Desarrollo y caracterización de nuevos derivados de cannabinoides no psicotrópicos para el tratamiento de enfermedades inflamatorias

Los fitocannabinoides son sustancias liposolubles presentes en la planta Cannabis sativa y entre sus dianas se encuentran los receptores cannabinoides y el receptor nuclear PPARγ. Los cannabinoides han mostrado ejercer potentes efectos antiinflamatorios e inmunomoduladores in vivo e in vitro a través de estos mecanismos, por lo que se consideran de especial interés para el tratamiento de enfermedades inflamatorias crónicas, tales como la esclerosis sistémica (SSc) o la enfermedad de Huntington (EH). Entre los fitocannabinoides destacan el cannabidiol y el cannabigerol debido a la falta de psicoactividad, las numerosas propiedades terapéuticas y su baja toxicidad. Con el fin de desarrollar nuevas terapias para el tratamiento de enfermedades inflamatorias, se han introducido modificaciones químicas para generar derivados cannabinoides más potentes y con un perfil farmacológico seguro. Nuestros resultados muestran que VCE-004.3 es un agonista dual PPARγ/CB2 y antagonista de CB1 y VCE-004.8 es un agonista dual PPARγ/CB2 . Ambos han mostrado inhibir los mecanismos fibróticos inducidos por TGFβ in vitro. Además, VCE-004.3 y VCE-004.8 han resultado efectivos como tratamientos sistémicos y también tópicos en modelos animales de SSc. La combinación de estas actividades (PPARγ/CB1/CB2) representa un avance importante en el desarrollo de nuevas terapias en enfermedades inflamatorias y fibróticas ya que todas estas dianas son críticas y complementarias. Por su parte, VCE-003.2 es un agonista de PPARγ con actividad neuroprotectora in vitro. Además hemos demostrado que los fitocannabinoides ácidos Δ9-THCA, CBDA y CBGA, presentan una mayor afinidad a PPARγ que sus correspondientes formas neutras. Entre ellos, Δ9-THCA es un agonista potente de PPARγ con actividad neuroprotectora in vitro. Es más, VCE-003.2 y Δ9-THCA previenen la neurodegeneración y la activación de las células de la glía en el modelo animal de EH. Estos datos nos llevan a considerar que VCE-003.2 y Δ9-THCA tienen un alto potencial para el tratamiento de la EH y otras enfermedades neurodegenerativas con rasgos neuroinflamatorios

Fine-tuning a brain spheroid model as a screening platform for neuroprotective therapies against ischemic stroke.

Motivation: Stroke is the second cause of death and a leading cause of disability worldwide. Ischemic stroke represents more than 80% of all events and it is due to the presence of a clot blocking the blood flow. European legislation has already assumed the goal of no longer needing to use animals for scientific research in the future. To improve current in vitro models, a two-stage 3D brain spheroid model was previously described to recreate the blood-brain barrier and serve as a tool to assess ischemic damage and neuroinflammation. Our objective is to fine tune this organoid model, so we can use it as a screening platform of different neuroprotective therapies against ischemic stroke. Methods: Six different human brain cells were cultured following manufacturer´s recommendations. In order to be able to visualize each cell type in the 3D model, immunocytochemistry protocols were stablished. Cells were fixed and immunolabeled using a suitable primary antibody for each type of cell followed by the appropriate Alexa 568 secondary antibody. Negative controls were also prepared following the same protocol without the addition of the primary antibody. As a fist approach for spheroid formation, astrocytes, microglia, oligodendrocytes and cortical neurons were co-cultured in round bottom 96 well plates building the spheroid core. After 48h microvascular cells and pericytes were added to the culture creating the surface area. Finally, spheroids images were taken employing bright field microscopy and diameter was measured. Results: Fluorescent images confirmed that the immunolabeling protocols for each cell type were set up appropriately. Red fluorescence staining indicated cell´s detection by their specific markers. Fluorescent signal was absent in negative controls, confirming the specific binding. Finally, two-stage spheroids (4.000 cells) were formed. After 48, one round-shaped spheroid per well was observed. Spheroid diameter was 313.9± 22 μm after 48 h and 690.9±64 μm after 4 days. Conclusions: We have fine-tuned the immunocytochemical technique to identify each cell line that will be used to analyse brain spheroids. The first trial for spheroid formation showed homogenous shape and size. Next step would be testing smaller spheroids (2.000 cells) and verifying that each cell type is correctly located in the 3D structure. If results are satisfactory these spheroids could be used as a tool to test potential neuroprotective therapies

Diet Supplementation with Polyphenol-Rich Salicornia ramosissima Extracts Protects against Tissue Damage in Experimental Models of Cerebral Ischemia

Salicornia; Ischemia; NeuroprotectionSalicornia; Isquemia; NeuroprotecciónSalicornia; Isquèmia; NeuroproteccióStrokes are the second most common cause of death worldwide and a leading cause of disability. Regular consumption of polyphenols has been shown to reduce the risk of suffering a cardiovascular event. For this reason, we have investigated the protective effect of Salicornia ramosissima, a seasonal halophyte that synthetizes high amounts of bioactive compounds, including polyphenols, in response to environmental stress. Aqueous, hydroalcoholic, and ethanolic extracts were prepared to investigate if dietary supplementation prior to ischemic challenge can prevent subsequent damage using two animal models. First, we screened the protective effect against hypoxia–reoxygenation in Drosophila melanogaster and observed that both ethanolic and hydroalcoholic extracts protected flies from the deleterious effects of hypoxia. Second, we confirmed the protective effect of S. ramosissima ethanolic extract against brain ischemia using the transient middle cerebral artery occlusion mice model. Four weeks of oral supplementation with the ethanolic extract before artery occlusion reduced infarct volume and lowered the plasma levels of the DNA peroxidant product 8-hydroxydeoxyguanosine. Phytochemical profiling of S. ramosissima ethanolic extract revealed 50 compounds. Thus, it represents a valuable source of bioactive compounds that show promising disease-modifying activities and could be further developed as an effective food supplement for the prevention or treatment of neurovascular disorders.The authors received financial support from “CSF-Proyectos estratégicos de I+D+i. Proyectos cofinanciados en un 80% por fondos del Programa Operativo FEDER de Andalucía 2014–2020”, grant number PE-0527-2019. This research was partially funded by “Consejería de Transformación Económica, Industria, Conocimiento y Universidades (CTEICU) y 80% cofinanciados por la UE, PO FEDER Andalucía 2014-2020”, grant number [PY20_01351]. C.d.R. received financial support from the Sara Borrell program funded by ISCIII, grant number [CD21/00148]

Gestión del conocimiento. Perspectiva multidisciplinaria. Volumen 2

El libro “Gestión del Conocimiento. Perspectiva Multidisciplinaria”, es una publicación internacional, seriada, continua, arbitrada de acceso abierto a todas las áreas del conocimiento, es el esfuerzo de investigadores de varios países del mundo, orientada a contribuir con procesos de gestión del conocimiento científico, tecnológico y humanístico que consoliden la transformación del conocimiento en diferentes escenarios, tanto organizacionales como universitarios, para el desarrollo de habilidades cognitivas del quehacer diario. En este sentido, partiendo de los aportes teóricos y prácticos que presentan los autores, resultados de sus trabajos de investigación, análisis de diversas teorías, propuestas, enfoques, así como experiencias que se han dado a nivel del tema de gestión del conocimiento, presentamos a la comunidad internacional el libro Gestión del conocimiento. Perspectivas multidisciplinarias, que permitirá un mayor conocimiento y aplicación de estos conceptos, traduciéndose en una mejor aplicación y posicionamiento de las organizaciones en la utilización del conocimiento, la apropiación y transformación del mismo

In vivo high throughput screening assay to test potential neuroprotective therapies for ischemic stroke

Stroke is the second cause of death worldwide and a leading cause of disability, with ischemic stroke accounting for 85% of all cases. Ischemic stroke is the result of the blockage of a cerebral artery by a clot and the only approved medical treatment for this condition is reperfusion therapy. However, reperfusion therapies are only applicable to a small percentage of patients. Since many patients remain untreated and the disease burden is expected to triple in the next decade, there is an urgent need for neuroprotective therapies. Among the possible causes of neuroprotectants failure are the simplistic approach of targeting a single pathway among the multiple ones affected in cerebral ischemia. Therefore, we thought to establish an in vivo screening model using Drosophila melanogaster where we can test multiple drug combinations already identified by in silico drug repositioning. This model organism has a short life cycle, the entire hypoxia signaling cascade conserved and a high degree of homology with approximately 75% of human disease-associated genes. Flies from the Oregon R strain were exposed to anoxia or severe hypoxia  (1% O2) for different periods of time. We observed a sexually dimorphic response with males being more vulnerable than female flies to hypoxia. Then, we established the standard conditions to induce hypoxia in D. melanogaster males under defined environment using a hypoxia chamber. During posthypoxic reoxygenation (21% O2), we assessed daily mortality, fly locomotor activity and analyzed molecular characteristics (reactive oxygen species (ROS) production and caspase activation) at various timepoints. Periods of severe hypoxia longer than 2 hours affected flies´ survival, impaired locomotor ability and increased caspase activation. Most of the flies died within 24 hours of exposure (early reoxygenation phase). In conclusion, we established a reliable and reproducible high throughput screening operation protocol to study the impact of different therapeutic approaches on hypoxia-reoxygenation injury using D. melanogaster as model system. Our protocol and the availability of mutants and transgenic strains may facilitate the study of stroke treatments and potentially, other neurodegenerative diseases.  ischemic stroke; neuroprotective therapies; Drosophila Melanogaste

Neuroprotection strategies towards neonatal stroke

Motivation: Neonatal hypoxia-ischemia, a type of stroke, is a cerebrovascular insult that affects the brain at early stages of development, from birth until postnatal day 28. It is the most important cause of death and disability in the pediatric population; long-term sequelae include behavioral and learning disabilities, cerebral palsy and motor dysfunction. Since brain injury is highly associated to oxidative stress and there is currently no effective treatment, antioxidant therapy has been proposed as a novel approach to prevent and reduce brain damage. Previous preclinical in vivo studies have shown that nutraceuticals, including polyphenols, have neuroprotective properties against hypoxia-ischemia induced damage. Thus, we are interested in evaluating the neuroprotective activity of a phenolic compound present in olive leaf extract on neonatal stroke. Methods: Our group uses the Rice-Vannucci hypoxic-ischemic mouse model by ligation of the left common carotid artery and hypoxia in 7 day old pups. We inject the phenolic compound intraperitoneally 20 min before the ligation and dissect the brain a week later to analyze the neuroprotective effects by histology and immunohistochemistry. In parallel, we are obtaining brain and plasma samples to analyze the concentration of the compound at different time points by mass spectrometry. We are also developing a screening model using Drosophila melanogaster by supplementing larvae diet as a way to optimize the analysis of the neuroprotective properties of this and other phenolic compounds. Results: Our lab has developed a histological scoring system based on histology images to categorize brain damage in different cerebral structures. We are optimizing the protocols to further evaluate differences in myelinization, astrocytes and microglia activation by immunohistochemistry. Plasma and brain samples have been collected, and the phenolic compound concentration over time  will be analyzed by collaborators from University of Seville. In the Drosophila model, we are studying mortality and motility changes after hypoxia. Conclusions: The hypoxic-ischemic mouse model is appropriate to study neuroprotection as it recapitulates brain damage in neonatal stroke. Preliminary results suggest that a phenolic compound present in olive leaf extract could protect against stroke-induced brain tissue loss. If the results are satisfactory, we will then test the neuroprotective effects of supplementing mothers during pregnancy

ARIA Mexico 2014 Adaptation of the Clinical Practice Guide ARIA 2010 for Mexico. Methodology ADAPTE

Background: The global prevalence of allergic rhinitis is high. International Study of Asthma and Allergies in Childhood (ISAAC) Phase III reports a total estimated prevalence of 4.6% in Mexico. There is evidence based on allergic rhinitis Clinical Practice Guidelines (CPG), but its promotion, acceptance and application is not optimal or adequate in Mexico. Objective: To generate a guideline for the treatment of allergic rhinitis and its impact on asthma by adaptating the 2010 ARIA Guideline to Mexican reality, through a transculturation process applying the ADAPTE methodology. Material and method: Using the ADAPTE Methodology, the original 2010 ARIA CPG recommendations were evaluated by the guideline development group (GDG) into which multiple medical specialities managing patients with allergic rhinitis were incoorporated. The GDG valorated the quality of 2010 ARIA, checked and translated key clinical questions. Moreover, the GDG adjusted recommendations, patient preferences and included comments in the context of the Mexican reality (safety, costs and cultural issues). To accomplish this, we ran Delphi panels with as many rounds as necessary to reach agreement. One extra question, not included in the original 2010 ARIA, on the use of Nasal Lavages for AR was created sustained by a systematic literature review. Results: A total of 45 questions from the original 2010 ARIA were included and divided into six groups covering prevention, medical treatment, immunotherapy and alternative medicine to treat patients with allergic rhinitis with or without asthma. Most of the questions reached agreement in one or two rounds; one question required three rounds. Conclusions: An easy-to-use, adaptated, up-to-date and applicable allergic rhinitis guideline for Mexico is now available

Coeducación : una escuela hacia la igualdad

Mención honorífica de la convocatoria de premios 'Irene: la paz empieza en casa 2008'. Incluye resumen con las actividades realizadas en el proyectoPresenta un proyecto de coeducación para la igualdad de sexos realizado en el IES 'Reyes de España' en Linares (Jaén), entre los cursos 2006 y 2008. A través del proyecto se han introducido cambios que incluyen una perspectiva de género en la práctica docente buscando favorecer prácticas correctoras de estereotipos sexistas; planifica objetivos y actuaciones para corregir desigualdades y discriminaciones sexistas, promueve la autoformación y el trabajo en equipo; fomenta un uso no sexista del lenguaje; utiliza una metodología activa, participativa, significativa y lúdica; utiliza las nuevas tecnologías como edición de video, carteles, página web, presentaciones didácticas en PowerPoint e Impress, revistas digitales, ejercicios informáticos en software libre realizados con Flash, Graffitis, campañas publicitarias, periódico escolar, etc.; incorpora procedimientos de evaluación para valorar el grado de consecución de los objetivos establecidos; establece mecanismos de difusión escrita, informática y audiovisual a través de una página web; y utiliza materiales y recursos en español, francés e inglés.AndalucíaBiblioteca de Educación del Ministerio de Educación, Cultura y Deporte; Calle San Agustín, 5 - 3 Planta; 28014 Madrid; Tel. +34917748000; Fax +34917748026; [email protected]

Espacios y destinos turísticos en tiempos de globalización y crisis

2 volúmenesXII Coloquio de Geografía del Turismo, Ocio y Recreación de la Asociación de Geógrafos Españoles. Colmenarejo (Madrid), del 17 al 19 de junio de 2010.Este libro ha sido editado con la colaboración económica del Ministerio de Ciencia e Innovación (ref. CS02010-10416-E)

Compilación de Proyectos de Investigación desde el año 2003 al 2012

Listado de Proyectos de investigación de UPIICSA desde 2003 a 201