Chapter 6: Characterization of TRPC Channels in a Heterologous System Using Calcium Imaging and the Patch-Clamp Technique

Transient receptor potential (TRP) ion channels are involved in a variety of fundamental physiological processes, and their malfunction produces numerous human diseases. Therefore, these proteins represent a class of attractive drug targets and a class of important off-targets for in vitro pharmacological profiling. In the past decades, the rapid progress in emerging functional assays and instrumentation has enabled to readily monitor thermoTRP channel activity, and to develop high throughput screening (HTS) assays for TPR drug discovery. Chronologically, functional methods for ion channels include the ligand binding assay, flux-based assay, electrophysiology, fluorescence-based assays, and, more recently, automated electrophysiological assays. Here we described the methodology used to monitor the functionality of two thermoTRPs, TRPV1 and TRPM8, based on Ca2+ microfluorography using a 96-well fluorescence plate reader that allows the implementation of a medium- to high-throughput format ideal for drug screening

Direct detection of the triphenylpyrylium-derived short-lived intermediates in the photocatalyzed degradation of acetaminophen, acetamiprid, caffeine and carbamazepine

[EN] Advanced oxidation processes are useful methodologies to accomplish abatement of contaminants; however, elucidation of the reaction mechanisms is hampered by the difficult detection of the short-lived primary key species involved in the photocatalytic processes. Nevertheless, herein the combined use of an organic photocatalyst such as triphenylpyrylium (TPP+) and photophysical techniques based on emission and absorption spectroscopy allowed monitoring the photocatalyst-derived short-lived intermediates. This methodology has been applied to the photocatalyzed degradation of different pollutants, such as acetaminophen, acetamiprid, caffeine and carbamazepine. First, photocatalytic degradation of a mixture of the pollutants showed that acetaminophen was the most easily photodegraded, followed by carbamazepine and caffeine, being the abatement of acetamiprid almost negligible. This process was accompanied by mineralization, as demonstrated by trapping of carbon dioxide using barium hydroxide. Then, emission spectroscopy measurements (steady-state and time-resolved fluorescence) allowed demonstrating quenching of the singlet excited state of TPP+. Laser flash photolysis experiments with absorption detection showed that oxidation of contaminants is accompanied by TPP+ reduction, with formation of a pyranyl radical (TPP center dot), that constituted a fingerprint of the redox nature of the occurring process. The relative amounts of TPP center dot detected was also correlated with the efficiency of the photodegradation process.Financial support from Spanish Government (Grants SEV-2016-0683, CTQ2012-38754-C03-02 and CTQ2015-69832-C4-4-R) and Generalitat Valenciana (Prometeo Program) is gratefully acknowledged. R. Martinez-Haya thanks financial support from Spanish Government (Grant SEV-2012-0267).Martínez-Haya, R.; Gomis Vicens, J.; Arqués Sanz, A.; Amat Payá, AM.; Miranda Alonso, MÁ.; Marín García, ML. (2018). Direct detection of the triphenylpyrylium-derived short-lived intermediates in the photocatalyzed degradation of acetaminophen, acetamiprid, caffeine and carbamazepine. Journal of Hazardous Materials. 356:91-97. https://doi.org/10.1016/j.jhazmat.2018.05.023S919735

Towards a Sustainable Campus: Working Together to Achieve the Green Campus Flag on the UDC Peripheral Campus of Ferrol

[Abstract] Purpose. This paper aims to present and describe the main actions carried out in six different faculties and common areas such as cultural and research centres and administrative buildings in the Ferrol campus at the University of A Coruña to achieve the second green flag on a Galician University. Design/methodology/approach. A case study describing the steps for implementing a green campus programme in a medium-size, young university campus integrated into a small city. An Environmental Campus Committee was created to assess the main factors that affect environmental footprint, discuss sustainability initiatives and develop a guide to action regarding different goals related to sustainable transport options, energy, water conservation and waste reduction. The actions included several fields such as education, circular economy and healthy life and involved the on and off-campus community. Findings. The programme achieved a decrease in water consumption and electrical energy. An important change in educational values and behaviours regarding sustainability was observed in and out of the campus community. The measurements adopted mainly in waste management, mobility and education led the Ferrol campus to achieve a green campus flag on November 2019. Originality/value. This experiment can serve as a guide to establish the Green Campus philosophy in other similar university campuses.The authors of this article want to thank the finantial and institutional support of Environmental Office of University of A Coruña to develop talks, activities and to implement different actions in Ferrol Campus. The authors of this article want to thank the finantial and institutional support of Vicerrectorado del Campus de Ferrol y Responsabilidad Social of University of A Coruña to achieve the Green Flag. Ana Ares-Pernas wants to thank the financial support of Vicerrectorado de Planificación Académica e Innovación Docente of University of A Coruña obtained in the I Edition of the Prizes for Teaching Innovation Projects in Service-Learning to develop Service-Learning activitie

La imagen que el profesor tiene de los alumnos y la aceptación de los alumnos por sus iguales en primero de primaria

Comunicación presentada en las I Congreso Internacional sobre Ciudadanía Crítica y Mejora Educativa: recursos, estrategias y experiencias, celebradas en la Universitat Jaume I (Castellón, España) los días 13 al 15 de octubre 2011El presente trabajo estudia la relación entre la imagen del alumno que tiene el profesor y la aceptación de los alumnos por sus iguales. La muestra está formada por 247 alumnos escolarizados en 11 aulas de primero de primaria, de cinco colegios públicos de Castellón de la Plana. A nivel metodológico se ha realizado un estudio de correlaciones, el procedimiento Anova de un factor, un análisis de tablas de contingencia r, y un análisis de regresión. Respecto a los resultados, las correlaciones son significativas, el Anova de un factor muestra que el profesor establece claras diferencias entre tipologías sociométricas y el análisis de regresión refleja cierto carácter predictivo de la variable preferencia del maestro sobre la aceptación de los iguales

Potenciación del contexto social escolar que promueve la convivencia, la inclusión, la ciudadanía y el aprendizaje

Comunicación presentada en las I Congreso Internacional sobre Ciudadanía Crítica y Mejora Educativa: recursos, estrategias y experiencias, celebradas en la Universitat Jaume I (Castellón, España) los días 13 al 15 de octubre 2011.El artículo trata sobre la implementación de un programa multinivel, multicomponente y multiagente. Se aplicó a 4 colegios de Castellón en las aulas de 1º de primaria (curso 2010-2011). Se basa en 3 grandes componentes (normas de convivencia y estrategias permanentes, aprendizaje cooperativo y desarrollo socioemocional) a partir de los cuales se capacitó a los profesores durante 6 sesiones de formación (30 horas). Ellos mismos fueron los que implementaron el programa adaptándolo al currículo, por lo que se pudo observar mejoras considerables, por una parte, aprendieron a ser más conscientes en la forma de dirigirse al alumnado; y por otra, en las relaciones interpersonales con su clase y entre los propios alumnos

Declaración de Chinchón: decálogo sobre eldulcorantes sin y bajos en calorías (ESBC)

Multidisciplinary experts in the areas of nutrition and health met in Chinchón, Madrid, on November 25-26, 2013 under the auspices of the Fundación para la Investigación Nutricional (Nutrition Research Foundation) and with the collaboration of the Madrid Regional Government’s Health Ministry, the International Sweeteners Association and the Carlos III Health Institute CIBER of Physiopathology of Obesity and Nutrition. They analyzed the current status of scientific knowledge on low- and no-calorie sweeteners (LNCS) and developed a consensus Decalogue on their use; this constitutes the Chinchón Declaration. Sweeteners, including sugar, represent a subject of undeniable interest and are currently a popular topic, although areas relating to their safety and benefits remain unknown to segments of academia and the general public. The nature of LNCS makes them vulnerable to biased and even contradictory information. They are food additives that are broadly used as sugar substitutes to sweeten foods, medicines and food supplements when non-nutritional or non-caloric alternatives are needed. The Chinchón Decalogue is the outcome of a meeting for reflection and consensus by a group of experts with backgrounds in different scientific disciplines (toxicology, clinical nutrition, community nutrition, physiology, food science, public health, pediatrics, endocrinology and nutrition, nursing, pharmaceutical care and food legislation). The Decalogue includes different aspects of LNCS related to regulation, use, benefits and safety. In general, benefits of LNCS have been traditionally neglected in comparison with the tendency for emphasising unexisting or unproven possible risks. The need to strengthen research on LNCS in Spain was emphasized, as well as the need to educate both professionals and the publicExpertos de carácter multidisciplinar de las áreas de conocimiento de la nutrición y la salud reunidos en Chinchón, Madrid, los días 25 y 26 de noviembre de 2013 , bajo los auspicios de la Fundación para la Investigación Nutricional y con la colaboración de la Consejería de Sanidad del Gobierno de la Comunidad de Madrid, la International Sweeteners Association y el CIBER de Fisiopatología de la Obesidad y la Nutrición del Instituto de Salud Carlos III, analizaron el estado actual del conocimiento científico en torno a los Edulcorantes sin y bajos en calorías (ESBC) y desarrollaron un Decálogo sobre su uso que constituye la Declaración de Chinchón. Los edulcorantes, incluido el azúcar, constituyen un elemento de indudable interés y actualidad, aunque no exento de desconocimiento por algunos sectores tanto académicos como de la población en general. La propia naturaleza de los ESBC los hace susceptibles de informaciones tergiversadas e incluso contradictorias. Son aditivos alimentarios ampliamente utilizados como sustitutivos del azúcar para endulzar alimentos, medicamentos y complementos alimenticios cuando se persiguen fines no nutritivos. El Decálogo de Chinchón es fruto de una reunión de reflexión y consenso por parte de un grupo de expertos procedentes de distintas disciplinas científicas (toxicología, nutrición clínica, nutrición comunitaria, fisiología, bromatología, salud pública, atención primaria, pediatría, endocrinología y nutrición, enfermería, atención farmacéutica y legislación alimentaria). El decálogo incluye diferentes aspectos de los EBSC relacionados con la legislación, uso, beneficios y seguridad. En general, los beneficios de los EBSC han sido tradicionalmente desatendidos en comparación con la tendencia de destacar posibles riesgos inexistentes o que no han sido probados. Hace especial hincapié en la necesidad de fortalecer la investigación de los EBSC en España, así como la necesidad de formar en este ámbito a los profesionales y a los consumidores en genera

Effects and mechanisms of mindfulness training and physical exercise on cognition, emotional wellbeing, and brain outcomes in chronic stroke patients: Study protocol of the MindFit project randomized controlled trial

Background: Post-stroke cognitive and emotional complications are frequent in the chronic stages of stroke and have important implications for the functionality and quality of life of those affected and their caregivers. Strategies such as mindfulness meditation, physical exercise (PE), or computerized cognitive training (CCT) may benefit stroke patients by impacting neuroplasticity and brain health. Materials and methods: One hundred and forty-one chronic stroke patients are randomly allocated to receive mindfulness-based stress reduction + CCT (n = 47), multicomponent PE program + CCT (n = 47), or CCT alone (n = 47). Interventions consist of 12-week home-based programs five days per week. Before and after the interventions, we collect data from cognitive, psychological, and physical tests, blood and stool samples, and structural and functional brain scans. Results: The effects of the interventions on cognitive and emotional outcomes will be described in intention-to-treat and per-protocol analyses. We will also explore potential mediators and moderators, such as genetic, molecular, brain, demographic, and clinical factors in our per-protocol sample. Discussion: The MindFit Project is a randomized clinical trial that aims to assess the impact of mindfulness and PE combined with CCT on chronic stroke patients' cognitive and emotional wellbeing. Furthermore, our design takes a multimodal biopsychosocial approach that will generate new knowledge at multiple levels of evidence, from molecular bases to behavioral changes. Clinical trial registration: www.ClinicalTrials.gov, identifier NCT04759950

Fatty liver index as a predictor for type 2 diabetes in subjects with normoglycemia in a nationwide cohort study

Our aim was to evaluate whether fatty liver index (FLI) is associated with the risk of type 2 diabetes (T2DM) development within the Spanish adult population and according to their prediabetes status; additionally, to examine its incremental predictive value regarding traditional risk factors. A total of 2260 subjects (Prediabetes: 641 subjects, normoglycemia: 1619 subjects) from the [email protected] cohort study were studied. Socio-demographic, anthropometric, clinical data and survey on habits were recorded. An oral glucose tolerance test was performed and fasting determinations of glucose, lipids and insulin were made. FLI was calculated and classified into three categories: Low ( 60). In total, 143 people developed diabetes at follow-up. The presence of a high FLI category was in all cases a significant independent risk factor for the development of diabetes. The inclusion of FLI categories in prediction models based on different conventional T2DM risk factors significantly increase the prediction power of the models when all the population was considered. According to our results, FLI might be considered an early indicator of T2DM development even under normoglycemic condition. The data also suggest that FLI could provide additional information for the prediction of T2DM in models based on conventional risk factors

Intrinsic Subtypes and Gene Expression Profiles in Primary and Metastatic Breast Cancer

Biological changes that occur during metastatic progression of breast cancer are still incompletely characterized. In this study, we compared intrinsic molecular subtypes and gene expression in 123 paired primary and metastatic tissues from breast cancer patients. Intrinsic subtype was identified using a PAM50 classifier and χ 2 tests determined the differences in variable distribution. The rate of subtype conversion was 0% in basal-like tumors, 23.1% in HER2-enriched (HER2-E) tumors, 30.0% in luminal B tumors, and 55.3% in luminal A tumors. In 40.2% of cases, luminal A tumors converted to luminal B tumors, whereas in 14.3% of cases luminal A and B tumors converted to HER2-E tumors. We identified 47 genes that were expressed differentially in metastatic versus primary disease. Metastatic tumors were enriched for proliferation-related and migration-related genes and diminished for luminal-related genes. Expression of proliferation-related genes were better at predicting overall survival in metastatic disease (OSmet) when analyzed in metastatic tissue rather than primary tissue. In contrast, a basal-like gene expression signature was better at predicting OSmet in primary disease compared with metastatic tissue. We observed correlations between time to tumor relapse and the magnitude of changes of proliferation, luminal B, or HER2-E signatures in metastatic versus primary disease. Although the intrinsic subtype was largely maintained during metastatic progression, luminal/HER2-negative tumors acquired a luminal B or HER2-E profile during metastatic progression, likely reflecting tumor evolution or acquisition of estrogen independence. Overall, our analysis revealed the value of stratifying gene expression by both cancer subtype and tissue type, providing clinicians more refined tools to evaluate prognosis and treatment. Cancer Res; 77(9); 1-9. ©2017 AACR