Search

La presente invención se refiere a materiales textiles fabricados a partir de fibra de algodón funcionalizada con nanopartículas de óxidos semiconductores que están distribuidas de forma homogénea en el material textil, los cuales le confieren doble funcionalidad de ser antimicrobiano y de protección a la radiación UV; la presente invención también se refiere a los métodos de funcionalización para la obtención de dichos materiales textiles. Vigente

Trabajo presentado en el VIII Symposia of medicinal chemistry young researchers, celebrado en Barcelona (España) el 22 de julio de 2022., Ion channels are macromolecular complexes present in the plasma membrane and in intracellular organelles of the cells, where they play important functions. The dysfunction of these channels results in several disorders named channelopathies, which represent a challenge for study and treatment.[1] We are focused on voltage-gated potassium channels, specifically on KV4.3. Within the heart, KV4.3 channels generate the transient outward potassium current (ITO). However, ITO characteristics are only observed when KV4.3 assembles with accessory subunits as KChIP2 and DPP6. KV4.3 channelosome play a key role in atrial fibrillation (AF), the most common cardiac arrhythmia, with an estimated prevalence in the general population of 1.5–2%. However, current antiarrhythmic drugs for AF prevention have limited efficacy and considerable potential for adverse effects.[2] KChIP2 (Potassium Channel Interacting Protein 2) belongs to the calcium binding protein superfamily. It is the KChIP member predominantly expressed in heart and a key regulator of cardiac action potential duration. Novel KChIP2 ligands could be a useful pharmacological tool to understand the role of KV4.3 channelosome in AF and it could help to discover new treatments for AF. [3] In this communication, we will describe a multidisciplinary approach that, starting with a structure-based virtual screening, followed by an iterative process of synthesis/biological evaluation/docking studies, has led to the identification of new KChIP2 ligands., PID2019-104366RB-C21, PID2019-104366RB-C22, PID2020-114256RB-I00 and PID2020-119805RB-I00 grants funded by MCIN/AEI/10.13039/501100011033; and PIE202180E073 and 2019AEP148 funded by CSIC. C.V.B. holds PRE2020-093542 FPI grant funded by MCIN/AEI/10.13039/501100011033. PGS was recipient of an FPU grant (FPU17/02731). AB-B holds BES-2017-080184 FPI grant and A.P-L.holds RYC2018-023837-I grant both funded by MCIN/AEI/ 10.13039/501100011033 and by “ESF Investing in your future”.

Resumen del trabajo presentado en el XX National Meeting of the Spanish Society of Medicinal Chemistry, celebrado en Santiago de Compostela (España) del 19 al 22 de junio de 2022., Ion channels are macromolecular complexes present in the plasma membrane and in intracellular organelles of the cells, where they play important functions, such as smooth muscle contraction or secretion of hormones. The dysfunction of these channels results in several disorders named channelopathies, which represent a challenge for study and treatment.[1] We are focused on voltage-gated potassium channels, specifically on KV4.3 and its role in atrial fibrillation (AF). KV4.3 channel is expressed in smooth muscle, heart and brain. Its activation generates outward currents operating at subthreshold membrane potentials as recorded from myocardial cells (ITO, transient outward current). KV4.3 plays a key role in AF, the most common cardiac arrhythmia, with an estimated prevalence in the general population of 1.5–2%. However, current antiarrhythmic drugs for AF prevention have limited efficacy and considerable potential for adverse effects.[2] To reproduce the ITO currents, KV4.3 channels need to assemble with other subunits, forming channelosome. KChIPs (Potassium Channel Interacting Proteins), which belong to the calcium binding protein superfamily, forms the KV4.3 channelosome, along with other proteins.[3] To study the complex interaction of KV4.3 channelosome and the development of modulators, the interplay of different techniques is central to advance knowledge. In this regard, the development of novel KChIPs modulators and fluorescent biosensors constitutes invaluable pharmacological tools to unravel the KV4 channelosome and its role in AF.[3] In this communication, we will describe preliminary results towards the identification of new pharmacological tools to study the KChIPs/KV4 interaction., This work was supported by the projects: PID2019-104366RB-C21, PID2019-104366RB-C22, PID2020-114256RB-I00 and PID2020-119805RB-I00 funded by MCIN/ AEI /10.13039/501100011033; and the projects PIE202180E073 and 2019AEP148 funded by CSIC. C. Viedma Barba holds the grant PRE2020-093542 and M. Valencia holds the grant PRE2020-093950, both funded by MCIN/AEI/10.13039/501100011033. A. Pérez-Lara holds the grant RYC2018-023837-I funded by MCIN/AEI/10.13039/501100011033 and by “ESF Investing in your future”.

Resumen del trabajo presentado en el VIII Congreso Red Española de Canales iónico, celebrado en Alicante (España) del 24 al 27 de mayo de 2022., Ion channels are macromolecular complexes present in the plasma membrane and in intracellular organelles of the cells, where they play important functions. The dysfunction of these channels results in several disorders named channelopathies, which represent a challenge for study and treatment.[1] We are focused on voltage-gated potassium channels, specifically on KV4.3. Kv4.3 is expressed in smooth muscle, heart and brain. Within the heart, Kv4.3 channels generate the transient outward potassium current (ITO). However, ITO characteristics are only observed when Kv4.3 assemble with accessory subunits as KChIP2 and DPP6. KV4.3 channelosome play a key role in atrial fibrillation (AF),the most common cardiac arrhythmia, with an estimated prevalence in the general population of 1.5–2%. However, current antiarrhythmic drugs for AF prevention have limited efficacy and considerable potential for adverse effects.[2] KChIP2 (Potassium Channel Interacting Protein 2) belongs to the calcium binding protein superfamily. It is the KChIP member predominantly expressed in heart and a key regulator of cardiac action potential duration. The identification of novel KChIP2 ligands could be useful to understand the role of KV4.3 channelosome in AF and it could help to discover new treatments for AF. [3] In this regard, structure-based virtual screening could be an important tool to accelerate the identification of novel KChIP2 ligands. In this communication, we will describe a multidisciplinary approach that, starting with a structurebased virtual screening, followed by an iterative process of synthesis/biological evaluation/docking studies, has led to the identification of new KChIP2 ligands., PID2019-104366RB-C21, PID2019-104366RB-C22, PID2020-114256RB-I00 and PID2020-119805RB-I00 grants funded by MCIN/AEI/10.13039/501100011033; and PIE202180E073 and 2019AEP148 funded by CSIC. C.V.B. holds PRE2020-093542 FPI grant funded by MCIN/AEI/10.13039/501100011033. PGS was recipient of an FPU grant (FPU17/02731). AB-B holds BES-2017-080184 FPI grant and A.P-L.holds RYC2018-023837-I grant both funded by MCIN/ AEI/ 10.13039/501100011033 and by “ESF Investing in your future”

Trabajo presentado en el VIII Congreso Red Española de Canales Sónicos, celebrado en Alicante (España) del 24 al 27 de mayo de 2022., The transient outward potassium current (Itof), responsible for the repolarization of cardiac action potential, is generated by the activation of KV4 channels assembled with KChIP2 and other accessory subunits, such as DPP6 and KCNE2. To test the hypothesis that these subunits modify 18 VIII Congreso Red Española de Canales Iónicos the pharmacological response of the channel, we have analyzed the electrophysiological efects of IQM-266 on the currents generated by KV4.3/KChIP2, KV4.3/KChIP2/DPP6, KV4.3/KChIP2/KCNE2 channels and on the total I K and I tof in cardiac myocytes. CHO cells were transiently transfected (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. In mice cardiac myocytes, potassium currents were recorded by using the perforated patch-clamp technique. Our results indicate that IQM-266 exerts an activating efect on the I K and I tof peak amplitude in cardiac myocytes. To decipher whichbeta subunits were involved in these efects, IQM-266 was studied in CHO cells transfected with the above-mentioned cDNAs. IQM-266, at 3 μM, induced an increase in the KV4.3/KChIP2 current charge, which augmented in the presence of DPP6, whereas it was abolished when KCNE2 was expressed. However, IQM-266 decreased the peak amplitude in transfected cells. In this study we concluded that: i) IQM-266 is a new activator of the I tof in mice cardiac myocytes, ii) DPP6 and KCNE2 modify the pharmacological response of KV4.3/KChIP2 channels to IQM-266, and iii) the presence of either DPP6 or KCNE2 is not enough to reproduce the activator efect observed when IQM-266 was tested on Ito recorded from mouse ventricular myocytes., Grants SAF2016-75021-R, RTI2018-097189-B-C22 funded by MCIN/AEI/10.13039/501100011033 and by ¿ERDF A way of making Europe¿; Grants PID2019-104366RB-C21, PID2019-104366RB-C22, PID2020-113238RB-I00 funded by MCIN/AEI/ 10.13039/501100011033; Grant CB/11/00222 funded by ISCIII CIBERCV; Grants PIE202180E073 and 2019AEP148 funded by CSIC. Grants BES-2017-080184, BES-2010-036573 and FPU17/02731 funded by MCIN/AEI/10.13039/501100011033 and by ¿ESF Investing in your future¿.

Additional file 1: Table S1. Tandem MS conditions.

El objetivo del estudio fue demostrar que las 3R de la ecología y medioambiente, usadas en la gestión de los residuos sólidos de manera exitosa, se pueden aplicar en la gestión de los efluentes textiles. Se empezó por la reducción bajando el consumo de agua con un manejo adecuado de la relación de baño en los procesos tintóreos. Para el reúso de baños se generaron efluentes textiles en el laboratorio realizando tinturas madres por en diversos colores a una relación de baño 1/8 en telas de poliéster y algodón. Se caracterizaron, reconstruyeron los baños madres, y luego se reutilizaron en nuevas tinturas. Los resultados obtenidos fueron: Para la R de reducción un ahorro de agua del 13,7% en el teñido reactivo del algodón y 11,8% en el teñido disperso del poliéster al bajar un punto en la relación de baño. Para segunda R, los teñidos con baños reutilizados presentan valores de diferencia de color DE CMC (2:1) promedio por debajo de la unidad, tolerancia usada en la industria textil, con valores de 0,44 y 0,62 para colores grises en poliéster y algodón, respectivamente. Se obtuvo un ahorro de agua de 32,4% para poliéster y de 12,1% para algodón y un ahorro promedio en insumos para el teñido disperso de 10,64% y 18,39% para el teñido reactivo, para este último el ahorro principal fue 56,25% en cloruro de sodio. Finalmente, para la tercera R, se evaluó la carga contaminante de los efluentes y se demostró la necesidad de un tratamiento centralizado.

Ion channels are macromolecular complexes present in the plasma membrane and intracellular organelles of cells. Dysfunction of ion channels results in a group of disorders named channelopathies, which represent an extraordinary challenge for study and treatment. In this review, we will focus on voltage-gated potassium channels (KV), specifically on the KV4-family. The activation of these channels generates outward currents operating at subthreshold membrane potentials as recorded from myocardial cells (ITO, transient outward current) and from the somata of hippocampal neurons (ISA). In the heart, KV4 dysfunctions are related to Brugada syndrome, atrial fibrillation, hypertrophy, and heart failure. In hippocampus, KV4.x channelopathies are linked to schizophrenia, epilepsy, and Alzheimer’s disease. KV4.x channels need to assemble with other accessory subunits () to fully reproduce the ITO and ISA currents. Subunits affect channel gating and/or the traffic to the plasma membrane, and their dysfunctions may influence channel pharmacology. Among KV4 regulatory subunits, this review aims to analyze the KV4/KChIPs interaction and the effect of small molecule KChIP ligands in the A-type currents generated by the modulation of the KV4/KChIP channel complex. Knowledge gained from structural and functional studies using activators or inhibitors of the potassium current mediated by KV4/KChIPs will better help understand the underlying mechanism involving KV4-mediated-channelopathies, establishing the foundations for drug discovery, and hence their treatments., This research was funded by Ministerio de Ciencia e Innovación (MICIU) Spain SAF2016- 75021-R and PID2019-104366RB-C21 (to CV), RTI2018-097189-B-C22 (to MMM), PID2019-104366RBC22 (to MG-R); the Instituto de Salud Carlos III CIBERCV program CB/11/00222 (to CV); and the Consejo Superior de Investigaciones Científicas grants: PIE 201820E104, 2019AEP148 (to CV), and 201880E109 (to MMM and MG-R). The cost of this publication was paid in part by funds from the European Fund for Economic and Regional Development (FEDER). PC held a postgraduate FPI fellowship from the Spanish Ministry of Economy, Industry, and Competitivity (MINECO). DAP held CSIC contracts. AdB-B holds a MICIU predoctoral contract (BES-2017-080184). PGS holds a MICIU predoctoral contract FPU2017/02731. AAN, DAE, EB, and EPA were sponsored by CUNY Research Scholars Program (CRSP) and Collegiate Science Technology Entry Program (CSTEP). YR is grateful to the Hostos Office of Academic Affairs for providing travel awards and the OpenEye Scientific Software for providing free academic licenses. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).

Textile effluents from reactive black dyes, decolorized using the advanced Fenton oxidation method, were reused in cellulosic fiber exhaustion bleaching processes. Textile black reactive effluents were collected, provided from two recipes of reactive blacks; Corafix Black SD (Black SD) and Corazol Black B 133 % (Black 5), simulated and real, due to being decolored by the Fenton treatment. The discoloration was monitored using UVvisible spectrophotometry, the optimal proportions of Fenton reagent (Fe (II) salt (g) / H2O2 (mL)) were determined: 0,0156 g / mL and 0,0251 g / mL with 90 minutes of reaction time, and optimal treatment pH of 3,70 and 3,88 for textile effluents from dyeing with Corafix Black SD and Corazol Black B 133 % dyestuffs respectively. The discolored effluents were characterized, adequated, and used in bleaching of cellulosic fabrics. Textiles with a degree of whiteness of 74,73 °Be and 73,41 °Be were obtained, similar to the substrate bleached with soft water (76,61 °Be). The reflectance curves were compared with the Datacolor SF550 spectrophotometer, hydrophilicity and capillarity according to standardized methods. Finally, it was concluded that the Fenton treatment is viable for this type of reactive black effluent since bleached substrates with properties similar to bleaching with soft water were obtained Se reusaron efluentes textiles provenientes de teñidos negros reactivos, decolorados por el método de oxidación avanzada Fenton en procesos de blanqueo de fibra celulósica por agotamiento. Se recolectaron efluentes textiles provenientes de dos teñidos reactivos negros: Corafix Black SD y Corazol Black B 133 %, simulados y reales, para ser decolorados con el tratamiento Fenton. Se monitoreó la decoloración usando espectrofotometría UV-visible, se determinaron las proporciones óptimas de reactivo Fenton (sal de Fe (II) (g)/H2O2 (mL)): 0,0156 g/mL y 0,0251 g/mL con un tiempo de reacción de 90 minutos y pH óptimos de tratamiento de 3,70 y 3,88 para los efluentes textiles provenientes de teñido con colorantes Corafix Black SD y Corazol Black B 133%, respectivamente. Los efluentes tratados se caracterizaron, adecuaron y usaron en el blanqueo de tejidos celulósicos. Se obtuvo textiles con un grado de blancura de 74,73 °Be y 73,41 °Be, similares al sustrato blanqueado con agua blanda (76,61°Berger). Se compararon: las curvas de reflectancia con el espectrofotómetro Datacolor SF550, hidrofilidad y capilaridad según métodos normalizados. Finalmente se concluyó que el tratamiento Fenton es viable para este tipo de efluentes negros reactivos, pues se obtuvieron sustratos blanqueados con propiedades similares al blanqueo con agua blanda.

El continuo crecimiento poblacional a nivel mundial, junto con la rápida urbanización e industrialización de los países en vías de desarrollo, sin un crecimiento en consonancia de las infraestructuras de tratamiento de vertidos y residuos, está generando contaminación ambiental sin precedentes. Las características de los efluentes industriales en cuanto a la naturaleza de los contaminantes, concentraciones, técnicas de tratamiento y métodos de eliminación requeridos, varían significativamente dependiendo del tipo de industria, por lo que la elección de una técnica de tratamiento de vertidos ha de regirse por múltiples parámetros, tales como la naturaleza físico-química de los contaminantes, concentración y volumen a tratar, toxicidad y el coste del tratamiento. La electrocoagulación es un método físico-químico para el tratamiento de aguas residuales preferentemente industriales, fundamentado en el empleo de una corriente eléctrica continua que promueve procesos de coagulación. Esta tecnología electroquímica se ha desarrollado en gran medida para su uso alternativo a la depuración mediante fangos activados, ofreciendo una alternativa prometedora para reducir los problemas de contaminación por ciertos efluentes industriales. Este trabajo pretende realizar una revisión de los fundamentos tecnológicos de la electrocoagulación y el estado del conocimiento de la aplicación de esta técnica para el tratamiento de aguas residuales industriales, con especial énfasis en el tratamiento de pigmentos textiles, problema de gran repercusión especialmente en países en vías de desarrollo, lugar de origen de la mayoría de prendas que vestimos. The continuous worldwide population growth, along with the rapid urbanization and industrialization of developing countries, without a growth in line with the waste and wastewater treatment infrastructures, is generating unprecedented problems of environmental pollution. The characteristics of industrial effluents in terms of the nature of the pollutants, concentrations, treatment techniques and disposal methods required, vary significantly depending on the type of industry, so the choice of a spill treatment technique must be governed by multiple parameters, such as the physical-chemical nature of the contaminants, concentration and volume to be treated, toxicity and the cost of treatment. Electrocoagulation is a physical-chemical method for the treatment of preferably industrial wastewater, based on the employment of a continuous electric current that promotes coagulation processes. This electrochemical technology has been largely developed for alternative use to water treatment by activated sludge, offering a promising alternative to reduce pollution problems by certain industrial effluents. This work aims to review the technological fundamentals of electrocoagulation and the state of knowledge of the application of this technique for the treatment of industrial wastewater, with special emphasis on the treatment of textile pigments, a problem of great impact especially in developing countries, place of origin of most of the clothes we wear.

RESUMEN: El estigma son las actitudes o creencias negativas que tiene una persona o grupo de personas hacia aquellos que perciben como diferentes. Por ello, el presente estudio tiene como objetivo principal el análisis de la relación entre el nivel de estigma, la inteligencia emocional y la actitud hacia la inclusión educativa de familiares con un hijo con discapacidad. El estudio es de carácter cuantitativo con un diseño cuasi- experimental en el que participa una muestra de familias con un hijo con discapacidad y otra de familias con un hijo sin discapacidad. La recopilación de datos se realiza a través de tres cuestionarios. Un cuestionario va dirigido a medir el nivel de estigma de familiares con un hijo con discapacidad. Otro cuestionario mide el nivel de inteligencia emocional de familiares con un hijo con discapacidad. El último cuestionario va dirigido a medir la actitud hacia la inclusión educativa tanto de familiares con un hijo con discapacidad como para familiares con un hijo sin discapacidad. Los resultados finales indican que los familiares con un hijo con discapacidad tienen niveles bajos de estigma hacia su hijo. Además, hay una relación estadísticamente significativa entre los niveles de estigma y la inteligencia emocional, es decir, cuando poseen un nivel de inteligencia emocional mayor, los niveles de estigma son bajos. Por último, en cuanto al nivel de actitud hacia la inclusión educativa, los familiares con un hijo sin discapacidad tienen un nivel más alto. ABSTRACT: Stigma is the negative attitudes or beliefs that a person or group of people has towards those they perceive as different. Therefore, the main purpose of this stud is to analyze the level of stigma, emotional intelligence and attitude towards the educational inclusion of family members with a child with disability. The study is quantitative in nature with a quasi-experimental design involving a sample of families with a child with disability and families with child with no disability. Data collection is carried out through three questionnaires. A questionnaire is aimed at measuring the level of stigma of family members with a child with disability. Another questionnaire measures the level of emotional intelligence of family members with a child with disability. The last questionnaire is aimed at measuring the attitude towards educational inclusion of both family members with a child with disability and family members with a child with no disability. The final results indicate that family members with a child with disabilityhave low levels of st igma towards their child. In addition, there is a statistically significant relationship between stigma levels and emotional intelligence, that is to say, when they have an adequate level of emotional intelligence, stigma levels are low. Finally, with regard to the level of attitude towards educational inclusion, family members with a child with no disability are at a higher level.

The present invention relates to a family of compounds which are capable of modulating KChIP proteins and are therefore useful for the treatment of heart diseases in which the expression levels of this protein are abnormal. These compounds have the following general formula: (I)., Consejo Superior de Investigaciones Científicas (España), Universidad Autónoma de Madrid, A1 Solicitud de patente con informe sobre el estado de la técnica

El uso de plaguicidas en el sector agrícola sinaloense es una de las prácticas más comunes en la actividad primaria de la región. La exposición a éstos representa un factor definitivo, a la vez para la salud ambiental y la de la sociedad, en este caso la de los jornaleros rurales, principalmente. El presente trabajo tiene como objetivo evidenciar las implicaciones que tiene sobre la salud pública y medio ambiente, la ineficacia en las formas de gobernar ambientalmente un sector tan importante como lo es el agrícola, para la sociedad mexicana, no solo por su aportación a la alimentación, sino a la economía. Para ello se utilizó una metodología mixta: información documental, entrevistas, así como la toma de muestras biológicas, analizadas mediante métodos diversos. Se observa que la exposición a plaguicidas, en jornaleros agrícolas, detona un daño genotóxico y ambiental considerable, por lo que se establece que la gobernanza ambiental y las políticas públicas no han sido aplicadas de manera apropiada, generándose una actividad y comportamiento incongruentes, para con las necesidades de la sociedad y del medio ambiente, por parte de los actores involucrados., L’utilisation de pesticides dans la filière agricole dans l’état de Sinaloa est l’une des pratiques les plus courantes de l’activité primaire de la région. L’exposition à ces produits représente un facteur définitif à la fois pour la santé environnementale et celle de la société, en particulier celle des ouvriers agricoles. Notre travail a pour but de mettre en évidence les incidences de l’inefficacité des actions gouvernementales en matière d’environnement sur la santé publique et sur l’environnement, dans un secteur, le secteur agricole, dont l’importance sociétale est reconnue non seulement pour son apport alimentaire, mais aussi pour celui de l’économie. Pour ce faire, nous avons utilisé une méthodologie mixte fondée sur des informations documentaires, des interviews, mais aussi à partir d'échantillons biologiques analysés selon divers procédés. On observera que l’exposition aux pesticides pour les travailleurs agricoles entraîne un dommage génotoxique et environnemental considérable. C’est la raison pour laquelle nous considérons que la gouvernance en matière environnementale et les politiques publiques n’ont pas été appliquées de manière appropriée par les différents responsables et ont généré une activité et un comportement incongrus, quant aux besoins de la société et de l’environnement., O uso dos pesticidas no setor agrícola de Sinaloa é uma das práticas mais comuns da região. A exposição a estes representa um fator definitivo, tanto para saúde ambiental como para a social -no caso a saúde dos trabalhadores rurais, principalmente-. O presente trabalho tem como objetivo evidenciar as implicações que tem sobre a saúde pública e médio ambiental, a ineficiência nas formas de governar ambientalmente um setor tão importante como é o setor agrícola, para a sociedade mexicana, não só pela contribuição na alimentação, mas também na economia. Para isso se utilizou uma metodologia mista: informação documental, entrevistas, assim como a toma de amostras biológicas, analisadas mediante diversos métodos. Observa-se que a exposição aos pesticidas em trabalhadores agrícolas detona um dano genotóxico e ambiental considerável, portanto, estabelece-se que a governança ambiental e as políticas públicas não tem sido aplicadas de maneira apropriada, gerando-se uma atividade e comportamento incongruente, para as necessidades da sociedade e do médio ambiente, por parte dos atores envolvidos., The use of pesticides in the Sinaloa agricultural sector is one of the most common practices in the primary activity of the region. Exposure to these represents a definitive factor, both for environmental health and for society - in this case, that of rural day laborers, mainly. The present work aims to highlight the implications it has on public health and the environment, the inefficiency in the ways of environmentally governing a sector as important as the agricultural sector, for Mexican society, not only for its contribution to food , but to the economy. For this, a mixed methodology was used: documentary information, interviews, as well as the taking of biological samples, analyzed by various methods. It is observed that exposure to pesticides in agricultural day laborers detonates considerable genotoxic and environmental damage, for which reason it is established that environmental governance and public policies have not been applied properly, generating an incongruent activity and behavior, for the needs of society and the environment, by the actors involved., Trayectorias Humanas Trascontinentales, HS N° 4 | 2019

Trabajo presentado en el 19th Meeting SEQT, celebrado en Vitoria-Gasteiz (España) del 8 al 11 de julio de 2019., DREAM (Downstream Regulatory Element Antagonist Modulator), also known as KChIP-3 or calsenilin, is a multifunctional calcium binding protein that controls the expression level and/or the activity of several proteins related to calcium homeostasis, neuronal excitability and neuronal survival. As an auxiliary protein in the plasma membrane, DREAM interacts with KV4 potassium channels, L- and T-type voltage-dependent calcium channels, NMDA receptors, presenilins and the transcriptor factor ATF6.1 The interaction between DREAM and KV4 potassium channels regulates of A-type outward potassium currents (IA) that is responsible of the fast repolarization of neuron action potentials and frequency of firing.2 Using a multidisciplinary approach that involves drug design, organic chemistry, surface plasmon resonance assays, electrophysiological recordings of KV4.3/DREAM channels, and IA recordings in rat dorsal root ganglion neurons, we have identified IQM-266.3 IQM-266 slows the inactivation kinetics, and this effect may explain why at concentrations lower than the IC50, IQM-266 augments the efflux of potassium ions resulting in an increase of the charge (activating effect). This effect could be the basis of a promising therapeutic strategy for the treatment of certain neuronal pathologies (epilepsy, Alzheimer disease or ataxia), in which a downregulation of KV4.3 or DREAM has been demonstrated.4 IQM-266 also modulated IA from rat DRG neurons. Overall, IQM-266 constitutes a novel small chemical tool suitable to modulate KV4.3 channels in native systems, that might allow a better understanding of DREAM physiological roles, as well as modulation of neuronal IA current in pathological processes.

Resumen del trabajo presentado al 6th European Chemical Biology Symposium ECBS/LS-EuChemS, celebrado en Madrid (España) del 3 al 5 de abril de 2019., DREAM (Downstream Regulatory Element Antagonist Modulator), also known as KChIP-3 or calsenilin, is a multifunctional calcium binding protein that controls the expression level and/or the activity of several proteins related to calcium homeostasis, neuronal excitability and neuronal survival. As an auxiliary protein in the plasma membrane, DREAM interacts with, and regulates, the gating of KV4 potassium channels, L- and T-type voltage-dependent calcium channels, NMDA receptors and the transcriptor factor ATF6, which is involved in the unfolding protein response machinary. Considering that altered neuronal calcium homeostasis and the accumulation of poorly folded proteins are common features of many neurodegenerative pathologies, the DREAM modulation could open new avenues for the treatment of neurodegenerative diseases. We recently showed that reduced DREAM expression or blockade of DREAM activity by repaglinide is neuroprotective in Huntington's disease (HD)[1]. However, up to now, only three DREAM binding molecules have been identified. Hence, there is a clear need for the development of chemical tools to modulate and characterize DREAM activity and its interactions. In this communication we will report the identification of novel DREAM modulators by following a multidisciplinary strategy that involves structure-based design, organic chemistry, site-directed mutagenesis and in vitro and in vivo experiments. Our findings open a new avenue in the search of effective treatments of neurodegenerative diseases., Spanish Ministery of Economy, Industry and Competitivity (AEI-FEDER, EU grants): BFU2015-67284-R, SAF2017-89554-R, SAF2016-75021-R, SAF2015-66275-C2-2-R; Instituto de Salud Carlos III CIBERNED and CIBERCV programs; the Madrid regional government/Neurodegmodels; and CSIC PIE 201580E073.

Resumen del trabajo presentado al VII Congreso Red Española Canales Iónicos, celebrado en Cáceres del 15 al 17 de mayo de 2019., The outward potassium current IKur is the main responsible of the atrial repolarization process and it is generated by the activation of KV1.5 channels, widely expressed in human atria. It is known that mutations in KCNA5 gene, which induce both gain- and loss-of-function in KV1.5 channel, enhance atrial fibrillation susceptibility. Thus, these channels represent a pharmacological target for the development of antiarrhythmic drugs useful in the treatment of supraventricular arrhythmias. KV1.5 channels assembly with several regulatory subunits such as KVβ and KChIPs (KV Channel Interacting Proteins). It has been described that KChIP2 physically interacts with KV1.5 and reduces KV1.5 cell surface expression levels. Our research group has demonstrated that IQM-266 inhibits the current generated by the activation of KV4.3 and KV4.3/KChIP2, being the effects more marked when KChIP2 is present. The aim of the present study is to analyze the effects of IQM-266 on KV1.5 channels. In order to achieve these objectives, HEK293 cells transiently expressing KV1.5 were used. Currents were recorded using the whole-cell configuration of the patch-clamp technique. The effects of IQM-266 on KV1.5 currents were concentration-dependent with an IC50 of 11 μM (n=24). Block induced by IQM-266 (20 μM) sharply increased within the membrane voltage range of the channel activation, arising a maximum degree of block at +20 mV that remained constant at more positive membrane potentials. This compound at 20 μM produced a timedependent block, inducing a: 1) faster inactivation (τ = 305.8±47.6 ms vs. 168.0±13.4 ms, in the absence and in the presence of IQM-266, respectively, n=9, p, Supported by SAF2016-75021-R and CSIC PIE201820E104 to CV, BFU2015-67284-R and CSIC PIE201880E109 to MGR.

Resumen del trabajo presentado al VII Congreso Red Española Canales Iónicos, celebrado en Cáceres del 15 al 17 de mayo de 2019., Neuropathic pain is a form of chronic pain arising from damage to the nerves that sense, transmit or process information about environmental stimuli. Given its growing prevalence and common refractoriness to conventional analgesics, the development of new drugs with pain relief effect constitute a prominent clinical need. In this respect, drugs that reduce activity of sensory neurons by modulating ion channels hold the promise to become effective analgesics. Here we have used two models of neuropathic pain, namely the chronic constriction injury of the sciatic nerve (CCI) and the streptozotocin-induced diabetic neuropathy, to evaluate the mechanical antiallodynic effect of IQM-PC332, a novel Downstream Regulatory Element Antagonist Modulator (DREAM)/KChIP3/calsenilin ligand potentially affecting DREAM modulated ionic currents like potassium IA and ICav. IQM-PC332 exerted a mechanical antiallodynic effect following intraperitoneal (I.P; DE50 of 0.06 μg/kg, Emax of 65%) and intraplantar (DE50 of 0.24 mg, Emax of 68%) administration in the CCI, model. Likewise, IQM-PC332 2 μg/kg I.P. significantly reduced mechanical allodynia in diabetic animals. Interestingly, no effect of IQM-PC332 on glucose levels from both CCI and diabetic animal could be observed at analgesic doses. The effects of IQM-PC332 on INav, ICav, IA, and the electrical excitability in neurons isolated from dorsal root ganglia (L4-L6; DRG) were also studied with the patch-clamp technique. IQM-PC332 reduced peak amplitudes of ICav from both Control and CCI animals with CI50 (@ +10 mV) of 78 μM y 40 μM, respectively. At variance, IQM-PC332 did not exert any effect on INav in the concentration range from 0.01 to 50 μM. PC332 10 μM also inhibited IA in DRG neurons from CCI animals, this effect being larger with membrane depolarization (≈ 50% inhibition at -10 mV). Interestingly, in about 55% of the neurons, IQM-PC332 slowed inactivation of IA. In accordance with its effects on ion currents, IQM-PC332 10 μM reduced the time to the first action potential (AP) evoked by current injection while decreasing the amplitude and duration of the after hyperpolarization that followed after APs within a train. However, no clear effect on instantaneous frequency of AP could be observed. It is suggested that inhibition of Ca2+ entry through Cav channels coupled to neurotransmitter release from peripheral and central terminals of nociceptive neurons may underlie the analgesic effect of IQM-PC332., Supported by Universidad Complutense, grant PR75/18-21593 to ARA, SAF2016-75021-R and PIE201820E104 to CV, BFU2015-67284-R and PIE201880E109 (CSIC grant) to MGR.

Resumen del trabajo presentado al VII Congreso Red Española Canales Iónicos, celebrado en Cáceres del 15 al 17 de mayo de 2019., Immune cells have an important role in the tumor-microenvironment. Macrophages may tune the immune response toward inflammatory or tolerance pathways. Tumor associated macrophages (TAM) have immunosuppressive functions and they are considered a therapeutic target in cancer. The aim of this study was to evaluate the effects of trabectedin, a new class of antitumor agent, on the tumor-microenvironment through the study of electrophysiological and molecular phenotype of macrophages. Experiments were performed using the whole-cell patchclamp configuration of the patch-clamp technique in resident peritoneal mouse macrophages under different types of polarization. Trabectedin decreased macrophages viability and increased ROS production. Trabectedin does not directly interact with KV1.5 and KV1.3 channels, but treatment (16h) of macrophages with sub-cytotoxic concentrations (0.1-5 nM) increased their KV current in a concentration-dependent manner due to an upregulation of KV1.3 channels. In vitro generated TAM (TAMiv), by a co-culture of ID8 cells and macrophages, exhibited a M2 phenotype. TAMiv generated a small KV current, similarly to M2 polarized macrophages, and expressed high levels of M2 markers. In this study, we demonstrated that TAMiv polarization could be re-educated by using sub-cytotoxic concentration of trabectedin. TAMiv treated with sub-cytotoxic concentrations of trabectedin exhibited an upregulation of KV1.3 channels and their M2 phenotype changed towards M1 pro-inflammatory one., Funded by PharmaMar, CSIC 201820E104, CIBERCV and SAF2016-75021-R.

Los ARN circulares (circRNAs) han sido identificados recientemente como una clase de isoformas de ARN que forman una molécula cerrada de forma covalente y que se expresan de manera estable, generalizada y abundante en tejidos. Existen evidencias de que pueden actuar como esponjas de micro ARNs, y de su implicación en diferentes tipos de cáncer. Los exosomas son pequeñas vesículas derivadas de las células y de origen endocítico que participan en los procesos celulares actuando como una vía de comunicación entre ellas. Se ha demostrado que las células de un tumor suelen producir más exosomas que las sanas y que pueden detectarse en la sangre periférica humana. Adicionalmente, en la secuenciación de ARN, tanto en sangre completa como en exosomas, se han detectado miles de circRNAs de forma reproducible. En este trabajo se han cuantificado los circRNAs presentes en exosomas de sangre periférica de individuos sanos y con tres tipos de cáncer: colorrectal, hepatocelular y pancreático. Usando la expresión de estos circRNAs se ha conseguido discriminar, con una alta precisión, entre personas sanas y con cáncer, usando técnicas de Machine Learning, reforzando así la hipótesis de que los circRNAs presentes en exosomas de sangre periférica son biomarcadores prometedores que se expresan de forma diferente para distintos tipos de cáncer. Adicionalmente se hace una revisión sobre los micro ARNs sobre los que los circRNAs más expresados podrían estar actuando como esponjas, encontrando evidencias de la implicación de la desregulación de estos micro ARNs en los tres tipos de cáncer estudiados. Covalently closed circular RNA molecules (circRNAs) have recently emerged as a class of RNA isoforms with widespread and tissue-specific expression. circRNAs are remarkably stable and highly expressed molecules. Emerging evidence reveals that they might act as micro RNAs sponges, and also there is evidence of their involvement in different types of cancer. Exosomes are small membrane vesicles of endocytic origin secreted by most cell types and they are thought to play important roles in intercellular communications. It has been shown that tumour cells tend to produce more exosomes than healthy cells and that they can be detected in human peripheral blood. Additionally, in RNA sequencing, in whole blood as well as in exosomes, thousands of circRNAs have been consistently detected. Here we have quantified the circRNAs present in exosomes of peripheral blood of healthy people and with three types of cancer: colorectal, hepatocellular and pancreatic. Using the expression of these circRNAs, we have been able to discriminate, with high precision, between healthy individuals and patients of each cancer group using Machine Learning techniques, reinforcing the hypothesis that circRNAs present in peripheral blood exosomes are very promising biomarkers since they are expressed differently for different types of cancer. Additionally, a review has been made about the micro RNAs on which the most expressed circRNAs could be acting as sponges, finding evidence of the implication of the dysregulation of these micro RNAs in the three types of carcinomas. Els ARN circulars (circRNAs) han estat identificats recentment com una classe de isoformas de ARN que formen una molècula tancada de forma covalent i que s'expressen de manera estable, generalitzada i abundant en teixits. Existeixen evidències que poden actuar com a esponges de micro ARNs, i de la seva implicació en diferents tipus de càncer. Els exosomas són petites vesícules derivades de les cèl·lules i d'origen endocítico que participen en els processos cel·lulars actuant com una via de comunicació entre elles. S'ha demostrat que les cèl·lules d'un tumor solen produir més exosomas que les sanes i que poden detectar-se en la sang perifèrica humana. Addicionalment, en la seqüenciació de ARN, tant en sang completa com en exosomas, s'han detectat milers de circRNAs de forma reproduïble. En aquest treball s'han quantificat els circRNAs presents en exosomas de sang perifèrica d'individus sans i amb tres tipus de càncer: colorrectal, hepatocelular i pancreàtic. Usant l'expressió d'aquests circRNAs s'ha aconseguit discriminar, amb una alta precisió, entre persones sanes i amb càncer, usant tècniques de Machine Learning, reforçant així la hipòtesi que els circRNAs presents en exosomas de sang perifèrica són biomarcadores prometedors que s'expressen de forma diferent per a diferents tipus de càncer. Addicionalment es fa una revisió sobre els micro ARNs sobre els quals els circRNAs més expressats podrien estar actuant com a esponges, trobant evidències de la implicació de la desregulación d'aquests micro ARNs en els tres tipus de càncer estudiats.

Resumen del trabajo presentado al VI Meeting de la Red Española de Canales Ióniocs (RECI), celebrado en Santiago de Compostela del 6 al 8 de septiembre de 2017., KChIP-3 (potassium channel interacting protein-3), also known as DREAM (Downstream Regulatory Element Antagonist Modulator) or calsenilin, is a multifunctional calcium binding protein that controls the expression level and/or the activity of several proteins related to calcium homeostasis, neuronal excitability and neuronal survival.1 As an auxiliary protein in the plasma membrane, KChIP-3 interacts with, and regulates, among others, the gating of KV4 potassium channels, L- and T-type voltage-dependent calcium channels, NMDA receptors and the transcriptor factor ATF6, which is involved in the unfolding protein response machinery. Considering that altered neuronal calcium homeostasis and the accumulation of poorly folded proteins are common features of many neurodegenerative pathologies, the KChIP-3 modulation could open new avenues for the treatment of different neurodegenerative diseases. However, up to now, only three KChIP-3 binding molecules have been identified. Hence, there is a clear need for the development of chemical tools to modulate and characterize KChIP-3 activity and its interactions. In this communication we report the rational design and synthesis of new KChIP-3 ligands as pharmacological tools to study its protein-protein interaction network, focusing on those interactions involved in neurodegenerative pathologies., This work was supported by Ministerio de Economia y Competitividad (BFU2015-67284-R) and CSIC (201580E073-PIE).

Resumen del trabajo presentado al VI Meeting de la Red Española de Canales Ióniocs (RECI), celebrado en Santiago de Compostela del 6 al 8 de septiembre de 2017., Macrophages play an important role in the inflammatory response, acting as antigen-presenting cells and modifying the cytokine milieu and the intensity of T-cell signaling. The electrophysiological properties of macrophages depend on their state of functional activation. Trabectedin induces rapid apoptosis exclusively in mononuclear phagocytes and this effect contributes directly to its antitumor activity. The aim of this study was to evaluate the effects of trabectedin on KV and Kir channels in resident peritoneal and bone marrow-derived macrophages (BMDM) under both short- and long-term incubation with the drug. Experiments were performed using the whole-cell configuration of the patchclamp technique. Following exposure (ca. 10 min) of peritoneal macrophages or BMDM to 100 nM trabectedin neither KV nor Kir were modified. The electrophysiological effects of prolonged treatment with trabectedin were studied after 16 h of incubation with the drug. Under these conditions, trabectedin produced a concentration-dependent effect that was qualitatively similar to the response observed after LPS challenge. Moreover, the increase in KV observed appeared in parallel with a use-dependent decline, typical of macrophages stimulated with LPS. Similarly, the degree of inactivation of the current was greater and faster at higher trabectedin concentrations. All these results suggest that: 1) Trabectedin does not interact with Kir or KV channel. 2) Trabectedin acts by re-educating mouse resting peritoneal macrophages into M1-subtype., Supported by PharmaMar, SAF2016-75021-R and CIBERCV FIS (CB/11/00222) Grants.

An interdisciplinary study of the geomorphology, sedimentology, stratigraphy and physical oceanography of the deep-sea environments of the Alboran Sea (south-western Mediterranean Sea) has been carried out with the purpose of evidencing and understanding the role of bottom currents in the sedimentary evolution of the Spanish and Moroccan continental margins and adjacent basins during the Pliocene and Quaternary. This study was conducted using swath bathymetry data, more than 1900 profiles consisting of parametric, single- and multi-channel seismic records, scientific and commercial wells, sediment cores, and hydrographic data comprising: Conductivity, Temperature and Depth (CTD) profiles, Acoustic Doppler Current (ADCP) profiles, and EK60 echograms. Here, for the first time, a morphosedimentary scenario with a wide spectrum of depositional (plastered, sheeted, channel-related, mounded confined, elongated and separated drifts) and erosional (terraces, escarpments, moats, channels and furrows) contourite features are described in the Alboran Sea, from the shelf break to the basin floor. Hydrographic data offers new insights into the distribution of the Mediterranean water masses, and reveals that the bottom circulation of the Western Intermediate Water (WIW) and the Levantine Intermediate Water (LIW) interact with the Spanish slope, and the Western Mediterranean Deep Water (WMDW) with the Moroccan slope, Spanish base- of-slope and deep basins. The integration of distinct datasets and approaches allow a new sedimentary model to be proposed for the Alboran Sea that underlines the significance of bottom current processes in shaping deep-sea morphology. This model suggests that the bottom circulation of water masses governs physiography that the interface positions of water-masses with contrasting densities sculpt terraces at a regional scale, and that morphological obstacles play an essential role in the local control of processes and water- mass distribution. An analysis of the seismic stratigraphy from the Pliocene and Quaternary sequences has enabled to update and rename the stratigraphic boundaries and establish a new seismic stratigraphy for the Alboran Sea, after relocating the base of the Quaternary from 1.8 to 2.6Ma. Additionally, the seismic analysis involves the presentation and discussion for the first time of the evidence for contourite features reaching the scale of the Alboran Basin. Contourite drifts (plastered, sheeted, elongated separated and confined mounded drifts) and erosive features (terraces, escarpments, moats, channels, furrows) were developed under the continuous influence of Mediterranean water masses after the opening of the Strait of Gibraltar (-5.33Ma). At least two primary factors have controlled the contourite features in this sea: i) tectonics, which has governed the relocation of the main Mediterranean flow pathways and their circulation patterns; and ii) climate, which has influenced both water-mass conditions (depth and density contrast of the interfaces) and hinterland sediment sources, conditioning the morphoseismic expression and growth pattern of drifts and terrace formation (dimensions). The distribution of contourite features through time and space allows to propose three main scenarios for ocean circulation since the opening of the Strait of Gibraltar: i) Atlantic Zanclean flooding; ii) the Pliocene sea, with two different stages for the dense circulation and characterised by poorly-defined and unstable interfaces for the Atlantic Waters (AW), light and dense Mediterranean waters and the presence of a strong countercurrent in the Western Basin; and iii) the Quaternary sea, characterised by tabular Mediterranean water masses with multiple current dynamics, increasingly important density contrasts, and climate shifts causing major vertical and horizontal displacement of the interfaces. These stages reflect variability in the bottom current regimes and related alongslope efficiency in terms of transport, deposition and erosion. The detailed seismic analysis of the units making up the Pliocene and Quaternary sequences allows for the first time, to make an in-depth analysis of the contourite features, turbidite systems and mass-movement deposits, and map them through time. These maps are enormously helpful when it comes to understanding the sedimentary architecture of the Spanish and Moroccan continental margins and basins, as well as for decoding the palaeoceanographic processes from a geological perspective. Two main contourite depositional systems are defined: the Intermediate Mediterranean Contourite System (IMCS), formed under the action of the Light Mediterranean Waters (LMW) on the Spanish margin, and the Deep Mediterranean Contourite System (DMCS), formed under the action of the Dense Mediterranean Waters (DMW) mainly on the Moroccan margin and basins. The characterisation of the terraces as contourite features that form under the combination of two water masses, has also led to the definition of the Atlantic Contourite System (ACS). The occurrence of several contourite depositional systems has led to the suggestion of a new term, not heretofore considered in the literature: Multiple Contourite Depositional System (MCDS), which refers to the set of different CDSs that occurs in the same area and evolving under the action of multiple water masses. In addition, twenty turbidite systems have been characterised, revealing that they are responsible for the different sedimentary architecture of the Spanish margin, where they coexist with contourites, as on the Moroccan margin the turbidite systems are less well developed. The mass-movement deposits are mainly related to the reworking of the contourites draping the highs. Mainly contourites but also turbidites, allowed to define from a geological perspective the basic oceanographic processes and to determine their occurrence, relative magnitude and energy, and time of action. This PhD thesis also explains the uneven development of the turbidite systems in the Alboran Sea, which is interpreted to be conditioned by the interaction of alongslope with downslope processes. Several morphological and sedimentary signatures produced by the interaction between both processes have been identified in the Pliocene and Quaternary records, as well as on the present-day seafloor of the Alboran Sea. The interaction scenarios move between two-end-members: from bottom currents dominating gravity flows to gravity flows dominating contour currents. In between these extreme cases, the alternation and mutual influence of both processes can occur. Two different conceptual models of interaction are proposed for the Spanish and Moroccan margins. i) On the Spanish margin, the alongslope and downslope interaction is especially complex and varied, with both regional and local effects on the turbidite systems. This is because here the turbidite systems are influenced at different water depths by Atlantic and Mediterranean water masses and their interfaces, with current flows that change across- and downslope. ii) On the Moroccan margin, the vigorous action of the WMDW primarily inhibits the formation of canyons and associated deposits. The findings of this PhD thesis suggest that the relevance of bottom-water processes in deep sea must be reevaluated. It is concluded that understanding the influence of bottom currents is not only essential for reconstructing present and past water mass circulation, but also for recognising sea floor morphologies and decoding the sedimentary stacking pattern and evolution of deposits, as well as global climate and periods of eustatic variation., En el presente trabajo se ha llevado a cabo un estudio multidisciplinar de la geomorfología, sedimentología, estratigrafía y oceanografía física de los ambientes profundos del Mar de Alborán (extremo sur-oeste del Mar Mediterráneo), con el propósito de comprender el papel que los procesos asociados a las corrientes de fondo juegan en la evolución sedimentaria de los márgenes continentales de España y Marruecos, así como de las cuencas adyacentes, durante el Plioceno y el Cuaternario. Este estudio se ha llevado a cabo empleando datos de batimetría multihaz, más de 1900 registros sísmicos de sonda paramétrica, mono- y multi-canal, sondeos científicos e industriales, testigos de sedimento, y varios tipos de datos hidrográficos que comprenden: perfiles de conductividad, temperatura y profundidad (CTD), perfiles obtenidos con un correntómetro acústico Doppler (ADCP), y ecogramas registrados con una ecosonda EK60. Por primera vez en el Mar de Alborán, se ha sido descrito un contexto morfosedimentario compuesto por un amplio espectro de rasgos contorníticos deposicionales (crestas adosadas, laminares, asociadas a canales, monticulares confinadas y elongadas separadas) y erosivos (terrazas, escarpes, fosas, canales y surcos erosivos), desde el borde de la plataforma continental hasta las cuencas. Los datos hidrográficos han ofrecido nueva informaci6n sobre la distribución de las masas de agua mediterráneas, y han revelado que la circulación de fondo del Agua Intermedia Occidental ("Winter Intermediate Water", WIW) y el Agua Intermedia Levantina ("Levantine Intermediate Water", LIW) interaccionan con el talud continental del margen español, mientras que el Agua Mediterránea Occidental Profunda ("Western Mediterranean Deep Water", WMDW) interactúa con el talud continental del margen marroquí, la base de talud del margen español y las cuencas profundas. La integraci6n de diversas bases de datos y de distintas disciplinas ha permitido proponer un nuevo modelo sedimentario para el Mar de Alborán el cual enfatiza la importancia de los procesos sedimentarios asociados a corrientes de fondo en el moldeado de los fondos marinos. Este modelo sugiere que la circulación de fondo de las masas de agua condiciona la fisiografía, que la posici6n de las interfaces de las masas agua con un importante contraste en sus densidades, es capaz de esculpir terrazas a escala regional, y que los altos morfológicos desempeñan un papel esencial en el control local de procesos y en la distribución de las masas de agua. El análisis de la estratigrafía sísmica de las secuencias Pliocena y Cuaternaria ha permitido actualizar y renombrar los límites estratigráficos del Mar de Alborán, tras la reubicación además de la base del Cuaternario de 1,8 a 2.6 Ma, así como establecer una aproximación cronológica a los mismos. Asimismo, este análisis sísmico ha permitido presentar y discutir evidencias de depósitos contorníticos a escala del Mar de Alborán. Los rasgos contorníticos de tipo deposicional (crestas adosadas, laminares, elongadas separadas y confinadas) y erosivo (terrazas, escarpes, fosas, canales y surcos) se desarrollaron bajo la influencia continua de las masas de agua mediterráneas tras la apertura del Estrecho de Gibraltar (-5.33 Ma). Al menos dos factores principales han controlado los rasgos contorníticos en esta cuenca: i) la tectónica, que rige la reubicación de los principales flujos mediterráneos y por tanto sus patrones de circulaci6n; y ii) el clima, que ha influido en las condiciones de las masas de agua (profundidad y contraste de densidad en las interfaces) y en las áreas fuentes de sedimento en tierra, condicionando la expresión morfo-sísmica y los patrones de crecimiento de los depósitos contorníticos así como la formación de terrazas (dimensiones). La distribución de los elementos contorníticos en el espacio y el tiempo, permite proponer tres escenarios principales para explicar la circulación de las masas de agua desde la apertura del Estrecho de Gibraltar: i) la inundación atlántica en el Zancliense; ii) el mar Plioceno, con dos etapas diferentes para la circulación profunda y que en general se caracteriza por la presencia de interfaces poco definidas e inestables entre las aguas atlánticas (AW), y las aguas mediterráneas ligeras y densas, así como por la presencia de una fuerte contracorriente en la cuenca occidental de Alborán; por último, iii) el mar Cuaternario, que se caracteriza por masas de agua mediterráneas con flujo mayormente tabular pero con múltiples dinámicas de flujo a nivel local, una creciente influencia de los contrastes de densidad, y grandes cambios climáticos que provocan desplazamientos verticales y horizontales de las interfaces. Estas tres grandes etapas reflejan la variabilidad en los regímenes de corrientes de fondo, y la eficiencia en el transporte de sedimento, sedimentación y erosión a lo largo del margen. El estudio sísmico detallado de las unidades que componen las secuencias Pliocena y Cuaternaria ha permitido por primera vez el análisis y la cartografía a lo largo del tiempo de los rasgos contorníticos, los sistemas turbidfticos y los depósitos de inestabilidad sedimentaria. Estos mapas han permitido comprender la arquitectura sedimentaria de los margenes continentales espanol y marroquí y de las cuencas, así como definir los procesos paleoceanograficos desde un enfoque geológico. Se han definido dos grandes sistemas deposicionales contorníticos: el Sistema Deposicional Contornítico Intermedio (SDCI), formado bajo la acci6n de las aguas mediterráneas de menor densidad en el margen español, y el Sistema Deposicional Contornítico Profundo (SDCP), formado bajo la acción de las aguas mediterráneas densas principalmente en el margen marroquf y en las cuencas. La caracterización de las terrazas como rasgos contorníticos formados bajo la acción combinada de dos masas de agua ha llevado tambien a definir el Sistema Deposicional Contornítico Atlántico (SDCA). El desarrollo de varios sistemas deposicionales contorníticos ha dado lugar a la definición de un nuevo termino no planteado previamente en la literatura: el Sistema Deposicional Contornftico Multiple (SDCM), referido a un conjunto de diferentes SDC que se forman en una misma zona bajo la acción de múltiples masas de agua. Asimismo, se han caracterizado veinte sistemas turbidítiicos que son la causa principal de las diferencias en la arquitectura sedimentaria que presentan el margen español, en el que coexisten con las contornitas, y el margen marroquí, donde están menos desarrollados. Con respecto a los depósiitos de inestabilidad sedimentaria, su formación esta relacionada principalmente con el retrabajamiento de las contornitas que recubren los altos. Las contornitas principalmente, pero tambien las turbiditas, han permitido definir desde una perspectiva geológica los principales procesos oceanográficos así como determinar su ocurrencia, magnitud y energía relativas, y su tiempo de acción. En esta Tesis se explica tambien el desarrollo desigual que presentan los sistemas turbidíticos en el mar de Alborán, y que ha sido interpretado como resultado de la interacción entre los procesos sedimentarios paralelos (asociados a las corrientes de fondo) y perpendiculares (asociados a los procesos de flujos gravitativos) al margen. En los registros Plioceno y Cuaternario, asf como en el fondo marino actual del mar de Alborán, se han identificado rasgos de tipo morfológico, sedimentario y sedimentológico producidas por la interacción entre ambos procesos. Los escenarios de interacción varfan entre dos situaciones extremas: las corrientes de fondo dominan sobre los flujos gravitativos, y los flujos gravitativos dominan sobre la acción de las corrientes de fondo. Entre ambos extremos puede producirse la alternancia y la influencia mutua de ambos procesos. Se han propuesto dos modelos de interacción conceptuales para los margenes español y marroquí. i) En el margen espanol la interacción es especialmente compleja y variada, con efectos regionales y locales sobre los sistemas turbidíticos. Esto ocurre debido a que los sistemas turbidíticos están influenciados a diferentes profundidades por las masas de agua atlánticas y mediterráneas y sus interfaces, con una dinámica además que varía longitudinal y perpendicularmente al margen. ii) En el margen marroquí, la acción intensa de la WMDW inhibe en gran parte la formación de canones y depósitos asociados. Los resultados de esta Tesis sugieren que la importancia de los procesos relacionados con corrientes de fondo en ambientes marinos profundos debería ser reevaluada. Se concluye que una mayor comprensión de su influencia es esencial no sólo para reconstruir los patrones de circulación recientes y pasados, sino también para identificar determinados rasgos morfosedimentarios, descifrar sus patrones de sedimentación y evolución, así como establecer tendencias en el clima y variaciones eustáticas a nivel global.

Resumen del póster presentado al 60th Annual Meeting Biophysical Society, celebrado en Los Angeles, California (USA) del 27 de febrero al 2 de marzo de 2016., KCNQ1 and KCNE1 encode Kv7.1 and KCNE1, respectively, the poreforming and the accessory subunits of the slow delayed rectifier potassium current, IKs. KCNQ1 mutations are associated with long and short QT syndrome. The aim of this study was to characterize the biophysical and cellular pheno-type of a KCNQ1 missense mutation, F279I, found in a 23 years old man with a QTc of 356 ms and a family history of sudden cardiac death (SCD). Experiments were performed using perforated patch-clamp, western blot, coimmunoprecipitation, biotinylation and immunocytochemistry techniques in HEK293, COS7 cells and in cardiomyocytes transfected with WT Kv7.1/KCNE1 or F279I Kv7.1/KCNE1 channels. In the absence of KCNE1, F279I Kv7.1 current exhibited less degree of inactivation than WT Kv7.1. Also, functional analysis of F279I Kv7.1 in the presence of KCNE1 revealed a negative shift in the activation curve and an acceleration of the activation kinetics leading to a gain of function in IKs. The coassembly between F279I Kv7.1 channels and KCNE1 was markedly decreased compared with WT Kv7.1 channels, as revealed by coimmunoprecipitation and FRET experiments. All these effects contribute to the increase of IKs when channels incorporate F279I Kv7.1 subunits, as shown by a computer model simulation of these data that predicts a shortening of the action potential consistent with the patient phenotype. In conclusion, the F279I mutation induces a gain of function of IKs due to an impaired gating modulation of Kv7.1 induced by KCNE1; leading to a shortening of the cardiac action potential.

Resumen del póster presentado al Spanish-Italian Medicinal Chemistry Congress, celebrado en Barcelona (España) del 12 al 15 de julio de 2015., Altered neuronal calcium homeostasis and early compensatory changes in transcriptional programs are common features of many neurodegenerative pathologies including Alzheimer's disease, Down syndrome and Huntington's disease. DREAM (Downstream Regulatory Element Antagonist Modulator, also known as calsenilin or KChIP-3 (potassium channel interacting protein-3), is a multifunctional calcium binding protein that controls the expression level and/or the activity of several proteins related to calcium homeostasis, neuronal excitability and neuronal survival. This protein is widely expressed in the brain and, depending on the cell type and physiological conditions, shows multiple subcellular localizations, in the nucleus, cytosol or cell membrane. The interest in DREAM is based on its key role in the regulation of intracellular calcium levels. As a calcium-dependent transcriptional repressor, DREAM is a master regulator of activity-dependent gene expression and controls genes important for calcium homeostasis. As an auxiliary protein in the plasma membrane, DREAM interacts with and regulates the gating of Kv4 potassium channels, L- and T-type voltage-dependent calcium channels and NMDA receptors. These findings suggest that DREAM could be a novel and versatile target for therapeutic intervention in neurodegeneration and that molecules able to bind to DREAM and modulate its physiological functions could be candidates for drugs to treat neurodegenerative diseases. Moreover, up to now, only two DREAMbinding molecules have been identified. In this communication we report the rational design and the synthesis of novel DREAM-binding molecules and their effects on the modulation of DREAM/protein interactions., This work was supported by Ministerio de Economia y Competitividad (SAF2012-32209 and SAF2013-45800-R) CSIC (201280E096-PIE) and Instituto de Salud Carlos III/CIBERNED . P. Cercós and A. Prieto hold a FPI fellowship from the Spanish Ministerio de Ciencia e Innovación. T. Gonzalez holds a Ramón y Cajal contract. This work has been awarded with the Janssen Cilag Award for Young Researchers from the Sociedad Española de Química Terapéutica (SEQT).

[ES] Compuestos moduladores del sensor neuronal de calcio dream y sus usos terapéuticos. La presente invención se refiere a un grupo de compuestos con un núcleo estructural derivado de fenilacetamida con la siguiente fórmula (I): que presentan capacidad de modulación del sensor neuronal de calncio DREAM, por lo que la presente invención también se refiere al uso de estos compuestos para el tratamiento o prevención de trastornos o enfermedades donde los niveles de DREAM están por encima o por debajo de los niveles considerados fisiológicamente normales., [EN] The invention relates to a group of compounds with a structural nucleus derived from phenylacetamide, having formula (I), which compounds can modulate the DREAM neuronal calcium sensor. Consequently, the invention also relates to the use of these compounds for the treatment or prevention of disorders or diseases in which DREAM levels are above or below physiologically normal levels., Consejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED) y Universidad Autónoma de Madrid, A1 Solicitud de patente con informe sobre el estado de la técnica

Resumen del póster presentado a la 5ª Reunión de la Red Española de Canales Iónicos: "Present and future in ion channel research", celebrada en Barcelona (España) del 4 al 6 de octubre de 2015., DREAM, also named calsenilin or KChIP3, is a neuronal calcium sensor that regulates multiple genes after binding to DRE sites in the nucleus. Outside nucleus, DREAM regulates diverse cellular functions by binding to several proteins, one of them the Kv4.3 channel. Kv4.3 channels, after assembling with different modulatory subunits, generate a fast and transient outward potassium current. In the heart, Kv4.3 binds with KChIP2 subunits, generating the Ito1, one of the main repolarizing cardiac currents. In neurons, they associate to KChIP1 or KChIP3 and generate the IA, which is implicated in very specialized functions as learning, memory and behavior. Here, we describe the electrophysiological effects of new compounds designed to bind with high affinity to KChIP3, in Kv4.3 and Kv4.3+KChIP3 channels expressed in CHO cells. PC342 inhibited Kv4.3 and Kv4.3+KChIP3 to a similar extent, whereas PC332 and PC358 inhibited Kv4.3+KChIP3 to a greater extent. Block induced by PC332 was time- and voltage-dependent, consistent with binding to a closed-active state. Therefore, PC332 and PC358 may represent good compounds to develop new drugs that selectively modulate the neuronal electric activity in distinct neuronal populations.

Los estudios de casos han sido usados en una variedad de disciplinas en las ciencias sociales y salud, al tener cualidades para comprender en profundidad un fenómeno en variados contextos y situaciones naturales. Sin embargo, su uso ha sido confuso por las diferentes visiones de los investigadores. Este artículo tiene como propósito hacer una distinción de los estudios de casos: con perspectiva cualitativa y como un diseño de investigación. Los estudios de casos cualitativos se originan por la forma particular de ver el caso como un todo: su contexto y sus límites, con análisis intensivo del caso o casos colectivos, y siempre bajo la concepción de su idiosincrasia y sin generalización. El diseño de estudios de casos como parte de una estrategia investigativa busca dar respuesta a una pregunta de investigación que permite usar diferentes métodos para hacer constantes comparaciones múltiples. En síntesis, los estudios de casos son usados en ambas formas por los investigadores y tienen un potencial de utilidad en situaciones y contextos de enfermería y salud. Case studies have been used into social sciences and health disciplines because of their properties to understand complex phenomena in a variety of contexts and situations. However, its use has been confusing because of the different researcher's perspectives. This article aims to distinguish two types of two types of case studies approaches: the one with a qualitative perspective and as a design research strategy. The qualitative case study are originated by the particular way of seing the case as a whole: its contexts and limits, intensive case analysis or collective cases, and always under the conception of their idiosyncrasy without generalization. In a research design, case study is a research strategy that tries to answer a research question by applying different methods for data collection and analyzing by using constant comparison. Currently, both approaches of case studies are used by researchers, having a potential benefit for nursing and health settings and contexts.

The present descriptive research is based on the analysis of the curriculum proposal for “Sector de Lengua Indígena” of “Programa de Educación Intercultural Bilingüe (PEIB)”, which was submitted to public consultation by the Ministry of Education of the Chilean government (Mineduc) in 2012. From a communicative approach of language teaching, the objective is to describe “the communicative competence” presented in these pedagogical proposal of the Mineduc for Sector de Lengua Indígena. The results show that the learning objectives of PEIB proposal are mainly oriented to develop sociocultural competence, rather than towards the development of grammatical morphosintactic, discursive and strategic competence.Keywords: Indigenous Language, Intercultural Education, communicative competence. El presente trabajo es de carácter descriptivo y analiza la Propuesta de Bases Curriculares para el Sector Lengua Indígena del Programa de Educación Intercultural Bilingüe (PEIB), sometido a consulta ciudadana por el Ministerio de Educación del Gobierno de Chile (Mineduc) el año 2012. Desde un enfoque comunicativo de enseñanza de lenguas, el objetivo es describir la “competencia comunicativa” comprometida en la propuesta pedagógica del Mineduc para el sector de Lengua Indígena. Los resultados muestran que los Objetivos de Aprendizaje en la Propuesta del PEIB se orientan principalmente hacia el desarrollo de la competencia sociocultural y, en menor medida, hacia el desarrollo de la competencia gramatical morfosintáctica, discursiva y estratégica.Palabras clave: Lengua Indígena, Educación Intercultural, competencia comunicativa. El presente trabajo es de carácter descriptivo y analiza la Propuesta de Bases Curriculares para el Sector Lengua Indígena del Programa de Educación Intercultural Bilingüe (PEIB), sometido a consulta ciudadana por el Ministerio de Educación del Gobierno de Chile (Mineduc) el año 2012. Desde un enfoque comunicativo de enseñanza de lenguas, el objetivo es describir la “competencia comunicativa” comprometida en la propuesta pedagógica del Mineduc para el sector de Lengua Indígena. Los resultados muestran que los Objetivos de Aprendizaje en la Propuesta del PEIB se orientan principalmente hacia el desarrollo de la competencia sociocultural y, en menor medida, hacia el desarrollo de la competencia gramatical morfosintáctica, discursiva y estratégica.Palabras clave: Lengua Indígena, Educación Intercultural, competencia comunicativa.

Resumen del trabajo presentado al 37th Congress of the Spanish Society of Physiological Sciences (SECF), celebrado del 24 al 26 de Septiembre de 2014 en Granada (España):-- et al., [Introduction]: Voltage-dependent K+ channels from Kv7 (KCNQ) family have well-established physiological roles in cardiovascular and nervous system, although functions in blood vessels remain unclear. Evidence suggests that Kv7.1, Kv7.4 and Kv7.5 control vascular reactivity. However, because controversial pharmacological results Kv7.1 is under intense investigation. In this scenario, the ability of Kv7 channels to form heterotetramers is of physiological relevance. Thus, Kv7.4/Kv7.5 heterotetramers paves the way for novel interaction that could shed light to controversial pharmacological results. We aim whether Kv7.1 and Kv7.5 form heterotetramers increasing the diversity of channel responses in vascular smooth muscle. [Methods and Results]: We demonstrated Kv7.1/Kv7.5 structures in heterologous system by many different approaches, such as electrophysiology, coimmunoprecipitation and FRET. Heteromeric channels are retained at the endoplasmatic reticulum and, unlike Kv7.1 channels, heteromers localize out of lipid rafts. These results are supported by experiments in isolated smooth muscle myocytes. Kv7.1 and Kv7.5 are expressed in aorta, cava and coronary myocytes. Electrophysiological and miography recordings using linopiridine, chromanol 293B and retigabine support Kv7.1/Kv7.5 heterotetramers that co-immunoprecipitation experiments further confirmed. Finally, lipid raft isolation from different tissues corroborated that expression of Kv7.5 releases Kv7.1/Kv7.5 channels out of raft structures. [Discussion]: Kv7.1 and Kv7.5 are differentially expressed in blood vessels where they contribute to control vascular reactivity. We prove that they do heterotetramerize increasing the diversity of their physiological response. These data may help to better understand the scenario of Kv7 channels and vascular physiology., Supported by BFU2011-23268 and CSD2008-00005 to AF (MINECO, Spain)

La presente invención se refiere a un grupo de compuestos con un núcleo estructural derivado de fenilacetamida con la siguiente fórmula (I): que presentan capacidad de modulación del sensor neuronal de calcio DREAM, por lo que la presente invención también se refiere al uso de estos compuestos para el tratamiento o prevención de trastornos o enfermedades donde los niveles de DREAM están por encima o por debajo de los niveles considerados fisiológicamente normales. [ES], The present invention relates to a group of compounds with a structural nucleus derived from phenylacetamide, having the following formula (I): that can modulate the DREAM neuronal calcium sensor. Consequently, the present invention also relates to the use of these compounds for the treatment or prevention of disorders or diseases in which DREAM levels are above or below physiologically normal levels. [EN], Consejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas, Universidad Autónoma de Madrid, T3 Traducción de patente europea

[ES] Compuestos moduladores del sensor neuronal de calcio dream y sus usos terapéuticos. La presente invención se refiere a un grupo de compuestos con un núcleo estructural derivado de fenilacetamida con la siguiente fórmula (I): que presentan capacidad de modulación del sensor neuronal de calncio DREAM, por lo que la presente invención también se refiere al uso de estos compuestos para el tratamiento o prevención de trastornos o enfermedades donde los niveles de DREAM están por encima o por debajo de los niveles considerados fisiológicamente normales., [EN] The invention relates to a group of compounds with a structural nucleus derived from phenylacetamide, having formula (I), which compounds can modulate the DREAM neuronal calcium sensor. Consequently, the invention also relates to the use of these compounds for the treatment or prevention of disorders or diseases in which DREAM levels are above or below physiologically normal levels., Consejo Superior de Investigaciones Científicas (España), Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED) y Universidad Autónoma de Madrid, A1 Solicitud de patente con informe sobre el estado de la técnica

[Backgroun adn Purpose]: The Kv ß1.3 subunit modifies the gating and pharmacology of Kv 1.5 channels in a PKC-dependent manner, decreasing channel sensitivity to bupivacaine- and quinidine-mediated blockade. Cardiac Kv 1.5 channels associate with receptor for activated C kinase 1 (RACK1), the Kv ß1.3 subunit and different PKC isoforms, resulting in the formation of a functional channelosome. The aim of the present study was to investigate the effects of PKC inhibition on bupivacaine and quinidine block of Kv 1.5 + Kv ß1.3 channels. [Experimental Approach]: HEK293 cells were transfected with Kv 1.5 + Kv ß1.3 channels, and currents were recorded using the whole-cell configuration of the patch-clamp technique. PKC inhibition was achieved by incubating the cells with either calphostin C or bisindolylmaleimide II and the effects of bupivacaine and quinidine were analysed. [Key Results]: The voltage-dependent inactivation of Kv 1.5 + Kv ß1.3 channels and their pharmacological behaviour after PKC inhibition with calphostin C were similar to those displayed by Kv 1.5 channels alone. Indeed, the IC50 values for bupivacaine were similar in cells whose PKC was inhibited with calphostin C or bisindolylmaleimide II. Similar results were also observed in the presence of quinidine. [Conclusions and Implications]: The finding that the voltage-dependence of inactivation and the pharmacology of Kv 1.5 + Kv ß1.3 channels after PKC inhibition resembled that observed in Kv 1.5 channels suggests that both processes are dependent on PKC-mediated phosphorylation. These results may have clinical relevance in diseases that are characterized by alterations in kinase activity., This study was supported by MINECO (SAF2010-14916 and SAF2013-45800-R), FIS-RECAVA (RD06/0014/0006) and FISRIC (RD12/0042/0019) grants to C. V. A. M. and A. P. hold JAE-Predoc and FPI fellowships respectively. A. dlC. and T. G. hold RIC and R&C contracts respectively

The post-Messinian tectonic activity has an important role on the physiographic configuration of the Alboran Sea basin. The compressive regime between the Eurasian and Nubian plates has produced the progressive narrowing of this basin and the formation sub-basins and highs while the surrounding Betic-Rif Mountains uplift. The major submarine reliefs include the Alboran Ridge, a 1000 m high NE-SW structural and volcanic high with steep slopes facing northwards and southwards and the Cape Tres Forcas promontory in the Moroccan margin, which are the northern and southern borders of the South Alboran Basin respectively. The bathymetric and high resolution study of South Alboran Basin highlights the presence of several mass movement deposits (MMDs) ranging from sliding to well-defined slope apron characterized by channelized and lobulated deposits coming from the basin margins and resting on the basin interior. Seismic stratigraphy study of this basin shows it has been filled partially since the Pliocene. These deposits show a major recurrence since the Late Pleistocene (0.92 M.y.), with a successive stack of at least five MMDs implying a recurrence interval o about 0.18 M.y. during this period. Many of them proceed from the Alboran Ridge, altough the largest one, the just discovered Montera Slide, comes from the Cape Tres Forcas promontory. This MMD is 50 ms average thick (maximum 180 ms), and covers an area around 90 km². The successive deposition of MMDs since the Pliocene in the SAB would be triggered by the interplay between the tectonic activity of the Alboran Ridge and the promontory of Cape Tres Forcas. In this sense the major number of MMDs appoints to a reactivation of tectonics since the Late Pleistocene.

Resumen del póster presentado al III SEQT Summer School: "Medicinal Chemistry in Drug Discovery: The Pharma Perspective", celebrado en Tres Cantos (Madrid) del 25 al 27 de junio de 2013., Altered neuronal calcium homeostasis and early compensatory changes in transcriptional programs are common features of many neurodegenerative pathologies including Alzheimer’s disease (AD), Down syndrome (DS) and Huntington’s disease (HD). DREAM (Downstream Regulatory Element Antagonist Modulator), also known as calsenilin or KChIP-3 (potasium channel interacting protein-3), is a multifunctional Ca2+ binding protein that controls the expression level and/or the activity of several proteins related to Ca2+ homeostasis, neuronal excitability and neuronal survival. This protein is widely expressed in the brain and, depending on the cell type and physiological conditions, shows multiple subcellular localizations, in the nucleus, cytosol or cell membrane. Initially, the interest in DREAM was based on its key role in the regulation of intracelular Ca2+ levels. An early reduction in DREAM levels is found in the pre-symptomatic phase of several neurodegenerative mouse models, including AD, DS and HD. These data support the idea that an early down regulation of the DREAM level in neurons during the pre-symptomatic phase of the AD, DS and HD might be part of its neuroprotective mechanism. These findings suggest that DREAM could be a novel and versatile target for therapeutic intervention in neurodegeneration and that molecules able to bind to DREAM and block its physiological functions could be candidates of drugs to treat neurodegenerative diseases. Up to know, low molecular weight molecules have not been described able to interact with DREAM and to modulate its action. In this communication we report the rational design, the synthesis and the biological evaluation of novel DREAM-binding molecules.

0

K(v)1.5 channels are the primary channels contributing to the ultrarapid outward potassium current (I(Kur)). The regulatory K(v)β1.3 subunit converts K(v)1.5 channels from delayed rectifiers with a modest degree of slow inactivation to channels with both fast and slow inactivation components. Previous studies have shown that inhibition of PKC with calphostin C abolishes the fast inactivation induced by K(v)β1.3. In this study, we investigated the mechanisms underlying this phenomenon using electrophysiological, biochemical, and confocal microscopy approaches. To achieve this, we used HEK293 cells (which lack K(v)β subunits) transiently cotransfected with K(v)1.5+K(v)β1.3 and also rat ventricular and atrial tissue to study native α-β subunit interactions. Immunocytochemistry assays demonstrated that these channel subunits colocalize in control conditions and after calphostin C treatment. Moreover, coimmunoprecipitation studies showed that K(v)1.5 and K(v)β1.3 remain associated after PKC inhibition. After knocking down all PKC isoforms by siRNA or inhibiting PKC with calphostin C, K(v)β1.3-induced fast inactivation at +60 mV was abolished. However, depolarization to +100 mV revealed K(v)β1.3-induced inactivation, indicating that PKC inhibition causes a dramatic positive shift of the inactivation curve. Our results demonstrate that calphostin C-mediated abolishment of fast inactivation is not due to the dissociation of K(v)1.5 and K(v)β1.3. Finally, immunoprecipitation and immunocytochemistry experiments revealed an association between K(v)1.5, K(v)β1.3, the receptor for activated C kinase (RACK1), PKCβI, PKCβII, and PKCθ in HEK293 cells. A very similar K(v)1.5 channelosome was found in rat ventricular tissue but not in atrial tissue.

Local anesthetics are useful probes of ion channel function and structure. Stereoselective interactions are especially interesting because they can reveal three-dimensional relationships between drugs and channels with otherwise identical biophysical and physicochemical properties. Furthermore, stereoselectivity suggests direct and specific receptor-mediated action, and identification of such stereospecific interactions may have important clinical consequences. The fact that drug targets are able to discriminate between the enantiomers present in a racemic drug is the consequence of the ordered asymmetric macromolecular units that form living cells. However, almost 25% of the drugs used in the clinical practice are racemic mixtures, and their individual enantiomers frequently differ in both their pharmacodynamic and pharmacokinetic profiles. Moreover, their effects can be similar to or different from the pharmacological effect of the drug and may contribute to the undesired effects of the drug. In other cases, the pharmacological effects induced by the two enantiomers on the molecular target are opposite. In the present manuscript, we will review the stereoselective effects of bupivacaine-like local anesthetics on cardiac sodium and potassium channels., This work has been funded by CICYT SAF2010-14916 and Red Cooperativa de Enfermedades Cardiovasculares RECAVA FIS RD06/0014/0006 grants to CV. CM and AP held an FPI grant. AC holds a UAM grant. AM and TG held a JAE-Predoc and a JAE-Doc grants, respectively.

Resumen del póster presentado al Biophysical Society 56th Annual Meeting, celebrado en San Diego-California (US) del 25 al 29 de febrero de 2012., Irvalec© (PM02734, Elisidepsin) is a marine-derived cytotoxic depsipeptide that is currently undergoing phase II clinical studies in non-small cell lung cancer. In vitro treatment of tumor cells with Irvalec© induces necrotic cell death, a process associated with rapid loss of membrane integrity and subsequent cell permeabilization. These results suggested an effect of the drug, either on an ion channel or in the plasma membrane. In order to explore this hypothesis patch-clamp experiments were performed in different tumor cell lines (HeLa, A549 and HCT116). Treated cells underwent rapid and dramatic morphological changes, including cell blebbing, severe swelling, plasma membrane permeabilization and cell lysis. Apart from the numerous small blebs, membranes from damaged cells also re-organized to form enormous bubbles surrounded by cell membrane. Using electrophysiological techniques, it was shown that Irvalec© induced an important increase in membrane conductance. The compound permeabilized the plasma membrane to ions, even when the cells were not pulsed, causing important changes in the holding current. It has been described that zinc attenuates the drastic effects of some membrane disrupting agents. Hence, to test if zinc exerted some protective effect against Irvalec© effects, cells were treated with this drug in the presence or absence of zinc salts and its membrane permeability was analyzed by using electrophysiology techniques, measuring the variations in the ion currents induced by the drug. Interestingly, in cells treated with zinc (10 mM), a decrease in the membrane permeability induced by Irvalec© was observed. Altogether, these results suggest that Irvalec© induces a rapid membrane permeabilization that lead to a necrotic cell death., Supported by CICYT SAF2010-14916 and FIS (RD06/0014/0006) Grants.

Kv1.5 channels are the primary channels contributing to the ultrarapid outward potassium current (IKur). The regulatory Kvβ1.3 subunit converts Kv1.5 channels from delayed rectifiers with a modest degree of slow inactivation to channels with both fast and slow inactivation components. Previous studies have shown that inhibition of PKC with calphostin C abolishes the fast inactivation induced by Kvβ1.3. In this study, we investigated the mechanisms underlying this phenomenon using electrophysiological, biochemical, and confocal microscopy approaches. To achieve this, we used HEK293 cells (which lack Kvβ subunits) transiently cotransfected with Kv1.5+Kvβ1.3 and also rat ventricular and atrial tissue to study native α-β subunit interactions. Immunocytochemistry assays demonstrated that these channel subunits colocalize in control conditions and after calphostin C treatment. Moreover, coimmunoprecipitation studies showed that Kv1.5 and Kvβ1.3 remain associated after PKC inhibition. After knocking down all PKC isoforms by siRNA or inhibiting PKC with calphostin C, Kvβ1.3-induced fast inactivation at +60 mV was abolished. However, depolarization to +100 mV revealed Kvβ1.3-induced inactivation, indicating that PKC inhibition causes a dramatic positive shift of the inactivation curve. Our results demonstrate that calphostin C-mediated abolishment of fast inactivation is not due to the dissociation of Kv1.5 and Kvβ1.3. Finally, immunoprecipitation and immunocytochemistry experiments revealed an association between Kv1.5, Kvβ1.3, the receptor for activated C kinase (RACK1), PKCβI, PKCβII, and PKCθ in HEK293 cells. A very similar Kv1.5 channelosome was found in rat ventricular tissue but not in atrial tissue., This work was supported by Ministerio de Ciencia e Innovación (MICINN) Grants SAF2010-14916, Ministerio de Educación y Ciencia-PR2003-0056, and Red Cooperativa de Enfermedades Cardiovasculares RECAVA FIS RD06/0014/0006 (to C. V.) and BFU2011-23268 and CSD2008-00005 (to A. F.). C. V. recipient of Grant MEC-PR2003-0056.

The polycyclic aromatic hydrocarbons (PAHs) dibenzo(a,h)anthracene, benzo(ghi)perylene, benzo(b)fluoranthene and benzo(a)pyrene have been identified in urban air from Mexico City and some of them are classified as human carcinogens. In the present study, human peripheral blood lymphocytes were exposed in vitro to different concentrations of PAHs with (+S9) or without (-S9) metabolic activation. The genotoxic and cytotoxic effects of each PAH were examined with an alkaline comet assay and trypan blue dye exclusion, and oxidative DNA damage was determined via the detection of 8-hydroxy-2'-deoxyguanosine (8-OhdG) adduct levels by enzyme-linked immunosorbent assay (ELISA). The DNA damage was evaluated with two genotoxicity parameters: the frequency of comets and the comet tail length. Concentrations of 20, 40, 80, 160 and 320 µM DB(a,h)A-S9; 20, 40, 80, 160 and 240 µM B(ghi)P-S9; 20, 30, 40, 60 and 80 µM B(b)F-S9; and 80 µM B(a)P-S9 for 24 h induced a small but significant increase in the means of comet frequency, in the tail length and in the 8-oHDg levels in relation to the control (0.5% DMSO-S9). However, all PAHs+S9 produced a more significant increase in DNA strand breaks and the level of 8-OHdG compared with the control (0.5% DMSO+S9), with a concentration-effect relationship. The viability of lymphocytes exposed to all PAHs-S9 and PAHs+S9 was not modified compared with the control. The results of this study demonstrate that the comet and ELISA are rapid, suitable and sensitive methods to detect in vitro PAH-induced DNA damage in human peripheral lymphocytes.

Kv1.5 channels are homotetramers of α-pore subunits mainly present in human atrium and pulmonary vasculature. Thus, Kv1.5 is a pharmacological target for cardiovascular diseases. Kvβ1.3 assemblies with Kvα1.5 and modifies its gating and pharmacology. A further knowledge of α-β interactions and pharmacology will lead a better design of new drugs.

Celecoxib is a COX-2 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets. The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels (Kv) involved on cardiac repolarization Kv1.5 (IKur), Kv4.3+KChIP2 (Ito1) and Kv7.1+KCNE1 (IKs) and to compare with another COX-2 inhibitor, rofecoxib. Currents were recorded in transfected mammalian cells by whole-cell patch-clamp. Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent, except on Kv4.3+KChIP2 channels. Kv1.5 block increased in the voltage range of channel activation, decreasing at potentials positive to 0mV. The drug modified the activation curve of the channels that became biphasic. Block was frequency-dependent, increasing at fastest frequencies. Celecoxib effects were not altered by TEAout in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with TEAin, indicating that celecoxib acts from the cytosolic side. We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells, where Kv1.5 are the main contributors (IC50≈7μM). Finally, we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes, suggesting that Kv block induced by celecoxib may be of clinical relevance. © 2010 Elsevier Ltd., This study was supported by Ministerio de Ciencia e Innovación (SAF2007-65868, SAF2008-03948, SAF2010-14916 and AGL2007-66108); Instituto de Salud Carlos III (RECAVA RD06/0014/0006); and Fundación Mutua Madrileña. T.G. holds a JAE-Doc contract from CSIC. A.M. is a JAE-Predoc fellow from CSIC. C.M. is an FPI fellow and J.M-S. is an FPU fellow from Ministerio de Ciencia e Innovación. M.D. holds a contract from RECAVA.

1. The goal of this study was to analyse the effects of propafenone and its major metabolite, 5-hydroxy-propafenone, on a human cardiac K+ channel (hKv1.5) stably expressed in Ltk- cells and using the whole-cell configuration of the patch-clamp technique. 2. Propafenone and 5-hydroxy-propafenone inhibited in a concentration-dependent manner the hKv1.5 current with K(D) values of 4.4+/-0.3 microM and 9.2+/-1.6 microM, respectively. 3. Block induced by both drugs was voltage-dependent consistent with a value of electrical distance (referenced to the cytoplasmic side) of 0.17+/-0.55 (n=10) and 0.16+/-0.81 (n=16). 4. The apparent association (k) and dissociation (l) rate constants for propafenone were (8.9+/-0.9) x 10(6) M(-1) s(-1) and 39.5+/-4.2 s(-1), respectively. For 5-hydroxy-propafenone these values averaged (2.3+/-0.3) x 10(6) M(-1) s(-1) and 21.4+/-3.1 s(-1), respectively. 5. Both drugs reduced the tail current amplitude recorded at -40 mV after 250 ms depolarizing pulses to +60 mV, and slowed the deactivation time course resulting in a 'crossover' phenomenon when the tail currents recorded under control conditions and in the presence of each drug were superimposed. 6. Both compounds induced a small but statistically significant use-dependent block when trains of depolarizations at frequencies between 0.5 and 3 Hz were applied. 7. These results indicate that propafenone and its metabolite block hKv1.5 channels in a concentration-, voltage-, time- and use-dependent manner and the concentrations needed to observe these effects are in the therapeutical range.

Capítulo 38 de la 3ª edición., Los fármacos antiarrítmicos forman un grupo muy heterogéneo de sustancias que se caracterizan por suprimir o prevenir las alteraciones del ritmo cardíaco a concentraciones a las que no ejercen efectos adversos sobre el latido sinusal normalmente propagado. En la actualidad, continúan siendo el tratamiento de elección en la mayoría de los pacientes con arritmias, aunque diversas estrategias eléctricas (desfibriladores, marcapasos y técnicas de ablación) y quirúrgicas pueden reemplazarlos en determinados grupos de pacientes. Las alteraciones del ritmo cardíaco son el resultado de anomalías en: a) la génesis del impulso cardíaco (alteraciones del automatismo), y b) la secuencia de activación del miocardio (alteraciones de la conducción o reentrada). Estas anomalías del automatismo o de la conducción del impulso cardíaco pueden ser desencadenadas bien por cambios en los mecanismos iónicos responsables de la génesis o el mantenimiento de los potenciales de acción cardíacos; bien por alteraciones de tipo anatómico-funcional (p. ej., cardiopatía isquémica, hipertrofia ventricular o fibrosis). En este capítulo se explicarán los conceptos básicos de la electrofisiología cardíaca del miocardio sano y enfermo, los mecanismos desencadenantes de las arritmias cardíacas y, por último, la farmacología de los agentes antiarrítmicos utilizados actualmente en la prevención y supresión de las arritmias cardíacas.