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The successful combination of highly sensitive mass spectrometry and pre-fractionation techniques has provided a powerful tool to detect dynamic changes in low abundant regulatory proteins at the organelle level. Subcellular fractionation, being flexible, adjustable (bo ...

Proteomic analysis of heart tissue is complicated by the large dynamic range of its proteins. The most abundant proteins are the myofilament proteins, which comprise the contractile apparatus. This chapter describes a protocol for fractionation of heart tissue that extracts the myof ...

Cardiac troponin I (cTnI) is a key regulator of cardiac muscle contraction. Upon myocardial cell injury, cTnI is lost from the cardiac myocyte and can be detected in serum, in some cases with specific disease-induced modifications, making it an important diagnostic marker for acute myocardi ...

Atherosclerosis is one of the most common causes of death in developed countries. Atherosclerosis is an inflammatory process that results in the development of complex lesions or plaques that protrude into the arterial lumen. Plaque rupture and thrombosis result in the acute clinical co ...

Endothelial cells form a continuous monolayer lining the inside face of all blood vessels, and present the ability to selectively control vascular permeability. The endothelium is involved in a wide variety of normal physiological and pathological processes. The endothelial dysf ...

High-throughput immunoblotting or Western array screening of tissue is a unique advancement in proteomics that may help researchers in their quest to elucidate the proteins and signaling pathways involved in complex human pathologies such as atherosclerosis. The technique enta ...

Atherosclerosis is one of the main causes of death in developed countries. Atheroma plaque formation is promoted by the interaction between the cells conforming the arterial wall, smooth muscle cells, and endothelial cells, together with lipoproteins and inflammatory cells (mainly ...

Adenoviruses are viruses with genomes of double-stranded DNA approx 36 kb in size. There are more than 100 different serotypes that have been identified, 47 of them of human origin. Human serotypes have been grouped into six subgenera (A-F) and whereas sequence similarity between genera is low, o ...

Adeno-associated virus (AAV) was discovered about 30 yr ago as a contaminant of adenovirus preparations. Since its discovery, researchers have described many unique characteristics of AAV biology that have made it attractive as a potential vector for gene therapy. For example, AAV is not pa ...

Nitric oxide (NO) produced in endothelial cells exerts important roles in the vascular system. In recent years, posttranslational modifications induced by NO have been increasingly studied and, among them, cysteine modification by S-nitrosylation (also called S-nitrosation) h ...

Smooth muscle cells (SMCs) play a crucial role in cardiovascular diseases. Proteomic analysis using two-dimensional gel electrophoresis (2DE) associated with mass spectrometry allows characterization of the proteome and secretome of human smooth muscle. The presence of a dist ...

Relaxation of smooth muscle can occur through agonists (such as nitric oxide) that activate guanylyl cyclase and stimulate the production of cGMP, activating its target, cGMP-dependent protein kinase (PKG). This kinase can raise the Ca2+ threshold for contraction, thus causing Ca2+ des ...

Apoptosis of vascular smooth muscle cells (VSMC) plays an important role in remodeling the vessel walls, one of the major determinants of long-term blood pressure elevation and an independent risk factor for cardiovascular morbidity and mortality. Apoptosis in VSMC can be inhibited by in ...

Cardiac myocytes are activated by hormonal and mechanical signals and respond in a variety of ways, from altering contractile function to inducing cardio-protection and growth responses. The use of genetic mouse models allows one to examine the role of cardiac-specific and other genes in c ...

Foam cells are characteristic pathological cells in the lesions of atherosclerosis. Previous works have established macrophage-derived foam cell model to study the central role of the foam cells, and analyzed the protein expression profiles in foam cells. The reported in vitro foam cell ...

Proteomics offers the opportunity to comprehensively investigate the anucleate platelet. Here, we present a detailed procedure for enrichment by immunoprecipitation, using the monoclonal antibody 4G10, of the dynamic phosphotyrosine proteome of human platelets. Such an ap ...

This chapter describes an approach to isolate, separate, and identify the contents of the platelet releasate, a fraction highly enriched for platelet granular and exosomal contents. Investigation into such a fraction will improve our understanding of platelet interactions with ot ...

We describe a simple method for isolation of human blood monocytes with the high purity required for proteomic analysis, which avoids contamination by other blood cells (platelets and lymphocytes) and the most abundant plasma proteins (albumin and immunoglobulins). Blood monocytes ...

Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (TOF-MS) allows for rapid differential proteomic analysis of various experimental conditions. In the first step of retention chromatography, the proteome to be analyzed is fractionated bas ...

With a mean weight of 1500 g containing around 10 billion neurons, the adult brain represents about 2% of the total body mass, but requires 20% of the total energy produced. It consumes continuously 150 g of glucose and 72 L of oxygen every 24 h. A few minutes interruption of this supply can lead to dramatic brain dam ...