Characterization of AtDGK2 in relation to Contact Sites

Contact Sites are conserved cellular regions where two membranes of different organelles are very close but not merged. Contact sites between the endoplasmic reticulum (ER) and the plasma membrane (ER-PM CS) play important roles in metabolic functions. We have identified AtDGK2 (Diacylglycerol kinase 2) as an interactor of SYT1 (Synaptotagmin1), which is a protein located at ER-PM CS. DGKs phosphorylate diacylglycerol to produce phosphatidic acid, both important signalling molecules. Arabidopsis thaliana has seven AtDGKs, but only AtDGK1 and AtDGK2 present an ER transmembrane domain, the rest are cytoplasmic. We have analysed the subcellular localization and functions of these two proteins.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. BIO2017-82609-

Undercovering the molecular mechanisms of lipid signalling at ER-PM contact sites in tomato (Solanum lycopersicum) under abiotic stress conditions

Abiotic stresses cause large reductions in crop production. Therefore, is important to understand how plants respond in order to develop varieties with increased resistance. Lipid-transport proteins (LTP) are emerging as key players of lipid signaling in response to numerous stresses. Specifically, SYT1, a protein first identified by its role in abiotic stress tolerance, is now recognized as an endoplasmic reticulum-plasma membrane contact site tether capable. Our recent data support that SYT1 in involved on non-vesicular lipid-transport of diacyl glycerol (DAG) through its SMP domain. This data together with the interaction of SYT1 with a diacyl glycerol kinase (DGK) suggest a lipid signaling pathway where the product of phospholipase C, diacylglycerol, might be simultaneously translocated from the plasma membrane to the endoplasmic-reticulum by SYT1 and phosphorylated to phosphatidic acid by DGK at the plasma membrane. Using in vitro biochemical approaches we are investigating the affinity of specific lipid species transported by SYT1 using lipid-competition assays, where a fluorescent lipid competes for SYT1 binding-pocket with different lipid species. Using bioinformatic we are obtaining insight into the lipid signal pathway involving PHOSPHOLIPASE C (PLC), DIACYLGLYCEROL KINASE (DGK) and SYNAPTOTAGMIN1 (SYT1) in tomatoUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Role of Arabidopsis DGK1 and DGK2 in cold stress

There are regions, present in all eukaryotic cells, where the membranes of two different organelles are very close (10-30nm), but without fusion, due to proteins which act as tether. These regions are named Membrane Contact Sites (MCS), as for example those formed by endoplasmic reticulum (ER) and plasma membrane (PM). ER-PM CS play important roles in communication between membranes, lipid homeostasis and Ca2+ influx. When a plant is challenge with a stress, phospholipase C is activated at PM producing diacylglycerol (DAG) and inositol phosphates from the hydrolysis of PIP(4,5)P2 or PI4P. DAG is then phosphorylated by diacylglycerol kinases (DGKs) producing phosphatidic acid (PA). Both, DAG and PA, are molecules involved in signalling (Arisz et al., 2009). The genome of Arabidopsis thaliana contain 7 genes encoding DGKs. Most of them are predicted to be cytosolic, with only DGK1 and DGK2 are anchored to the ER due a transmembrane domain. Using different approaches (Co-Immunoprecipitation, confocal microscopy, FRET, Tap-tag…), our group has uncover, that AtDGK1 and AtDGK2 (Diacylglycerol kinase 1, AT5G07920 and Diacylglycerol kinase 2, At5g63770) form a complex with the lipid transport protein located at ER-PM CS known as Synaptotagmin1 (SYT1, At2g20990) (Pérez-Sancho et al., 2015). SYT1 is able to bind preferentially DAG (Ruiz-Lopez et al., 2021), which support the idea that SYT1, DGK1, and DGK2 function coordinated to regulate the levels of DAG at the PM. DGK1 and DGK2 are induced upon low temperatures and found that dgk2 mutants show reduced root growth in low temperature and have reduced freezing tolerance. Our studies suggest that DGK1 and DGK2 act in concert with SYT1 to regulate the production of DAG and PA at ER-PM CS and highlight the importance of these proteins for the correct response to stress.The authors acknowledge the support by: (1) Ministerio de Ciencia e Innovación BIO2017-82609-R, PGC2018-098789-B-100; (2) Ministerio de Economía, Industria y Competitividad (BES-2015-071256, RyC-2013-12699). Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

How are DGK1 and DGK2 involved in membrene contact sites?

Eukaryotic cells have regions of interaction between two organelles where some proteins, which act as tether, bring both membranes closer (10-30 nm) without fusion, named membrane contact sites (MCS). Two organelles that can form MCS are endoplasmic reticulum (ER) and plasma membrane (PM). ER-PM CS play important metabolic functions such as communication between both membranes, lipid homeostasis and Ca2+ influx. Our group has identified that AtDGK1 and AtDGK2 (Diacylglycerol kinase 1, AT5G07920 and Diacylglycerol kinase 2, At5g63770) form a complex with a well-known protein located at ER-PM CS, Synaptotagmin1 (SYT1, At2g20990). Upon perception of stress, phospholipase C (PLC) is activated at the plasma membrane to hydrolyse PIP(4,5)P2 or PI4P in order to generate DAG and inositol phosphates. Diacylglycerol (DAG) is phosphorylated by diacylglycerol kinases (DGKs) to produce phosphatidic acid (PA). DAG and PA are important cell signalling molecules. There are seven DGKs encoded in Arabidopsis thaliana genome, but only DGK1 and DGK2 have a transmembrane domain that anchors them to the endoplasmic reticulum, the rest are cytoplasmic. DGK1 and DGK2 appear to play a role in stress response as both are induced by exposure to low temperatures and wounding. Also, we found that dgk2 knockout mutant produces lower resistance to freezing. Using confocal microscopy, we have analysed the subcellular localization of these two proteins and investigated their interaction with SYT1 and between them using FRET and co-immunoprecipitation studies. Additionally, we report that the mutation of DGK1 is lethal in homozygosity. Our studies suggest that DGK1 and DGK2 act in concert with SYT1 to regulate the production of PA at ER-PM CS and highlight the importance of these proteins for the correct response to stress tolerance.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Reflexiones teóricas de preparación táctica ofensiva del espadista

Introducción: La preparación táctica ofensiva de los espadistas 13 - 15 años debe enfocarse a desarrollar la capacidad para analizar contrarios, anticipar ataques sorpresivos y crear planes tácticos emergentes. Objetivo: Se realizó una valoración teórica sobre consideraciones y definiciones para determinar la preparación táctica ofensiva de espadistas categoría 13-15 años. Métodos: Se realizó una búsqueda acerca de la literatura científica publicada en español e inglés (Base de datos del Google Académico) relacionada con la preparación táctica ofensiva del deportista. Se inició la clasificación y organización según tema principal. Resultados: El análisis de consideraciones y definiciones de términos relacionados con la táctica ofensiva facilitó elaborar la matriz metodológica de la variable dependiente preparación táctica ofensiva, las dimensiones (perceptivas y cognitivas), se establecieron indicadores, subindicadores, escalas y categorías para su evaluación. Conclusión: El análisis de consideraciones y definiciones sobre el tema principal, facilitó la determinación de dimensiones para desarrollar la operatividad del pensamiento táctico ofensivo e incrementar la preparación de espadistas categoría 13-15 años

Student Reciprocal Peer Teaching as a Method for Active Learning: An Experience in an Electrotechnical Laboratory

Active learning is one of the most efficient mechanisms for learning, according to the psychology of learning. When students act as teachers for other students, the communication is more fluent and knowledge is transferred easier than in a traditional classroom. This teaching method is referred to in the literature as reciprocal peer teaching. In this study, the method is applied to laboratory sessions of a higher education institution course, and the students who act as teachers are referred to as ‘‘laboratory monitors.’’ A particular way to select the monitors and its impact in the final marks is proposed. A total of 181 students participated in the experiment, experiences with laboratory monitors are discussed, and methods for motivating and training laboratory monitors and regular students are proposed. The types of laboratory sessions that can be led by classmates are discussed. This work is related to the changes in teaching methods in the Spanish higher education system, prompted by the Bologna Process for the construction of the European Higher Education Are

Flora of the tropical deciduous forest in a gypseous and calcareous area, Colima, Mexico

Antecedentes y Objetivos: La microcuenca La Salada, ubicada al centro-sur de Colima, cubierta por bosque tropical caducifolio (BTC) sobre sustratos yesosos (74.5%) y calcáreos (25.5%), fue explorada por el eminente botánico Rogers McVaugh para la Flora Novo-Galiciana. Sin embargo, solo parte del material colectado ha estado disponible. El objetivo del trabajo fue documentar la diversidad de plantas vasculares de la zona, para evidenciar su relevancia biológica. Métodos: Se realizaron una revisión bibliográfica y de material de herbario (IBUG y MICH) y colectas de campo para integrar la lista florística. Complementariamente se revisaron bases de datos electrónicas (IBdata y GBIF) para obtener registros de colectas en sitios de Colima con características ecológicas similares al área muestreada. Para comparar la diversidad del área con la de otros sitios con BTC se calculó el índice de diversidad taxonómica (IDT). Resultados clave: Se registraron 408 taxones, 248 géneros y 78 familias de la flora de BTC de sustratos yesoso-calcáreos de Colima. Se presentan dos apéndices, uno se basa en registros del área muestreada por los autores y otro en registros de otras áreas yesoso-calcáreas de Colima obtenidos de bases de datos. Del área muestreada se registraron 368 taxones (incluyendo 16 infraespecíficos) de 233 géneros y 75 familias. Fabaceae, Euphorbiaceae, Asteraceae, Malvaceae, Poaceae y Convolvulaceae son las familias más representadas (48% de los taxones). Los géneros más diversos son Euphorbia (13), Bursera (ocho), Ipomoea (siete) y Tillandsia (seis). De las especies registradas 39% (144) son endémicas de México y el área constituye la localidad tipo de 23. Se anotan 11 taxones no registrados antes para el estado de Colima. El valor IDT fue de 330.35 especies/ln área. Conclusiones: La alta diversidad florística y endemismo de la zona, y la singularidad del sustrato yesoso, merecen la conservación de estos ecosistemas que están siendo impactados en Colima.Background and Aims: The study area of La Salada basin is situated in the southern-central area of Colima, characterized by tropical dry forest vegetation on gypseous (74.5%) and calcareous (25.5%) soils. This site was visited by the eminent botanist Rogers McVaugh and is represented in the Flora Novo-Galiciana. However, only a part of his collections has been available for consultation. The objective here is to document the diversity of vascular plants in this site providing evidence which sustains its biological relevance. Methods: Revisions of bibliography and material in the herbaria IBUG and MICH were made. Specimens from recent fieldwork were integrated into the list. Additionally, electronic databases (IBdata and GBIF) were reviewed to obtain records of collections in sites of Colima with ecological characteristics similar to the sampled area. The Index of Taxonomic Diversity (IDT) was calculated facilitating comparisons of values for other areas with tropical dry forests. Key results: A total of 408 taxa, 248 genera and 78 families of vascular plants of BTC in gypseous-calcareous soils of Colima were registered. Two appendices are provided: the first with authors´ collections and those in bibliography; and the second with citations of species in other gypseous-calcareous soils in Colima from databases. In the first, 368 taxa (including 16 infraspecific taxa) in 233 genera and 75 families were recorded. The families Fabaceae, Euphorbiaceae, Asteraceae, Malvaceae, Poaceae and Convolvulaceae total 48% of the reported flora. The genera with greatest diversity were Euphorbia (13), Bursera (eight), Ipomoea (seven) and Tillandsia (six). Of the species recorded 39% (144) are endemic to Mexico and this site is the type location for 23 species. The IDT calculated is 330.35 species/ln area. Conclusions: The high floristic diversity and endemism of the area with gypseous and calcareous soils, substantiate conservation of these ecosystems which are being impacted in Colima

Estrategia pedagógica para compensar las adicciones tecnológicas desde la Educación Física en Secundaria Básica (Original)

New technologies hide a potential health risk. In recent years, several so-called technological diseases or addictions have arisen, derived from the inappropriate or indiscriminate use of the computer, mobile phone, video games and others. In this sense, this research was developed in the basic secondary education of the city of Holguín, with the aim of designing a pedagogical strategy based on a set of sports activities that allows compensating the technological addictions present in seventh and eighth grade schoolchildren. By applying the strategy, it was possible to increase the teachers' knowledge about the meaning of ICTs. The mode of action of teachers and students, their interpersonal relationships, and motivation towards the efficient and appropriate use of time were improved. The conclusions constitute generalizations for the achievement of the proposed objective. The recommendations are actions based on raising awareness among teachers and students of Basic Secondary School about the importance of sports activities to compensate for the manifestations caused by technological addictions.Las nuevas tecnologías esconden un riesgo potencial para la salud, en los últimos años han surgido varias enfermedades denominadas, tecnológicas o adicciones, por derivarse del uso inadecuado o indiscriminado de la computadora, el teléfono móvil, los videojuegos y otros. En este sentido se desarrolló esta investigación en la enseñanza secundaria básica de la ciudad de Holguín, con el objetivo de diseñar una estrategia pedagógica basada en un conjunto de actividades deportivas que permita compensar las adicciones tecnológicas presentes en los escolares de séptimo y octavo grado. Mediante la aplicación de la estrategia fue posible aumentar el conocimiento de los profesores sobre el significado de las Tic. Se modificó el modo de actuación de docentes y alumnos, sus relaciones interpersonales y se mejoró la motivación hacia el uso eficiente y adecuado del tiempo. Las conclusiones constituyen generalizaciones para la consecución del objetivo propuesto. Las recomendaciones son acciones en función de concientizar a los profesores y estudiantes de Secundaria Básica sobre la importancia que tienen las actividades deportivas para compensar las manifestaciones provocadas por las adicciones tecnológicas

Impact of the MIC of Piperacillin-Tazobactam on the Outcome of Patients with Bacteremia Due to Extended-Spectrum-B-Lactamase- Producing Escherichia coli

We investigated the impact of the piperacillin-tazobactam MIC in the outcome of 39 bloodstream infections due to extended- spectrum-B-lactamase-producing Escherichia coli. All 11 patients with urinary tract infections survived, irrespective of the MIC. For other sources, 30-day mortality was lower for isolates with a MIC of <2 mg/liter than for isolates with a higher MIC (0% ver- sus 41.1%; P = 0.02

Aportaciones a la flora del sureste ibérico

New records to the flora of SE Spain.Palabras clave: Corología, plantas vasculares, Murcia, Albacete, España.Key words: Chorology, vascular plants, Murcia, Albacete, Spain