Data on the generation of two Nr2e3 mouse models by CRISPR / Cas9D10A nickase

NR2E3 encodes an orphan nuclear receptor that plays a dual function as both transcriptional activator and repressor in photoreceptors, being necessary for cone fate inhibition as well as rod differentiation and homeostasis. Mutations in this gene cause retinitis pigmentosa (RP), enhanced S cone syndrome (ESCS) and Goldmann-Favre syndrome (GFS). There is one reported Nr2e3 isoform that contains all 8 exons and a second -previously unreported- shorter isoform, which only spans the first 7 exons and whose function is still unknown. In this data article, we designed and generated two new mouse models by targeting exon 8 of Nr2e3 using the CRISPR/Cas9-D10A nickase in order to dissect the role of the two isoforms in Nr2e3 function and elucidate the different disease mechanisms caused by NR2E3 mutations. This strategy generated several modified alleles that altered the coding sequence of the last exon thereby affecting functional domains of the transcription factor. Allele Δ27 is an in-frame deletion of 27 bp that ablates the dimerization domain, whereas allele ΔE8 (full deletion of exon 8), produces only the short isoform that lacks the dimerization and repressor domains. Morphological and functional alterations of both Δ27 and ΔE8 mutants are reported in the associated research article "Nr2e3 functional domain ablation by CRISPR-Cas9D10A identifies a new isoform and generated Retinitis Pigmentosa and Enhanced S-cone Syndrome models" (Aísa-Marín et al., 2020)

On the Specific Adsorption of 7-methylguanine on Au(111) Surfaces for the Electroanalytical Sensing of Methylation Levels

The electrochemical determination of 7-methylguanine (7-mG) may result of interest because its presence can serve as probe of cytosine methylation of which is known as the most relevant epigenetic modification of DNA. This work explores the electrochemical response of 7-mG on different gold surfaces, both poly and single crystalline surfaces (Au (110), Au (111) and Au (100)). The results show that the adsorption-desorption process of the 7-mG is sensitive to the surface structure of the gold electrodes. Particularly, 7-mG adsorption-desorption profile on a Au (111) electrode exhibits some specific contributions which are found sensitive to the 7-mG concentration and, thereby could allow its quantification. These results may shed light on the future development of an electrochemical sensor for the diagnosis of the methylation degree in DNA.Authors would like to acknowledge funding obtained through the Spanish Ministry of Science and Innovation (MICINN) CTQ2013-48280-C3-3-R project

The integrated stress response releases the oncoprotein in TP53

Introduction: The TP53 gene is surrounded by a great duality: it is a critical tumour suppressor gene, however, its protective nature is frequently lost in tumours, where it becomes a powerful oncogene. Numerous studies attribute the oncogenic profile of TP53 to the missense mutations that commonly occur in cancer. We propose that this duality is intrinsic to TP53 instead of a consequence of mere somatic mutations, which could not have been evolutionarily selected for. This gene encodes for a set of protein isoforms with distinct and complementary functions, from which the full-length p53 (FLp53) is the best characterized. FLp53 is a transcription factor that mediates stress responses by promoting cell cycle arrest, DNA repair or apoptosis. In stark contrast to FLp53, the shorter isoform Δ160p53 promotes cell survival, proliferation, and invasion, and it is commonly overexpressed in tumours. Here we identify a disruption in the normal balance of p53 isoforms upon induction of the Integrated Stress Response (ISR), with the translation of Δ160p53 being favoured. Materials and Methods: Different cell lines were used to verify the expression of endogenous p53 isoforms during ISR, and the internal translation of Δ160p53 was tested with bicistronic constructs. The interaction of Δ160p53 with FLp53 was assessed by co-immunoprecipitation followed by western blot, and its effect in the expression of target genes was measured by RT-qPCR. Cells were treated with thapsigargin and tunicamycin to induce ISR when required. Results: The induction of ISR in a group of cell lines led to increased levels of endogenous Δ160p53 protein, as well as increased luminescence signal in the bicistronic system. The FLp53-Δ160 interaction was confirmed, and the role of Δ160p53 in the selective regulation of p53 target genes was uncovered. Conclusions: These data uncover a mode of activation of the oncogenic Δ160p53 and how this isoform can work together with FLp53. In the future, we aim to explore the clinical potential of these discoveries.This research was funded by grants 18K07229 (KAKENHI) from Japan Society for the Promotion of Science (JSPS) and PTDC/MED-ONC/32048/2017 from Fundação para a Ciência e a Tecnologia of Portugal (FCT). This work was supported by FCT through the individual research grant 2020.06982.BD.N/

Congreso online como herramienta docente para estudiantes de tercer ciclo en Electroquímica

Los congresos científicos son una buena herramienta para que los estudiantes de tercer ciclo puedan ampliar sus conocimientos. Sin embargo, las intervenciones donde los estudiantes plantean dudas o preguntas son prácticamente nulas. Con el objetivo de fomentar la participación de los estudiantes en los congresos científicos, la nueva red continua con el trabajo anteriormente realizado, llevando a cabo la II edición del Congreso online de estudiantes dentro del programa interuniversitario “Electroquímica. Ciencia y Tecnología”. La red busca concienciar acerca de la importancia que para un investigador tiene un congreso científico y a su vez, incrementar y mejorar su participación. Para ello, se utiliza un formato más atractivo que en la edición anterior, que mejora el entorno de trabajo y favorece la interacción entre los estudiantes. Asimismo, se emplean estrategias de comunicación más desarrolladas para hacer crecer el número de participantes. Finalmente, se establecerán diferentes parámetros para evaluar la actividad durante el congreso y se entregarán premios para motivar la participación

ATP release in allotransplantation

©2017. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This document is the Accepted, version of a Published Work that appeared in final form in Cell Report. To access the final edited and published work see 10.1016/j.celrep.2017.11.079Immune cells are equipped with a number of receptors that recognize sterile injury and pathogens. We find that host immune cells release ATP as an inflammatory signal in response to allogeneic transplantation. ATP then acts via a feedback mechanism on the P2X7 channel to activate the NLRP3 inflammasome and subsequently process and release interleukin (IL)-18. This process is a necessary stage in the deleterious Th1 response against allotransplantation via interferon-g production. Lack of IL-18 resulted in a decrease in graft- infiltrated CD8 cells, but an increase in regulatory T cells. In human liver transplant patients subjected to progressive immunosuppressive drug withdrawal, we found that patients suffering acute rejection had higher levels of the P2X7 receptor in circulating inflammatory monocytes compared to tolerant patients. These data suggest that the pharmacological inhibition of the P2X7 receptor or the NLRP3 inflammasome will aid in inducing transplant tolerance without complete immunoparalysis

Effect of different media additives on capacitation of frozen-thawed ram spermatozoa as a potential replacement for estrous sheep serum

Capacitation is a key process through which spermatozoa acquire their fertilizing ability. This event is required for the successful application of assisted reproductive technologies such as IVF. The aim of the present study was to investigate the effect of using a synthetic oviductal fluid medium supplemented with either heparin–hypotaurine alone, in combination with progesterone (P4), 17β-estradiol (E2), or BSA, or just β-cyclodextrin, in replacement for estrous sheep serum (ESS) for ram sperm capacitation. After incubation in the corresponding media for 15 (time 0) or 60 minutes, sperm function was evaluated by computerized sperm motility analysis and flow cytometry (plasma membrane status and fluidity). Treatments rendering the best results in regards to sperm function parameters related to capacitation were used for an IVF test. Herein, neither heparin–hypotaurine (alone), or in combination with P4, or E2, nor β-cyclodextrin induced capacitation-related changes in frozen–thawed ram spermatozoa. Only the medium supplemented with heparin–hypotaurine–BSA was able to induce changes compatible with in vitro capacitation relating to sperm motility pattern and plasma membrane fluidity, comparable to those in ESS-containing medium. Both media yielded sperm parameter values that differed (P < 0.05) from those obtained in the rest of the media tested. However, after the IVF trial, BSA was unable to support cleavage rates (21.80%) comparable to those obtained with ESS (52.60%; P < 0.05). We conclude that heparin–hypotaurine, P4, E2, β-cyclodextrin, or BSA is not suitable for replacing ESS in capacitation and fertilization media for ram spermatozoa.M. Ramón was supported by the Research Recruitment Program from the National Institute for Agricultural and Food Research program.Peer Reviewe

Retos de la universidad frente a la diversidad funcional (Dis-capacidad)

Existe en la sociedad actual un creciente interés por conocer la respuesta que ofrecen los distintos servicios de apoyo de las universidades españolas a sus estudiantes con diversidad funcional. Se han producido avances en cuanto a la integración de las personas en los niveles superiores y a pesar de que con el tiempo las universidades españolas han desarrollado servicios de orientación. A pesar de los avances en la supresión de barreras arquitectónicas, se siguen encontrando barreras sociales, actitudinales, de comunicación, técnicas y económicas que impiden el acceso a la educación a dichos alumnos o que les empujan hacia el abandono de sus estudios antes de finalizarlos. Comprobamos que muchas de las universidades españolas poseen un servicio de apoyo que garantiza la atención al alumnado con necesidades educativas especiales, ofreciendo medidas de atención individualizada, recursos adaptados, orientación, etc.., pero concluimos afirmando que siguen necesitando un cierto impulso en la formación y concienciación del personal docente, en las creación de adaptaciones, un aumento en el nivel de comunicación y en el establecimiento de líneas comunes de actuación

Implementación de un sistema piloto de telemedicina en Mendoza

La telemedicina se caracteriza por utilizar las nuevas tecnologías informáticas y de comunicaciones para ofrecer acceso a la salud, independientemente del lugar donde se encuentre quien lo necesite. Este informe hace referencia a la utilización de dichas tecnologías al plantear el desarrollo e implementación de un sistema de telemedicina, para la transmisión y monitoreo de señales cardíacas, con la finalidad de suplir atención sanitaria en comunidades rurales, donde la distancia constituye un factor crítico. Los actores principales que conforman la arquitectura tecnológica y la red son: 1) Sistemas Embebidos, 2) Adquisidores de Señales Cardíacas Inalámbricos, 3) Computadoras Personales PC, 4) Asistentes Digitales Personales PDA 5) Software y 6) Infraestructura de Comunicación. Según se agrupen estos elementos es posible distinguir tres alternativas o unidades, que de acuerdo con sus ventajas y desventajas serían aplicables a diferentes usos. En todos los casos la transmisión es realizada hacia un servidor, donde los datos correspondientes a las variables biológicas son almacenados en bases de datos, pudiendo ser luego objeto de análisis y diagnóstico. Hasta el momento los resultados obtenidos en ambas alternativas han sido satisfactorios, tanto en la fabricación del adquisidor de señales y como en la transmisión de los datos.Telemedicine is characterized for offering access to healthcare by using the new computer and communication technologies, independently of where the individual in need is. This report refers to the use of these technologies while outlining the development and implementation of a system for the monitoring and transmission cardiac signals with the purpose of replacing sanitary services in rural communities, where distance constitutes a critical factor. The main elements that make up the technological architecture and the network are: 1) the embedded systems, 2) Wireless Cardiac signal data loggers 3) Personal Computers PCs, 4) Personal Digital Assistants PDA, 5) Software and 6) Communication Infrastructure. Depending on the way we group these elements it is possible to distinguish three different alternatives, each one with its pros and cons for different applications. In all these alternatives, the data is sent to a server where the data of the biological variables are stored in a database, subject to later analysis and diagnosis. So far, the results obtained have been satisfactory both in the manufacturing of the data logger as well as the data transmissionWorkshop de Arquitecturas, Redes y Sistemas Operativos (WARSO)Red de Universidades con Carreras en Informática (RedUNCI

Aplicación del aprendizaje basado en problemas como método de enseñanza en el Master Universitario de Uso sostenible y Protección del suelo en Ambientes Mediterráneos. Póster

[SPA]El método del aprendizaje basado en problemas (ABP) se caracteriza en un aprendizaje centrado en el alumno que desarrolla una serie de habilidades y competencias indispensables en el entorno profesional. El problema consiste en la descripción de una situación concreta con finalidades pedagógicas para aprender o perfeccionarse en algún campo determinado. Al utilizar el método ABP se pretende que los alumnos estudien la situación planteada, definan los problemas, lleguen a sus propias conclusiones sobre las acciones que habría que emprender, contrasten ideas, las defiendan y realicen sus propias aportaciones a la solución del caso. Con este método se pretende formar a futuros profesionales para que sean capaces de identificar problemas concretos y proponer las soluciones más adecuadas al mismo, aplicando para ello los conocimientos teóricos y prácticos adquiridos en diferentes asignaturas del Máster Universitario de Uso Sostenible y Protección del Suelo en Ambientes Mediterráneos de la Universidad de Murcia. [ENG]The method called “problem based learning” (PBL) is characterized by learning centered in students. Pupils learn to develop a range of skills and competencies needed in a professional environment. A problem based in specific situation is possed to students and they must solv it. This method allows students to learn and specialize in a particular subject. The PBL method is used to students analize the situation in question, define problems, draw their own conclusions about actions that should be undertaken, contrasting and defend ideas and make their own contributions to solve the problem they are been raised. This method aims to train future professionals to be able to identify specific problems and propose appropriate solutions. This method pretends that students are able to apply the knowledge and the skills acquired in the Master at the University of Murcia entitled “Sustainable Use of Soil Protection in Mediterranean Environments”.Campus Mare Nostrum, Universidad Politécnica de Cartagena, Universidad de Murcia, Región de Murci