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Trabajo fin de máster presentado en la Universidad de La Rioja, Máster Universitario en Tecnología, Gestión e Innovación Vitivinícola, [ES] Dentro de la industria enológica, la composición fenólica de las uvas es un parámetro de gran relevancia debido al impacto que estos metabolitos tienen en la generación de vinos con diferentes perfiles organolépticos. Además, dentro de los fenoles, los antocianos son unos de los compuestos con mayor importancia en la producción de vinos, ya que son los metabolitos implicados en la obtención de vinos con diferentes matices de color, como rosados o tintos. En el presente estudio se trabajó con dos variedades de uva blanca, Albariño y Verdejo, de mucho prestigio en el sector del vino, y sus dos variantes somáticas rojas (Albariño rojo y Verdejo Rojo), que fueron generadas a partir de una mutación somática del locus del color en los genes VvMybA, relacionados con la síntesis de antocianos en bayas. En este trabajo, se determinó el contenido fenólico en los momentos de madurez y envero, para determinar las rutas de síntesis de compuestos fenólicos activadas durante el desarrollo fenológico de las uvas. También se analizaron los compuestos fenólicos durante la fermentación para ver qué efecto tiene la recuperación del color en baya en la producción de vinos a partir de estas nuevas variantes. La variedad que alcanzó el mayor contenido en antocianos, a lo largo de todas las fases del estudio fue Albariño Rojo, un contenido mucho mayor significativamente respecto al Verdejo Rojo. Mientras que en las variedades blancas el contenido de antocianos no fue resaltable. Durante la fermentación también se acumularon compuestos fenólicos, aunque con un contenido menor respecto al detectado en piel En las variantes coloreadas se observó, una mayor presencia de antocianos dihidroxilados, principalmente cianidina, mientras que los antocianos trihidroxilados se acumularon en menor medida. También se encontró una mayor concentración de antocianos 3-glucóxidos en todas las variedades. Todas estas características se vieron a lo largo de todas las fases del estudio. En relación con los fenoles no coloreados, las mayores diferencias en los distintos perfiles de acumulación de metabolitos parecen estar más relacionadas con cuestiones intravarietales, así como con las condiciones y el manejo de cultivo. Se pudo observar que el flavonol mayoritario fue quercetina-3-glucoxido en todas las variedades y a lo largo del estudio. Este compuesto, se obtiene a través de la ruta de síntesis en la que está implicada la enzima flavonoide-3’- hidrolasa, que conduce a la obtención de derivados dihidroxilados, lo que concuerda con lo obtenido en los compuestos fenólicos coloreados, en detrimento de la rama de la ruta en la que está involucrada la enzima flavonoide-3’,5’-hidrolasa, que conduce a los derivados trihidroxilados. Así, los resultados obtenidos permiten evaluar el potencial vitivinícola de las nuevas variedades de uva coloreada rojas y abrir una puerta a futuros estudios orientados a evaluar su calidad organoléptica, enológica y su aplicación comercial., [EN] In the wine-making -industry, phenolic composition in grapes is a highly relevant parameter, due to the impact those metabolites have in the wine making with different organoleptic profiles. In addition, within the phenolic compounds, anthocyanins are one of the most important in wine making with different sensitive’s profiles, because they are metabolites involved in the development of wine with different colors, such as rosé or red. Thus, the present study analyzed two white varieties, Albariño and Verdejo, both elite varieties in the wine sector, and two red bud sports (Albariño Rojo and Verdejo Rojo) that come from a somatic mutation of the color locus in the VvMybA genes, that are involved in the anthocyanin biosynthesis in grapes. In this study, phenolic content was measured at veraison and ripening, to identify the activated biosynthesis pathway of phenolic compounds along phenological development of grapes. Phenolic composition was measured during alcoholic fermentation to determinate the effect of the color recovery in wine making from these new varieties. The variety that reached the highest anthocyanin concentration was Albariño Rojo, along all studied stages, a higher significant concentration compared to Verdejo Rojo. While in white varieties the anthocyanin content was not remarkable. Phenolic compounds were also accumulated during alcoholic fermentation. Although the content was lower than in the skin. In colored varieties, a higher presence of dihydroxylated anthocyanins, mainly cyaniding was quantified, although trihydroxylated anthocyanins accumulation was lower. A higher concentration of -3-glucoxides anthocyanins was found in all varieties. All these characteristics were shown along this study. In relation to non-colored phenols, the greatest differences in the different metabolites accumulation profiles seem to be more influenced by intra-varietal characteristics, and with crop conditions and management. Quercetin-3-glucoxide was the mainly flavonol identified in all varieties and along the study. This compound is obtained through the biosynthesis pathway in which the flavonoid 3’-hydroxylase enzyme is involved, that leads to the production of dihydroxylated derivatives, in the same way with colored phenolic compounds, to the detriment of the branch of the pathway in which the enzyme flavonoid 3’,5’-hydroxylase is involved, leading to trihydroxylated derivatives. Thus, the results obtained allow evaluating the viticultural potential of the new red varieties and open a door to future research to evaluate its organoleptic and oenological quality and its commercial application

Natural Killer (NK) cells belong to the innate lymphoid lineage and are highly present in the human skin. NK cells can produce a range of pro-inflammatory mediators, including cytokines and chemokines. The role of NK(-T) cells in the immune response towards Borrelia burgdorferi infection was studied. The production of interleukin (IL)-6, IL-1β and interferon-gamma (IFN-γ) by human primary peripheral blood mononuclear cells (PBMCs) exposed to B. burgdorferi was assessed. Interestingly, CD56

AIMS: To assess tolerability and optimal time point for initiation of sacubitril/valsartan in patients stabilised after acute heart failure (AHF). METHODS AND RESULTS: TRANSITION was a randomised, multicentre, open-label study comparing two treatment initiation modalities of sacubitril/valsartan. Patients aged ≥ 18 years, hospitalised for AHF were stratified according to pre-admission use of renin-angiotensin-aldosterone system inhibitors and randomised (n = 1002) after stabilisation to initiate sacubitril/valsartan either ≥ 12-h pre-discharge or between Days 1-14 post-discharge. Starting dose (as per label) was 24/26 mg or 49/51 mg bid with up- or down-titration based on tolerability. The primary endpoint was the proportion of patients attaining 97/103 mg bid target dose after 10 weeks. Median time of first dose of sacubitril/valsartan from the day of discharge was Day -1 and Day +1 in the pre-discharge group and the post-discharge group, respectively. Comparable proportions of patients in the pre- and post-discharge initiation groups met the primary endpoint [45.4% vs. 50.7%; risk ratio (RR) 0.90; 95% confidence interval (CI) 0.79-1.02]. The proportion of patients who achieved and maintained for ≥ 2 weeks leading to Week 10, either 49/51 or 97/103 mg bid was 62.1% vs. 68.5% (RR 0.91; 95% CI 0.83-0.99); or any dose was 86.0% vs. 89.6% (RR 0.96; 95% CI 0.92-1.01). Discontinuation due to adverse events occurred in 7.3% vs. 4.9% of patients (RR 1.49; 95% CI 0.90-2.46). CONCLUSIONS: Initiation of sacubitril/valsartan in a wide range of heart failure with reduced ejection fraction patients stabilised after an AHF event, either in hospital or shortly after discharge, is feasible with about half of the patients achieving target dose within 10 weeks. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT02661217. © 2019 The Authors. European Journal of Heart Failure published by John Wiley & Sons peerReviewed

[eng] The mammary gland develops postnatally and is remodeled, during each estrous cycle and pregnancy, through very dynamic expansions and involutions of its polarized epithelial tree. Moreover, the mammary gland is hierarchically organized, where the two main epithelial populations may arise from a common bipotent mammary stem cell (MaSC). The principal epithelial cell types in the mammary gland are luminal cells and myoepithelial cells (also named basal cells), which are found surrounding luminal cells and in contact with the extracellular matrix. Further, the luminal compartment has two main lineages: the alveolar lineage in charge of producing milk during lactation, and the ductal lineage that include the cells that express receptors of the ovarian hormones like estrogen and progesterone. Interestingly, each of these luminal lineages is sustained by their own progenitors. Thus, the mammary gland requires a tightly regulated balance between proliferation and differentiation, which is disrupted in breast tumors. Distinct transcriptional circuits orchestrating such finely tuned processes have been described; however, how translational control contributes to mammary gland homeostasis and tumorigenesis remains largely unexplored. The CPEB-family (Cytoplasmic Polyadenylation Element Binding Proteins) of RNA- binding proteins (RBPs) is composed of four members in vertebrates (CPEB1-4); differentially, in Drosophila Melanogaster, there are two CPEB orthologs (Orb and Orb2). Within the CPEB-family, there are two subfamilies based on sequence identity of the RNA recognition motifs: CPEB1 and CPEB2-4. CPEB proteins share the same RNA- binding C-terminal domain (CTD), while they differ on their unstructured regulatory N- terminal domain (NTD). This fact has two significant implications: CPEBs can bind overlapping target mRNAs by recognizing CPEs in the 3’UTR; however, they are regulated by different post-transcriptional modifications at the NTD that will be triggered by distinct signaling pathways. Therefore, CPEB proteins could compete against one another for the binding, act sequentially on the same target mRNA or even compensate the loss of one member of the CPEB-family. CPEBs regulated cytoplasmic polyadenylation by recognizing a motif called Cytoplasmic Polyadenylation Element (CPE) present in the 3’UTR of certain mRNAs. Remarkably, CPEBs can potentially regulate up to 25% of the genome and they can control translation of CPEB-bound mRNAs in time and subcellular space. Furthermore, CPEB proteins play pivotal roles in cell proliferation and lineage-specification. Therefore, the study of the CPEBs in a hierarchically organized tissue undergoing a lot of proliferation and remodeling like the mammary epithelia may shed light on novel functions of this family of RBPs. The main goal of this study is to analyze in vivo the contribution of the four members of the CPEB-family of RNA-binding proteins in mammary epithelial morphodynamics. Here we present a systematic study of the four members of the CPEB family (CPEB1-4) in the context of the adult mammary gland in vivo, using Knock-out (KO) models for all four CPEBs. By mammary wholemounts and flow cytometry analysis, we discovered that the lack of CPEB2 resulted in defects in mammary gland branching and lineage specification. Interestingly, CPEB2 depletion also had consequences for breast tumorigenesis. Moreover, were able to identify the target mRNAs bound by CPEB2 in mammary epithelial cells and to establish a molecular mechanism by which CPEB2 regulates mammary gland homeostasis and breast cancer. Altogether, this work unravels a novel translational mechanism regulating cell fate in the mammary gland and breast tumor development., [spa] La glándula mamaria es el único órgano que se desarrolla principalmente después del nacimiento y, además, se remodela durante cada ciclo menstrual y embarazo a través de expansiones e involuciones muy dinámicas del epitelio polarizado que la constituye. Por lo tanto, la glándula mamaria requiere un equilibrio finamente regulado entre proliferación y diferenciación. Este equilibro se encuentra perturbado en casos de cáncer de mama. De qué manera el control tradicional contribuye a la homeostasis y tumorigenesis de la glándula mamaria es un terreno por explorar. La familia de proteínas CPEBs (Cytoplasmic Polyadenylation Element Binding) incluye cuatro proteínas de unión al RNA mensajero que regulan, de manera temporal y espacial, la traducción y la localización sub-celular de los mRNAs que unen. Las CPEBs podrían regular hasta el 25% del genoma. Aquí presentamos un estudio sistemático de los cuatro miembro de la familia de las CPEBs (CPEB1-4) en el contexto de la glándula mamaria adulta, utilizando modelos knock-out (KO) en ratón. Durante esta investigación hemos descubierto que la falta de CPEB2 resulta en defectos en las ramificaciones de la glándula mamaria, y también en diferenciación. De manera muy relevante, la depleción de CPEB2 tiene consecuencias en cáncer de mama. En conclusión, es trabajo descifra un nuevo mecanismo a nivel de traducción responsable de la regulación de la homeostasis y el desarrollo de tumores en la glándula mamaria.