Metabolomic Analysis of Pediatric Patients with Idiosyncratic Drug-Induced Liver Injury According to the Updated RUCAM

Hepatotoxicity, a common adverse drug effect, has been extensively studied in adult patients. However, it is equally important to investigate this condition in pediatric patients to develop personalized treatment strategies for children. This study aimed to identify plasma biomarkers that characterize hepatotoxicity in pediatric patients through an observational case–control study. Metabolomic analysis was conducted on 55 pediatric patients with xenobiotic liver toxicity and 88 healthy controls. The results revealed clear differences between the two groups. Several metabolites, including hydroxydecanoylcarnitine, octanoylcarnitine, lysophosphatidylcholine, glycocholic acid, and taurocholic acid, were identified as potential biomarkers (area under the curve: 0.817; 95% confidence interval: 0.696–0.913). Pathway analysis indicated involvement of primary bile acid biosynthesis and the metabolism of taurine and hypotaurine (p < 0.05). The findings from untargeted metabolomic analysis demonstrated an increase in bile acids in children with hepatotoxicity. The accumulation of cytotoxic bile acids should be further investigated to elucidate the role of these metabolites in drug-induced liver injuryInstitute of Health Carlos III grants (PI17/01989) and Servicio Andaluz de Salud (F2-0071-2015

Exploring the Role of Sclerostin as a Biomarker of Cardiovascular Disease and Mortality: A Scoping Review

Sclerostin is most recognized for its role in controlling bone formation; however, it is also expressed in the heart, aorta, coronary, and peripheral arteries. Human studies have associated high circulating sclerostin levels with the presence of different cardiovascular diseases (CVD), surrogate CVD markers, and a high risk of cardiovascular events in some populations. However, this is still a matter of scientific debate, as the results have been very heterogeneous among studies. In the present review, the association between serum sclerostin levels and CVD and/or cardiovascular mortality was analyzed. For this purpose, a scoping review was performed in which articles measuring serum sclerostin levels and cardiovascular risk in patients were selected. Eleven articles answered the research question; of these articles, 8/11 evaluated the association between sclerostin and CVD, of which 4/8 found a positive association, 2/8 found a negative association, and 2/8 found no association between variables. Five (5/11) of the articles included in the study evaluated cardiovascular mortality, of which 3/5 found a positive association, 1/5 found a negative association, and 1/5 found no association between variables. In conclusion, we did not find sufficient results to be able to demonstrate an association between elevated sclerostin levels and the development of CVD and/or cardiovascular mortality in the general population due to heterogeneity in the results. However, there seems to be a tendency to consider increased sclerostin levels as a risk factor for both the development of cardiovascular events and cardiovascular mortality in specific populations. Further studies in this field will help to solve some of the inconsistencies found during this scoping review and allow for the future use of sclerostin measurement as a strategy in the prevention and diagnosis of CVD and/or cardiovascular mortality.Instituto de Salud Carlos III European Commission PI18-00803 PI21/01069 PI18-01235 CD20/00022European CommissionJunta de Andalucia PI-0268-2019 RH-0069-2021Instituto de Salud Carlos IIIUniversity of Granada FI19/00118European Commission 811

The Contribution of Wnt Signaling to Vascular Complications in Type 2 Diabetes Mellitus

Vascular complications are the leading cause of morbidity and mortality among patients with type 2 diabetes mellitus (T2DM). These vascular abnormalities result in a chronic hyperglycemic state, which influences many signaling molecular pathways that initially lead to increased oxidative stress, increased inflammation, and endothelial dysfunction, leading to both microvascular and macrovascular complications. Endothelial dysfunction represents the initial stage in both types of vascular complications; it represents “mandatory damage” in the development of microvascular complications and only “introductory damage” in the development of macrovascular complications. Increasing scientific evidence has revealed an important role of the Wnt pathway in the pathophysiology of the vascular wall. It is well known that the Wnt pathway is altered in patients with T2DM. This review aims to be an update of the current literature related to the Wnt pathway molecules that are altered in patients with T2DM, which may also be the cause of damage to the vasculature. Both microvascular complications (retinopathy, nephropathy, and neuropathy) and macrovascular complications (coronary artery disease, cerebrovascular disease, and peripheral arterial disease) are analyzed. This review aims to concisely concentrate all the evidence to facilitate the view on the vascular involvement of the Wnt pathway and its components by highlighting the importance of exploring possible therapeutic strategy for patients with T2DM who develop vascular pathologies.Instituto de Salud Carlos III European Commission PI18-00803 PI21-01069 PI18-01235 CD20/00022 FI19/00118European CommissionJunta de Andalucia PI-0268-2019University of GranadaEuropean Commission 811

Undercarboxylated Osteocalcin: A Promising Target for Early Diagnosis of Cardiovascular and Glycemic Disorders in Patients with Metabolic Syndrome: A Pilot Study

Lifestyle changes are causing an exponential increase in the prevalence of obesity and metabolic syndrome (MetS) worldwide. The most frequent complications of these are the development of diabetes (T2D) and cardiovascular disease (CVD). Accurate tools are needed to classify the cardiovascular risk (CVR) in the MetS population. In recent years, numerous biomarkers of bone metabolism have been associated with CVR. The aim of this study was to determine the levels of undercarboxylated osteocalcin (ucOC) in a cohort of patients with MetS and to analyse its association with MetS parameters and CVR as well as with T2D prevalence. A longitudinal study was conducted in which a MetS population was followed for one year. Weight change, adherence to the Mediterranean diet (MedDiet), ucOC levels, MetS parameters and CVR were analysed and CVR was calculated using different scores. Our results showed a decrease of CVR associated with a better adherence to the MetDiet resulting in higher HDL-C and ucOC levels though the improvement of MetS risk factors. This bone protein appeared as a potential biomarker to classify CVR in the MetS population, especially for MetS patients without prevalent T2D. Furthermore, ucOC serum levels could be good predictors of T2D prevalence.Instituto de Salud Carlos III European Commission PI18-00803 PI21/01069 PI18-01235 FI19/00118 CD20/00022European CommissionJunta de Andalucia CD20/00022 PI-0268-2019 RH-0069202

Systemic effects of hypophosphatasia characterization of two novel variants in the ALPL gene

IntroductionHypophosphatasia (HPP) is an inborn metabolic error caused by mutations in the ALPL gene encoding tissue non-specific alkaline phosphatase (TNSALP) and leading to decreased alkaline phosphatase (ALP) activity. Although the main characteristic of this disease is bone involvement, it presents a great genetic and clinical variability, which makes it a systemic disease.MethodsPatients were recruited based on biochemical assessments. Diagnosis was made by measuring serum ALP and pyridoxal 5-phosphate levels and finally by Sanger sequencing of the ALPL gene from peripheral blood mononuclear cells. Characterization of the new variants was performed by transfection of the variants into HEK293T cells, where ALP activity and cellular localization were measured by flow cytometry. The dominant negative effect was analyzed by co-transfection of each variant with the wild-type gene, measuring ALP activity and analyzing cellular localization by flow cytometry.ResultsTwo previously undescribed variants were found in the ALPL gene: leucine 6 to serine missense mutation (c.17T&gt;C, L6S) affecting the signal peptide and threonine 167 deletion (c.498_500delCAC, T167del) affecting the vicinity of the active site. These mutations lead mainly to non-pathognomonic symptoms of HPP. Structural prediction and modeling tools indicated the affected residues as critical residues with important roles in protein structure and function. In vitro results demonstrated low TNSALP activity and a dominant negative effect in both mutations. The results of the characterization of these variants suggest that the pleiotropic role of TNSALP could be involved in the systemic effects observed in these patients highlighting digestive and autoimmune disorders associated with TNSALP dysfunction.ConclusionsThe two new mutations have been classified as pathogenic. At the clinical level, this study suggests that both mutations not only lead to pathognomonic symptoms of the disease, but may also play a role at the systemic level

Breeding vegetables with increased content in bioactive phenolic acids

[EN] Vegetables represent a major source of phenolic acids, powerful antioxidants characterized by an organic carboxylic acid function and which present multiple properties beneficial for human health. In consequence, developing new varieties with enhanced content in phenolic acids is an increasingly important breeding objective. Major phenolic acids present in vegetables are derivatives of cinnamic acid and to a lesser extent of benzoic acid. A large diversity in phenolic acids content has been found among cultivars and wild relatives of many vegetable crops. Identification of sources of variation for phenolic acids content can be accomplished by screening germplasm collections, but also through morphological characteristics and origin, as well as by evaluating mutations in key genes. Gene action estimates together with relatively high values for heritability indicate that selection for enhanced phenolic acids content will be efficient. Modern genomics and biotechnological strategies, such as QTL detection, candidate genes approaches and genetic transformation, are powerful tools for identification of genomic regions and genes with a key role in accumulation of phenolic acids in vegetables. However, genetically increasing the content in phenolic acids may also affect other traits important for the success of a variety. We anticipate that the combination of conventional and modern strategies will facilitate the development of a new generation of vegetable varieties with enhanced content in phenolic acids.This research has been funded by Ministerio de Economia y Competividad and FEDER [grant AGL2012-34213]. Prashant Kaushik is thankful to Indian Council of Agricultural Research [ICAR] for providing an International Fellowship for doctoral studies. Isabel Andujar and Pietro Gramazio are grateful to Universitat Politecnica de Valencia [Programa de Ayudas de Investigacion y Desarrollo, PAID] for a postdoctoral and a predoctoral contract, respectively.Kaushik, P.; Andújar, I.; Vilanova Navarro, S.; Plazas Ávila, MDLO.; Gramazio, P.; Herraiz García, FJ.; Brar, NS.... (2015). Breeding vegetables with increased content in bioactive phenolic acids. Molecules. 20(10):18464-18481. https://doi.org/10.3390/molecules201018464S1846418481201

Desarrollo de un método de predicción de actividad proteica a partir de su secuencia aminoacídica

La revolución en cuanto a la obtención de datos biológicos procedentes de la secuenciación de alto rendimiento genera un distanciamiento importante entre los datos secuenciados y los datos analizados. Para reducir esta distancia, y poder llegar a un mayor entendimiento de la información que disponemos, se hace necesaria la aparición de herramientas que permitan digerir tanta información generada y ponerla en situación disponible para la comunidad científica para su comprensión y utilización en las diferentes áreas. Es por ello, por lo que hoy en día se trabaja concienzudamente en la realización de métodos informáticos, que permitan en cierta medida, completar el trabajo realizado por los métodos experimentales costosos y laboriosos, aunque necesarios, puesto que constituyen, en la mayoría de las ocasiones, la base sobre la que trabajar con los métodos informáticos. Con este trabajo se pretende proponer un método sencillo de predicción de función proteica basándose en la homología de secuencias de proteínas con función conocida y anotadas mediante el sistema de ontología de genes.The revolution in obtaining biological data from the high-throughput sequencing generates a significant gap between sequenced data and analyzed data. In order to reduce this problem, and to find a better understanding of the information we have, it is necessary the emergence of tools to digest the enormous quantity of information generated and make it available to the scientific community for their understanding and use in different fields. That is why nowadays, the bioinformaticians are hard working to develop computational methods that allow in this context, complete the work done by the expensive and laborious experimental methods, which are necessary, since they are in most cases, the basis on which to work using bioinformatic tools. This work aims to propose a simple method for predicting protein function based on sequence homology of proteins with known and annotated functions by gene ontology system.La revolució pel que fa a l'obtenció de dades biològiques procedents de la seqüenciació d'alt rendiment genera un distanciament important entre les dades seqüenciades i les dades analitzades. Per reduir aquesta distància, i poder arribar a un major enteniment de la informació que disposem, es fa necessària l'aparició d'eines que permetin digerir tanta informació generada i posar-la en situació disponible per a la comunitat científica per la seva comprensió i utilització en les diferents àrees. És per això, pel que avui en dia es treballa a consciència en la realització de mètodes informàtics, que permetin en certa mesura, completar el treball realitzat pels mètodes experimentals costosos i laboriosos, encara que necessaris, ja que constitueixen, en la majoria de les ocasions, la base sobre la qual treballar amb els mètodes informàtics. Amb aquest treball es pretén proposar un mètode senzill de predicció de funció proteica basant-se en l'homologia de seqüències de proteïnes amb funció coneguda i anotades mitjançant el sistema d'ontologia de gens

Molecular cloning, tissue distribution and daily expression of cry1 and cry2 clock genes in European seabass (Dicentrarchus labrax).

Biological rhythms are driven by circadian oscillators, which are ultimately controlled by the cyclic expression of clock genes. Cryptochromes (CRY), blue light photoreceptors, belong to the negative elements of the transcriptional feedback loop into the molecular clock. This paper describes the cloning and characterization of two cryptochromes (cry1 and 2) in European seabass, which is considered an interesting chronobiology model due to its dual (diurnal/nocturnal) behavior. The cloned cDNA fragments encoded for two proteins of 567 and 668 amino acids, which included the FAD-binding and the DNA-photolyase domains. Moreover, both proteins had a high homology with cryptochrome proteins (Cry) of other teleost fish. These cry1 and 2 genes were expressed in several tissues of seabass (brain, liver, heart, retina, muscle, spleen, gill and intestine). In addition, the daily expression of cry1 was rhythmic in brain, heart and liver with the acrophase around ZT 03:15 h (after the onset of lights). Similarly, the cry2 daily expression was rhythmic in liver, peaking at ZT 03:28 h, whereas in brain the acrophase was at ZT 11:08 h (shortly prior to the offset of lights). These findings provide new elements to help understanding the functioning of the molecular clock of seabass

Identification of Potential Targets Linked to the Cardiovascular/Alzheimer’s Axis through Bioinformatics Approaches

This research was funded by Instituto de Salud Carlos III grant (PI18-00803), co-funded by the European Regional Development Fund (FEDER) and by Junta de Andalucía grant (PI-0268- 2019). In addition, C.G.-F. and S.G.-S. are funded by postdoctoral and predoctoral fellowships from Instituto de Salud Carlos III with co-funding by FEDER (CD20/00022 and FI19/00118 respectively). R.S.d.l.T. was funded by the University of Granada with co-funding by FEDER, by grant number 8110 (Research investigator call in the framework of the youth guarantee Program)The identification of common targets in Alzheimer's disease (AD) and cardiovascular disease (CVD) in recent years makes the study of the CVD/AD axis a research topic of great interest. Besides aging, other links between CVD and AD have been described, suggesting the existence of common molecular mechanisms. Our study aimed to identify common targets in the CVD/AD axis. For this purpose, genomic data from calcified and healthy femoral artery samples were used to identify differentially expressed genes (DEGs), which were used to generate a protein-protein interaction network, where a module related to AD was identified. This module was enriched with the functionally closest proteins and analyzed using different centrality algorithms to determine the main targets in the CVD/AD axis. Validation was performed by proteomic and data mining analyses. The proteins identified with an important role in both pathologies were apolipoprotein E and haptoglobin as DEGs, with a fold change about +2 and -2, in calcified femoral artery vs healthy artery, respectively, and clusterin and alpha-2-macroglobulin as close interactors that matched in our proteomic analysis. However, further studies are needed to elucidate the specific role of these proteins, and to evaluate its function as biomarkers or therapeutic targets.Instituto de Salud Carlos III grant (PI18-00803) European Regional Development Fund (FEDER) Junta de Andalucía grant (PI-0268- 2019)Instituto de Salud Carlos III FEDER (CD20/00022 and FI19/00118 respectively)University of Granada FEDER 811

Analysis of the Genetic Relationship between Atherosclerosis and Non-Alcoholic Fatty Liver Disease through Biological Interaction Networks

Non-alcoholic fatty liver disease (NAFLD) seems to have some molecular links with atherosclerosis (ATH); however, the molecular pathways which connect both pathologies remain unexplored to date. The identification of common factors is of great interest to explore some therapeutic strategies to improve the outcomes for those affected patients. Differentially expressed genes (DEGs) for NAFLD and ATH were extracted from the GSE89632 and GSE100927 datasets, and common up- and downregulated DEGs were identified. Subsequently, a protein–protein interaction (PPI) network based on the common DEGs was performed. Functional modules were identified, and the hub genes were extracted. Then, a Gene Ontology (GO) and pathway analysis of common DEGs was performed. DEGs analysis in NAFLD and ATH showed 21 genes that were regulated similarly in both pathologies. The common DEGs with high centrality scores were ADAMTS1 and CEBPA which appeared to be down- and up-regulated in both disorders, respectively. For the analysis of functional modules, two modules were identified. The first one was oriented to post-translational protein modification, where ADAMTS1 and ADAMTS4 were identified, and the second one mainly related to the immune response, where CSF3 was identified. These factors could be key proteins with an important role in the NAFLD/ATH axis