Cystic Fibrosis Methods and Protocols
(Sprache: Englisch)
Since the cloning of the cystic fibrosis transmembrane conductance re- lator (CFTR) nearly a decade ago, cystic fibrosis (CF) research has witnessed a dramatic expansion into new scientific areas. Basic researchers, clinicians, and patients increasingly...
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Klappentext zu „Cystic Fibrosis Methods and Protocols “
Since the cloning of the cystic fibrosis transmembrane conductance re- lator (CFTR) nearly a decade ago, cystic fibrosis (CF) research has witnessed a dramatic expansion into new scientific areas. Basic researchers, clinicians, and patients increasingly rely on fundamental techniques of genetics, molecular biology, electrophysiology, biochemistry, cell biology, microbiology, and immunology to understand the molecular basis of this complex disease. Research into the pathophysiology of CF has established numerous paradigms of ion channel dysfunction that extend from inflammation and infection in the airways of patients to basic mechanisms of protein processing and regulation in intracellular components. With these rapid advances has come an increasing need for research scientists to understand and utilize a growing array of basic laboratory tools. This volume of Methods in Molecular Medicine, Cystic Fibrosis Methods and Protocols satisfies that need by providing detailed protocols for the laboratory techniques used throughout CF research. From electrophysiology and cell biology, to animal models and gene therapy, the comprehensive set of methods covered here provide step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Contributions have been chosen to reflect the rich diversity of techniques and to provide a cohesive framework for understanding challenges that are currently at the forefront of CF research. It is hoped that this volume will serve as a valuable reference that will not only foster interdisciplinary investigations into current problems encountered in CF, but also facilitate the translation of new scientific discoveries into clinical solutions.
Inhaltsverzeichnis zu „Cystic Fibrosis Methods and Protocols “
Genetics of Cystic Fibrosis.- CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel.- cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis.- Natural Animal Models of Human Genetic Diseases.- CFTR structure and Function:.- Electrophysiological Approach to Studying CFTR.- Quantitative Analysis of ATP-Dependent Gating of CFTR.- CFTR Regulation by Phosphorylation.- Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells.- Transepithelial Impedance Analysis of Chloride Secretion.- Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein.- Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method.- Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function.- Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells.- Immunolocalization of CFTR in Intact Tissue and Cultured Cells.- Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy.- CFTR Folding and Maturation in Cells.- Isolation of CFTR.- CFTR Expression and ER-Associated Degradation in Yeast.- Manipulating the Folding Pathway of ?F508 CFTR Using Chemical Chaperones.- CFTR Degradation and Aggregation.- In Vitro Reconstitution of CFTR Biogenesis and Degradation.- In Vitro CFTR Folding Assays.- Analysis of CFTR Endocytosis by Cell Surface Biotinylation.- CFTR Regulation of ENaC.- Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR.- Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton.- CFTR-Associated ATP Transport and Release.- Pathophysiology of Cystic Fibrosis.- Inflammatory Mediators in CF Patients.- Bacterial Colonization andInfection in the CF Lung.- Antimicrobial Peptides and Proteins in the CF Airway.- Bacterial-Epithelial Interactions.- Thin-Film Measurements of Airway Surface Liquid Volume/Composition and Mucus Transport
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Rates In Vitro.- Murine Models of CF Airway Infection and Inflammation.- Analysis of Lipid Abnormalities in CF Mice.- Bioelectric Measurement of CFTR Function in Mice.- Xenograft Model of the CF Airway.- Development of Conditionally Immortalized Epithelial Cell Lines from CF and Non-CF Mice.- Technical Approaches to Analyze the In Vivo Ion Composition of Airway Surface Liquid.- Novel Therapeutic Approaches for Cystic Fibrosis.- Design of Gene Therapy Clinical Trials in CF Patients.- Formulation of Synthetic Vectors for Cystic Fibrosis Gene Therapy.- Adeno-Associated Viral Vectors for CF Gene Therapy.
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Bibliographische Angaben
- 2011, XVI, 615 Seiten, Maße: 15,2 x 22,8 cm, Kartoniert (TB), Englisch
- Herausgegeben: William R. Skach
- Verlag: Springer, Berlin
- ISBN-10: 1617372447
- ISBN-13: 9781617372445
Sprache:
Englisch
Pressezitat
"The coverage of methodologies is extremely broad and will be useful for those wanting to find out more about some of the less routine techniques used in the analysis of the CFTR protein/chloride channel. The style of protocol-based chapters enables in manuscripts due to considerations of space or potential repetition of previously published techniques. As such many of the chapters are valuable." -Molecular Biotechnology"Undoubtedly this book will be regarded as essential reading for scientists in the field. It provides details of research methodology, which are rarely published in peer reviewed journals. It will also enable researchers to identify subtle differences in techniques in their own laboratories from those described in these chapters. As such, it will be a very important book for all those who conduct laboratory-based research into cystic fibrosis." -Journal of Paediatrics and Child Health
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