Activation of Endothelial Nitric Oxide (eNOS) Occurs through Different Membrane Domains in Endothelial Cells.

Endothelial cells respond to a large range of stimuli including circulating lipoproteins, growth factors and changes in haemodynamic mechanical forces to regulate the activity of endothelial nitric oxide synthase (eNOS) and maintain blood pressure. While many signalling pathways have been mapped, the identities of membrane domains through which these signals are transmitted are less well characterized. Here, we manipulated bovine aortic endothelial cells (BAEC) with cholesterol and the oxysterol 7-ketocholesterol (7KC). Using a range of microscopy techniques including confocal, 2-photon, super-resolution and electron microscopy, we found that sterol enrichment had differential effects on eNOS and caveolin-1 (Cav1) colocalisation, membrane order of the plasma membrane, caveolae numbers and Cav1 clustering. We found a correlation between cholesterol-induced condensation of the plasma membrane and enhanced high density lipoprotein (HDL)-induced eNOS activity and phosphorylation suggesting that cholesterol domains, but not individual caveolae, mediate HDL stimulation of eNOS. Vascular endothelial growth factor (VEGF)-induced and shear stress-induced eNOS activity was relatively independent of membrane order and may be predominantly controlled by the number of caveolae on the cell surface. Taken together, our data suggest that signals that activate and phosphorylate eNOS are transmitted through distinct membrane domains in endothelial cells.

Plataforma web para la promociónde contenidos sobre la Salud Mentalde la Comunidad Valenciana / A web platform of the Valencia local government for the promotion of contents on Mental Health

El desarrollo de aplicaciones telemáticas como soporte para la divulgaciónde contenidos sobre Salud Mental en España se ha convertidoen la actualidad en un nuevo ámbito de investigación socialcentrado en el desarrollo de plataformas digitales de informaciónque permitan ofrecer asesoramiento e información directa, no sóloa pacientes, sino a la comunidad de profesionales sanitarios.Hasta este momento, la web como medio y los contenidos digitales decarácter sanitario, como base, han demostrado su uso y eficiencia ennumerosas investigaciones internacionales constituyendo una mejoraindiscutible en la difusión del conocimiento global de la medicina.A nivel nacional, la mayoría de los contenidos digitales y de las webde Salud Mental se hallan inmersas en escenarios digitales de caráctersanitario globalizado, plataformas digitales de instituciones y asociacionesde profesionales sanitarios o bien iniciativas privadas conun fin determinado. Hay muy pocas plataformas a nivel nacional quetrabajen en la difusión de contenidos sobre Salud Mental y cuya gestiónsea realizada por los agentes sanitarios, pacientes y/o familiaresque lo deseen, aportando contenidos de forma desinteresada.El objetivo de la plataforma web para la promoción de contenidos sobrela Salud Mental de la Comunidad Valenciana, como iniciativa delÁrea 12 de Salud Mental del Hospital San Francisco de Borja, es servircomo referente para la consulta y gestión de la información digital (..) (AU)

The development of telematic applications to support the disseminationof mental health contents in Spain has become a new area ofsocial research focused on the development of digital information platformsthat offer the possibility of providing advice and direct informationnot only to patients, but to the community of health careprofessionals as well.Up until now, the internet as a means and health care-related digitalcontents as a basis, have demonstrated their use and efficiency in numerousinternational research projects, resulting in undeniable improvementof the worldwide dissemination of medical knowledge.At an international level, most of these digital contents and MentalHealth websites are immersed in globalized health care-related digitalstages, digital platforms of institutions and associations of healthcare professionals or private initiatives with a specific objective. Thereare very few platforms at a national level that work to disseminateMental Health contents and the management of which is carriedout by health care agents, patients and/or family members that sodesire it, contributing contents in a non-lucrative manner.The objective of this Valencian internet-based platform for the promotionof Mental Health contents, as an initiative of Area 12 of MentalHealth of the San Francisco de Borja Hospital, is to serve as a referencefor consulting and management of digital information onthe latest news and developments in the field of research and healthcare in Mental Health, at a regional and national level.This article presents the Mental Health platform that has been createdand offers the possibility of including general information on thedifferent topics related with documentation of the latest researchworks, events and courses, etc., as well as a participative environmentfor users who register, where they will be able to share experiencesacquired within professional and social settings (..) (AU)

Sterol regulatory element binding proteins downregulate LDL receptor-related protein (LRP1) expression and LRP1-mediated aggregated LDL uptake by human macrophages.

OBJECTIVE: In the extracellular intima, extracellular matrix proteoglycans favor LDL retention and aggregation (agLDL). In contrast to native LDL (nLDL), agLDL induces high intracellular cholesteryl ester (CE) accumulation in macrophages. It has been suggested that LDL receptor-related protein (LRP1) is involved in agLDL binding and internalization by macrophages. The aim of this work was to analyze whether sterol regulatory element binding proteins (SREBPs) modulate LRP1 expression and LRP1-mediated agLDL uptake by human monocyte-derived macrophages (HMDM). METHODS AND RESULTS: The treatment of HMDM with small anti-LRP1 interfering RNA (siRNA-LRP1) led to the specific inhibition of LRP1 mRNA expression and also to the inhibition of LRP1 protein expression in these cells. In siRNA-LRP1-treated HMDM, CE accumulation from agLDL uptake (84.66+/-5 microg CE/mg protein) was reduced by 95.76+/-5.22%. This suggests that LRP1 plays a pivotal role in agLDL uptake by HMDM. N-acetyl-leucyl-leucyl-norleucinal (ALLN), an inhibitor of SREBP catabolism, maintained high levels of active SREBP-2 and SREBP-1 even in the presence of nLDL and agLDL. Therefore, ALLN induced LDL receptor (LDLR) upregulation. Concomitantly, a strong downregulation of LRP1 mRNA and LRP1 protein was observed in ALLN-treated macrophages. By decreasing LRP1 expression levels, ALLN reduced CE accumulation from agLDL at all tested concentrations. CONCLUSIONS: These results suggest that high levels of active SREBPs downregulate LRP1 expression and intracellular CE accumulation in HMDM.