Guido Dentesano

Modelling Neuroinflammation In Vitro: A Tool to Test the Potential Neuroprotective Effect of Anti-Inflammatory Agents

Neuron-microglia co-cultures treated with pro-inflammatory agents are a useful tool to study neuroinflammation in vitro, where to test the potential neuroprotective effect of anti-inflammatory compounds. However, a great diversity of experimental conditions can be found in the literature, making difficult to select the working conditions when considering this approach for the first time. We compared the use of neuron-primary microglia and neuron-BV2 cells (a microglial cell line) co-cultures, using different neuron:microglia ratios, treatments and time post-treatment to induce glial activation and derived neurotoxicity. We show that each model requires different experimental conditions, but that both neuron-BV2 and neuron-primary microglia LPS/IFN-γ-treated co-cultures are good to study the potential neuroprotective effect of anti-inflammatory agents. The contribution of different pro-inflammatory parameters in the neurotoxicity induced by reactive microglial cells was determined. IL-10 pre-treatment completely inhibited LPS/IFN-γ-induced TNF-α and IL-6 release, and COX-2 expression both in BV2 and primary microglial cultures, but not NO production and iNOS expression. However, LPS/IFN-γ induced neurotoxicity was not inhibited in IL-10 pre-treated co-cultures. The inhibition of NO production using the specific iNOS inhibitor 1400 W totally abolished the neurotoxic effect of LPS/IFN-γ, suggesting a major role for NO in the neurotoxic effect of activated microglia. Consequently, among the anti-inflammatory agents, special attention should be paid to compounds that inhibit NO production.

Pro-inflammatory gene expression and neurotoxic effects of activated microglia are attenuated by absence of CCAAT/enhancer binding protein β

BackgroundMicroglia and astrocytes respond to homeostatic disturbances with profound changes of gene expression. This response, known as glial activation or neuroinflammation, can be detrimental to the surrounding tissue. The transcription factor CCAAT/enhancer binding protein β (C/EBPβ) is an important regulator of gene expression in inflammation but little is known about its involvement in glial activation. To explore the functional role of C/EBPβ in glial activation we have analyzed pro-inflammatory gene expression and neurotoxicity in murine wild type and C/EBPβ-null glial cultures.MethodsDue to fertility and mortality problems associated with the C/EBPβ-null genotype we developed a protocol to prepare mixed glial cultures from cerebral cortex of a single mouse embryo with high yield. Wild-type and C/EBPβ-null glial cultures were compared in terms of total cell density by Hoechst-33258 staining; microglial content by CD11b immunocytochemistry; astroglial content by GFAP western blot; gene expression by quantitative real-time PCR, western blot, immunocytochemistry and Griess reaction; and microglial neurotoxicity by estimating MAP2 content in neuronal/microglial cocultures. C/EBPβ DNA binding activity was evaluated by electrophoretic mobility shift assay and quantitative chromatin immunoprecipitation.ResultsC/EBPβ mRNA and protein levels, as well as DNA binding, were increased in glial cultures by treatment with lipopolysaccharide (LPS) or LPS + interferon γ (IFNγ). Quantitative chromatin immunoprecipitation showed binding of C/EBPβ to pro-inflammatory gene promoters in glial activation in a stimulus- and gene-dependent manner. In agreement with these results, LPS and LPS+IFNγ induced different transcriptional patterns between pro-inflammatory cytokines and NO synthase-2 genes. Furthermore, the expressions of IL-1β and NO synthase-2, and consequent NO production, were reduced in the absence of C/EBPβ. In addition, neurotoxicity elicited by LPS+IFNγ-treated microglia co-cultured with neurons was completely abolished by the absence of C/EBPβ in microglia.ConclusionsThese findings show involvement of C/EBPβ in the regulation of pro-inflammatory gene expression in glial activation, and demonstrate for the first time a key role for C/EBPβ in the induction of neurotoxic effects by activated microglia.

Inhibition of CD200R1 expression by C/EBP beta in reactive microglial cells

BackgroundIn physiological conditions, it is postulated that neurons control microglial reactivity through a series of inhibitory mechanisms, involving either cell contact-dependent, soluble-factor-dependent or neurotransmitter-associated pathways. In the current study, we focus on CD200R1, a microglial receptor involved in one of these cell contact-dependent mechanisms. CD200R1 activation by its ligand, CD200 (mainly expressed by neurons in the central nervous system),is postulated to inhibit the pro-inflammatory phenotype of microglial cells, while alterations in CD200-CD200R1 signalling potentiate this phenotype. Little is known about the regulation of CD200R1 expression in microglia or possible alterations in the presence of pro-inflammatory stimuli.MethodsMurine primary microglial cultures, mixed glial cultures from wild-type and CCAAT/enhancer binding protein β (C/EBPβ)-deficient mice, and the BV2 murine cell line overexpressing C/EBPβ were used to study the involvement of C/EBPβ transcription factor in the regulation of CD200R1 expression in response to a proinflammatory stimulus (lipopolysaccharide (LPS)). Binding of C/EBPβ to the CD200R1 promoter was determined by quantitative chromatin immunoprecipitation (qChIP). The involvement of histone deacetylase 1 in the control of CD200R1 expression by C/EBPβ was also determined by co-immunoprecipitation and qChIP.ResultsLPS treatment induced a decrease in CD200R1 mRNA and protein expression in microglial cells, an effect that was not observed in the absence of C/EBPβ. C/EBPβ overexpression in BV2 cells resulted in a decrease in basal CD200R1 mRNA and protein expression. In addition, C/EBPβ binding to the CD200R1 promoter was observed in LPS-treated but not in control glial cells, and also in control BV2 cells overexpressing C/EBPβ. Finally, we observed that histone deacetylase 1 co-immunoprecipitated with C/EBPβ and showed binding to a C/EBPβ consensus sequence of the CD200R1 promoter in LPS-treated glial cells. Moreover, histone deacetylase 1 inhibitors reversed the decrease in CD200R1 expression induced by LPS treatment.ConclusionsCD200R1 expression decreases in microglial cells in the presence of a pro-inflammatory stimulus, an effect that is regulated, at least in part, by C/EBPβ. Histone deacetylase 1 may mediate C/EBPβ inhibition of CD200R1 expression, through a direct effect on C/EBPβ transcriptional activity and/or on chromatin structure.

CD200R1 and CD200 expression are regulated by PPAR-γ in activated glial cells.

The mechanisms that control microglial activation are of interest, since neuroinflammation, which involves reactive microglia, may be an additional target in the search for therapeutic strategies to treat neurodegenerative diseases. Neuron‐microglia interaction through contact‐dependent or independent mechanisms is involved in the regulation of the microglial phenotype in both physiological and pathological conditions. The interaction between CD200, which is mainly present in neurons but also in astrocytes, and CD200R1, which is mainly present in microglia, is one of the mechanisms involved in keeping the microglial proinflammatory phenotype under control in physiological conditions. Alterations in the expression of CD200 and CD200R1 have been described in neurodegenerative diseases, but little is known about the mechanism of regulation of these proteins under physiological or pathological conditions. The aim of this work was to study the modulation of CD200 and CD200R1 expression by peroxisome proliferator‐activated receptor gamma (PPAR‐γ), a transcription factor involved in the control of the inflammatory response. Mouse primary neuronal and glial cultures and neuron‐microglia cocultures were treated with the PPAR‐γ endogenous ligand 15‐deoxy‐Δ12, 14‐prostaglandin J2 (15d‐PGJ2) in the presence and absence of lipopolysaccharide plus interferon‐γ (LPS/IFN‐γ)‐induced glial activation. We show that 15d‐PGJ2 inhibits the pro‐inflammatory response and prevents both CD200R1 downregulation and CD200 upregulation in reactive glial cells. In addition, 15d‐PGJ2 abrogates reactive‐microglia induced neurotoxicity in neuron‐microglia cultures through a CD200‐CD200R1 dependent mechanism. These results suggest that PPAR‐γ modulates CD200 and CD200R1 gene expression and that CD200‐CD200R1 interaction is involved in the anti‐inflammatory and neuroprotective action of PPAR‐γ agonists. GLIA 2014;62:982–998

CCAAT/enhancer binding protein β regulates prostaglandin E synthase expression and prostaglandin E2 production in activated microglial cells.

The eicosanoid prostaglandin E2 (PGE2) plays important roles in neuroinflammation and it is produced by the sequential action of the enzymes cyclooxygenase‐2 (COX‐2) and prostaglandin E synthase (PTGES). The expression of both enzymes and the production of PGE2 are increased in neuroinflammation. The objective of this study was to elucidate whether the transcription factor CCAAT/enhancer binding protein β (C/EBPβ) regulates the expression of prostaglandin synthesis enzymes in neuroinflammation. To this aim, the expression of these enzymes in wild‐type and C/EBPβ‐null mice was analyzed in vitro and in vivo. In mixed glial cultures, lipopolysaccharide (LPS) ± interferon γ (IFN‐γ) induced C/EBPβ binding to COX‐2 and PTGES promoters. LPS ± IFN‐γ‐induced increases in PTGES expression and in PGE2 production in mixed glial and microglial cultures were abrogated in the absence of C/EBPβ. Also, increased brain PTGES expression induced by systemic LPS administration was markedly reduced in C/EBPβ‐null mice. In contrast to PTGES, the induction of COX‐2 expression in vitro or in vivo was not markedly affected by the absence of C/EBPβ. These results demonstrate that C/EBPβ regulates PTGES expression and PGE2 production by activated microglial cells in vitro and point to C/EBPβ as a regulator of PTGES expression in vivo in the inflamed central nervous system. Altogether, these findings strengthen the proposed role of C/EBPβ as a key player in the orchestration of neuroinflammatory gene response. GLIA 2013;61:1607–1619

Modulation of neuroinflammation by the microglial inhibitory receptor CD200R1

Resumen del poster presentado al 50th Inner Ear Biology Workshop, celebrado en Alcala de Henares-Madrid (Espana) del 10 al 13 de septiembre de 2013.Resumen del trabajo presentado al 15o Congreso Nacional de la Sociedad Espanola de Neurociencia (SENC) celebrado en Oviedo del 25 al 27 de septiembre de 2013.Resumen del poster presentado al CIBERDEM Annual Meeting, celebrado en Cerdanyola del Valles, Barcelona (Espana) del 11 al 13 de mayo de 2016.-- et al.Resumen del trabajo presentado al XXXVIII Congreso de la Sociedad Espanola de Ciencias Fisiologicas (SECF), celebrado en Zaragoza del 13 al 16 de septiembre de 2016.Poster presentado en el XI European Meeting on Glial Cells in Health and Disease, celebrado los dias 3 al 6 de julio de 2013 en Berlin (Alemania)Memoria presentada para optar al grado de Doctor por la Licenciada en Biologia Angela Prieto Folgado y realizada en el Instituto de Investigaciones Biomedicas Alberto Sols.La realizacion de este trabajo ha sido posible gracias a la financiacion otorgada por el FIS al proyecto de investigacion 96/1803.Grant Funding Source: Supported by the Fondo de Investigaciones Sanitarias (PI0011406) to MF.The chemotherapeutic study of a limited series of steroidal sapogenins from several endemic species of the flora of the Canary Islands is presented here. On the whole, they possess a very weak antibacterial activity, a slight antifungal effect and one of them, vespertilin, displays interesting cytostatic activity (ID50 = 5 micrograms/ml). A pharmacodynamic screening carried out on this product mainly revealed very slight toxicity, antihistaminic activity and a light tranquilizing effect. The data obtained justify further research.The purpose of this study was to characterize the role of ions other than Ca2+ in hepatic responses to alpha 1-adrenergic stimulation. We report that the alpha 1-adrenoreceptor activation of hepatic functions is accompanied by extracellular acidification and an increase in intracellular pH. These effects are dependent on extracellular Na+ concentration and are inhibited by the Na+/H+ antiporter blocker 5-(N-ethyl-N-isopropyl) amiloride under conditions that preclude antagonistic effects on agonist binding. Thus, the activation of plasma membrane Na+/H+ exchange is an essential feature of the hepatic alpha-adrenoreceptor-coupled signaling pathway. The following observations indicate that the sustained hepatic alpha 1-adrenergic actions rely on a functional coupling between the plasma membrane Na+/H+ and Na+/Ca2+ exchangers, resulting in the stimulation of Ca2+ influx. 1) Inhibition of the Na+/K(+)-ATPase does not prevent the alpha 1-adrenergic effects. However, alpha 1-adrenoreceptor stimulation fails to induce intracellular alkalinization and to acidify the extracellular medium in the absence of extracellular Ca2+. 2) A non-receptor-induced increase in intracellular Na+ concentration, caused by the ionophore monensin, stimulates Ca2+ influx and increases vascular resistance. 3) Inhibition of Na+/Ca2+ exchange prevents, in a concentration-dependent manner, most of the alpha 1-agonist-induced responses. 4) The actions of Ca(2+)-mobilizing vasoactive peptide receptors or alpha 2-adrenoreceptors, which produce neither sustained extracellular acidification nor release of Ca2+, are insensitive to Na+/H+ exchange blockers.Poster presentado en la VII Reunion Anual de la Red Tematica de Investigacion Cooperativa en Cancer (RTICC), celebrada en Salamanca el 24 de septiembre de 2014Resumen del trabajo presentado al VI Meeting de la Red Espanola de Canales Ioniocs (RECI), celebrado en Santiago de Compostela del 6 al 8 de septiembre de 2017.Tesis Doctoral presentada por Laura Jimenez Perez para optar al grado de doctor por la Universidad de Valladolid, Departamento de Bioquimica y Biologia Molecular y FisiologiaPoster presentado en la VII Reunion Anual de la Red Tematica de Investigacion Cooperativa en Cancer (RTICC), celebrada en Salamanca el 24 de septiembre de 2014Resumen del trabajo presentado al XXXXVIII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular (SEBBM), celebrado en Valencia del 7 al 10 de septiembre de 2015.Esta Tesis Doctoral fue realizada en el Centro Andaluz de Biologia del Desarrollo por la licenciada Briseida Beli Cacho Valadez para optar al grado de Doctor por la Universidad Pablo de Olavide.Rat liver S-adenosylmethionine (AdoMet) synthetase appears as high-M(r) (tetramer) and low-M(r) (dimer) forms. Both are inhibited in the presence of GSSG at pH 8. The calculated Ki values are 2.14 and 4.03 mM for the high- and low-M(r) forms, respectively. No effect on enzyme activity was observed in the presence of GSH, but modulation of inhibition by GSSG can be obtained by addition of GSH. At a total glutathione concentration (GSH + GSSG) of 10 mM, a KOX of 1.74 was calculated for the high-M(r) form, whereas this constant was 2.85 for the low-M(r) AdoMet synthetase. No incorporation of [35S]GSSG was observed in either of the enzyme forms, and inhibition of enzyme activity was correlated with dissociation of both AdoMet synthetases to a monomer. The data obtained in the presence of GSSG seem to suggest that oxidation leads to the formation of an intrasubunit disulfide. The possible regulation of AdoMet synthetase activity by the GSH/GSSG ratio is discussed, as well as its in vivo significance.Trabajo presentado en el XI Simposi de Neurobiologia: Future technical advances, organizado por la Socitat Catalana de Biologia, en Barcelona, los dias 12 y 13 de noviembre de 2018El estudio de la relacion entre componentes de la dieta y la salud/enfermedad utiliza metodos de valoracion de la ingesta dietetica, del estatus nutricional y de marcadores de funcion o de efecto. En concreto, en el estudio de los carotenoides y la salud ocular, interesa el estudio de dos carotenoides sin actividad provitamina A, la luteina y la zeaxantina, por su posible papel en la optimizacion de la funcion visual y en la prevencion de enfermedades cronicas asociadas a la edad, y de tres carotenoides con actividad provitamina A: -caroteno, -caroteno y -criptoxantina, por ser precursores de retinol, nutriente del que depende el ciclo visual para una vision normal. En el presente trabajo se ha llevado a cabo el estudio de los carotenoides de la dieta mas relevantes para la salud ocular humana considerando de forma simultanea parametros relacionados con la ingesta, el estatus y la funcion visual, asi como diversas variables que pueden modificar el estatus nutricional, como son la concentracion de lipidos en sangre, y la bioaccesibilidad de los carotenoides a partir de alimentos de amplio consumo...Fetal rat hepatocytes treated with transforming growth factor beta (TGF-beta) die by apoptosis. However, a subpopulation of them survives and undergoes an epithelial mesenchymal transition (EMT). This transition also occurs upon incubation with fetal bovine serum. We have isolated the subpopulations that undergo EMT (TGF-beta-treated-fetal hepatocytes: TbetaT-FH; serum-treated-fetal hepatocytes: ST-FH) and show that they present high levels of vimentin and Snail expression and lack cytokeratin 18 and E-cadherin. Both TbetaT-FH and ST-FH cells require mitogens to grow and maintain the response to TGF-beta in terms of growth inhibition. However, they lack differentiation markers such as the liver-enriched transcription factors hepatocyte nuclear factor 4 (HNF-4) or HNF-1alpha and express the progenitor marker OV-6. Interestingly, the EMT process confers them resistance to the apoptotic effect of TGF-beta, with cells showing higher levels of active AKT and Bcl-x(L) than fetal hepatocytes. In summary, these cells are refractory to the apoptotic effects of TGF-beta, showing characteristics of liver progenitors and of some hepatocellular carcinoma cells.Memoria de tesis presentada por Luis Vazquez Fonseca, Licenciado en Bioquimica para optar al grado de Doctor. Esta Tesis Doctoral ha sido realizada bajo el programa de doctorado de Biotecnologia y Tecnologia Quimica en el grupo de investigacion del CIBERER U729 en el Centro Andaluz de Biologia del Desarrollo, Area de Biologia Celular del Departamento de Fisiologia, Anatomia y Biologia Celular de la Universidad Pablo de Olavide y bajo la direccion del Dr. Carlos Santos Ocana y el Dr. Placido NavasResumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Poster presentado al 17o Congreso Nacional de la Sociedad Espanola de Neurociencia, celebrado en Alicante del 27 al 30 de septiembre de 2017.The mutations at the bithorax locus produce a transformation of anterior haltere into anterior wing. The bx1 allele presents unusual features when compared with other bx alleles. The phenotype of bx1 homozygotes is temperature sensitive but only with regard to the distal and not to the proximal transformation, thus suggesting two different components in the bithorax transformation. The phenotype of bx1 homozygotes is stronger than that of bx1 over the deletion of the gene, suggesting a trans interaction of the bx1 chromosomes which results in mutual partial inactivation. We show by temperature shift and clonal analysis experiments that the decision on whether to differentiate haltere or wing structures is taken at the end of the proliferation period of the mutant disc.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Poster presentado al XXVII Congreso Nacional de la Asociacion Espanola de Genetica Humana celebrado en Madrid del 10 al 12 de abril de 2013.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Poster presentado en el XI European Meeting on Glial Cells in Health and Disease, celebrado los dias 3 al 6 de julio de 2013 en Berlin (Alemania)Resumen del trabajo presentado al Spanish Society of Biochemistry and Molecular Biology (SEBBM), celebrado en Madrid del 16 al 19 de julio de 2019.Poster presentado en el XII European Meeting on Glial Cells in Health and Disease, celebrado los dias 15 a 18 de julio de 2015 en Bilbao (Espana)Trabajo presentado en el XL Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular. FEBS3+1st Joint Meeting of the French-Portuguese-Spanish Biochemical and Molecular, celebrado en Barcelona (Espana), del 23 al 26 de octubre de 2017Resumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Trabajo presentado en el XII GEIRLI Meeting: New trends in redox biology: a multidisciplinary approach, celebrado en Barcelona (Espana), los dias 4 y 5 de julio de 2019Treatment of nucleosomal particles with dimethylmaleic anhydride, a reagent for protein amino groups, is accompanied by a biphasic release of histones H2A plus H2B; one H2A.H2B dimer is more easily released than the other. This behavior allows the preparation of nucleosomal particles containing only one H2A.H2B dimer, which were complemented with 125I-labeled H2A.H2B. These reconstituted particles, which contain one labeled and one unlabeled H2A.H2B dimer, were treated with the amount of reagent needed to release one of the two H2A.H2B dimers. Radioactivity was equally distributed between residual particles and released proteins, which is consistent with equivalent binding sites in the nucleosomal particle for H2A.H2B dimers, rather than with intrinsically different sites. The asymmetric release of H2A.H2B dimers would be caused by a change in the binding site of one dimer following the release of the other. This behavior might be related to the structural dynamics of nucleosomes.Resumen del trabajo presentado al European Society of Cardiology (ESC) Congress, celebrado en Barcelona (Espana) del 26 al 30 de agosto de 2017.Resumen del poster presentado al 49th European Association for the Study of Diabetes Annual Meeting, celebrado en Barcelona (Espana) del 23 al 27 de septiembre de 2013.-- et al.Trabajo doctoral realizado por Da Rebeca Lapresa Ruiz de Gauna, para optar al grado de doctor por la Universidad de Salamanca.Rationale: Several animal models have been developed to study acute lung injury (ALI); however the majority of these studies are focused on different mechanisms within the acute phase. These models do not allow studying the mechanisms in the later phases or testing any possible long-term treatment. The aim of this study was to develop an experimental ALI model simulating bronchial aspiration of gastric contents with bacterial superinfection with alveolar epithelial damage persisting over time. Methods: Sprague-Dawley rats (200-250g) were anesthetized with isofluorane. ALI was induced by intratracheal instillation of HCl (1 µl/g, 0.1 mol/L pH=1.4) followed by instillation of LPS from Escherichia coli O55:B5 (0, 10, 20, 30 or 40µg/g b.w.) two hours later. Control rats were treated with intratracheal instillations of saline. After 72h, the animals were sacrificed and bronchoalveolar lavage fluid (BALF) was sampled for further analysis of total protein concentration by bicinchoninic acid method. Results: At 72 h, rats suffered a significant loss of weight proportional to the administered dose of LPS (5.6% with 10µg/g b.w, 12.6% with 20µg/g b.w, 14.2% with 30µg/g b.w and 17.7% with 40µg/g b.w). Control rats gained in weight at 72h. LPS at 10, 20, 30 and 40µg/g b.w induced a 1.7, 2.5, 2.9 and 3.4 fold increase in total protein concentration in BAL fluid, respectively, reflecting a substantial increase proportional to the LPS dose. Conclusion: The degree of weight loss and the increase of total protein concentration in BAL fluid in the current model may reflect disease severity and progression. This model would be useful in future for new therapeutical options. Grant acknowledgements: FIS-PI12/02548 and Fundacio Parc Tauli.Resumen del trabajo presentado al European Respiratory Society (ERS) International Congress, celebrado en Paris (Francia) del 15 al 19 de septiembre de 2018.Resumen del trabajo presentado a las 5as Jornadas de Formacion del CIBERES celebradas en Bunyola (Mallorca) del 18 al 19 de octubre de 2012.Resumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Resumen del trabajo presentado al XIII Congreso de la Sociedad Espanola del Dolor, celebrado en Pamplona del 2 al 4 de junio de 2016.This work was supported by grants FIS-01/1048 and FIS-02/1199 from the Fondo de Investigacion Sanitaria and grant SA-087/01 from Junta de Castilla y Leon.Resumen del poster presentado al Joint Meeting of the American Physiological Society and the Physiological Society, celebrado en Dublin (Irlanda) del 29 al 31 de julio de 2016.Trabajo presentado al 5th International Conference on Phospholipase A2 Mediated Signaling in Translational Medicine celebrado en New Orleans (US) del 20 al 21 de mayo de 2013.Tesis Doctoral presentada por Rebeca Torres Merino para optar al grado de Doctora por la Universidad de Valladolid, Facultad de Medicina: Dpto. de Bioquimica y Biologia Molecular y Fisiologia.Poster presentado al Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), celebrado en Seattle, Washington (US) del 1 al 5 de mayo de 2016.Resumen del trabajo presentado al 63rd Annual Meeting Biophysical Society, celebrado en Baltimore, Maryland (USA) del 2 al 6 de marzo de 2019.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Resumen del poster presentado a la 5th Conference on Advances in Molecular Mechanisms Underlying Neurological Disorders (Joint conference of the European Society for Neurochemistry and the Biochemical Society) en la University of Bath (UK) del 23 al 26 de junio de 2013.-- Tambien presentado al 15o Congreso Nacional de la Sociedad Espanola de Neurociencia (SENC) celebrado en Oviedo del 25 al 27 de septiembre de 2013.Resumen del trabajo presentado al XXXVI Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular celebrado en Madrid del 4 al 6 de septiembre de 2013.Resumen del trabajo presentado a la 5th Conference on Advances in Molecular Mechanisms Underlying Neurological Disorders (Joint conference of the European Society for Neurochemistry and the Biochemical Society) en la University of Bath (UK) del 23 al 26 de junio de 2013.Resumen del poster presentado al XXVIII Congreso Nacional de la Sociedad Espanola de diabetes, celebrado en Bilbao del 20 al 22 de abril de 2016.SAF2016-77703-C2-2-R of the Ministerio de Economia y Competitividad, Spain and the European Regional Development Fund (ERDF); AGAUR 2017-SGR106 and the CERCA Programme of the Generalitat de Catalunya; C. Sanfeliu belong to Group 05 of CIBER Epidemiologia y Salud Publica (CIBERESP) of the Instituto de Salud Carlos III, Ministerio de Sanidad y Consumo, Spain

The microglial inhibitory receptor CD200R1 as a candidate target to control neuroinflammation

Trabajo presentado en el European Congress on Thrombosis and Haemostasis (ECTH), celebrado en Glasgow (Reino Unido), del 2 al 4 de octubre de 2019

Temporal and regional pattern of expression of pro-inflammatory and anti-inflammatory genes in mouse experimental autoimmune encephalomyelitis

Resumen del poster presentado al 50th Inner Ear Biology Workshop, celebrado en Alcala de Henares-Madrid (Espana) del 10 al 13 de septiembre de 2013.Resumen del trabajo presentado al 15o Congreso Nacional de la Sociedad Espanola de Neurociencia (SENC) celebrado en Oviedo del 25 al 27 de septiembre de 2013.Resumen del poster presentado al CIBERDEM Annual Meeting, celebrado en Cerdanyola del Valles, Barcelona (Espana) del 11 al 13 de mayo de 2016.-- et al.Resumen del trabajo presentado al XXXVIII Congreso de la Sociedad Espanola de Ciencias Fisiologicas (SECF), celebrado en Zaragoza del 13 al 16 de septiembre de 2016.Poster presentado en el XI European Meeting on Glial Cells in Health and Disease, celebrado los dias 3 al 6 de julio de 2013 en Berlin (Alemania)Memoria presentada para optar al grado de Doctor por la Licenciada en Biologia Angela Prieto Folgado y realizada en el Instituto de Investigaciones Biomedicas Alberto Sols.La realizacion de este trabajo ha sido posible gracias a la financiacion otorgada por el FIS al proyecto de investigacion 96/1803.Grant Funding Source: Supported by the Fondo de Investigaciones Sanitarias (PI0011406) to MF.The chemotherapeutic study of a limited series of steroidal sapogenins from several endemic species of the flora of the Canary Islands is presented here. On the whole, they possess a very weak antibacterial activity, a slight antifungal effect and one of them, vespertilin, displays interesting cytostatic activity (ID50 = 5 micrograms/ml). A pharmacodynamic screening carried out on this product mainly revealed very slight toxicity, antihistaminic activity and a light tranquilizing effect. The data obtained justify further research.The purpose of this study was to characterize the role of ions other than Ca2+ in hepatic responses to alpha 1-adrenergic stimulation. We report that the alpha 1-adrenoreceptor activation of hepatic functions is accompanied by extracellular acidification and an increase in intracellular pH. These effects are dependent on extracellular Na+ concentration and are inhibited by the Na+/H+ antiporter blocker 5-(N-ethyl-N-isopropyl) amiloride under conditions that preclude antagonistic effects on agonist binding. Thus, the activation of plasma membrane Na+/H+ exchange is an essential feature of the hepatic alpha-adrenoreceptor-coupled signaling pathway. The following observations indicate that the sustained hepatic alpha 1-adrenergic actions rely on a functional coupling between the plasma membrane Na+/H+ and Na+/Ca2+ exchangers, resulting in the stimulation of Ca2+ influx. 1) Inhibition of the Na+/K(+)-ATPase does not prevent the alpha 1-adrenergic effects. However, alpha 1-adrenoreceptor stimulation fails to induce intracellular alkalinization and to acidify the extracellular medium in the absence of extracellular Ca2+. 2) A non-receptor-induced increase in intracellular Na+ concentration, caused by the ionophore monensin, stimulates Ca2+ influx and increases vascular resistance. 3) Inhibition of Na+/Ca2+ exchange prevents, in a concentration-dependent manner, most of the alpha 1-agonist-induced responses. 4) The actions of Ca(2+)-mobilizing vasoactive peptide receptors or alpha 2-adrenoreceptors, which produce neither sustained extracellular acidification nor release of Ca2+, are insensitive to Na+/H+ exchange blockers.Poster presentado en la VII Reunion Anual de la Red Tematica de Investigacion Cooperativa en Cancer (RTICC), celebrada en Salamanca el 24 de septiembre de 2014Resumen del trabajo presentado al VI Meeting de la Red Espanola de Canales Ioniocs (RECI), celebrado en Santiago de Compostela del 6 al 8 de septiembre de 2017.Tesis Doctoral presentada por Laura Jimenez Perez para optar al grado de doctor por la Universidad de Valladolid, Departamento de Bioquimica y Biologia Molecular y FisiologiaPoster presentado en la VII Reunion Anual de la Red Tematica de Investigacion Cooperativa en Cancer (RTICC), celebrada en Salamanca el 24 de septiembre de 2014Resumen del trabajo presentado al XXXXVIII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular (SEBBM), celebrado en Valencia del 7 al 10 de septiembre de 2015.Esta Tesis Doctoral fue realizada en el Centro Andaluz de Biologia del Desarrollo por la licenciada Briseida Beli Cacho Valadez para optar al grado de Doctor por la Universidad Pablo de Olavide.Rat liver S-adenosylmethionine (AdoMet) synthetase appears as high-M(r) (tetramer) and low-M(r) (dimer) forms. Both are inhibited in the presence of GSSG at pH 8. The calculated Ki values are 2.14 and 4.03 mM for the high- and low-M(r) forms, respectively. No effect on enzyme activity was observed in the presence of GSH, but modulation of inhibition by GSSG can be obtained by addition of GSH. At a total glutathione concentration (GSH + GSSG) of 10 mM, a KOX of 1.74 was calculated for the high-M(r) form, whereas this constant was 2.85 for the low-M(r) AdoMet synthetase. No incorporation of [35S]GSSG was observed in either of the enzyme forms, and inhibition of enzyme activity was correlated with dissociation of both AdoMet synthetases to a monomer. The data obtained in the presence of GSSG seem to suggest that oxidation leads to the formation of an intrasubunit disulfide. The possible regulation of AdoMet synthetase activity by the GSH/GSSG ratio is discussed, as well as its in vivo significance.Trabajo presentado en el XI Simposi de Neurobiologia: Future technical advances, organizado por la Socitat Catalana de Biologia, en Barcelona, los dias 12 y 13 de noviembre de 2018El estudio de la relacion entre componentes de la dieta y la salud/enfermedad utiliza metodos de valoracion de la ingesta dietetica, del estatus nutricional y de marcadores de funcion o de efecto. En concreto, en el estudio de los carotenoides y la salud ocular, interesa el estudio de dos carotenoides sin actividad provitamina A, la luteina y la zeaxantina, por su posible papel en la optimizacion de la funcion visual y en la prevencion de enfermedades cronicas asociadas a la edad, y de tres carotenoides con actividad provitamina A: -caroteno, -caroteno y -criptoxantina, por ser precursores de retinol, nutriente del que depende el ciclo visual para una vision normal. En el presente trabajo se ha llevado a cabo el estudio de los carotenoides de la dieta mas relevantes para la salud ocular humana considerando de forma simultanea parametros relacionados con la ingesta, el estatus y la funcion visual, asi como diversas variables que pueden modificar el estatus nutricional, como son la concentracion de lipidos en sangre, y la bioaccesibilidad de los carotenoides a partir de alimentos de amplio consumo...Fetal rat hepatocytes treated with transforming growth factor beta (TGF-beta) die by apoptosis. However, a subpopulation of them survives and undergoes an epithelial mesenchymal transition (EMT). This transition also occurs upon incubation with fetal bovine serum. We have isolated the subpopulations that undergo EMT (TGF-beta-treated-fetal hepatocytes: TbetaT-FH; serum-treated-fetal hepatocytes: ST-FH) and show that they present high levels of vimentin and Snail expression and lack cytokeratin 18 and E-cadherin. Both TbetaT-FH and ST-FH cells require mitogens to grow and maintain the response to TGF-beta in terms of growth inhibition. However, they lack differentiation markers such as the liver-enriched transcription factors hepatocyte nuclear factor 4 (HNF-4) or HNF-1alpha and express the progenitor marker OV-6. Interestingly, the EMT process confers them resistance to the apoptotic effect of TGF-beta, with cells showing higher levels of active AKT and Bcl-x(L) than fetal hepatocytes. In summary, these cells are refractory to the apoptotic effects of TGF-beta, showing characteristics of liver progenitors and of some hepatocellular carcinoma cells.Memoria de tesis presentada por Luis Vazquez Fonseca, Licenciado en Bioquimica para optar al grado de Doctor. Esta Tesis Doctoral ha sido realizada bajo el programa de doctorado de Biotecnologia y Tecnologia Quimica en el grupo de investigacion del CIBERER U729 en el Centro Andaluz de Biologia del Desarrollo, Area de Biologia Celular del Departamento de Fisiologia, Anatomia y Biologia Celular de la Universidad Pablo de Olavide y bajo la direccion del Dr. Carlos Santos Ocana y el Dr. Placido NavasResumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Poster presentado al 17o Congreso Nacional de la Sociedad Espanola de Neurociencia, celebrado en Alicante del 27 al 30 de septiembre de 2017.The mutations at the bithorax locus produce a transformation of anterior haltere into anterior wing. The bx1 allele presents unusual features when compared with other bx alleles. The phenotype of bx1 homozygotes is temperature sensitive but only with regard to the distal and not to the proximal transformation, thus suggesting two different components in the bithorax transformation. The phenotype of bx1 homozygotes is stronger than that of bx1 over the deletion of the gene, suggesting a trans interaction of the bx1 chromosomes which results in mutual partial inactivation. We show by temperature shift and clonal analysis experiments that the decision on whether to differentiate haltere or wing structures is taken at the end of the proliferation period of the mutant disc.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Poster presentado al XXVII Congreso Nacional de la Asociacion Espanola de Genetica Humana celebrado en Madrid del 10 al 12 de abril de 2013.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Poster presentado en el XI European Meeting on Glial Cells in Health and Disease, celebrado los dias 3 al 6 de julio de 2013 en Berlin (Alemania)Resumen del trabajo presentado al Spanish Society of Biochemistry and Molecular Biology (SEBBM), celebrado en Madrid del 16 al 19 de julio de 2019.Poster presentado en el XII European Meeting on Glial Cells in Health and Disease, celebrado los dias 15 a 18 de julio de 2015 en Bilbao (Espana)Trabajo presentado en el XL Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular. FEBS3+1st Joint Meeting of the French-Portuguese-Spanish Biochemical and Molecular, celebrado en Barcelona (Espana), del 23 al 26 de octubre de 2017Resumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Trabajo presentado en el XII GEIRLI Meeting: New trends in redox biology: a multidisciplinary approach, celebrado en Barcelona (Espana), los dias 4 y 5 de julio de 2019Treatment of nucleosomal particles with dimethylmaleic anhydride, a reagent for protein amino groups, is accompanied by a biphasic release of histones H2A plus H2B; one H2A.H2B dimer is more easily released than the other. This behavior allows the preparation of nucleosomal particles containing only one H2A.H2B dimer, which were complemented with 125I-labeled H2A.H2B. These reconstituted particles, which contain one labeled and one unlabeled H2A.H2B dimer, were treated with the amount of reagent needed to release one of the two H2A.H2B dimers. Radioactivity was equally distributed between residual particles and released proteins, which is consistent with equivalent binding sites in the nucleosomal particle for H2A.H2B dimers, rather than with intrinsically different sites. The asymmetric release of H2A.H2B dimers would be caused by a change in the binding site of one dimer following the release of the other. This behavior might be related to the structural dynamics of nucleosomes.Resumen del trabajo presentado al European Society of Cardiology (ESC) Congress, celebrado en Barcelona (Espana) del 26 al 30 de agosto de 2017.Resumen del poster presentado al 49th European Association for the Study of Diabetes Annual Meeting, celebrado en Barcelona (Espana) del 23 al 27 de septiembre de 2013.-- et al.Trabajo doctoral realizado por Da Rebeca Lapresa Ruiz de Gauna, para optar al grado de doctor por la Universidad de Salamanca.Rationale: Several animal models have been developed to study acute lung injury (ALI); however the majority of these studies are focused on different mechanisms within the acute phase. These models do not allow studying the mechanisms in the later phases or testing any possible long-term treatment. The aim of this study was to develop an experimental ALI model simulating bronchial aspiration of gastric contents with bacterial superinfection with alveolar epithelial damage persisting over time. Methods: Sprague-Dawley rats (200-250g) were anesthetized with isofluorane. ALI was induced by intratracheal instillation of HCl (1 µl/g, 0.1 mol/L pH=1.4) followed by instillation of LPS from Escherichia coli O55:B5 (0, 10, 20, 30 or 40µg/g b.w.) two hours later. Control rats were treated with intratracheal instillations of saline. After 72h, the animals were sacrificed and bronchoalveolar lavage fluid (BALF) was sampled for further analysis of total protein concentration by bicinchoninic acid method. Results: At 72 h, rats suffered a significant loss of weight proportional to the administered dose of LPS (5.6% with 10µg/g b.w, 12.6% with 20µg/g b.w, 14.2% with 30µg/g b.w and 17.7% with 40µg/g b.w). Control rats gained in weight at 72h. LPS at 10, 20, 30 and 40µg/g b.w induced a 1.7, 2.5, 2.9 and 3.4 fold increase in total protein concentration in BAL fluid, respectively, reflecting a substantial increase proportional to the LPS dose. Conclusion: The degree of weight loss and the increase of total protein concentration in BAL fluid in the current model may reflect disease severity and progression. This model would be useful in future for new therapeutical options. Grant acknowledgements: FIS-PI12/02548 and Fundacio Parc Tauli.Resumen del trabajo presentado al European Respiratory Society (ERS) International Congress, celebrado en Paris (Francia) del 15 al 19 de septiembre de 2018.Resumen del trabajo presentado a las 5as Jornadas de Formacion del CIBERES celebradas en Bunyola (Mallorca) del 18 al 19 de octubre de 2012.Resumen del poster presentado al Joint FEPS & XXXVI Spanish Physiological Society Congress (Sociedad Espanola de Ciencias Fisiologicas) celebrado en Santiago de Compostela (Espana) del 8 al 11 de septiembre de 2012.Resumen del trabajo presentado al XIII Congreso de la Sociedad Espanola del Dolor, celebrado en Pamplona del 2 al 4 de junio de 2016.This work was supported by grants FIS-01/1048 and FIS-02/1199 from the Fondo de Investigacion Sanitaria and grant SA-087/01 from Junta de Castilla y Leon.Resumen del poster presentado al Joint Meeting of the American Physiological Society and the Physiological Society, celebrado en Dublin (Irlanda) del 29 al 31 de julio de 2016.Trabajo presentado al 5th International Conference on Phospholipase A2 Mediated Signaling in Translational Medicine celebrado en New Orleans (US) del 20 al 21 de mayo de 2013.Tesis Doctoral presentada por Rebeca Torres Merino para optar al grado de Doctora por la Universidad de Valladolid, Facultad de Medicina: Dpto. de Bioquimica y Biologia Molecular y Fisiologia.Poster presentado al Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO), celebrado en Seattle, Washington (US) del 1 al 5 de mayo de 2016.Resumen del trabajo presentado al 63rd Annual Meeting Biophysical Society, celebrado en Baltimore, Maryland (USA) del 2 al 6 de marzo de 2019.Poster presentado al XXXVII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular, celebrado en Granada del 9 al 12 de septiembre de 2014.Resumen del poster presentado a la 5th Conference on Advances in Molecular Mechanisms Underlying Neurological Disorders (Joint conference of the European Society for Neurochemistry and the Biochemical Society) en la University of Bath (UK) del 23 al 26 de junio de 2013.-- Tambien presentado al 15o Congreso Nacional de la Sociedad Espanola de Neurociencia (SENC) celebrado en Oviedo del 25 al 27 de septiembre de 2013.Resumen del trabajo presentado al XXXVI Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular celebrado en Madrid del 4 al 6 de septiembre de 2013.Resumen del trabajo presentado a la 5th Conference on Advances in Molecular Mechanisms Underlying Neurological Disorders (Joint conference of the European Society for Neurochemistry and the Biochemical Society) en la University of Bath (UK) del 23 al 26 de junio de 2013.Resumen del poster presentado al XXVIII Congreso Nacional de la Sociedad Espanola de diabetes, celebrado en Bilbao del 20 al 22 de abril de 2016.SAF2016-77703-C2-2-R of the Ministerio de Economia y Competitividad, Spain and the European Regional Development Fund (ERDF); AGAUR 2017-SGR106 and the CERCA Programme of the Generalitat de Catalunya; C. Sanfeliu belong to Group 05 of CIBER Epidemiologia y Salud Publica (CIBERESP) of the Instituto de Salud Carlos III, Ministerio de Sanidad y Consumo, Spain