A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake

Producción CientíficaMitochondrial Ca2+ homeostasis is crucial for regulating vital functions such as respiration or apoptosis. Targeted aequorins are excellent probes to measure subcellular Ca2+. Ca2+ concentration in mitochondria ([Ca2+]M) is low at rest (about 10−7 M) and can increase to the micromolar or even approach the millimolar range, upon cell activation. Here we describe a new quantitative luminescent protocol to directly measure mitochondrial Ca2+ uptake, optimized for high throughput. The sensitivity of the method allows detection of changes in either the capacity or the affinity of mitochondrial Ca2+ transport.Ministerio de Economía, Industria y Competitividad (Project BFU2014-534698P)Instituto de Salud Carlos III (RD16/0011/0003

Using aequorin probes to measure Ca2+ in intracellular organelles

Producción CientíficaAequorins are excellent tools for measuring intra-organellar Ca2+ and assessing its role in physiological and pathological functions. Here we review targeting strategies to express aequorins in various organelles. We address critical topics such as probe affinity tuning as well as normalization and calibration of the signal. We also focus on bioluminescent Ca2+ imaging in nucleus or mitochondria of living cells. Finally, recent advances with a new chimeric GFP-aequorin protein (GAP), which can be used either as luminescent or fluorescent Ca2+ probe, are presented. GAP is robustly expressed in transgenic flies and mice, where it has proven to be a suitable Ca2+ indicator for monitoring physiological Ca2+ signaling ex vivo and in vivo.Ministerio de Economía, Industria y Competitividad (Project BFU2014-53469P)Instituto de Salud Carlos III (TerCel, RD16/0011/0003

Measuring Ca2 + inside intracellular organelles with luminescent and fluorescent aequorin-based sensors

Producción CientíficaGFP-Aequorin Protein (GAP) can be used to measure [Ca2 +] inside intracellular organelles, both by luminescence and by fluorescence. The low-affinity variant GAP3 is adequate for ratiometric imaging in the endoplasmic reticulum and Golgi apparatus, and it can be combined with conventional synthetic indicators for simultaneous measurements of cytosolic Ca2 +. GAP is bioorthogonal as it does not have mammalian homologues, and it is robust and functionally expressed in transgenic flies and mice, where it can be used for Ca2 + measurements ex vivo and in vivo to explore animal models of health and disease. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.Ministerio de Economía, Industria y Competitividad (Project BFU2014-53469P)Instituto de Salud Carlos III (TerCel; RD16/0011/0003

GAP, an aequorin-based fluorescent indicator for imaging Ca2+ in organelles

Producción CientíficaGenetically encoded calcium indicators allow monitoring subcellular Ca2+ signals inside organelles. Most genetically encoded calcium indicators are fusions of endogenous calcium-binding proteins whose functionality in vivo may be perturbed by competition with cellular partners.We describe here a novel family of fluorescent Ca2+ sensors based on the fusion of two Aequorea victoria proteins, GFP and apo-aequorin (GAP). GAP exhibited a unique combination of features: dual-excitation ratiometric imaging, high dynamic range, good signal-to-noise ratio, insensitivity to pH and Mg2+, tunable Ca2+ affinity, uncomplicated calibration, and targetability to five distinct organelles. Moreover, transgenic mice for endoplasmic reticulum-targeted GAP exhibited a robust long-term expression that correlated well with its reproducible performance in various neural tissues. This biosensor fills a gap in the actual repertoire of Ca2+ indicators for organelles and becomes a valuable tool for in vivo Ca2+ imaging applications

Nuevas formas de interacción social: comunicación virtual y salud

It is examined if the relationship empirically demonstrated between health and face-to-face social interactions is confirmed when it is virtual. The objective is to know if the perception of psychological well-being and receiving social support would explain the frequency of participation in virtual social networks (VSN). An ex-post-facto design was applied to a sample of 510 university Internet users (age, M = 22.89, SD = 5.67, 78.9% women). The level of psychological well-being (Spanish adaptation of the Ryff Psychological Well-Being Scale), social support (Family Social Support and Friends questionnaire - AFA) and the frequency of use of VSN are evaluated. To determine the relationships between the variables, correlation and hierarchical regression analyzes were carried out. The relative contribution of the perception of psychological well-being and of obtaining social support in the frequency of VSN use is confirmed, controlling the influences of age and gender. Specifically, the perception of having less ability to dominate the social environment and to obtain social support for friendships through VSN explains and allows predicting the increase in the frequency of use of such networks.Se examina si la relación demostrada empíricamente entre salud e interacciones sociales presenciales se confirma cuando ésta es virtual. El objetivo es conocer si la percepción de bienestar psicológico y de recibir apoyo social explicaría la frecuencia de participación en redes sociales virtuales (RSV). Se aplica un diseño ex-post-facto sobre una muestra de 510 internautas universitarios (edad; M =22,89; DT= 5,67; 78,9% mujeres). Se evalúa el nivel de bienestar psicológico (adaptación española de la Escala de Bienestar Psicológico de Ryff), de apoyo social (cuestionario Apoyo Social Familiar y Amigos - AFA) y la frecuencia de uso de las RSV. Para determinar las relaciones entre las variables se realizaron análisis de correlación y de regresión jerárquica Se confirma la contribución relativa de la percepción de bienestar psicológico y de obtención de apoyo social en la frecuencia de uso de RSV, controlando las influencias de la edad y el género. Concretamente, la percepción de poseer menor capacidad de dominar el ambiente social y la de obtener apoyo social por las amistades a través de las RSV, explica y permite predecir el incremento en la frecuencia de uso de dichas redes

Psychological factors and wellbeing in breast cancer patients compared to healthy women

Breast cancer is the most common cancer in women worldwide, with nearly 1.7 million new cases diagnosed every year. This represents about 12% of all new cancer cases and 25% of all cancers in women (World Cancer Research Fund International, 2015). Also, it is estimated an increase of between 22.000-25.000 new cases per year. Even more, in spite of being considered a multi-causal disease, we must consider that between the 70% and the 80% breast cancer occurs in women without a likely risk factor

Ejercicio físico y aspectos psicosociales y emocionales en mujeres con cáncer de mama

Breast cancer is one of the most common types of cancer in Spain. Most studies in psychooncology focus on the approach of commorbid psychopathology, such as anxiety and depression. Several authors have reported the relationship between variables such as dispositional optimism and resilience, regarding psychological and subjective well-being in these patients. The practice of physical exercise seems to improve certain variables related to the mood, being able to be relevant in the achievement of an adequate emotional health. On the basis of these evidences, we propose to explore if the women that are in adjuvant treatment for breast cancer perform less exercise than other women without this pathology, as well as the existence of possible natural clusters that allow us to differentiate, if within these women, we found different typologies in relation to their perceived psychological quality of life levels (anxiety, depression, satisfaction with life, dispositional optimism and resilience). For this, the semi-structured interview of their own elaboration was administered, in addition to the sociodemographic data, if they performed physical exercise and frequency with which they performed it, as well as the questionnaires used to evaluate state / trait anxiety (STAI E / R), Depression (BDI), satisfaction with life (SWL), dispositional optimism (LOT-R), and resilience to a sample of 60 women, 30 of whom were being treated by the Oncology Unit of the Hospital de Jerez de la Border, For being treated for breast cancer. The results indicate that women with breast cancer who perform physical exercise, present lower levels of anxiety and depression and higher satisfaction with life, dispositional optimism and resilience.El cáncer de mama es una de las tipologías de cáncer más frecuente en España. La mayoría de los estudios en Psicooncología se centran en el abordaje de la psicopatología commorbida, como la ansiedad y la depresión. Varios autores han informado de relación entre variables como el optimismo disposicional y la resiliencia, respecto al bienestar psicológico y subjetivo en estas pacientes. La práctica del ejercicio físico parece mejorar ciertas variables relacionadas con el estado de ánimo, pudiendo ser relevante en el logro de una adecuada salud emocional. Sobre la base de estas evidencias, nos proponemos explorar si la mujeres en tratamiento adyuvante por cáncer de mama realizan menos ejercicio que otras mujeres sin esta patología, así como la existencia de posibles agrupaciones naturales o conglomerados que nos permitan diferenciar, si dentro de estas mujeres, encontramos diversas tipologías en relación a sus niveles psicológicos percibidos de calidad de vida (ansiedad, depresión , satisfacción con la vida, optimismo disposicional y resiliencia). Para ello, se administró una entrevista semiestructurada de elaboración propia, donde se recogían, además de los datos sociodemográficos, si realizaban ejercicio físico y frecuencia con que lo realizaban, así como los cuestionarios empleados para evaluar la ansiedad estado/rasgo (STAI E/R), depresión (BDI), satisfacción con la vida (SWL), optimismos disposicional (LOT-R) y resiliencia, a una muestra de 60 mujeres, 30 de las cuales estaban siendo atendidas por la Unidad de Oncología del Hospital de Jerez de la Frontera, por estar recibiendo tratamiento para el cáncer de mama. Los resultados indican que las mujeres con cáncer de mama que realizan ejercicio físico, presentan menores niveles de ansiedad y depresión y mayores de satisfacción con la vida, optimismo disposicional y resiliencia

Procedimiento de detección molecular de bacterias ácido lácticas productoras de β-glucanos.

Esta invención presenta un nuevo método de detección e identificación rápida de bacterias ácido lácticas productoras de exopolisacáridos por amplificación de su gen gtf codificante de una glicosil transferasa. La utilización de este método podría permitir la prevención del ahilamiento de bebidas alcohólicas y el aislamiento de nuevas estirpes que podrían ser utilizadas para la producción de alimentos fermentados.Solicitud: 200402176 (11.09.2004)Nº Pub. de Solicitud: ES2315038A1 (16.03.2009)Nº de Patente: ES2315038B1 (29.12.2009

Neuronal and astrocytic tetraploidy is increased in drug-resistant epilepsy

Aims: Epilepsy is one of the most prevalent neurological diseases. A third of patients with epilepsy remain drug-resistant. The exact aetiology of drug-resistant epilepsy (DRE) is still unknown. Neuronal tetraploidy has been associated with neuropathology. The aim of this study was to assess the presence of tetraploid neurons and astrocytes in DRE. Methods. For that purpose, cortex, hippocampus and amygdala samples were obtained from patients subjected to surgical resection of the epileptogenic zone. Post-mortem brain tissue of subjects without previous records of neurological, neurodegenerative or psychiatric diseases was used as control. Results: The percentage of tetraploid cells was measured by immunostaining of neurons (NeuN) or astrocytes (S100β) followed by flow cytometry analysis. The results were confirmed by image cytometry (ImageStream X Amnis System Cytometer) and with an alternative astrocyte biomarker (NDRG2). Statistical comparison was performed using univariate tests. A total of 22 patients and 10 controls were included. Tetraploid neurons and astrocytes were found both in healthy individuals and DRE patients in the three brain areas analysed: cortex, hippocampus and amygdala. DRE patients presented a higher number of tetraploid neurons (p = 0.020) and astrocytes (p = 0.002) in the hippocampus than controls. These results were validated by image cytometry. Conclusions: We demonstrated the presence of both tetraploid neurons and astrocytes in healthy subjects as well as increased levels of both cell populations in DRE patients. Herein, we describe for the first time the presence of tetraploid astrocytes in healthy subjects. Furthermore, these results provide new insights into epilepsy, opening new avenues for future treatment