Regulación de la reparación de los cortes de doble cadena en el ADN: papel de la nedilación de proteínas

Las roturas de doble cadena del ADN son las lesiones más citotóxicas que pueden ocurrir en el ADN. Pueden ser reparadas por diferentes mecanismos y para la supervivencia celular se requiere un control preciso de ellos. En esta tesis se ha caracterizado y validado un nuevo sistema de reparación denominado See-Saw reporter (SSR) que mide el balance entre la ruta de reparación por unión de extremos no homólogos (NHEJ) y la reparación por recombinación homóloga (HR), siendo por tanto una herramienta que permite encontrar y caracterizar nuevos factores implicados en la regulación de las rutas de reparación de los DBSs. De hecho, se ha determinado el flujo de trabajo para la realización de un escrutinio con una librería de esiRNA del genoma completo usando este sistema. Así mismo, se ha analizado el efecto en el balance NHEJ/HR de una minilibrería de inhibidores con potencial terapéutico y se ha puesto de manifiesto el papel principal que tiene la nedilación de proteínas en la elección de la ruta de reparación mediante la utilización del inhibidor de la nedilación MLN4924. La inhibición de la nedilación cambia el perfil normal de reparación hacia un aumento en la reparación por recombinación homóloga. Además hemos comprobado que la nedilación dependiente de RNF111 y UBE2M inhibe la resección del ADN mediada por CtIP y BRCA1 y, por tanto, la recombinación homóloga. Por otro lado, hemos comprobado que las denedilasas COP9 signalosoma y NEDP1 son necesarias para la resección del ADN y también modulan el equilibrio NHEJ/HR. Finalmente hemos concluido que la nedilación no sólo afecta al equilibrio entre la reparación por NHEJ y la HR sino que también controla el balance entre las diferentes rutas de reparación por HR. Por tanto, el estado de nedilación de las proteínas tiene un gran impacto en la manera en la que las células humanas responden a las roturas en el ADN

Occupational Safety and Health 5.0—A Model for Multilevel Strategic Deployment Aligned with the Sustainable Development Goals of Agenda 2030

The concept of Industry 4.0 (I4.0) is evolving towards Industry 5.0 (I5.0), where the human factor is the central axis for the formation of smart cyber-physical socio-technical systems that are integrated into their physical and cultural host environment. This situation generates a new work ecosystem with a radical change in the methods, processes and development scenarios and, therefore, in the occupational risks to which safety science must respond. In this paper, a historical review of the evolution of work as a complex socio-technical system formalised through Vygostky’s theory of Activity and the contributions of safety science is carried out, for its projection in the analysis of the future of complex systems as an opportunity for safety research linked to the current labour context in transformation. Next, the Horizon 2020 strategies for Occupational Safety and Health (OSH) at the European level are analysed to extract the lessons learned and extrapolate them towards the proposed model, and subsequently the conceptual frameworks that are transforming work and Occupational Risk Prevention (ORP) in the transition to Industry 4.0 are identified and reviewed. Finally, a model is formulated that formalises the deployment of public policies and multi-level and multi-scale OSH 5.0 strategies within the framework of the Sustainable Development Goals (SDGs) of the United Nations (UN) for Horizon 2030

New Tools to Study DNA Double-Strand Break Repair Pathway Choice

A broken DNA molecule is difficult to repair, highly mutagenic, and extremely cytotoxic. Such breaks can be repaired by homology-independent or homology-directed mechanisms. Little is known about the network that controls the repair pathway choice except that a licensing step for homology-mediated repair exists, called DNA-end resection. The choice between these two repair pathways is a key event for genomic stability maintenance, and an imbalance of the ratio is directly linked with human diseases, including cancer. Here we present novel reporters to study the balance between both repair options in human cells. In these systems, a double-strand break can be alternatively repaired by homology-independent or -dependent mechanisms, leading to the accumulation of distinct fluorescent proteins. These reporters thus allow the balance between both repair pathways to be analyzed in different experimental setups. We validated the reporters by analyzing the effect of protein downregulation of the DNA end resection and non-homologous end-joining pathways. Finally, we analyzed the role of the DNA damage response on double-strand break (DSB) repair mechanism selection. Our reporters could be used in the future to understand the roles of specific factors, whole pathways, or drugs in DSB repair pathway choice, or for genome-wide screening. Moreover, our findings can be applied to increase gene-targeting efficiency, making it a beneficial tool for a broad audience in the biological sciences. © 2013 Gomez-Cabello et al.This work has been funded by a R+D+I grant from the Spanish Ministry of Economy and Competitivity (SAF2010-14877) and an European Research Council (ERC) Starting Grant (DSBRECA).Peer Reviewe

New tools to study DNA double-strand break repair pathway choice

A broken DNA molecule is difficult to repair, highly mutagenic, and extremely cytotoxic. Such breaks can be repaired by homology-independent or homology-directed mechanisms. Little is known about the network that controls the repair pathway choice except that a licensing step for homology-mediated repair exists, called DNA-end resection. The choice between these two repair pathways is a key event for genomic stability maintenance, and an imbalance of the ratio is directly linked with human diseases, including cancer. Here we present novel reporters to study the balance between both repair options in human cells. In these systems, a double-strand break can be alternatively repaired by homology-independent or -dependent mechanisms, leading to the accumulation of distinct fluorescent proteins. These reporters thus allow the balance between both repair pathways to be analyzed in different experimental setups. We validated the reporters by analyzing the effect of protein downregulation of the DNA end resection and non-homologous end-joining pathways. Finally, we analyzed the role of the DNA damage response on double-strand break (DSB) repair mechanism selection. Our reporters could be used in the future to understand the roles of specific factors, whole pathways, or drugs in DSB repair pathway choice, or for genome-wide screening. Moreover, our findings can be applied to increase gene-targeting efficiency, making it a beneficial tool for a broad audience in the biological sciences

Planes de Emergencia Exterior en accidentes graves de industrias químicas. Formulación de un modelo de referencia

In this document, a complete methodological proposal on the realization of an Emergency Action Plan (EAP) is developed, to serve as guide to industries in which dangerous chemical substances are present and affected by the Directive 2012/18/UE and the Royal Decree 840/2015. First, a search on the best available techniques and current legislation is realized, pointing all the necessary steps previous to the realization of the actual EAP, and the variety of documents apart from the EAP that must be generated to complement it. Once all the previous information has been gathered, the methodology on the elaboration of an EAP is proposed, including information related to the industry, the chemicals present in it, the risks and accidents probabilities, the installation organization for emergencies, the Administration cooperation, the population information, the actuation, etc. This document, as mentioned, comprises every necessary step to achieve in an optimal way a complete EAP. This is advantageous related to this prevention field, as it includes information and documents that complement the EAP and eases its elaboration and comprehension, which is not covered currently in the Best Available TechniquesEn el presente trabajo se plantea una propuesta metodológica de realización de un Plan de Emergencia Exterior (PEE), para servir como guía a industrias donde haya presentes sustancias químicas peligrosas y afectadas por la Directiva 2012/18/UE y el Real Decreto 840/2015. En primer lugar se realiza un exhaustivo estado del arte relativo a las mejores técnicas disponibles referentes a planes de emergencia para accidentes graves en industrias químicas, así como el estudio de la legislación vigente, indicando los documentos y procedimientos a tener en cuenta en un PEE. Con la información especificada se desarrolla una guía de implantación que incluye una propuesta metodológica para la elaboración de un PEE completo, incluyendo información sobre la industria, sustancias presentes, riesgos y probabilidades de accidentes, organización de la instalación frente a emergencias, cooperación con la administración, información a la población, actuación, etc. Este trabajo realizado engloba todos los pasos necesarios para facilitar y optimizar la elaboración de los Planes de Emergencia Exterior, suponiendo un importante avance en este campo de la prevención, pues incluye también información y documentos complementarios al PEE que facilitan su elaboración y comprensión, lo cual no queda reflejado en la actualidad dentro de las Mejores Técnicas Disponibles

Neddylation inhibits CtIP-mediated resection and regulates DNA double strand break repair pathway choice

DNA double strand breaks are the most cytotoxic lesions that can occur on the DNA. They can be repaired by different mechanisms and optimal survival requires a tight control between them. Here we uncover protein deneddylation as a major controller of repair pathway choice. Neddylation inhibition changes the normal repair profile toward an increase on homologous recombination. Indeed, RNF111/UBE2M-mediated neddylation acts as an inhibitor of BRCA1 and CtIP-mediated DNA end resection, a key process in repair pathway choice. By controlling the length of ssDNA produced during DNA resection, protein neddylation not only affects the choice between NHEJ and homologous recombination but also controls the balance between different recombination subpathways. Thus, protein neddylation status has a great impact in the way cells respond to DNA breaks.España , Ministerio de Economía y Competitividad SAF2010-1487

Diferencias en el sustrato endo y epicárdico de las taquicardias ventriculares posinfarto analizado mediante resonancia magnética cardiaca con realce tardío

Algunas taquicardias ventriculares (TV) posinfarto se localizan en el epicardio. La identificación de diferencias en el sustrato de las TV endocárdicas y epicárdicas permitiría definir una mejor estrategia de ablación

The Helicase PIF1 Facilitates Resection over Sequences Prone to Forming G4 Structures

DNA breaks are complex lesions that can be repaired either by non-homologous end joining (NHEJ) or by homologous recombination (HR). The decision between these two routes of DNA repair is a key point of the DNA damage response (DDR) that is controlled by DNA resection. The core machinery catalyzing the resection process is well established. However, little is known about the additional requirements of DNA resection over DNA structures with high complexity. Here, we found evidence that the human helicase PIF1 has a role in DNA resection, specifically for defined DNA regions, such as those prone to form G-quadruplexes. Indeed, PIF1 is recruited to the site of DNA damage and physically interacts with proteins involved in DNA resection, and its depletion causes DNA damage sensitivity and a reduction of HR efficiency. Moreover, G4 stabilization by itself hampers DNA resection, a phenomenon suppressed by PIF1 overexpressio

Evolución temporal de las características de la escara isquémica en un modelo experimental de infarto agudo de miocardio: estudio basado en resonancia magnética

En los pacientes con infarto de miocardio, las características de la escara isquémica, incluyendo la extensión de tejido heterogéneo (TH) y los canales intraescara, se relacionan con la mortalidad arrítmica. Sin embargo, aún no se conoce bien la evolución temporal de la estructura de la escara en los primeros meses tras el infarto agudo de miocardio (IAM)