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Hospital de Órbigo, Spain

Velez P.,University of Santiago de Compostela | Ocaranza-Sanchez R.,Institute Investigacion Sanitaria Of Santiago Of Compostela Idis Santiago Of Compostela Spain | Lopez-Otero D.,Institute Investigacion Sanitaria Of Santiago Of Compostela Idis Santiago Of Compostela Spain | Grigorian-Shamagian L.,Cedars Sinai Medical Center | And 4 more authors.
Proteomics - Clinical Applications | Year: 2016

Purpose: Platelets play a fundamental role in the atherothrombotic events that lead to an acute myocardial infarction. In the present study we compared the proteome of intracoronary and peripheral arterial platelets from ST-elevation myocardial infarction (STEMI) patients in the search for potential platelet biomarkers/drug targets related to what is happening at the culprit site. Experimental design: Ten STEMI patients were recruited and blood collected from the occluded coronary artery, at the culprit site, in the moment of reperfusion. Systemic blood obtained from the radial artery of the same patients was used as control. Proteome analysis was based on high-resolution 2D-DIGE and mass spectrometry. Validations were by western blotting in a group of 11 patients. Results: Sixteen differentially regulated protein features were identified, corresponding to 15 ORFs, mostly related to cytoskeletal and signaling proteins. We demonstrate the up-regulation of integrin αIIb (ITA2B), the adapter Src kinase-associated phosphoprotein-2 (SKAP2), and thrombospondin-1 isoforms in intracoronary platelets. Conclusion and clinical relevance: This study constitutes the first analyzing in detail the proteome of arterial intracoronary platelets from STEMI patients. We show variations in the platelet proteome when comparing intracoronary and peripheral platelets. Observed differences might be related to platelet activation events at the culprit site. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source

De La Cuesta F.,Laboratorio Of Fisiopatologia Vascular | Alvarez-Llamas G.,Institute Investigacion Sanitaria Fundacion Jimenez Diaz IIS FJD | Gil-Dones F.,Hospital Nacional de Paraplejicos | Darde V.M.,Proteomic Unit | And 5 more authors.
Methods in Molecular Biology | Year: 2013

One of the major challenges in cardiovascular medicine is to identify candidate biomarker proteins. Between different proteomics studies, the secretome is a valuable tool in the search for biomarkers locally released by a studied tissue and remains particularly important while working with vascular tissues, since the secreted proteins would be probably shed to the blood. In this chapter, we described a method to validate the origin of secreted proteins in human aortic valves, demonstrating their synthesis and release by the tissue and ruling out blood origin. © Springer Science+Business Media New York 2013. Source

de la Torre-Escudero E.,CSIC - Institute of Natural Resources and Agriculture Biology of Salamanca | Valero L.,Proteomic Unit | Perez-Sanchez R.,CSIC - Institute of Natural Resources and Agriculture Biology of Salamanca | Manzano-Roman R.,CSIC - Institute of Natural Resources and Agriculture Biology of Salamanca | Oleaga A.,CSIC - Institute of Natural Resources and Agriculture Biology of Salamanca
Journal of Proteomics | Year: 2012

Schistosomes are blood parasites adapted to their intravascular habitat that have evolved mechanisms to evade the immune and hemostatic responses of their hosts. It has been observed that the schistosome can regulate the endothelium function along the infection, which contributes to modulation of host defensive responses and parasite survival. The purpose of this work was the analysis of the changes induced by Schistosoma bovis adult worms in the proteome expressed by infected mice on the endothelial surface of their portal vein. With this aim, we have utilized a methodology that allows the purification, identification and relative quantification of endothelial cell surface proteins after their selective in vivo labeling with biotin. Trypsin digestion of the biotinylated proteins and subsequent liquid chromatography and tandem mass spectrometry analysis (LC-MS/MS) resulted in the identification of a total 127 non-redundant proteins. All these proteins have been classified according to their function and cellular location, and the differences between S. bovis-infected and non-infected mice in their endothelial surface proteomes have been analyzed. The present work provides the first data on the proteome of the endothelial surface of the portal vein, and identifies some of the changes induced in it after an infection by S. bovis. © 2012 Elsevier B.V. Source

Villacorta P.J.,University of Granada | Salmeron-Garcia A.,Hospitales Universitarios Of Granada | Pelta D.A.,University of Granada | Cabeza J.,Hospitales Universitarios Of Granada | And 2 more authors.
Analyst | Year: 2015

We evaluated the use of the peptide mass fingerprint (PMF) obtained by matrix assisted laser desorption and ionization (MALDI) time-of-flight mass spectrometry (TOF-MS) to track changes in the structure of a protein. The first problem we had to overcome was the inherent complexity of the PMF, which makes it difficult to compare. We dealt with this problem by developing a cluster-based comparison algorithm which takes into account the proportional error made by the mass spectrometer. This procedure involves grouping together similar masses in an intelligent manner, so that we can determine which data correspond to the same peptide (any slight differences can be explained as experimental errors), and which of them are too different and thus more likely to represent different peptides. The proposed algorithm was applied to track changes in a commercially available monoclonal antibody (mAb), namely rituximab (RTX), prepared under the usual hospital conditions and stored refrigerated (4°C) and frozen (-20°C) for a long term study. PMFs were obtained periodically over three months. For each checked time, five replicates of the PMFs were obtained in order to evaluate the similarities between them by means of the occurrences of the particular peptides (m/z). After applying the algorithm to the PMF, different approaches were used to analyse the results. Surprisingly, all of them suggested that there were no differences between the two storage conditions tested, i.e. the RTX samples were almost equally well preserved when stored refrigerated at 4°C or frozen at -20°C. The cluster-based methodology is new in protein mass spectrometry and could be useful as an easy test for major changes in proteins and biopharmaceutics for diverse applications in industry and other fields, and could provide additional stability data in relation to the practical use of anticancer drugs. This journal is © The Royal Society of Chemistry. Source

Gil-Dones F.,Hospital Nacional de Paraplejicos | Darde V.M.,Proteomic Unit | Vivanco F.,IIIS Fundacion Jimenez Diaz | Vivanco F.,Complutense University of Madrid | Barderas M.G.,Hospital Nacional de Paraplejicos
Methods in Molecular Biology | Year: 2013

Calcified aortic valve disease is a slowly progressive disorder that ranges from mild valve thickening with no obstruction of bloodflow, known as aortic sclerosis, to severe calcification with impaired leaflet motion or aortic stenosis. Until now, aortic stenosis (AS) was thought to result from aging and "wear and tear" of the aortic valve, but nowadays, it is known that it presents the same risk factors as atherosclerosis and cardiovascular diseases. A proteomic analysis of plasma could permit to identify the changes in protein expression induced by AS in this biological sample. However, the characterization of human plasma proteome is a very complicated task, due to the wide dynamic range of concentration that separates the most abundant proteins and the less common ones (10-12 orders of magnitude). For this reason, plasma analysis requires pre-fractionation methods, and several such techniques are currently used to deplete albumin and other abundant plasma proteins. In this work we describe two different and optimized protocols to decrease the plasma proteome complexity for proteomic analysis. With this, comprehensive and systematic characterization of the plasma proteome in the healthy and diseased aortic stenosis (AS) state will greatly facilitate the development of "useful" biomarkers for early disease detection, clinical diagnosis, and therapy. © Springer Science+Business Media New York 2013. Source

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