Report on the 24th meeting of the ECCR 8th-9th October 2021.

The 24th meeting of the European Council for Cardiovascular Research (ECCR) was virtual and held online on October 8th and 9th, 2021. Over 130 participants including trainees, early career researchers (ECR) and established investigators from eleven European countries (Austria, Denmark, France, Germany, Italy, Netherlands, Poland, Slovenia, Spain, Turkey, U.K.), and participants also from Canada, Chile, Saudi Arabia, and the U.S.A. connected to enjoy two days of outstanding research. The meeting was opened by its president, Professor Marisol Fernandez-Alfonso from the Complutense University in Madrid and covered several topics of cardiovascular research: from vascular and metabolic aspects to novel immunological mechanisms of cardiovascular disease.

Rapid, Automated, and Specific Immunoassay to Directly Measure Matrix Metalloproteinase-9-Tissue Inhibitor of Metalloproteinase-1 Interactions in Human Plasma Using AlphaLISA Technology: A New Alternative to Classical ELISA.

The protocol describes a novel, rapid, and no-wash one-step immunoassay for highly sensitive and direct detection of the complexes between matrix metalloproteinases (MMPs) and their tissue inhibitor of metalloproteinases (TIMPs) based on AlphaLISA® technology. We describe two procedures: (i) one approach is used to analyze MMP-9-TIMP-1 interactions using recombinant human MMP-9 with its corresponding recombinant human TIMP-1 inhibitor and (ii) the second approach is used to analyze native or endogenous MMP-9-TIMP-1 protein interactions in samples of human plasma. Evaluating native MMP-9-TIMP-1 complexes using this approach avoids the use of indirect calculations of the MMP-9/TIMP-1 ratio for which independent MMP-9 and TIMP-1 quantifications by two conventional ELISAs are needed. The MMP-9-TIMP-1 AlphaLISA® assay is quick, highly simplified, and cost-effective and can be completed in less than 3 h. Moreover, the assay has great potential for use in basic and preclinical research as it allows direct determination of native MMP-9-TIMP-1 complexes in circulating blood as biofluid.

Impacto de la evaluación formativa en el aprendizaje de la Farmacología: un seguimiento de tres años / No disponible

Objetivo: Evaluar el impacto de la introducción de la evaluación formativa (EF) en los resultados obtenidos por los alumnos en la asignatura de Farmacología General, impartida en el tercer curso del Grado de Farmacia. Material y Métodos: Se han comparado las notas obtenidas en los cursos 2011/12 (sin EF) con las de los cursos 2012/13 y 2013/14. La EF se realizó dos veces/semana y consistió en i) preguntas orales a toda la clase, ii) preguntas abiertas de respuesta corta (al principio o al final de la clase)y iii) trabajos individuales o en pequeños grupos (casos farmacoterapéuticos) realizados en clase o fuera del horario de clase. Las correcciones (únicamente comentarios de mejora y progreso) se hicieron por la profesora, bien en clase o por escrito. El trabajo no se calificó y sólo se tuvo en cuenta si se había realizado. Las notas se obtuvieron como resultado de un examen que fue similar en los tres cursos analizados y que combinaba preguntas tipo test, preguntas de respuesta corta y casos farmacoterapéuticos. Resultados: La suma de alumnos no presentados y suspensos disminuyó significativamente de 48,5% (2011/13) hasta 23,4% (2012/12) y 20,5% (2013/14) tras la introducción de la EF. El número de alumnos aprobados aumentó significativamente de 51,5% (2011/12) hasta 76,6% (2012/12) y 79,5% (2013/14). También hay que destacar el desplazamiento de las notas hacia valores superiores(aumento significativo de la proporción de notables y de sobresalientes). Conclusión: La introducción de la EF como actividad de aprendizaje de la Farmacología tiene un gran impacto en la mejora de la calidad del aprendizaje de los alumnos (AU)

Aim: To evaluate the impact of formative assessment (FA) in the results obtained by students in the subject General Pharmacology, given in the third year of the Bachelor of Pharmacy. Material and Methods: We compared the grades reached in years 2011/12 (without FA) with those in years 2012/13 and 2013/14 (with FA). FA was performed twice a week consisting in i) oral questions to the whole student group, ii) short-answer open questions (at the beginning or end of class) and iii) individual work or collaborative work in small groups (pharmacotherapeutic cases) answered in the classroom or at home. Corrections were made by the teacher, either in class or by written feedback (only opinions for improvement and progress were indicated). The work was not scored, only taking into account whether or not it had been performed. The marks were obtained as a result of an exam, which was similar in the three courses, combining multiple choice questions, short-answer questions and pharmacotherapeutic cases. Results: The sum of failing students and students not taking exam significantly decreased after the introduction of the FA from 48.5 % (2011/12) to 23.4 % (2012/12) and 20.5 % (2013/14). The number of passing students significantly increased from 51.5 % (2011/12) to 76.6 % (2012/12) and 79.5 %(2013/14). We also noticed a shift in marks to higher values (significant increase in the proportion of B and A). Conclusion: The introduction of FA as a learning strategy in General Pharmacology has a great impact on improving the quality of student learning (AU)

Remodeling of energy metabolism and absence of electrophysiological changes in the heart of obese hyperleptinemic mice. New insights into the pleiotropic role of leptin.

Dietary treatment with high-fat diets (HFD) triggers diabetes and hyperleptinemia, concomitantly with a partial state of leptin resistance that affects hepatic and adipose tissue but not the heart. In this context, characterized by widespread steatosis, cardiac lipid content remains unchanged. As previously reported, HFD-evoked hyperleptinemia could be a pivotal element contributing to increase fatty-acid (FA) metabolism in the heart and to prevent cardiac steatosis. This metabolic adaptation might theoretically reduce energy efficiency in cardiomyocytes and lead to cardiac electrophysiological remodeling. Therefore the aim of the current study has been to investigate the impact of long-term HFD on cardiac metabolism and electrophysiological properties of the principal ionic currents responsible of the action potential duration in mouse cardiomyocytes. Male C57BL/6J mice were fed a control (10 kcal% from fat) or HFD (45 kcal% from fat) during 32 weeks. Quantification of enzymatic activities regulating mitochondrial uptake of pyruvate and FA showed an increase of both carnitine-palmitoyltransferase and citrate synthase activities together with a decrease of lactate dehydrogenase and pyruvate dehydrogenase activities. Increased expression of uncoupling protein-3, Mn-, and Cu/Zn-superoxide dismutases and catalase were also detected. Total glutathione/oxidized glutathione ratios were unaffected by HFD. These data suggest that HFD triggers adaptive mechanisms aimed at (i) facilitating FA catabolism, and (ii) preventing oxidative stress. All these changes did not affect the duration of action potentials in cardiomyocytes and only slightly modified electrocardiographic parameters.

Leptin drives fat distribution during diet-induced obesity in mice

Objective Desensitization of leptin receptors is a process that specifically occurs in some tissues. We have hypothesized that during the development of obesity tissue lipids would increase gradually in particular organs depending on leptin responsiveness. Our aim was to establish a relationship between leptin resistance and lipid deposition by using a model of diet-induced obesity (DIO) and we have characterized, in mice undergoing a dietary treatment with a high-fat (HF) diet, the evolution of lipid content and leptin responsiveness in white adipose tissue and liver. Methods Four-week-old male C57BL/6J mice were divided into two groups and assigned either to a low-fat or to a high-fat diet. Dietary treatment lasted 8, 20 or 32 weeks. The last day animals received 1mg/kg leptin and then tissues were weighed and processed for Western-blotting and lipid determination. Results We observed an initial increase of the relative weight of adipose pads that was blunted after 32-week HF. In contrast, liver size exhibited an initial decrease followed by a progressive increase, which was coincident with the increase of hepatic triglycerides and with the impairment of leptin receptor signalling. Conclusion Our data show that leptin resistance within white adipose tissue does not deal with an increase of the size of adipose pads and suggest that consequences of leptin resistance, in terms of fat accumulation, are tissue-dependent (AU)

Objetivo La desensibilización de los receptores de la leptina es un proceso que ocurre de forma específica en algunos tejidos. Comprobamos la hipótesis de si durante el desarrollo de la obesidad, aumentarían los lípidos en tejido de forma progresiva en órganos concretos y en función de la capacidad de respuesta a la leptina. Nuestro objetivo fue establecer una relación entre la resistencia a la leptina y la deposición de lípidos mediante el uso de un modelo de obesidad inducida por dieta (OID) y caracterizamos, en ratones sometidos a un tratamiento dietético con una dieta elevada en grasas (DEG), la evolución del contenido lipídico y la capacidad de respuesta a la leptina en tejido adiposo blanco y en el hígado. Métodos Ratones C57BL/6J machos de cuatro semanas de edad fueron divididos en dos grupos y asignados a una dieta de bajo o de elevado contenido en grasas. El tratamiento dietético duró 8, 20 o 32 semanas. El último día, los animales recibieron 1mg / kg de leptina y luego se pesaron y se procesaron los tejidos para transferencia de tipo Western y la determinación de lípidos. Resultados Se observó un aumento inicial en el peso relativo del tejido adiposo, que se redujo después de 32 semanas con DEG. Por otro lado, el tamaño del hígado mostró una descenso inicial, seguido de un aumento progresivo que coincidió con un aumento de los triglicéridos hepáticos y un deterioro en la señalización del receptor de la leptina. Conclusión Nuestros datos muestran que la resistencia a la leptina en el tejido adiposo blanco no aborda un aumento de tamaño del tejido adiposo y sugiere que las consecuencias de la resistencia a la leptina, en términos de acumulación de grasa, dependen del tejido (AU)

Leptin drives fat distribution during diet-induced obesity in mice.

OBJECTIVE: Desensitization of leptin receptors is a process that specifically occurs in some tissues. We have hypothesized that during the development of obesity tissue lipids would increase gradually in particular organs depending on leptin responsiveness. Our aim was to establish a relationship between leptin resistance and lipid deposition by using a model of diet-induced obesity (DIO) and we have characterized, in mice undergoing a dietary treatment with a high-fat (HF) diet, the evolution of lipid content and leptin responsiveness in white adipose tissue and liver. METHODS: Four-week-old male C57BL/6J mice were divided into two groups and assigned either to a low-fat or to a high-fat diet. Dietary treatment lasted 8, 20 or 32 weeks. The last day animals received 1mg/kg leptin and then tissues were weighed and processed for Western-blotting and lipid determination. RESULTS: We observed an initial increase of the relative weight of adipose pads that was blunted after 32-week HF. In contrast, liver size exhibited an initial decrease followed by a progressive increase, which was coincident with the increase of hepatic triglycerides and with the impairment of leptin receptor signalling. CONCLUSION: Our data show that leptin resistance within white adipose tissue does not deal with an increase of the size of adipose pads and suggest that consequences of leptin resistance, in terms of fat accumulation, are tissue-dependent.

Circadian feeding drive of metabolic activity in adipose tissue and not hyperphagia triggers overweight in mice: is there a role of the pentose-phosphate pathway?

High-fat (HF) diets trigger an increase in adipose tissue and body weight (BW) and disordered eating behavior. Our study deals with the hypothesis that circadian distribution of energy intake is more relevant for BW dynamics than diet composition. Four-week-old mice were exposed for 8 wk to a HF diet and compared with animals receiving control chow. HF mice progressively increased BW, decreased the amount of nocturnal (1800-0900 h) calories (energy or food intake) (30%) and increased diurnal (0900-1800 h) caloric intake (energy or food intake), although total daily intake was identical between groups. Animals were killed at 3-h intervals and plasma insulin, leptin, corticosterone, glucose, and fatty acid levels quantified. Adipose tissue was weighed, and enzymatic activities integral to the pentose phosphate pathway (PPP) assayed in lumbar adipose tissue. Phosphorylated AMP-dependent protein kinase and fatty acid synthase were quantified by Western blotting. In HF mice, there was a shift in the circadian oscillations of plasma parameters together with an inhibition of PPP activity and a decrease in phosphorylated AMP-dependent protein kinase and fatty acid synthase. In a second experiment, HF mice were forced to adhere to a circadian pattern of food intake similar to that in control animals. In this case, BW, adipose tissue, morning plasma parameters and PPP activity appeared to be normal. These data indicate that disordered feeding behavior can trigger BW gain independently of food composition and daily energy intake. Because PPP is the main source of reduced nicotinamide adenine dinucleotide phosphate, we suggest that PPP inhibition might be an early marker of adipose dysfunction in diet-induced obesity.

Resistencia a leptina inducida por fructosa: ¿un camino rápido hacia la esteatosis hepática? / No disponible

No disponible