Identification and characterization of Syndapin I, Vacuolar protein sorting 35 and Neurobeachin as new interaction partners of the glycine receptor

The glycine receptor (GlyR) is the major inhibitory neurotransmitter receptor in spinal cord and brainstem. Heteropentameric GlyRs are clustered and anchored at inhibitory postsynaptic sites by the binding of the large intracellular loop between transmembrane domains 3 and 4 of the GlyRbeta subunit (GlyRbeta-loop) to the cytoplasmic scaffolding protein gephyrin. GlyRs are also cotransported with gephyrin along microtubules in the anterograde and retrograde direction due to the binding of gephyrin to microtubule-associated motor proteins. Additionally, GlyRs undergo lateral diffusion in the plasma membrane from extrasynaptic to synaptic sites and vice versa. Since its discovery, gephyrin has remained for many years the only binding partner interacting directly with the GlyRbeta subunit. In an attempt to elucidate further mechanisms involved in GlyR function and regulation at inhibitory postsynaptic sites, a proteomic screen for putative binding partners to the GlyRbeta loop was performed. Three proteins were identified as putative interactors. In this thesis, the interaction between these putative binding proteins and the GlyRbeta subunit was analyzed and characterized. Binding studies with glutathione-S-transferase fusion proteins revealed that all putative binding proteins, Syndapin (Sdp), Vacuolar Protein Sorting 35 (Vps35) and Neurobeachin (Nbea), interact specifically with the GlyRbeta loop. The Sdp family of proteins are F-BAR and SH3 domain containing proteins. Inmmunocytochemical experiments showed that SdpI as well as the isoforms SdpII-S and SdpIIL colocalize with the full-length GlyRbeta subunit in a mammalian cell expression system. In cultured spinal cord neurons, a partial colocalization of endogenous SdpI with several excitatory and inhibitory synaptic markers was demonstrated. Mapping experiments using deletion mutants narrowed the SdpI binding site down to 22 amino acids. Peptide competition experiments confirmed the specificity of the interaction between SdpI and this sequence of the GlyRbeta subunit. Point mutation analysis revealed a SH3-proline rich domain dependent interaction between SdpI and the GlyRbeta subunit, respectively. In addition, binding studies in mammalian cells showed that both splice variants of SdpII as well as SdpI interact with the GlyR scaffolding protein gephyrin. Although the SdpI and gephyrin binding sites do not overlap, protein competition studies revealed that interaction of the E-domain of gephyrin with the GlyRbeta loop interferes with SdpI binding. Since SdpI is a dynamin binding protein involved in vesicle endocytosis and recycling pathways, a possible function of SdpI in the regulation of GlyR synaptic distribution was investigated. Co-immunoprecipitation experiments confirmed a SdpI-GlyR association in the vesicle-enriched fraction of rat spinal cord tissue. Immunocytochemical studies of SdpI knock out mice showed that the clustering and distribution of GlyRs in the brain stem is unchanged. However, acute down-regulation of SdpI in rat spinal cord neurons by viral shRNA expression led to a reduction in the number and size of GlyR clusters, an effect that could be rescued upon shRNA-resistant SdpI overexpression. Further immunocytochemical analysis of the localization of gephyrin, the gamma2 subunit of the type A gamma-aminobutyric acid receptor (GABAARgamma2 subunit) and the vesicular inhibitory amino acid transporter (VIAAT) under SdpI knock-down conditions showed that both the number and average size of the gamma2-subunit containing GABAA receptor clusters were significantly reduced in spinal cord neurons. In contrast to GlyR and GABAARgamma2 immunoreactivity, the number and average size of gephyrin and VIAAT clusters were barely reduced upon SdpI downregulation. These results suggest that SdpI has a role in GlyR trafficking that can be compensated by other syndapin isoforms or other trafficking pathways. Furthermore, SdpI might be required for the clusters of GlyRs and gamma2-subunit containing GABAARs in spinal cord and brainstem. Vps35 is the core protein of the retromer complex, which mediates the endosome to Golgi apparatus retrieval of different types of receptors in mammals and yeast. Here, protein-protein interaction assays revealed for the first time that Vps35 interacts directly with the GlyRbeta loop as well as with gephyrin. The generation of specific Vps35 antibodies allowed to determine the distribution of this protein in the central nervous system. Immunocytochemical analyses revealed the presence of Vps35 in the somata and neurites of spinal cord neurons, suggesting a possible interaction of Vps35 with the GlyR under physiological conditions. Nbea is a BEACH domain containing, neuron-specific protein. Binding studies revealed a direct interaction between two regions of Nbea and the GlyRbeta loop. Immunocytochemical experiments confirmed a somatic and synaptic distribution of Nbea in primary cultures. In spinal cord neurons, a partial colocalization of Nbea with excitatory and inhibitory synaptic markers suggests a possible interaction of Nbea with the GlyR at inhibitory synaptic sites

Teoría de Juegos: una introducción

En este trabajo se realiza una introducción a la teoría de juegos, área de la matemática aplicada que estudia una gran diversidad de situaciones entre diferentes actores que compiten o colaboran para conseguir un fin determinado. Comenzamos introduciendo la teoría general (juego, jugadores, estrategias, pagos, etc.) en el capítulo 1, para luego centrarnos en los juegos para dos jugadores. Entre estos, comenzamos examinando, ya en el capítulo 2 un caso particular más sencillo, los juegos de suma cero, que son aquellos en los que las ganancias de un jugador coinciden con las pérdidas del otro. Aquí estudiamos primero las estrategias individuales que pueden tomar los jugadores o estrategias puras, analizando cuáles deben elegir los jugadores buscando su propio beneficio. El resultado más importante en esta parte es el lema minimax. A continuación consideramos el caso en el que el juego se puede repetir un número de veces, de manera que a los jugadores les pueda convenir la elección de distintas estrategias puras cada vez que se juegue. Este hecho lleva a la introducción de las llamadas estrategias mixtas. En este caso se prueba que, bajo la suposición de que los jugadores buscarán obtener racionalmente el mejor resultado posible, siempre existe una situación de equilibrio que resuelve, por así decir, el juego. El resultado que lleva a esta conclusión es el teorema minimax de Von Neumann, resultado que también tiene numerosas aplicaciones fuera del ámbito de la teoría de juegos. A continuación, introducimos distintos métodos de resolución de juegos de suma cero, que se completan unos a otros y proporcionan distintos puntos de vista del problema. Posteriormente, en el capítulo 3, se aborda la situación general, en la que los juegos pueden ser de suma no nula. Análogamente a como se hizo con los juegos de suma cero, introducimos primero el concepto de estrategia pura. Una situación deseable será que el juego tenga al menos un equilibrio de Nash, que es una situación estable para el juego. Sin embargo, probamos que estos equilibrios solo se dan necesariamente cuando se utilizan estrategias mixtas. Este resultado, que es el principal de este trabajo, es el llamado teorema de Nash. Recogemos también diferentes métodos de resolución de juegos de suma no nula, esto es, con el fin de encontrar equilibrios de Nash. Además, para cuando el juego tenga más de un equilibrio de Nash, se introducen algunos criterios para elegir el que sea mejor en cierto sentido. Los conceptos y resultados de estos tres primeros capítulos se ilustran con ejemplos. Finalmente, en el capítulo 4 se presentan las conclusiones de este trabajo. Abstract: In this work, we present an introduction to game theory, an area of applied mathematics that studies a great diversity of situations among different actors who compete or collaborate to achieve a particular goal. We begin by introducing the general theory (game, players, strategies, payments, etc.) in the chapter 1, and then focus on two-player games. Among these, in chapter 2, we examine a particular case, the zero-sum games, which are those in which one player wins whatever the other player loses. Here we first study the individual strategies that players can take (pure strategies), analyzing which ones players should choose for their own benefit. The most important result in this part will be the minimax lemma. Then we consider the case where the game can be repeated a number of times, so that the players can be persuaded to choose different pure strategies each time it is played. This leads to the introduction of so-called mixed strategies. In this case it is proven that, under the assumption that the players will rationally seek to obtain the best possible result, there is always a balance situation that solves, so to speak, the game. The result that leads to this conclusion is Von Neumann’s minimax theorem, a result that also has numerous applications outside the scope of game theory. Next, we introduce different methods of solving zero-sum games, which complement each other and provide different views of the problem. Later, in chapter 3, we address the general situation where games can be non-zerosum games. As with zero-sum games, we first introduce the concept of pure strategy. A desirable situation will be that the game has at least a Nash equilibrium, which is a stable situation for the game. However, we prove that these equilibria only necessarily occur when using mixed strategies. This result, which is the main one in this work, is the so-called Nash theorem. We also collect different methods of solving non-zero-sum games, in order to find Nash equilibria. In addition, we introduce some criteria to choose the best (in some sense) Nash equilibrium, when there are more than one. The concepts and results of these first three chapters are illustrated with examples. Finally, the conclusions of this work are presented in chapter 4

Plan de Apoyo a Personas Refugiadas de la Universidad de Málaga y de 16 ONG

Esta investigación tiene como finalidad conocer en profundidad el Plan de Apoyo a Personas Refugiadas de la Universidad de Málaga y de 16 ONG. El estudio pretende conocer las debilidades del plan y poder trabajar en la mejora de éstas. Los resultados servirán para seguir avanzando en la inclusión de las personas refugiadas y en la mejora de su calidad de vida.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Shifting Developmental Trajectories During Critical Periods of Brain Formation

Critical periods of brain development are epochs of heightened plasticity driven by environmental influence necessary for normal brain function. Recent studies are beginning to shed light on the possibility that timely interventions during critical periods hold potential to reorient abnormal developmental trajectories in animal models of neurological and neuropsychiatric disorders. In this review, we re-examine the criteria defining critical periods, highlighting the recently discovered mechanisms of developmental plasticity in health and disease. In addition, we touch upon technological improvements for modeling critical periods in human-derived neural networks in vitro. These scientific advances associated with the use of developmental manipulations in the immature brain of animal models are the basic preclinical systems that will allow the future translatability of timely interventions into clinical applications for neurodevelopmental disorders such as intellectual disability, autism spectrum disorders (ASD) and schizophrenia.The work was supported by the Australian National University to ND and by the Spanish Ministry of Science, Innovation and Universities (Ministerio de Ciencia, Innovación y Universidades, RTI2018-100872-J-I00) as well as the CIDEGENT excellence research program of the Valencian regional government (CIDENGENT/2019/044) to ID

Too Many Males or Too Many Females? Classroom Sex Ratio, Life History Strategies and Risk-Taking Behaviors

Prior research finds that sex ratio, defined as the proportion of males and females in a given context, is related to engagement in risk-taking behaviors. However, most research operationalizes sex ratio at a local context (e.g., regional or county), which fails to reflect with precision the sex ratios contexts of individuals at a closer level. Furthermore, the relationship between sex ratio and risk-taking behaviors may be affected by individuals’ life history strategy, with previous studies showing fast life history strategies linked to risk-taking behaviors, compared to slow life history strategies. The present study analyzes the relationship between classroom sex ratio and risk-taking behaviors and the interaction between classroom sex ratio and life history strategy in adolescents. The sample comprised 1214 participants nested in 57 classrooms, 49.75% females, 91.5% Spanish and a mean age of 16.15 years (SD = 1.23, range 14–21). Results from multilevel modeling showed a negative relation between classroom sex ratio and risk-taking behaviors in female adolescents with faster life history strategy. By contrast, classroom sex ratio in male adolescents related positively to risk-taking behaviors but did not interact with life history strategy. These findings underscore the importance of studying proximate sex ratio on risk-taking behaviors in adolescents and underline its potential influence in the development and expression of life history strategiesThis research was supported by the Andalucía ERDF 2014-20 OP [UMA18-FEDERJA-071] and the Research Grants of the Reina Sofia Centre on Adolescence and Youth. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Funding for open access charge: Universidad de Málaga/CBUA

Defining a roadmap for harmonizing quality indicators in Laboratory Medicine: A consensus statement on behalf of the IFCC Working Group "laboratory Error and Patient Safety" and EFLM Task and Finish Group "performance specifications for the extra-analytical phases"

The improving quality of laboratory testing requires a deep understanding of the many vulnerable steps involved in the total examination process (TEP), along with the identification of a hierarchy of risks and challenges that need to be addressed. From this perspective, the Working Group “Laboratory Errors and Patient Safety” (WG-LEPS) of International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) is focusing its activity on implementation of an efficient tool for obtaining meaningful information on the risk of errors developing throughout the TEP, and for establishing reliable information about error frequencies and their distribution. More recently, the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) has created the Task and Finish Group “Performance specifications for the extraanalytical phases” (TFG-PSEP) for defining performance specifications for extra-analytical phases. Both the IFCC and EFLM groups are working to provide laboratories with a system to evaluate their performances and recognize the critical aspects where improvement actions are needed. A Consensus Conference was organized in Padova, Italy, in 2016 in order to bring together all the experts and interested parties to achieve a consensus for effective harmonization of quality indicators (QIs). A general agreement was achieved and the main outcomes have been the release of a new version of model of quality indicators (MQI), the approval of a criterion for establishing performance specifications and the definition of the type of information that should be provided within the report to the clinical laboratories participating to the QIs project

Defining a roadmap for harmonizing quality indicators in Laboratory Medicine: A consensus statement on behalf of the IFCC Working Group "laboratory Error and Patient Safety" and EFLM Task and Finish Group "performance specifications for the extra-analytical phases"

The improving quality of laboratory testing requires a deep understanding of the many vulnerable steps involved in the total examination process (TEP), along with the identification of a hierarchy of risks and challenges that need to be addressed. From this perspective, the Working Group \u201cLaboratory Errors and Patient Safety\u201d (WG-LEPS) of International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) is focusing its activity on implementation of an efficient tool for obtaining meaningful information on the risk of errors developing throughout the TEP, and for establishing reliable information about error frequencies and their distribution. More recently, the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) has created the Task and Finish Group \u201cPerformance specifications for the extraanalytical phases\u201d (TFG-PSEP) for defining performance specifications for extra-analytical phases. Both the IFCC and EFLM groups are working to provide laboratories with a system to evaluate their performances and recognize the critical aspects where improvement actions are needed. A Consensus Conference was organized in Padova, Italy, in 2016 in order to bring together all the experts and interested parties to achieve a consensus for effective harmonization of quality indicators (QIs). A general agreement was achieved and the main outcomes have been the release of a new version of model of quality indicators (MQI), the approval of a criterion for establishing performance specifications and the definition of the type of information that should be provided within the report to the clinical laboratories participating to the QIs project

Cineclub: una experiencia en el barrio Cuba. Guayaquil, Ecuador

El artículo que se presenta recoge las experiencias de la aplicación de un proyecto de vinculación con la sociedad de la carrera de Comunicación Social de la Universidad Politécnica Salesiana, sede Guayaquil, Ecuador. El objetivo de su aplicación responde a facilitar el acceso a la cultura a los pobladores del barrio Cuba, emblemático de la ciudad de Guayaquil, mediante la difusión de obras clásicas de la cinematografía Ecuatoriana. Se anima la investigación apoyados en la premisa de que por lo general, en las comunidades de Guayaquil, la oferta cinematográfica se reduce a la más elemental selección de cine comercial y no existe ninguna oferta de valiosas obras cinematográficas que forman parte de la cultura universal. Se utiliza una metodología de trabajo de matriz cualitativa de tipo exploratorio-descriptivo. Como parte de las conclusiones se evidencia el cumplimiento del objetivo planteado y el hecho que la inclusión social, de este sector poblacional es, todavía, un problema por resolver. Como principal resultado se expone que la aplicación del proyecto fortaleció la unión de los pobladores del barrio, su interés por la cinematografía y la proyección de la universidad hacia la construcción, en el barrio, de una sala de cine con todo su equipamiento tecnológico. Palabras claves: Barrio Cuba, cineclub, vinculación, cinematografía, cultura