Cross-linking mass spectrometry uncovers protein interactions and functional assemblies in synaptic vesicle membranes

Synaptic vesicles are storage organelles for neurotransmitters. They pass through a trafficking cycle and fuse with the pre-synaptic membrane when an action potential arrives at the nerve terminal. While molecular components and biophysical parameters of synaptic vesicles have been determined, our knowledge on the protein interactions in their membranes is limited. Here, we apply cross-linking mass spectrometry to study interactions of synaptic vesicle proteins in an unbiased approach without the need for specific antibodies or detergent-solubilisation. Our large-scale analysis delivers a protein network of vesicle subpopulations and functional assemblies including an active and an inactive conformation of the vesicular ATPase complex as well as non-conventional arrangements of the luminal loops of SV2A, Synaptophysin and structurally related proteins. Based on this network, we specifically target Synaptobrevin-2, which connects with many proteins, in different approaches. Our results allow distinction of interactions caused by ‘crowding’ in the vesicle membrane from stable interaction modules.Projekt DEA

An overview of the synaptic vesicle lipid composition

This work was funded by a Ramon y Cajal grant to A.P-L. (RYC2018-023837-I) and by a grant from the National Institutes of Health to R.J. (P01 GM072694) .Chemical neurotransmission is the major mechanism of neuronal communication. Neurotransmitters are released from secretory organelles, the synaptic vesicles (SVs) via exocytosis into the synaptic cleft. Fusion of SVs with the presynaptic plasma membrane is balanced by endocytosis, thus maintaining the presynaptic membrane at steadystate levels. The protein machineries responsible for exo-and endocytosis have been extensively investigated. In contrast, less is known about the role of lipids in synaptic transmission and how the lipid composition of SVs is affected by dynamic exo-endocytotic cycling. Here we summarize the current knowledge about the composition, organization, and function of SV membrane lipids. We also cover lipid biogenesis and maintenance during the synaptic vesicle cycle.Spanish Government RYC2018-023837-IUnited States Department of Health & Human Services National Institutes of Health (NIH) - USA P01 GM07269

Evaluation of quality in higher education: content validity

El cambio de perspectiva sobre la educación superior en España ha posibilitado el desarrollo de nuevos marcos jurídicos que dan mayor autonomía a las universidades. Al mismo tiempo los gobiernos, tanto nacional como autonómicos, necesitan recabar información sobre cómo se gestiona dicha autonomía. Esta situación ha impulsado la necesidad de definir el constructo de calidad en la universidad y de generar procedimientos adecuados para su evaluación. El objetivo de este trabajo es por una parte, justificar cómo los sistemas de indicadores suponen una posible solución a esta problemática, y por otra analizar empíricamente la validez de contenido de un conjunto de indicadores seleccionados a partir de una revisión bibliográfica. El proceso seguido en el estudio empírico fue el siguiente: se seleccionaron indicadores que hacen referencia a distintos aspectos relacionados con la educación superior (enseñanza, investigación y gestión); se consultó a un grupo de expertos sobre la adecuación de dichos indicadores y se utilizó el índice de Osterlind (1989) como una forma de operativizar el consenso entre los expertos respecto a la idoneidad de los indicadores seleccionados para evaluar la calidad universitaria. Se presentan los indicadores obtenidos en el análisis como una alternativa posible para el estudio de la calidad universitariaThe new conception of higher education in Spanish society and in its national, regional and local governments has generated laws that give more independence to universities. Because of this situation, governments need information about how universities use that independence. This context has stressed the need of defining the construct of quality in higher education and the difficulty that implementing an appropriate evaluation procedure entails. This paper has two aims: first, to justify that systems of indicators are a plausible alternative to both problems, and second, to study the content validity of a group of indicators chosen from the literature. We have carried out the following process in the empirical study: we selected some indicators related to higher education (about teaching, research, and services); we consulted experts about the congruence of these indicators, and finally we used the Osterlind´s index (1989) to operationalize the consensus between experts about their judgments. We describe the quality indicators obtained in this analysis as a possible alternative to study quality in higher education

Modulation of KV4.3-KChIP2 Channels by IQM-266: Role of DPP6 and KCNE2

The transient outward potassium current (Itof) is generated by the activation of KV4 channels assembled with KChIP2 and other accessory subunits (DPP6 and KCNE2). To test the hypothesis that these subunits modify the channel pharmacology, we analyzed the electrophysiological effects of (3-(2-(3-phenoxyphenyl)acetamido)-2-naphthoic acid) (IQM-266), a new KChIP2 ligand, on the currents generated by KV4.3/KChIP2, KV4.3/KChIP2/DPP6 and KV4.3/KChIP2/KCNE2 channels. CHO cells were transiently transfected with cDNAs codifying for different proteins (KV4.3/KChIP2, KV4.3/KChIP2/DPP6 or KV4.3/KChIP2/KCNE2), and the potassium currents were recorded using the whole-cell patch-clamp technique. IQM-266 decreased the maximum peak of KV4.3/KChIP2, KV4.3/KChIP2/DPP6 and KV4.3/KChIP2/KCNE2 currents, slowing their time course of inactivation in a concentration-, voltage-, time- and use-dependent manner. IQM-266 produced an increase in the charge in KV4.3/KChIP2 channels that was intensified when DPP6 was present and abolished in the presence of KCNE2. IQM-266 induced an activation unblocking effect during the application of trains of pulses to cells expressing KV4.3/KChIP2 and KV4.3/KChIP2/KCNE2, but not in KV4.3/KChIP2/DPP6 channels. Overall, all these results are consistent with a preferential IQM-266 binding to an active closed state of Kv4.3/KChIP2 and Kv4.3/KChIP2/KCNE2 channels, whereas in the presence of DPP6, IQM-266 binds preferentially to an inactivated state. In conclusion, DPP6 and KCNE2 modify the pharmacological response of KV4.3/KChIP2 channels to IQM-266.MCIN/AEI SAF2016-75021-R RTI2018-097189-B-C22 BIO2017-89523-R PID2019-104366RB-C21 PID2019-104366RB-C22 PID2020-114256RB-I00 PID2020-119805RB-I00 BES-2017-080184 BES-2010-036573 PRE2018-083280 RYC2018-023837-IERDF A way of making Europe SAF2016-75021-R RTI2018-097189-B-C22 BIO2017-89523-RFEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento A-FQM-386-UGR20Instituto de Salud Carlos III CIBERCV CB/11/00222Consejo Superior de Investigaciones Cientificas PIE202180E073 PIE201820E104 2019AEP148ESF Investing in your future BES-2017-080184 BES-2010-036573 PRE2018-083280 RYC2018-023837-IInstituto de Salud Carlos IIISpanish GovernmentEuropean Commission FPU17/0273

Dynamic Excimer (DYNEX) Imaging of Lipid Droplets

This work was funded by grants CTQ2017-85658-R (MICIU/AEI/ERDF), FQM-337 (Junta de Andalucía), and PIUJA 2019-20 (Universidad de Jaén). We acknowledge the Universidad de Granada (Spain) microscopy central facilities (CIC-UGR) and computing time from CSIRC-UGR. M.C.G.-G. thanks MICIU/AEI for a predoctoral fellowship. Funding for open access charge: Universidad de Granada/CBUA. We are indebted to Prof. Reinhard Jahn for his generous support.Unraveling cellular physiological processes via luminescent probes that target specific cellular microenvironments is quite challenging due to the uneven distribution of probes. Herein, we designed a new dynamic excimer (DYNEX) imaging method that involves the sensitive detection of nanosecond-scale dynamic molecular contacts of a fluorescent acridone derivative and reveals the cell microenvironment polarity. Using our method, we specifically tracked cell lipid droplets in fibroblast colon carcinoma cells. These organelles play a central role in metabolic pathways, acting as energy reservoirs in regulatory processes. DYNEX imaging provides the inner polarity of cell lipid droplets, which can be related to lipid contents and metabolic dysfunctions. This new methodology will inspire development of novel multidimensional fluorescent sensors that are able to provide target-specific and orthogonal information at the nanosecond scale.CBUAMICIUUniversidad de GranadaUniversidad de JaénAgencia Estatal de InvestigaciónEuropean Regional Development Fund FQM-337Junta de Andalucía PIUJA 2019-2

Mountain waves analysis in the vicinity of the Madrid-Barajas Airport using the WRF model

Turbulence and aircraft icing associated with mountain waves are weather phenomena potentially affecting aviation safety. In this paper, these weather phenomena are analysed in the vicinity of the Adolfo Su´arez Madrid-Barajas Airport (Spain). Mountain waves are formed in this area due to the proximity of the Guadarrama mountain range. Twenty different weather research and forecasting (WRF) model configurations are evaluated in an initial analysis. .is shows the incompetence of some experiments to capture the phenomenon. .e two experiments showing the best results are used to simulate thirteen episodes with observed mountain waves. Simulated pseudosatellite images are validated using satellite observations, and an analysis is performed through several skill scores applied to brightness temperature. Few differences are found among the different skill scores. Nevertheless, the .ompson microphysics scheme combined with the Yonsei university PBL scheme shows the best results. .e simulations produced by this scheme are used to evaluate the characteristic variables of the mountain wave episodes at windward and leeward and over the mountain. .e results show that north-northwest wind directions, moderate wind velocities, and neutral or slightly stable conditions are the main features for the episodes evaluated. In addition, a case study is analysed to evidence the WRF ability to properly detect turbulence and icing associated with mountain waves, even when there is no visual evidence available..is work was partially supported by the following research projects: PID2019-105306RB-I00, PCIN-2014-013-C07-04, and PCIN2016-080 (UE ERA-NET Plus NEWA Project), CGL2016-78702-C2-1-R and CGL2016-78702-C2-2-R (SAFEFLIGHT Project), FEI-EU-17-16 and SPESMARTand SPESVALE (ECMWF Special Projects)

El debate académico de competición como instrumento para el aprendizaje de cuestiones de derecho, ciencia política y criminología

Memoria ID-0032. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2016-2017

Synthesis, characterization, and antibacterial activities of a heteroscorpionate derivative platinum complex against methicillin-resistant Staphylococcus aureus

Staphylococcus aureus is one of the species with the greatest clinical importance and greatest impact on public health. In fact, methicillin-resistant S. aureus (MRSA) is considered a pandemic pathogen, being essential to develop effective medicines and combat its rapid spread. This study aimed to foster the translation of clinical research outcomes based on metallodrugs into clinical practice for the treatment of MRSA. Bearing in mind the promising anti-Gram-positive effect of the heteroscorpionate ligand 1,1’-(2-(4-isopropylphenyl)ethane-1,1-diyl)bis(3,5-dimethyl-1H-pyrazole) (2P), we propose the coordination of this compound to platinum as a clinical strategy with the ultimate aim of overcoming resistance in the treatment of MRSA. Therefore, the novel metallodrug 2P-Pt were synthetized, fully characterized and its antibacterial effect against the planktonic and biofilm state of S. aureus evaluated. In this sense, three different strains of S. aureus were studied, one collection strain of S. aureus sensitive to methicillin and two clinical MRSA strains. To appraise the antibacterial activity, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), minimum biofilm inhibitory concentration (MBIC), and minimum biofilm eradication concentration (MBEC) were determined. Moreover, successful outcomes on the development of biofilm in a wound-like medium were obtained. The mechanism of action for 2P-Pt was proposed by measuring the MIC and MBC with EDTA (cation mediated mechanism) and DMSO (exogenous oxidative stress mechanism). Moreover, to shed light on the plausible antistaphylococcal mechanism of this novel platinum agent, additional experiments using transmission electron microscopy were carried out. 2P-Pt inhibited the growth and eradicated the three strains evaluated in the planktonic state. Another point worth stressing is the inhibition in the growth of MRSA biofilm even in a wounded medium. The results of this work support this novel agent as a promising therapeutic alternative for preventing infections caused by MRSA

SARS-CoV-2 Infection in Multiple Sclerosis

To understand COVID-19 characteristics in people with multiple sclerosis (MS) and identify high-risk individuals due to their immunocompromised state resulting from the use of disease-modifying treatments. Retrospective and multicenter registry in patients with MS with suspected or confirmed COVID-19 diagnosis and available disease course (mild = ambulatory; severe = hospitalization; and critical = intensive care unit/death). Cases were analyzed for associations between MS characteristics and COVID-19 course and for identifying risk factors for a fatal outcome. Of the 326 patients analyzed, 120 were cases confirmed by real-time PCR, 34 by a serologic test, and 205 were suspected. Sixty-nine patients (21.3%) developed severe infection, 10 (3%) critical, and 7 (2.1%) died. Ambulatory patients were higher in relapsing MS forms, treated with injectables and oral first-line agents, whereas more severe cases were observed in patients on pulsed immunosuppressors and critical cases among patients with no therapy. Severe and critical infections were more likely to affect older males with comorbidities, with progressive MS forms, a longer disease course, and higher disability. Fifteen of 33 patients treated with rituximab were hospitalized. Four deceased patients have progressive MS, 5 were not receiving MS therapy, and 2 were treated (natalizumab and rituximab). Multivariate analysis showed age (OR 1.09, 95% CI, 1.04-1.17) as the only independent risk factor for a fatal outcome. This study has not demonstrated the presumed critical role of MS therapy in the course of COVID-19 but evidenced that people with MS with advanced age and disease, in progressive course, and those who are more disabled have a higher probability of severe and even fatal diseas

Insectos como plagas potenciales del cacao (Theobroma cacao L.) en El Salvador

The research was carried out in cocoa trees in El Salvador, identifying in the study more than 250 species of arthropods, of which few insect species can become cocoa pests (Theobroma cacao L.), when inappropriate use of synthetic agrochemicals is made and poor management of the crop, soil and environment. This article describes and provides photographs taken in the field and laboratory of insects that can become pests of the cocoa crop in El SalvadorEl trabajo de investigación fue desarrollado en cacaotales de El Salvador, identificando más de 250 especies de artrópodos, durante el estudio, de los cuales pocas especies de insectos se pueden convertir en plagas del cacao (Theobroma cacao L.), cuando se hace un uso inadecuado de agroquímicos sintéticos y mal manejo del cultivo, suelo y ambiente. En este artículo se describen y se proporcionan fotografías tomadas en campo y laboratorio, de los insectos que pueden convertirse en plagas del cultivo de cacao en El Salvado