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The K/BxN Mouse Model of Inflammatory Arthritis: Theory and Practice

Mice expressing the KRN T cell receptor transgene and the MHC class II molecule Ag7 (K/B�N mice) develop severe inflammatory arthritis, and serum from these mice causes similar arthritis in a wide range of mouse strains, owing to pathogenic autoantibodies to glucose-6-phosphate isomerase ...

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Analysis of Arthritic Lesions in the Del1 Mouse: A Model for Osteoarthritis

Osteoarthritis (OA) is characterised by progressive erosion of articular cartilage with a number of associated degenerative processes within the joint. Animal models of OA provide the only feasible way to systematically study the development and progression of OA, in order to unders ...

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Gene Expression Profiling in Rheumatology

In the last decade, the analysis of gene expression in tissues and cells has evolved from the analysis of a selected set of genes to an efficient high throughput whole-genome screening approach of potentially all genes expressed. Development of sophisticated methodologies such as microa ...

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Differential Display Reverse Transcription-Polymerase Chain Reaction to Identify Novel Biomolecules in Arthritis Research

Differential display is one of the simplest techniques for discovering novel transcripts when comparing gene expression in biological systems. The method can be carried out on small amounts of total RNA and permits the simultaneous comparison of multiple independent samples in a sing ...

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Two-Dimensional Electrophoresis of Proteins Secreted from Articular Cartilage

Two-dimensional electrophoresis (2DE) is a powerful method for separation of complex mixtures of proteins. The standard procedure is not, however, well suited to analysis of articular cartilage, which contains high concentrations of proteoglycans, the polyanionic glycosami ...

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Mapping Lymphocyte Plasma Membrane Proteins: A Proteomic Approach

The pathological importance of tumor necrosis factor (TNF)-α in rheumatoid arthritis (RA) is now widely accepted. Ex vivo data from synovial cell cultures suggest that direct cell contact between activated T-cells and macrophages may be an important driver of macrophage TNF-α producti ...

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In Vivo Phage Display Selection in the Human/SCID Mouse Chimera Model for Defining Synovial Specific Determinants

Phage display has represented a phenomenal technological advance of the last two decades. This technique is a very effective way of producing large numbers (up to 1012) of diverse peptides and proteins (including antibodies), presented as fusion proteins on the viral capsid, that can be used f ...

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Adenoviral Targeting of Signal Transduction Pathways in Synovial Cell Cultures

Methods for high efficiency gene transfer into primary cells of various lineages and disease states are desirable, as they remove the uncertainties associated with using transformed cell lines. Adenoviruses have evolved to deliver their genes into cells with high efficiency and in rec ...

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Overview of Adipose Tissue and Its Role in Obesity and Metabolic Disorders

As the result of its apparent structural and histological simplicity, adipose tissue (AT) functions initially were limited to energy storage, insulation, and thermoregulation. Only decades later was the extraordinarily dynamic role of AT recognized, revealing its participati ...

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Study of Adipose Tissue Gene Expression by In Situ Hybridization

Adipose tissue synthesizes factors involved in the body's homeostasis. Thus, measurements of messenger ribonucleic acid (mRNA) concentrations are important to study the involvement of adipose tissue in various physiological and pathophysiological conditions, in partic ...

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Application of Immunocytochemistry and Immunofluorescence Techniques to Adipose Tissue and Cell Cultures

When isolated from tissue, white adipose cells are round, and their interior is filled with a large (80–120 μm) droplet of stored triglyceride, leaving a thin (1–2-μm) layer of cytoplasm between the lipid droplet and the plasma membrane. Their three-dimensional architecture, together with t ...

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Determination of Lipolysis in Isolated Primary Adipocytes

Lipolysis involves the sequential breakdown of triglycerides into free fatty acids and glycerol. The extent of lipolysis is therefore a key determinant of the energy status of an individual and also dictates insulin resistance. Here, we describe a protocol for estimating lipolysis in mu ...

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Study of Glucose Uptake in Adipose Cells

Glucose is the main metabolic fuel in mammalian cells. Glucose entry into cells is facilitated by a family of ubiquitously expressed glucose transporter proteins. Typically, glucose transporters are localized on the plasma membrane. One notable exception is the glucose transporter ...

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Measurement of Phosphoinositide 3-Kinase and Its Products to Study Adipogenic Signal Transduction

Adipogenesis is an important component of adipose tissue development and growth. Alterations in adipogenesis may promote adipose tissue insulin resistance and inflammation. The ability of preadipocytes to differentiate into mature adipocytes depends on the activation of ph ...

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Generation of Adipose Tissue-Specific Transgenic Mouse Models

Adipose tissue plays a critical role in energy homeostasis, not only in storing triglycerides, but also in responding to nutrient, neural, and hormonal signals, and secreting adipokines that control feeding, thermogenesis, immunity, and neuroendocrine function. It is conceivable ...

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Application of Imaging and Other Noninvasive Techniques in Determining Adipose Tissue Mass

In vivo adipose tissue quantification is an important tool to characterize phenotypes of obesity, especially in the human. The amount and distribution of adipose tissue is associated with many of the adverse consequences of obesity. Recent studies suggest that adipose tissue is not a sing ...

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Adipose Organ Nerves Revealed by Immunohistochemistry#

Brown and white adipose tissue have recently gained prominence as key players in obesity and related health problems, such as type-2 diabetes and cardiovascular disease. Brown adipose tissue-dependent nonshivering thermogenesis significantly affects the body's energy bala ...

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Angiogenesis in Adipose Tissue

Angiogenesis is required for the growth and expansion of both healthy and pathological tissues. The plasticity of the adipose tissue is reflected by its remarkable ability to expand or to reduce in size throughout the adult lifespan. We, and others, have recently shown that expansion of fat mass is ...

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Human Adipose Tissue Blood Flow and Micromanipulation of Human Subcutaneous Blood Flow

Regulation of blood flow in tissues such as skeletal muscle, liver, and adipose tissue is needed to meet the changing local metabolic and physiological demands under varying conditions. In healthy individuals, adipose tissue blood flow (ATBF) is remarkably responsive to meal ingestion, ...

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Studies of Thermogenesis and Mitochondrial Function in Adipose Tissues

Brown and white adipose tissues in mammals have a number of similar properties, such as lipid storage and adipokine production, but also distinctive properties. The energy-storing white adipose tissue has few mitochondria and low oxidative capacity. The heat-producing brown adipose ...

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