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Isolation, Culture, and Functional Analysis of Adult Mouse Cardiomyocytes

The pathogenesis of heart disease and the development of myocardial failure are highly dependent upon the cardiomyocyte—the basic contractile cell within the heart. Understanding and elucidating the complex networks that regulate cardiomyocyte function are central to the dev ...

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Mechanical Measurement of Contractile Function of Isolated Ventricular Myocytes

Isolation of ventricular myocytes from all species of animals has revolutionized the field of cardiovascular research, allowing the assessment of true cardiac effects of drugs, treatments, and so on. With recent advances in physiology at the cellular level, direct assessment of isola ...

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Studying Ischemia and Reperfusion in Isolated Neonatal Rat Ventricular Myocytes Using Coverslip Hypoxia

In vitro experimental models designed to study the effects of hypoxia and ischemia typically employ oxygen-depleted media and/or hypoxic chambers. These approaches, however, allow for metabolites to diffuse away into a large volume and may not replicate the local buildup of metabolic b ...

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Isolation and Functional Studies of Rat Aortic Smooth Muscle Cells

Migration, proliferation, and collagen synthesis by vascular smooth muscle cells are thought to be key events involved in the pathogenesis of cardiovascular disease. Following endothelial injury, smooth muscle cells (SMCs) in the intima of the blood vessels assume a synthetic, pro-m ...

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Rat Carotid Artery Balloon Injury Model

Numerous and diverse experimental animal models have been used over the years to examine reactions to various forms of blood vessel disease and/or injury across species and in multiple vascular beds in a cumulative effort to relate these findings to the human condition. In this context, the rat c ...

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Detection of Reactive Oxygen Species and Nitric Oxide in Vascular Cells and Tissues: Comparison of Sensitivity and Specificity

Reactive oxygen and nitrogen species are thought to contribute to pathogenesis of many cardiovascular diseases including hypertension, atherosclerosis, restenosis, heart failure, and diabetic vascular complications. Some of these reactive oxygen species also play an imp ...

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Assessment of Protein Glycoxidation in Ventricular Tissues

Advanced glycation end products are permanently modified protein derivatives formed in the presence of reducing sugars, such as glucose, fructose, hexose-phosphates, trioses, and triose-phosphates by non-enzymatic glycation and oxidation (“glycoxidation”) reactions a ...

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Two-Dimensional Gel Electrophoresis in Platelet Proteomics Research

Proteomics technology allows a comprehensive and efficient analysis of the proteome and has become an indispensable tool in biomedical research. Since the late 80s, advances on mass spectrometry (MS) instrumentation and techniques have revolutionized the way proteins can be anal ...

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Assessment of PI-3 Kinase and Akt in Ischemic Heart Diseases in Diabetes

Diabetes mellitus is the most common disease in Westernized countries in large part because of the rising prevalence of obesity and physical inactivity. In addition, diabetes mellitus is an important risk factor for both heart failure and ischemic heart disease. As insulin resistance is k ...

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Histological and Morphometric Analyses for Rat Carotid Balloon Injury Model

Experiments aimed at analyzing the response of blood vessels to mechanical injury and ensuing remodeling responses often employ the highly characterized carotid artery balloon injury model in laboratory rats. This approach utilizes luminal insertion of a balloon embolectomy c ...

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Stem Cell Therapy in the Heart and Vasculature

Stem cell therapy is a progressive approach to a pervasive clinical problem; cardiovascular disease is the number 1 killer in the USA and other developed countries, and aspects of it are amenable to stem cell therapy. Many types of stem cells have been used in treating the heart during myocardial inf ...

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Plaque Rupture Model in Mice

It is widely believed that rupture of a vulnerable atherosclerotic plaque leads to acute coronary events and stroke. However, the exact mechanisms involved in the plaque rupture remain unknown. Pathological animal models are valuable in the research on human disease mechanism, their th ...

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Immunostaining of Mouse Atherosclerotic Lesions

Atherosclerotic lesions develop through interactions with diverse cell types whose functions are determined by a complex array of regulators. Immunostaining is now a commonly applied technique to identify these numerous cell types and regulators in lesions. The principle of the te ...

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Cytogenetics Analysis

The establishment of reliable and meaningful chromosomal (cytogenetic, karyotypic) changes in hematological disorders, primarily the leukemias and lymphomas, must be based on the examination of the involved cells or tissues. Thus, in the case of the leukemias bone marrow (BM) aspira ...

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FISH Analysis

In situ hybridization of specific DNA or RNA sequences to cellular targets was developed over 20 yr ago (1,2). The early techniques employed isotopically labeled probes and subsequent autoradiographic detection using a photographic emulsion overlying the metaphase chromosomes, ...

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Comparative Genomic Hybridization for the Analysis of Leukemias and Lymphomas

Cytogenetic methods have become increasingly important tools for both research in hematological malignancies and for the diagnostic workup of leukemias and lymphomas. The knowledge about specific chromosomal aberrations has been an essential prerequisite for the identif ...

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Spectral Karyotyping (SKY) of Hematologic Malignancies

The discovery of the Philadelphia chromosome in chronic myeloid leukemia (CML) by Novell and Hungerford in 1960 (1), the subsequent clarification of this chromosomal abnormality as areciprocal translocation t(9;22)(q34;q1 1) by Rowley in 1973 (2), the identification of the genes invol ...

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Molecular Genetic Methods in Diagnosis and Treatment

During the past 20 yr, molecular biology and molecular genetics have provided techniques and procedures that allow the precise diagnoses of leukemias and lymphomas (for a review, see ref. 1). These methodologies have provided powerful, robust, and precise characterizations of genetic ...

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The Blood-CSF Barrier in Culture: Development of a Primary Culture and Transepithelial Transport Model from Choroidal Epithelial Cells

The chemical stability of the central nervous system (CNS) is safeguarded by two major barrier systems that separate the systemic circulation from the cerebral compartment. Within the cerebral compartment, the interstitial fluid (ISF) flows between neurons and the cerebrospinal f ...

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An In Vitro Model of Differentiated Human Airway Epithelia: Methods for Establishing Primary Cultures

The human airway epithelium forms a barrier between the external and internal environments, separating air from the interstitial space. However, it also serves many other functions. By active transepithelial transport of electrolytes, it controls the composition and quantity of t ...

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