severity parameters in euthyroid and levothyroxine-treated patients

Metabolic reprogramming is required to fight infections and thyroid hormones are key regulators of metabolism. We have analyzed in hospitalized COVID-19 patients: 40 euthyroid and 39 levothyroxine (LT4)-treated patients in the ward and 29 euthyroid and 9 LT4-treated patients in the intensive care unit (ICU), the baseline characteristics, laboratory data, thyroid-stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), the FT3/FT4 ratio, 11 antiviral cytokines and 74 metabolomic parameters. No evidence for significant differences between euthyroid and LT4-treated patients were found in the biochemical, metabolomic and cytokines parameters analyzed. Only TSH (p=0.009) and ferritin (p=0.031) showed significant differences between euthyroid and LT4-treated patients in the ward, and TSH (p=0.044) and FT4 (p=0.012) in the ICU. Accordingly, severity and mortality were similar in euthyroid and LT4-treated patients. On the other hand, FT3 was negatively related to age (p=0.012), independently of sex and body mass index in hospitalized COVID-19 patients. Patients with low FT3 and older age showed a worse prognosis and higher levels of the COVID-19 severity markers IL-6 and IL-10 than patients with high FT3. IL-6 negatively correlated with FT3 (p=0.023) independently of age, body mass index and sex, whereas IL-10 positively associated with age (p=0.035) independently of FT3, body mass index and sex. A metabolomic cluster of 6 parameters defined low FT3 ward patients. Two parameters, esterified cholesterol (p=4.1x10) and small HDL particles (p=6.0x10) correlated with FT3 independently of age, body mass index and sex, whereas 3-hydroxybutyrate (p=0.010), acetone (p=0.076), creatinine (p=0.017) and high-density-lipoprotein (HDL) diameter (p=8.3x10) were associated to FT3 and also to age, with p-values of 0.030, 0.026, 0.017 and 8.3x10, respectively. In conclusion, no significant differences in FT3, cytokines, and metabolomic profile, or in severity and outcome of COVID-19, were found during hospitalization between euthyroid patients and hypothyroid patients treated with LT4. In addition, FT3 and age negatively correlate in COVID-19 patients and parameters that predict poor prognosis were associated with low FT3, and/or with age. A metabolomic cluster indicative of a high ketogenic profile defines non-critical hospitalized patients with low FT3 levels.PID2020-116146RB-I00 from the Ministerio de Ciencia e Innovación with European Regional Development Funds (FEDER), BMD-3724 from the Comunidad de Madrid, 202020E169 from the CSIC, 2020PANDE00082 from the Generalitad de Cataluña and Fundación Hay Esperanza

Determinación de marcas epigenéticas específicas de tejido para la identificación del origen de las vesículas circulantes

El presente Trabajo Final de Máster (TFM) pretende dar respuesta a la necesidad de identificar el origen de las vesículas extracelulares circulantes. Así teniendo en cuenta que las vesículas extracelulares contienen material genético de la célula progenitora, incluyendo ADN genómico, y que cada tipo celular tiene un patrón epigenético específico, el objetivo principal de este TFM es la aplicación de métodos y herramientas bioinformáticas en datos epigenéticos depositados en bases de datos públicos, para el diseño y el desarrollo de un algoritmo clasificador de marcadores epigenéticos capaz de diferenciar entre tejidos. Para alcanzar el objetivo principal se han analizado los datos referentes a la metilación de DNA de un total de 252 experimentos, agrupados en 24 clústers específicos de tejido o tipo celular, y mediante la creación de una función (BiosourceMapper) se han identificado posiciones o loci diferencialmente metilados para el tejido o tipo celular seleccionado, así como las regiones enriquecidas en posiciones diferencialmente metiladas. Además para mejorar la visualización e interpretación de los datos se ha creado la aplicación shiny Biosource Mapper App, que permite poden acceder y analizar los resultados generados durante este TFM a cualquier usuario. La principal conclusión obtenida es que mediante el análisis bioinformático del patrón de metilación con la función BiosourceMapper implementada en la aplicación Biosource Mapper App es posible obtener marcas específicas de tejido.The aim of this Final Master's Project (FMP) is to identify the origin of circulating extracellular vesicles. Taking into account that the extracellular vesicles contain genetic material from the progenitor cell, including genomic DNA, and that each cell type has a specific epigenetic pattern, the main objective of this FMP is the application of bioinformatic methods and tools on epigenetic data deposited in public repositories, for the design and development of an algorithm based on epigenetic marks to differentiate between tissues. To achieve this objective, the DNA methylation data from a total of 252 experiments, grouped in 24 tissuespecific or cell-type clusters, were analyzed and positions or loci were identified by creating a function (BiosourceMapper). Differentially methylated positions specific for the tissue or cell type selected, as well as regions enriched in differentially methylated positions, were found. Furthermore, to improve the visualization and interpretation of the data, the shiny Biosource Mapper App has been created, allowing users to access and analyze the results generated within this TFM. The main conclusion obtained is that bioinformatic analysis of the methylation pattern with the BiosourceMapper function implemented in the Biosource Mapper App identify specific tissue marks.El present Treball Final de Màster (TFM) pretén donar resposta a la necessitat d'identificar l'origen de les vesícules extracel·lulars circulants. Així tenint en compte que les vesícules extracel·lulars contenen material genètic de la cèl·lula progenitora, incloent ADN genómico, i que cada tipus cel·lular té un patró epigenètic específic, l'objectiu principal d'aquest TFM és l'aplicació de mètodes i eines bioinformátiques en dades epigenètiques dipositades en bases de dades públics, per al disseny i el desenvolupament d'un algorisme classificador de marcadors epigenètics capaç de diferenciar entre teixits. Per aconseguir l'objectiu principal s'han analitzat les dades referents a la metilació de DNA d'un total de 252 experiments, agrupats en 24 clústers específics de teixit o tipus cel·lular, i mitjançant la creació d'una funció (BiosourceMapper) s'han identificat posicions o loci diferencialment metilados per al teixit o tipus cel·lular seleccionat, així com les regions enriquides en posicions diferencialment metiladas. A més per millorar la visualització i interpretació de les dades s'ha creat l'aplicació shiny Biosource Mapper App, que permet podin accedir i analitzar els resultats generats durant aquest TFM a qualsevol usuari. La principal conclusió obtinguda és que mitjançant l'anàlisi bioinformático del patró de metilación amb la funció BiosourceMapper implementada en l'aplicació Biosource Mapper App és possible obtenir marques específiques de teixit

Data on the circulating levels of endothelial microparticles are elevated in patients with bicuspid aortic valve and are related to aortic dilation

The data included here support the research article “Circulating endothelial microparticles are elevated in bicuspid aortic valve (BAV) disease and related to aortic dilation” (Alegret et al., 2016 [1]) where circulating levels of platelet endothelial cell adhesion molecule (PECAM+) endothelial microparticles (EMPs) were identified as a biological variable related to aortic dilation in patients with BAV disease. The data presented in this article are composed by four tables and one figure containing the clinical and echocardiographic characteristics of the patients (Alegret et al., 2016 [1]) included in this study, and summarize the results of multivariate linear analyses. Furthermore, is also included a figure showing a representative flow cytometry dot plots and histograms used in PECAM+ EMPs quantification is also included

MicroRNAs Clustered within the 14q32 Locus Are Associated with Endothelial Damage and Microparticle Secretion in Bicuspid Aortic Valve Disease

Background: We previously described that PECAM+ circulating endothelial microparticles (EMPs) are elevated in bicuspid aortic valve (BAV) disease as a manifestation of endothelial damage. In this study, we hypothesized that this endothelial damage, is functionally related to the secretion of a specific pattern of EMP-associated miRNAs.Methods: We used a bioinformatics approach to correlate the PECAM+ EMP levels with the miRNA expression profile in plasma in healthy individuals and BAV patients (n = 36). In addition, using the miRNAs that were significantly associated with PECAM+ EMP levels, we inferred a miRNA co-expression network using a Gaussian graphical modeling approach to identify highly co-expressed miRNAs or miRNA clusters whose expression could functionally regulate endothelial damage.Results: We identified a co-expression network composed of 131 miRNAs whose circulating expression was significantly associated with PECAM+ EMP levels. Using a topological analysis, we found that miR-494 was the most important hub within the co-expression network. Furthermore, through positional gene enrichment analysis, we identified a cluster of 19 highly co-expressed miRNAs, including miR-494, that was located in the 14q32 locus on chromosome 14 (p = 1.9 × 10−7). We evaluated the putative biological role of this miRNA cluster by determining the biological significance of the genes targeted by the cluster using functional enrichment analysis. We found that this cluster was involved in the regulation of genes with various functions, specifically the “cellular nitrogen compound metabolic process” (p = 2.34 × 10−145), “immune system process” (p = 2.57 × 10−6), and “extracellular matrix organization” (p = 8.14 × 10−5) gene ontology terms and the “TGF-β signaling pathway” KEGG term (p = 2.59 × 10−8).Conclusions: Using an integrative bioinformatics approach, we identified the circulating miRNA expression profile associated with secreted PECAM+ EMPs in BAV disease. Additionally, we identified a highly co-expressed miRNA cluster that could mediate crucial biological processes in BAV disease, including the nitrogen signaling pathway, cellular activation, and the transforming growth factor beta signaling pathway. In conclusion, EMP-associated and co-expressed miRNAs could act as molecular effectors of the intercellular communication carried out by EMPs in response to endothelial damage

Identification of a nutrient-sensing transcriptional network in monocytes by using inbred rat models on a cafeteria diet

Obesity has reached pandemic levels worldwide. The current models of diet-induced obesity in rodents use predominantly high-fat based diets that do not take into account the consumption of variety of highly palatable, energy-dense foods that are prevalent in Western society. We and others have shown that the cafeteria (CAF) diet is a robust and reproducible model of human metabolic syndrome with tissue inflammation in the rat. We have previously shown that inbred rat strains such as Wistar Kyoto (WKY) and Lewis (LEW) show different susceptibilities to CAF diets with distinct metabolic and morphometric profiles. Here, we show a difference in plasma MCP-1 levels and investigate the effect of the CAF diet on peripheral blood monocyte transcriptome, as powerful stress-sensing immune cells, in WKY and LEW rats. We found that 75.5% of the differentially expressed transcripts under the CAF diet were upregulated in WKY rats and were functionally related to the activation of the immune response. Using a gene co-expression network constructed from the genes differentially expressed between CAF diet-fed LEW and WKY rats, we identified acyl-CoA synthetase short-chain family member 2 (Acss2) as a hub gene for a nutrient-sensing cluster of transcripts in monocytes. The Acss2 genomic region is significantly enriched for previously established metabolism quantitative trait loci in the rat. Notably, monocyte expression levels of Acss2 significantly correlated with plasma glucose, triglyceride, leptin and non-esterified fatty acid (NEFA) levels as well as morphometric measurements such as body weight and the total fat following feeding with the CAF diet in the rat. These results show the importance of the genetic background in nutritional genomics and identify inbred rat strains as potential models for CAF-diet-induced obesity

The impact of high Zn degrees concentrations on the application of AGNES to determine free Zn(II) concentration

AGNES (Absence of Gradients and Nernstian Equilibrium Stripping) determination of free Zn(II) in a solution can be affected by the reaching of high Zn° concentrations inside the amalgam. At concentrations about the solubility limit of Zn° in mercury, the formation of dendrites and powders around the mercury surface can be seen with an optical microscope. At concentrations of Zn° quite below the solubility limit, an anomalous stripping current appears which increases with decreasing supporting electrolyte concentration. The current along the stripping time exhibits a convex shape, which is labelled here as "anomalous convex behaviour" (acb). The origin of acb is tentatively ascribed to different kinetic reasons (amongst which the electroneutrality limitation due to low ionic strength outstands), but more than one cause is necessary for a full account of the experimental observations. With various strategies, like monitoring the charge as response function or by application of lower gains, AGNES can successfully probe these high concentrations