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Hypersecretion of airway mucus is characteristic of several severe lung diseases, particularly those involving chronic inflammation such as asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis (CF) (1). Mucins are the major macromolecular component of mu ...

In chronic inflammatory diseases, the expression of multiple genes, including those for cytokines, chemokines, adhesion molecules, receptors, and inflammatory enzymes, is often upregulated. The problem for many academic or industrial scientists is to elucidate the mechanisms ...

Flow cytometry is an invaluable tool for the analysis of leukocyte populations in inflammation. Flow cytometers, particularly those designed purely for analysis rather than cell sorting, have now become user-friendly machines and are commonplace in many laboratories. Any cell type ...

Airway inflammation is currently the subject of intense research interest concerning both the nature of the inflammatory cells, proteins, and cytokines present. These data are being used to define and assess the severity, cause, prognosis, and response to treatment of airway inflammat ...

During the Phase I/II assessment of new therapies with the potential to suppress eosinophil and neutrophil inflammation, there is a need to assess the peripheral blood pharmacokinetic (PK) and pharmacodynamic (PD) profiles of the drug. This has relevance in respiratory disease since dr ...

A large component of airway inflammatory disease is the recruitment of activated leukocytes (primarily eosinophils and T lymphocytes) from the lung vasculature into the bronchial walls resulting in lung edema. Ultimately, many of the infiltrating leukocytes progress across the a ...

In this chapter, we will describe two different techniques used to trace storage and mobilization of intracellular granule-derived mediator proteins in eosinophils. The first is confocal laser scanning microscopy (CLSM) used to investigate immunofluorescence labeling in cyt ...

Matrix metalloproteinases (MMPs) (1,2) comprise a family of over 20 matrix degrading enzymes believed to be essential for normal development and physiologic tissue remodeling and repair. Abnormal expression of metalloproteinases has been implicated in many destructive proce ...

Immunohistochemistry allows the specific histochemical localization of many diverse classes of organic substances, including inflammatory markers ranging from biogenic amines to macromolecular cytokines and their receptors. Corresponding ligands and their recep ...

Metabolism of arachidonic acid results in a host of biologically active compounds with profound effects on airway inflammation (1). After activation of cellular phospholipases and release of free arachidonic acid, catalyzed insertion of oxygen occurs enzymatically via action of ...

The cysteinyl leukotrienes, comprising leukotriene (LT) C4 and its major metabolites LTD4 and LTE4, are inflammatory lipid mediators derived from metabolism of arachidonic acid by 5-lipoxygenase. These leukotrienes have received considerable attention for their potential ...

The discoveries by Jack Roberts and Jason Morrow of the nonenzymatic oxidation of cell membrane phospholipids to form isoprostanes has revolutionized the field of eicosanoids (1). Prior to their discoveries, it was dogma that the important biologically active eicosanoids were form ...

The regulation of chondrocytes in osteoarthritic cartilage and the expression of specific gene products by these cells during early-onset and late-stage osteoarthritis are not well characterized. With the introduction of cDNA array technology, the measurement of thousands of di ...

The introduction of foreign DNA into mammalian cells is an essential investigative tool in molecular biology. Nonviral approaches to transfection offer the advantage of relatively simple vector design, production, and purification and, for tissue engineering applications, a ...

The major requirement of a successful gene transfer is the efficient delivery of an exogenous therapeutic gene to the appropriate cell type with subsequent high or regulated levels of expression. In this context, viral systems are more efficient than nonviral systems, giving higher leve ...

Changes in chondrocyte metabolism in vitro using different support systems and under different culture conditions were studied with a proteomic approach. Qualitative and quantitative modifications in the synthesis of chondrocyte proteins were investigated using two-dim ...

Flow cytometry has been used as a procedure to characterize the phenotype and function of human articular cartilage cells cultured as monolayers or in gelled artificial matrices. Procedures allowing intact cells with their cell-associated matrix, to be obtained have been described. A ...

A simple and reliable method to measure proteoglycan synthesis by chondrocytes in culture is described. Confluent chondrocytes in 24-well plates are labeled for 24–72 h with 35SO4 2− in the presence of stimulating agents. At the end of treatment, the secretion medium containing radiolabel ...

Cultures of cartilage explants have long been used to study the effects of modulators of extracellular matrix degradation. We present a simple and rapid assay system, based on culture of rabbit cartilage explants, which permits study of the effects of protease inhibitors on proteoglycan d ...

The use of synthetic peptides to generate rabbit polyclonal anticatabolic neoepitope antibodies that can be used to study the presence of defined proteolytic cleavage sites in aggrecan is described. Principles of peptide design and methods for preparation and characterization of ...