Efecto de la administración posnatal de Epigalocatequina-3-galato sobre la función cardíaca y la respuesta psicomotora y cognitiva

[spa] INTRODUCCIÓN: El consumo de alcohol durante el embarazo puede tener graves consecuencias para el desarrollo fetal, dando lugar a los Trastornos del Espectro Alcohólico Fetal (TEAF). Dicho consumo varía según regiones y patrones destacando el patrón "Binge" que consiste en el consumo a dosis altas por atracones, y el patrón Mediterráneo, dónde se consumen niveles moderados de alcohol de forma crónica. El TEAF está relacionado con cardiopatías congénitas. Además, en modelos animales se ha observado que la exposición prenatal al alcohol induce alteraciones en la función y fisiología cardíaca que pueden persistir hasta la edad adulta. Se han estudiado compuestos como el ácido fólico y la betaína para contrarrestar estos efectos, pero se requiere más investigación para comprender su impacto en la especie humana. Durante la gestación, la exposición a alcohol también genera daños en el sistema nervioso central, causando cambios estructurales y alteraciones cognitivo-conductuales. Por ello, el tratamiento del TEAF requiere un enfoque multidisciplinario, incluyendo terapias conductuales y psicofármacos. Se han descrito compuestos como la colina, el cannabidiol y los ácidos grasos omega 3 como posibles agentes contra el TEAF estudiados mayoritariamente en modelo animal, pero su investigación en humanos no está desarrollada. Por este motivo persiste la necesidad de encontrar un compuesto que sea seguro y que pueda mejorar la función neurológica de las personas con TEAF. La epigalocatequina-3-galato (EGCG), es un flavonoide que se ha asociado al tratamiento de enfermedades cardiovasculares, neurodegenerativas y cáncer. En el TEAF la EGCG parece mejorar la restricción del crecimiento prenatal y el neurodesarrollo en modelos animales, pero todavía se desconocen los efectos de su administración posnatal sobre el daño cardíaco, así como sobre las alteraciones cognitivas y conductuales presentes en el TEAF. HIPÓTESIS Y OBJETIVOS: La hipótesis de este trabajo es que la exposición prenatal al alcohol afecta negativamente en la función cardiovascular y neuromotora que perdura hasta la edad adulta, y que puede ser diferente en función del tipo de consumo Binge o Mediterráneo. La EGCG, administrada durante la infancia y adolescencia en la población expuesta prenatalmente a alcohol, puede atenuar las alteraciones derivadas de dicha exposición. Nuestro objetivo es evaluar en un modelo experimental de ratón si el tratamiento posnatal con EGCG mejora la función cardiovascular, reduce el daño cardiaco y mejora el desempeño neuroconductual de adultos jóvenes expuestos prenatalmente a alcohol en los dos patrones de consumo de alcohol mencionados

Selective EP2 agonism attenuates HDM-induced murine airway pathology and mast cell activity, and triggers intracellular inhibitory signaling in mast cells

Descripció del recurs: el 01 setembre 2012L'asma al lèrgica és una malaltia respiratòria crònica amb una alta prevalença en els països desenvolupats. Els tractaments actuals no aturen el procés al lèrgic subjacent ni aconsegueixen el control dels símptomes de la malaltia. El tractament més eficaç és l'ús de corticoesteroids, que es basen en modificacions químiques de potents hormones endògenes antiinflamatòries. L'estudi de vies antiinflamatòries endògenes és una estratègia experimental eficaç per descobrir noves dianes terapèutiques potencials contra l'asma. Una d'aquestes vies endògenes és la de la ciclooxigenasa (COX). Prostaglandina (PG) PGE2, un producte de la COX, s'ha suggerit que exerceixen un efecte protector en els pulmons. En particular, els estudis experimentals en pacients amb asma van revelar que la PGE2 inhalada redueix la hiperreactivitat bronquial i la inflamació. Aquest efecte de protecció de la PGE2 també s'ha demostrat, directament i indirectament, en ratolins sensibilitzats amb OVA o HDM. Els mecanismes subjacents als efectes beneficiosos de la PGE2 en l'asma no es coneixen. Una de les característiques més constants de la PGE2 és la seva capacitat de modular l'activitat dels mastòcits in vitro. Els nostres darrers estudis in vivo van mostrar que la PGE2 també inhibeix l'activitat dels mastòcits en ratolins sensibilitzats a HDM i que aquest efecte modulador sobre els mastòcits és paral lel a la sobreexpressió del receptor EP2. D'aquests resultats sorgeix la hipòtesi que la PGE2 podria interactuar amb els receptors EP2 dels mastòcits de la superfície bronquials i així exercir una acció protectora enfront la patologia impulsada pels al lèrgens en contactar amb les vies respiratòries. El coneixement precís d'aquests mecanismes sens dubte ajudarà a descobrir potencials molècules diana contra l'asma. L'objectiu general d'aquesta tesi és establir preclínicament la rellevança del receptor EP2 mastocitari en l'efecte beneficiós de la PGE2 en l'asma al lèrgica, i descobrir els mecanismes moleculars que resulten d'aquesta activació selectiva del receptor. Per aconseguir aquest objectiu hem dut a terme diversos abordatges in vitro i in vivo. En primer lloc, determinar el patró d'expressió dels receptors EP de la PGE2 en diferents poblacions de mastòcits humans i murins, i es va avaluar a partir de llavors (a) si aquestes diferències en l'expressió relativa dels receptors de EP 1-4 influïa en la capacitat de PGE2 per modular la degranulació dels mastòcits i la mobilització del calci i (b) si els mastòcits humans es van comportar de manera similar als murins en diferents escenaris d'expressió dels receptors EP. Els resultats van apuntar a EP2 com el principal contribuent a l'efecte inhibitori de la PGE2 sobre els mastòcits murins i humans. Quan EP2 es va suggerir com a el receptor primari de protecció, vam abordar la importància de l'activació selectiva d'EP2 (a) en la protecció de la patologia de les vies respiratòries induïda per HDM en ratolins, i (b) la correlació d'aquesta patologia amb la capacitat de l'agonista selectiu EP2 en prevenir l'activitat de mastòcits in vivo. Hem demostrat que un agonista selectiu d'EP2 impedia el desenvolupament de l'AHR i la inflamació, i que aquest efecte estava relacionat amb la capacitat d'aquesta acció de l'agonista selectiu per atenuar l'activitat dels mastòcits de les vies respiratòries. A continuació, es van estudiar possibles mecanismes inhibitoris de senyalització implicats en aquest efecte de bloqueig intervingut per EP2. Hem observat que l'agonisme d'EP2 inhibeix in vivo i in vitro, l'activitat dels mastòcits. Hem descrit que la interacció amb PGE2-EP2 inhibeix la degranulació mastocitària a través de la supressió de la mobilització de calci intervinguda per la inhibició de la via Src-Fyn, i cAMP/PKA. Les nostres observacions ressalten que l'eix "la PGE2" - "l'EP2 mastocitari" - "les vies respiratòries" és una via endògena que condueix a una protecció natural contra la patologia de les vies respiratòries induïda pels aeroal lergens i ajuda a dilucidar els mecanismes precisos que descobreixen molècules diana objectiu de possibles nous tractaments antiasmàtics.Allergic asthma is a chronic respiratory disease with a high prevalence in developed countries. Current treatments do not halt the underlying allergic process and do not always control the symthomps of the disease. The most effective treatment is the use of glucocorticoids, which are based on chemical modifications of potent natural endogenous anti-inflammatory hormones. Studying endogenous anti-inflammatory pathways to explore new therapeutic targets is an efficient experimental strategy to uncover potential novel targets against asthma. One of such endogeneous pathways are cyclooxygenase (COX)-mediated. Prostaglandin (PG) PGE2, a COX product, has been suggested to exert a protective effect in the lungs. Notably, experimental studies with asthma patients revealed that inhaled PGE2 reduces airway hyperresponsiveness and inflammation. This protective PGE2 effect has also been demonstrated, directly and indirectly, in mice sensitized to OVA or HDM. The mechanisms underlying the beneficial effect of PGE2 in asthma are not understood. One of the most consistent features of PGE2 is its ability to modulate mast cell activity in vitro. Our recent in vivo studies showed that PGE2 also prevents mast cell activity in HDM sensitized mice and that this mast cell modulatory effect was paralleled by EP2 receptor overexpression. These results brought up the hypothesis that PGE2 might interact with EP2 receptor on the bronchial mast cells surface to exert a protective action against allergen-driven airway pathology. The precise understanding of such mechanisms will certainly help uncover potential anti-asthma target molecules along the way. The general objective of this thesis was to establish preclinically the relevance of the mast cell EP2 receptor to PGE2 beneficial effect in allergic asthma, and to uncover molecular mechanisms resulting from this receptor selective activation. To achieve this objective we have undertaken several in vitro and in vivo approaches. We first determined the PGE2 EP receptors expression pattern on different human and murine mast cell population, and thereafter assessed (a) whether such differences in the relative expression of EP receptors 1 to 4 influenced the ability of PGE2 to modulate mast cells degranulation and calcium mobilization, and (b) whether human mast cells behaved similarly to murine mast cells under different EP receptors expression scenarios. The results pointed at EP2 as the main contributor to mediate the inhibitory effect of PGE2 on both murine and human mast cells. Once EP2 had been suggested to be the primary protective receptor, we addressed the relevance of selective EP2 activation to (a) protection against HDMinduced airway pathology in mice, and (b) correlation of such pathology to the ability of selective EP2 agonism to prevent mast cells activity in vivo. We showed that a selective EP2 agonist prevented AHR and inflammation from developing, and that such effect was linked to the ability of such selective agonistic action to attenuate airway mast cell activity. We then studied potential inhibitory signaling mechanisms involved in such EP2-mediated blocking effect. We observed that EP2 agonism inhibited in vivo and in vitro, mast cell activity. We described that the PGE2-EP2 interaction on mast cells inhibiting mast cell degranulation through the supression of calcium influxes mediated by an inhibition of the Src-Fyn pathway, and cAMP/PKA. Our observations highlight that the "PGE2"-"mast cells EP2"-"airway" axis is an endogeneous pathway leading to natural protection against aeroallergens-induced airway pathology, and helps elucidate the precise mechanisms that will uncover clue molecules to be targeted by potential novel antiasthma treatments

Activity of the cyclooxygenase 2-prostaglandin-E prostanoid receptor pathway in mice exposed to house dust mite aeroallergens, and impact of exogenous prostaglandin E2

<p>Abstract</p> <p>Background</p> <p>Prostaglandin E₂(PGE₂), experimentally administered to asthma patients or assayed in murine models, improves allergen-driven airway inflammation. The mechanisms are unknown, but fluctuations of the endogenous cyclooxygenase (COX)-2/prostaglandin/E prostanoid (EP) receptor pathway activity likely contribute to the clinical outcome. We analyzed the activity of the pathway in mice sensitized to aeroallergens, and then studied its modulation under exogenous PGE₂.</p> <p>Methods</p> <p>Mice were exposed to house dust mite (HDM) aeroallergens, a model that enable us to mimic the development of allergic asthma in humans, and were then treated with either subcutaneous PGE₂or the selective EP1/3 receptor agonist sulprostone. Simultaneously with airway responsiveness and inflammation, lung COX-2 and EP receptor mRNA expression were assessed. Levels of PGE₂, PGI₂, PGD₂were also determined in bronchoalveolar lavage fluid.</p> <p>Results</p> <p>HDM-induced airway hyperreactivity and inflammation were accompanied by increased COX-2 mRNA production. In parallel, airway PGE₂and PGI₂, but not PGD₂, were upregulated, and the EP2 receptor showed overexpression. Subcutaneous PGE₂attenuated aeroallergen-driven airway eosinophilic inflammation and reduced endogenous PGE₂and PGI₂production. Sulprostone had neither an effect on airway responsiveness or inflammation nor diminished allergen-induced COX-2 and PGE₂overexpression. Finally, lung EP2 receptor levels remained high in mice treated with PGE₂, but not in those treated with sulprostone.</p> <p>Conclusion</p> <p>The lung COX-2/PGE₂/EP2 receptor pathway is upregulated in HDM-exposed mice, possibly as an effort to attenuate allergen-induced airway inflammation. Exogenous PGE₂downregulates its endogenous counterpart but maintains EP2 overexpression, a phenomenon that might be required for administered PGE₂to exert its protective effect.</p

Homogeneous Fe2O3 coatings on carbon nanotube structures for supercapacitors

The combination of carbon nanotubes with transition metal oxides can exhibit complementary charge storage properties for use as electrode materials for next generation energy storage devices. One of the biggest challenges so far is to synthesize homogeneous oxide coatings on carbon nanotube structures preserving their integrity. Here we present the formation of conformal coatings of Fe2O3 on vertically aligned carbon nanotubes obtained by atomic layer deposition. We investigate the effect of pristine, nitrogen plasma and water plasma treated carbon nanotube surfaces on the ALD-growth of Fe2O3 using ferrocene and ozone precursors. The surface morphology, coating thickness, microstructure and surface chemistry of iron oxide-carbon nanotube composites and their ultimate influence on the electrochemical behavior of the composites are evaluated. The most effective surface functionalization is that achieved by H2O plasma treatment, whereas untreated carbon nanotubes, despite the lack of active sites in the starting pristine surface, can be coated with an inhomogeneous Fe2O3 film

Enhancing hepatic fatty acid oxidation as a strategy for reversing metabolic disorders programmed by maternal undernutrition during gestation

Background/Aims: Moderate maternal calorie-restriction during gestation programmes offspring for a major propensity to develop metabolic alterations in adulthood. We aimed to assess whether increased hepatic fatty-acid oxidation (FAO), at early ages, by gene transfer of Cpt1am (active mutant of carnitine palmitoyltransferase-1a), may be a strategy for reversing metabolic disturbances associated to maternal calorie-restriction during gestation in rats. Methods: AAV-Gfp (control) and AAV-Cpt1am vectors were administered by tail vein injection in 18-day-old control-pups and the offspring of 20% calorie-restricted rats during gestation (CR). After weaning, animals were fed with normal-fat diet. At the age of 4 months, they were moved to HF-diet and sacrificed at the age of 6 months to collect tissues. Locomotive activity, energy expenditure and blood pressure were measured. Results: Under HF-diet, CR-animals showed higher HOMA-IR, adipocyte diameter and hepatic triglyceride accumulation than controls; these alterations were reverted in Cpt1am-injected animals. In liver, this treatment ameliorated inflammatory state, decreased expression of lipogenesis-related genes and partially restored the decreased expression of leptin-receptor occurring in CR-animals. Treatment also reverted the decreased energy expenditure and the increased blood pressure of CR-animals. Conclusion: Increasing hepatic FAO through AAV-Cpt1am injection at juvenile ages prevents some metabolic disorders associated to gestational maternal calorie-restriction

Murine models for the study of fetal alcohol spectrum disorders: An overview

Prenatal alcohol exposure is associated to different physical, behavioral, cognitive, and neurological impairments collectively known as fetal alcohol spectrum disorder. The underlying mechanisms of ethanol toxicity are not completely understood. Experimental studies during human pregnancy to identify new diagnostic biomarkers are difficult to carry out beyond genetic or epigenetic analyses in biological matrices. Therefore, animal models are a useful tool to study the teratogenic effects of alcohol on the central nervous system and analyze the benefits of promising therapies. Animal models of alcohol spectrum disorder allow the analysis of key variables such as amount, timing and frequency of ethanol consumption to describe the harmful effects of prenatal alcohol exposure. In this review, we aim to synthetize neurodevelopmental disabilities in rodent fetal alcohol spectrum disorder phenotypes, considering facial dysmorphology and fetal growth restriction. We examine the different neurodevelopmental stages based on the most consistently implicated epigenetic mechanisms, cell types and molecular pathways, and assess the advantages and disadvantages of murine models in the study of fetal alcohol spectrum disorder, the different routes of alcohol administration, and alcohol consumption patterns applied to rodents. Finally, we analyze a wide range of phenotypic features to identify fetal alcohol spectrum disorder phenotypes in murine models, exploring facial dysmorphology, neurodevelopmental deficits, and growth restriction, as well as the methodologies used to evaluate behavioral and anatomical alterations produced by prenatal alcohol exposure in rodents.This work was supported by Red de Salud Materno-Infantil y del Desarrollo (SAMID) (RD12/0026/0003 and RD16/0022/0002) from Instituto de Salud Carlos III and the PI15/01179 grant from Instituto de Salud Carlos II

Models for the Study of Fetal Alcohol Spectrum Disorders: An Overview

Prenatal alcohol exposure is associated to different physical, behavioral, cognitive, and neurological impairments collectively known as fetal alcohol spectrum disorder. The underlying mechanisms of ethanol toxicity are not completely understood. Experimental studies during human pregnancy to identify new diagnostic biomarkers are difficult to carry out beyond genetic or epigenetic analyses in biological matrices. Therefore, animal models are a useful tool to study the teratogenic effects of alcohol on the central nervous system and analyze the benefits of promising therapies. Animal models of alcohol spectrum disorder allow the analysis of key variables such as amount, timing and frequency of ethanol consumption to describe the harmful effects of prenatal alcohol exposure. In this review, we aim to synthetize neurodevelopmental disabilities in rodent fetal alcohol spectrum disorder phenotypes, considering facial dysmorphology and fetal growth restriction. We examine the different neurodevelopmental stages based on the most consistently implicated epigenetic mechanisms, cell types and molecular pathways, and assess the advantages and disadvantages of murine models in the study of fetal alcohol spectrum disorder, the different routes of alcohol administration, and alcohol consumption patterns applied to rodents. Finally, we analyze a wide range of phenotypic features to identify fetal alcohol spectrum disorder phenotypes in murine models, exploring facial dysmorphology, neurodevelopmental deficits, and growth restriction, as well as the methodologies used to evaluate behavioral and anatomical alterations produced by prenatal alcohol exposure in rodents

Industrial By-Products As a Novel Circular Source of Biocompatible Extracellular Vesicles

Extracellular vesicles (EVs) constitute an intricate system of molecular exchange that has recently gained tremendous interest. However, sustainable sources of safe biological EVs remain scarce and elusive. This study explores and defines the use of food industry by-products (BP) as a circular source of safe biocompatible EVs. Averaged diameter and molecular compositions indicate a large yield of exosomes and high abundancy of membrane lipids with signaling capacity in these vesicles. Complex proteomes mimicking those circulating in human blood plasma are also identified. Furthermore, BP-EVs do not show relevant cytotoxicity and display excellent oral and intravenous bioavailability together with specific organ targeting capacity. Collectively, it is believed that the novel findings reported here will open substantial venues for the use of BP as an optimal source of biocompatible nanovesicles in manifold applications of the biotechnological and biomedical fields.The authors sincerely thank Gemma Plaza, oenologist at the Castell del Remei winery in Penelles, Lleida, Spain and Juan Carlos Blanco, production manager at Mahou San Miguel in Alovera, Madrid, Spain for their kind and altruistic help on the obtention of their respective industry by-product samples. The authors also thank Dr. Hector Peinado and his research group at the National Center for Oncology Research (CNIO) in Madrid (Spain) for their support on the morphometric characterization of BP-EVs. Support for this work was provided by the Research and Education Council of the Community of Madrid, Spain (2018-T1/ BIO-10633), Ministry of Science and Innovation, Spain (PID2020- 114885RB-C21) and a FIS project by Carlos III Institute of Health (ISCIII), Spain (PI20/00623). A.S. acknowledges a grant from the Talento Program 2018 of the Community of Madrid. X.G.-P. acknowledges grants from Sara Borrell postdoctoral program (CD19/00243) and Miguel Servet tenure track program (CP21/00096) of the Instituto de Salud Carlos III (ISCIII, Spain), respectively awarded on the 2019 and 2021 calls under ISCIII-Health Strategy Actions [These grants are cofunded with European Union Funds (ISCIII Miguel Servet Program 2021 is cofunded by Fondo Social Europeo Plus, FSE+)]. M.V.C. acknowledges a Miguel Servet program contract (CPII20/00007). C.L.’s Ph.D. was funded by the Regional Ministry of Science, Universities and Innovation of the Community of Madrid and the European Social Fund for the recruitment of predoctoral researchers (PEJD-2019-PRE/BIO-16475). IRBLLEIDA and X.G.-P. are co-funded by CERCA Program/Generalitat de Catalunya

Evaluation of bacterial biomarkers to aid in challenging inflammatory bowel diseases diagnostics and subtype classification

Supported by the Spanish Ministry of Education and Science, No. SAF2010-15896, No. SAF2013-43284-P and No. SAF2017-82261-P.Peer reviewedPostprintPublisher PD

Murine Models for the Study of Fetal Alcohol Spectrum Disorders: An Overview.

Prenatal alcohol exposure is associated to different physical, behavioral, cognitive, and neurological impairments collectively known as fetal alcohol spectrum disorder. The underlying mechanisms of ethanol toxicity are not completely understood. Experimental studies during human pregnancy to identify new diagnostic biomarkers are difficult to carry out beyond genetic or epigenetic analyses in biological matrices. Therefore, animal models are a useful tool to study the teratogenic effects of alcohol on the central nervous system and analyze the benefits of promising therapies. Animal models of alcohol spectrum disorder allow the analysis of key variables such as amount, timing and frequency of ethanol consumption to describe the harmful effects of prenatal alcohol exposure. In this review, we aim to synthetize neurodevelopmental disabilities in rodent fetal alcohol spectrum disorder phenotypes, considering facial dysmorphology and fetal growth restriction. We examine the different neurodevelopmental stages based on the most consistently implicated epigenetic mechanisms, cell types and molecular pathways, and assess the advantages and disadvantages of murine models in the study of fetal alcohol spectrum disorder, the different routes of alcohol administration, and alcohol consumption patterns applied to rodents. Finally, we analyze a wide range of phenotypic features to identify fetal alcohol spectrum disorder phenotypes in murine models, exploring facial dysmorphology, neurodevelopmental deficits, and growth restriction, as well as the methodologies used to evaluate behavioral and anatomical alterations produced by prenatal alcohol exposure in rodents