ABSTRACT
BACKGROUND: The creation of the Biobank, a resource pertaining to the Spanish Renal Research Network (REDinREN) promotes advances in clinical research on kidney disease in Spain. The Biobank's aims are to generate an archive of clinical samples and associated data, furnish those samples to research teams, and coordinate with European biobanks. METHOD: Applicable legislation had to be complied with in order to launch the Biobank project (Biomedical Research Law, Data Protection Law and Biological Sample Transport Regulations). A strict work protocol and a new database for the Network's clinical data were also implemented. RESULTS: Over time, the Biobank has acquired additional infrastructure and qualified personnel. In 2010, 2953 new patient samples were collected, giving a total of 37,043 stored vials containing different types of samples. Furthermore, the Biobank is currently participating in eleven research projects. DISCUSSION: Although the Biobank was originally designed for REDinREN use, we must take joint action to make this biological sample storage system and the many possibilities it offers available to the entire nephrological community with a view to promoting kidney disease research.
Subject(s)
Biological Specimen Banks/organization & administration , Kidney Diseases , Biomedical Research , Humans , SpainABSTRACT
Antecedentes: La creación del Biobanco, como una plataforma dentro de la Red de Investigación Renal (REDinREN), impulsa el avance de la investigación clínica de la enfermedad renal en España. Los objetivos del Biobanco son la generación del archivo de muestras clínicas y de datos asociados, para su cesión a los grupos de investigación, y la coordinación con biobancos europeos. Métodos: Para su puesta en marcha, fue necesaria la implementación de la normativa vigente (Ley de Investigación Biomédica y de Protección de Datos y la Normativa del Transporte de Sustancias Biológicas), un estricto protocolo de trabajo y la creación de una base de datos clínicos de la Red. Resultados: En su evolución, el Biobanco ha adquirido infraestructura y personal cualificado, lo que permitió que en el año 2010 se obtuviera un total de 2.953 pacientes, lo que hace un total de 37.043 viales almacenados con muestras de diferentes naturalezas. Además, hasta la fecha, el Biobanco está incluido en 11 proyectos de investigación. Discusión: Aunque el Biobanco fue diseñado como una plataforma de soporte de la REDinREN, es necesario con una acción conjunta poner a disposición de toda la comunidad científica nefrológica las posibilidades que otorga este sistema de almacenamiento de muestras biológicas para potenciar la investigación de la enfermedad renal (AU)
Background: The Biobank creation in Network for the Kidney Research (REDinREN) promotes the advance of kidney disease clinic research in Spain. The Biobank's aims are to generate an archive of clinical samples and associated data, furnish those samples to research teams, and coordinate with European biobanks. Methods: In the beginning were indispensable the implementation of in force normative (Bio-medic Investigation Law, Dates Protect Law and Biologic Samples Transport Normative), a work protocol and the creation of medical database of the Red. Results: The Biobank growth included infastructures and qualified personnel. In 2010 year, the patien samples increased in 2953 achieved 37,042 vials of differect nature stored. Mereover in this moment the Biobank participates in eleventh research projects. Discussion: Even if the Biobank was designed for the support to REDinREN is neccesary to opening his biologic samples to all scientific nephrology community for promote the kidney disease research (AU)
Subject(s)
Humans , Biological Specimen Banks/organization & administration , Renal Insufficiency, Chronic , Specimen Handling/methods , Science and Technology Information NetworksABSTRACT
RATIONALE: Atherosclerotic lesions develop in regions of disturbed flow, whereas laminar flow protects from atherogenesis; however, the mechanisms involved are not completely elucidated. Integrins are mechanosensors of shear stress in endothelial cells, and integrin-linked kinase (ILK) is important for blood vessel integrity and cardiovascular development. OBJECTIVES: To explore the role of ILK in vascular function by studying conditionally ILK-deficient (cKO) mice and human atherosclerotic arteries. RESULTS: ILK expression was detected in the endothelial cell layer of nonatherosclerotic vessels but was absent from the endothelium of atherosclerotic arteries. Live ultrasound imaging revealed that acetylcholine-mediated vasodilatation was impaired in cKO mice. These mice exhibited lowered agonist-induced nitric oxide synthase (NOS) activity and decreased cyclic guanosine monophosphate and nitrite production. ILK deletion caused endothelial NOS (eNOS) uncoupling, reflected in reduced tetrahydrobiopterin (BH4) levels, increased BH2 levels, decreased dihydrofolate reductase expression, and increased eNOS-dependent generation of superoxide accompanied by extensive vascular protein nitration. ILK reexpression prevented eNOS uncoupling in cKO cells, whereas superoxide formation was unaffected by ILK depletion in eNOS-KO cells, indicating eNOS as a primary source of superoxide anion. eNOS and ILK coimmunoprecipitated in aortic lysates from control animals, and eNOS-ILK-shock protein 90 interaction was detected in human normal mammary arteries but was absent from human atherosclerotic carotid arteries. eNOS-ILK interaction in endothelial cells was prevented by geldanamycin, suggesting heat shock protein 90 as a binding partner. CONCLUSIONS: Our results identify ILK as a regulatory partner of eNOS in vivo that prevents eNOS uncoupling, and suggest ILK as a therapeutic target for prevention of endothelial dysfunction related to shear stress-induced vascular diseases.
Subject(s)
Arteries/physiopathology , Atherosclerosis/physiopathology , Endothelium, Vascular/physiopathology , Nitric Oxide Synthase Type III/physiology , Protein Serine-Threonine Kinases/physiology , Vasomotor System/physiology , Aged , Aged, 80 and over , Animals , Cattle , Cells, Cultured , Cyclic GMP/physiology , Disease Models, Animal , Down-Regulation/physiology , Female , HSP90 Heat-Shock Proteins/physiology , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Middle Aged , Nitric Oxide/physiology , Signal Transduction/physiologyABSTRACT
OBJECTIVE: Hydrogen peroxide (H(2)O(2)) is an important mediator in the vasculature, but its role in the regulation of soluble guanylate cyclase (sGC) activity and expression is not completely understood. The aim of this study was to test the effect of H(2)O(2) on sGC expression and function and to explore the molecular mechanism involved. METHODS AND RESULTS: H(2)O(2) increased sGCß1 protein steady-state levels in rat aorta and aortic smooth muscle cells (RASMCs) in a time- and dose-dependent manner, and this effect was blocked by catalase. sGCα2 expression increased along with ß1 subunit, whereas α1 subunit remained unchanged. Vascular relaxation to an NO donor (sodium nitroprusside) was enhanced by H(2)O(2), and it was prevented by ODQ (sGC inhibitor). cGMP production in both freshly isolated vessels and RASMCs exposed to H(2)O(2) was greatly increased after sodium nitroprusside treatment. The H(2)O(2)-dependent sGCß1 upregulation was attributable to sGCß1 mRNA stabilization, conditioned by the translocation of the mRNA-binding protein HuR from the nucleus to the cytosol, and the increased mRNA binding of HuR to the sGCß1 3' untranslated region. HuR silencing reversed the effects of H(2)O(2) on sGCß1 levels and cGMP synthesis. CONCLUSIONS: Our results identify H(2)O(2) as an endogenous mediator contributing to the regulation of vascular tone and point to a key role of HuR in sGCß1 mRNA stabilization.
