Site-to-site interdomain communication may mediate different loss-of-function mechanisms in a cancer-associated NQO1 polymorphism

Disease associated genetic variations often cause intracellular enzyme inactivation, dysregulation and instability. However, allosteric communication of mutational effects to distant functional sites leading to loss-of-function remains poorly understood. We characterize here interdomain site-to-site communication by which a common cancer-associated single nucleotide polymorphism (c.C609T/p.P187S) reduces the activity and stability in vivo of NAD(P)H:quinone oxidoreductase 1 (NQO1). NQO1 is a FAD-dependent, two-domain multifunctional stress protein acting as a Phase II enzyme, activating cancer pro-drugs and stabilizing p53 and p73a oncosuppressors. We show that p.P187S causes structural and dynamic changes communicated to functional sites far from the mutated site, affecting the FAD binding site located at the N-terminal domain (NTD) and accelerating proteasomal degradation through dynamic effects on the C-terminal domain (CTD). Structural protein:protein interaction studies reveal that the cancer-associated polymorphism does not abolish the interaction with p73a, indicating that oncosuppressor destabilization largely mirrors the low intracellular stability of p.P187S. In conclusion, we show how a single disease associated amino acid change may allosterically perturb several functional sites in an oligomeric and multidomain protein. These results have important implications for the understanding of loss-of-function genetic diseases and the identification of novel structural hot spots as targets for pharmacological intervention

Cannabidiol skews biased agonism at cannabinoid CB1 and CB2 receptors with smaller effect in CB1-CB2 heteroreceptor complexes

Currently, biased agonism is at the center stage of drug development approaches. We analyzed effects of a battery of cannabinoids plus/minus cannabidiol (CBD) in four functional parameters (cAMP levels, phosphorylation of extracellular signal–regulated kinases (ERK1/2), β-arrestin recruitment and label-free/DMR) in HEK-293T cells expressing cannabinoid receptors, CB or CB, or CB-CB heteroreceptor complexes. In all cases two natural agonists plus two selective synthetic agonists were used. Furthermore, the effect of cannabidiol, at a dose (100 nM) that does not allow significant binding to the orthosteric center of either receptor, was measured. From the huge amount of generated data, we would like to highlight that the two psychotropic molecules (Δ-tetrahydrocannabinol/THC and CP-55940) showed similar bias in CBR and that the bias of THC was particularly relevant toward MAPK pathway. Furthermore, THC did not activate the G protein coupled to CBR. Interestingly, the biased agonism was reduced when assays were performed in cells expressing the two receptors, thus suggesting that the heteromer allows less functional selectivity. In terms of cannabidiol action, the phytocannabinoid altered the functional responses, likely by allosteric means, and modified potency, agonist IC/EC values and biased agonism in qualitative and/or quantitative different ways depending on the agonist. The effect of cannabidiol on anandamide actions on both cannabinoid receptors was particularly noteworthy as was significantly different from that of other compounds. Results are a compendium of data on biased agonism on cannabinoid receptors in the absence and presence of cannabidiol. In addition, for the first time, GPCR biased agonism is characterized in an heteromeric context.This work was partially supported by grants from the Spanish Ministry of Economy and Competitiveness (Ref. no. BFU2015-64405-R and SAF2017-84117-R; they may include FEDER funds) and by grant 201413-30 from: Fundació la Marató de TV3Peer Reviewe

Educación Deportiva versus Enseñanza Tradicional: Influencia sobre la regulación motivacional en alumnado de Bachillerato

El objetivo de esta investigación fue analizar el impacto de una intervención basada en el Modelo de Educación Deportiva (MED), en comparación con el Modelo de Enseñanza Tradicional (MET), sobre la regulación motivacional en alumnado de bachillerato en clase de Educación Física. Participó un total de 44 estudiantes (22 hombres y 22 mujeres, Medad = 16.32, DT = 0.57) de bachillerato. El diseño fue un estudio cuasi-experimental con medidas pre- y post-test y comparaciones intra- e inter-grupo. El programa de intervención consistió en 12 sesiones para la enseñanza del baloncesto, siguiéndose las directrices recomendadas por la literatura tanto para el grupo MED como para el grupo MET. Los resultados han indicado que el MED, respecto al MET, mejoró signicativamente el nivel de motivación intrínseca y de regulación identicada. Los hallazgos también han mostrado como el MED redujo signicativamente el nivel de regulación externa y desmotivación comparado con el MET. Como conclusión, el MED es un modelo de enseñanza deportiva que favorece el desarrollo de las regulaciones de motivación más autodeterminadas (motivación intrínseca y regulación identicada) en el proceso de enseñanza-aprendizaje en el contexto escolar, lo que podría suscitar el interés en el alumnado por la práctica deportiva de modo regular en el tiempo libre.

Genome assembly and transcriptomic analysis to elucidate the ability of Nasonovia ribisnigri to break host plant resistance

Aphid genomic resources enable the study of complex life history traits and provide information on vector biology, host adaption and speciation. The currant–lettuce aphid (Nasonovia ribisnigri (Hemiptera: Aphididae) (Mosley)) is a cosmopolitan pest of outdoor lettuce (Lactuca sativa (Asterales: Asteraceae) (Linnaeus)). Until recently, the use of resistant cultivars was an effective method for managing N. ribisnigri. A resistant cultivar containing a single gene (Nr‐locus), introduced in the 1980s, conferred complete resistance to feeding. Overreliance of this Nr‐locus in lettuce resulted in N. ribisnigri's ability to break resistance mechanism, with first reports during 2003. Our work attempts to understand which candidate gene(s) are associated with this resistance‐breaking mechanism. We present two de novo draft assembles for N. ribisnigri genomes, corresponding to both avirulent (Nr‐locus susceptible) and virulent (Nr‐locus resistant) biotypes. Changes in gene expression of the two N. ribisnigri biotypes were investigated using transcriptomic analyses of RNA‐sequencing (RNA‐seq) data to understand the potential mechanisms of resistance to the Nr‐locus in lettuce. The draft genome assemblies were 94.2% and 91.4% complete for the avirulent and virulent biotypes, respectively. Out of the 18,872 differentially expressed genes, a single gene/locus was identified in N. ribisnigri that was shared between two resistant‐breaking biotypes. This locus was further explored and validated in Real‐Time Quantitative Reverse Transcription PCR (qRT‐PCR) experiments and has predicted localisations in both the cytoplasm and nucleus. This is the first study to provide evidence that a single gene/locus is likely responsible for the ability of N. ribisnigri to overcome the Nr‐locus resistance in the lettuce host

Structural basis of the pleiotropic and specific phenotypic consequences of missense mutations in the multifunctional NAD(P)H:quinone oxidoreductase 1 and their pharmacological rescue

The multifunctional nature of human flavoproteins is critically linked to their ability to populate multiple conformational states. Ligand binding, post-translational modifications and disease-associated mutations can reshape this functional landscape, although the structure-function relationships of these effects are not well understood. Herein, we characterized the structural and functional consequences of two mutations (the cancer-associated P187S and the phosphomimetic S82D) on different ligation states which are relevant to flavin binding, intracellular stability and catalysis of the disease-associated NQO1 flavoprotein. We found that these mutations affected the stability locally and their effects propagated differently through the protein structure depending both on the nature of the mutation and the ligand bound, showing directional preference from the mutated site and leading to specific phenotypic manifestations in different functional traits (FAD binding, catalysis and inhibition, intracellular stability and pharmacological response to ligands). Our study thus supports that pleitropic effects of disease-causing mutations and phosphorylation events on human flavoproteins may be caused by long-range structural propagation of stability effects to different functional sites that depend on the ligation-state and site-specific perturbations. Our approach can be of general application to investigate these pleiotropic effects at the flavoproteome scale in the absence of high-resolution structural models. © 202

Impact of the resignification of the research pedagogical practice and the curriculum of pedagogy degree of higher education institutions in Barranquilla-Colombia

Este trabajo tiene como objetivo indagar sobre cómo el graduado universitario de pedagogía asume desde el ejercicio de su rol profesional la resignificación de la práctica pedagógica investigativa. Se evalúa el impacto de los graduados del programa de Especialización en Estudios Pedagógicos de la Universidad de la Costa (Colombia) durante el periodo 2014-2017, frente a la resignificación de la práctica pedagógica investigativa. El estudio se fundamentó los preceptos y lineamentos planteados por el Consejo Nacional de Acreditación y el proyecto Tuning. Se trabajó bajo un enfoque cuantitativo con una población de 247 egresados, utilizando la técnica de la observación y el cuestionario de auto reporte como instrumento para la recolección de información. El estudio muestra que el trabajo curricular desarrollado en el programa impacta positivamente el desempeño de los egresados en el mercado laboral. Se evidencia así la pertinencia del programa en su formación profesional y desempeño laboral al ajustarse los planes curriculares acorde a las necesidades nacionales e internacionales.This work has the objective of investigating how the pedagogy university graduate assume, from the exercise of his professional role, the resignification of the pedagogical research practice. The impact of graduates of the Specialization Program in Pedagogical Studies of the University of the Coast during the 2014-2017 period, on the resignification of the research pedagogical practice is evaluated. The study was based on the precepts and guidelines proposed by the National Accreditation Council and the Tuning project. A quantitative approach with a population of 247 graduates was considered, using the observation technique and the self-report questionnaire as the instrument for gathering information. The study shows that the curricular work developed in the program positively impacts the performance of graduates in the labor market. This demonstrates the relevance of the program in their professional training and work performance by adjusting the curricular plans according to national and international needs

Estudo piloto em cuidadores de pacientes com enfermidades neurológicas, sobre o significado e conhecimento de cuidados paliativos

Aim. To describe the meaning attributed to palliative care (PC) by caregivers, as well as their preference with respect to informing the patient and their satisfaction in their caring role. Methodology: Study descriptive transversal, sample intended of 40 caregivers of neurologic patients needing PC. A questionnaire about knowledge of the topic and their burnout status was applied. A descriptive-comparative analysis was carried out with 95% (pObjetivo. Describir el significado que atribuyen cuidadores a los cuidados paliativos (CPS), así como su preferencia respecto a informar al paciente y su satisfacción con el papel de cuidador. Metodología. Estudio descriptivo transversal, muestra intencional de 40 cuidadores de enfermos neurológicos con necesidad de CPS. Se aplicó un cuestionario de conocimientos sobre el tema y sobrecarga. Se realizó análisis descriptivo-comparativo con un intervalo de confianza de 95% (pObjetivo. Descrever o significado que atribuem cuidadores aos cuidados paliativos (CPS), assim como sua preferência com respeito a informar ao paciente e sua satisfação com o papel de cuidador. Metodologia. Estudo descritivo transversal, amostra intencional de 40 cuidadores de enfermos neurológicos com necessidade de CPS. Aplicou-se um questionário de conhecimentos sobre o tema e sobrecarga. Realizou-se análise descritivo-comparativa com um intervalo de confiança de 95% (

Critical properties of the optical field localization in a three-dimensional percolating system: Theory and experiment

We systematically study the optical field localization in an active three-dimensional (3D) disordered percolating system with light nanoemitters incorporated in percolating clusters. An essential feature of such a hybrid medium is that the clusters are combined into a fractal radiation pattern, in which light is simultaneously emitted and scattered by the disordered structures. Theoretical considerations, based on systematic 3D simulations, reveal nontrivial dynamics in the form of propagation of localized field bunches in the percolating material. We obtain the length of the field localization and dynamical properties of such states as functions of the occupation probability of the disordered clusters. A transition between the dynamical states and narrow point-like fields pinned to the emitters is found. The theoretical analysis of the fractal field properties is followed by an experimental study of the light generation by nanoemitters incorporated in the percolating clusters. The experimental results corroborate theoretical predictions.Comment: 10 pages, 14 figures, to be published Chaos, Solitons & Fractal