Full Description
Asthma has rapidly become one of the most common chronic illnesses of the Western world, and its prevalence continues to rise, with the proportion of patients with more severe diseases also increasing. Faced with this problem, more researchers are focusing on the causes, mechanisms, and pathophysiology of asthma. The major hopes are that more effective drugs will become available and that preventive measures can be instituted. Increasingly, molecular and cell bi- ogy approaches are being used to characterize and understand the mechanisms of the inflammatory process that is typical of the asthmatic airway. This volume on Asthma: Mechanisms and Protocols in the Methods in Molecular Medicineseries provides an overview of the molecular mechanisms involved in asthma by p- viding extensive protocols that are being used in asthma research. Briefly, it c- ers details of methods for obtaining cells from the airways, analysis of gene and protein expression in the limited clinical samples from asthmatic airways, use of molecular and cellular tools for studying cytokine expression and release, studies of asthma-related genes and genetic polymorphisms, and understanding the - fects of asthma treatments. With such coverage, the volume ties in several dis- plines, including allergy and immunology, cell biology, pharmacology, and histology. We have continued in the spirit of the series to provide a bench book for day-to-day use.
Contents
Asthma: Application of Cell and Molecular Biology Techniques to Unravel Causes and Pathophysiological Mechanisms.- Culture of Normal Human Airway Epithelial Cells and Measurement of Mucin Synthesis and Secretion.- Brush Biopsy and Culture of Airway Epithelial Cells: ?-Adrenergic Receptor System Function.- Isolation and Characterization of Human Airway Fibroblasts in Culture.- Flow Cytometric Analysis of Blood Monocytes and Alveolar Macrophages.- Expression of IL-10 and GM-CSF in Blood Monocytes and Alveolar Macrophages.- MAP Kinase Expression in Eosinophils.- Human Eosinophil Isolation and the Measurement of Apoptosis.- Adhesion of T-Cells to Airway Smooth Muscle Cells.- Proliferation of Circulating T-Cells.- Differential Display Analysis of Inflammatory Genes in Airway Epithelium.- Analysis of Transcription Factor Activation: NF?B as a Regulator of Inflammatory Genes in Epithelial Cells.- Transient Transgenic Approaches for Investigating the Role of GM-CSF in Pulmonary Inflammation and Immune Diseases.- Quantitative Analysis of RANTES mRNA in Endobronchial Biopsies Using Polymerase Chain Reaction.- Intracellular Cytokine Staining for Analysis by Flow Cytometry.- Localization of IL-4 and IL-5 mRNA in Bronchial Biopsies.- Immunohistochemical Analysis of Adhesion Molecules in Airway Biopsies.- Site-Directed Mutagenesis in the Investigation of ?-Adrenoreceptor Exosite.- Methods for Determining ?2-Adrenoreceptor Genotype.- Modulation of ?-Adrenoreceptor Expression in the Airways.- Chemical Mutational Analysis of Glucocorticoid Receptor.- Histone Acetylation and Deacetylation.- Genome-Wide Search for Asthma Genes.
