Cystic Fibrosis Methods and Protocols


Book Description

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.




Molecular Diagnosis of Genetic Diseases


Book Description

This completely revised and updated second edition to integrates the many new technologies and insights now available for the diagnosis of genetic diseases. The authors use such methodologies as PCR optimization dosage analysis, mutation scanning, and quantitative fluorescent PCR for aneuploidy analysis, Neurofibromatosis type 1, and Duchenne muscular dystrophy. These largely generic methodologies may be adapted to most genetic conditions for which a molecular diagnosis is relevant, no matter how frequent or rare their incidence. Molecular Diagnosis of Genetic Diseases, Second Edition offers diagnostic molecular geneticists a unique opportunity to sharpen their scientific skills in the design of assays, their execution, and their interpretation.




Lung Cancer


Book Description

Leading physician scientists and noted researchers review novel methods for determining the etiology of a variety of lung cancers and present readily reproducible techniques for examining the associated multitude of genetic abnormalities. The methods make it possible to detect these alterations at the cellular, DNA, and protein levels, to study the development of lung cancer in vitro and in vivo-either in situ or in the form of metastases, and to test targeted therapies with detailed model systems. An animal models section gives explicit instructions for setting up and testing these systems, which are rarely described in the literature.




Psychiatric Genetics


Book Description

Psychiatric Genetics provides the reader with a complete view of the methodological problems encountered in psychiatry genetics and proposes solutions to commonly occurring questions. The best European and American specialists have given a thorough review on the advantages and disadvantages of genetic epidemiological methods, the way to choose a genetic marker or a clinical interview and how to ascertain patients, unaffected relatives and controls and what should be the criteria to include a case or a control. New phenotypic methods are described focusing on candidate symptom and endophenotype approaches. Examples coming from cognitive neurosciences, biochemistry, electrophysiology and brain imaging techniques are reviewed. This book will serve as an essential handbook for psychiatrists, psychologists, and geneticists involved in the genetics of psychiatric disorders.




Molecular Analysis of Cancer


Book Description

Over the past 20 years, technological advances in molecular biology have proven invaluable to the understanding of the pathogenesis of human cancer. The application of molecular technology to the study of cancer has not only led to advances in tumor diagnosis, but has also provided markers for the assessment of prognosis and disease progression. The aim of Molecular Ana- sis of Cancer is to provide a comprehensive collection of the most up-to-date techniques for the detection of molecular changes in human cancer. Leading researchers in the field have contributed chapters detailing practical pro- dures for a wide range of state-of-the-art techniques. Molecular Analysis of Cancer includes chapters describing techniques for the identification of chromosomal abnormalities and comprising: fluor- cent in situ hybridization (FISH), spectral karyotyping (SKY), comparative genomic hybridization (CGH), and microsatellite analysis. FISH has a pro- nent role in the molecular analysis of cancer and can be used for the detection of numerical and structural chromosomal abnormalities. The recently described SKY, in which all human metaphase chromosomes are visualized in specific colors, allows for the definition of all chromosomal rearrangements and marker chromosomes in a tumor cell. Protocols for the detection of chromosomal re- rangements by PCR and RT-PCR are described, as well as the technique of DNA fingerprinting, a powerful tool for studying somatic genetic alterations in tumorigenesis.




Diabetes Mellitus


Book Description

Diabetes mellitus is the collective name for a group of diseases associated with hyperglycemia (high levels of blood glucose) caused by defects in insulin p- duction, insulin action, or both. About 6. 2% of the US population (17 million people) have diabetes mellitus. It is the leading cause of kidney failure, bli- ness, and amputations. It is also a major risk factor for heart diseases, stroke, and birth defects. Diabetes Mellitus: Methods and Protocols provides a state-of-the-art account of the experimental methodology for studying the molecular defects leading to diabetes mellitus, both at the molecular and biochemical levels. The chapters cover a wide range of topics written by experts in their respective fields and are organized in two sections: Insulin Production and Insulin Action. The detailed experimental protocols presented, including the notes of interest, provide a very useful tool for basic researchers and clinicians for investigating and treating this disease. Each chapter starts with an introduction to a specific technique and explains its application in the field of diabetes research. Following the list of materials, a detailed description of the technique is presented in the methods section in a way that enables the successful execution of the protocol. The “Notes” section at the end discusses the pitfalls of the technique and alternative approaches. I am grateful to the numerous scientists who have contributed to this volume by writing both highly detailed and understandable chapters.




Suicide Gene Therapy


Book Description

Gene therapy has expanded rapidly over the last decade. The number of clinical trials reported by 2001 included 532 protocols and 3436 patients. Phase I trials predominate with 359 trials of 1774 patients versus Phase II (57 trials with 507 patients) and Phase III (3 trials of 251 patients). The disease overwhelmingly targeted by gene therapy is cancer: involving 331 trials with 2361 patients. Despite the somewhat disappointing results of clinical trials to date, gene therapy offers tremendous promise for the future of cancer therapy. The area of gene therapy is vast, and both malignant and nonmalignant cells can be targeted. Suicide Gene Therapy: Methods and Reviews covers gene therapy that targets malignant cells in a treatment that has become known as “suicide gene therapy. ” Basically, this approach uses the transduction of cancer cells with a gene for a foreign enzyme that, when expressed, is able to activate a nontoxic prodrug into a highly cytotoxic drug able to kill the cancer cell population. This is a major area in cancer gene therapy—in 2001 this technique was represented by 52 clinical protocols with a total of 567 patients. Additional trials used multiple gene therapy protocols that also involved suicide gene therapy (83 with 497 patients), indicating that the interest in this area is considerable. Suicide Gene Therapy: Methods and Reviews aims to cover comprehensively, both in theoretical and practical terms, the rapidly evolving area of suicide gene therapy for cancer.




Anticoagulants, Antiplatelets, and Thrombolytics


Book Description

This collection of review articles highlights the latest development of antithrombotics and provides proven experimental methods for the further development of new and improved anticoagulants. Among the cutting-edge developments reviewed are the novel usage of low molecular weight heparins, such antithrombin agents as the hirudin, and such antiplatelet drugs as the GPIIb/IIIa inhibitors and ADP receptor antagonists.




Molecular Diagnosis of Infectious Diseases


Book Description

Populations of the western world are now healthier and enjoying higher life expectancy than ever. They are beginning to benefit from an array of costly new therapies made possible through recent rapid advances in medical science and technology, and their demands on modern medicine are rising. Meanwhile, healthcare systems are struggling with their outdated legacy models of the m- th 20 century and are experiencing ever-increasing financial pressure from g- ernments and health insurance organizations. The equation is no longer in balance, and this predicament is forcing societies to explore new approaches to managing healthcare in the future. Since the first edition of Molecular Diagnosis of Infectious Diseases was published, we have witnessed the sequencing of the (almost) complete human genome and a shift in medical research from an emphasis on genetics to the advancement and useful application of proteomics. Bioinformatics has become the key tool for managing and analyzing the upsurge of data, and faster and more effective test methods and technologies have opened up new prospects for ind- try and academia. The tools of modern genomics and proteomics are now being utilized to specifically guide the discovery of drugs for the prevention, diagnosis, and treatment of human disease. They may also help us to find a way out of the current healthcare calamity.




Autogenic Drainage


Book Description

This work does not provide "recipes" or standardized solutions for the treatment of patients affected hypersecretion. The reader will find the book necessary ingredients to adapt to drain bronchial disease and the patient's condition, their needs and possibilities. Find relevant information ontology, anatomy, physiology and pulmonary ventilatory mechanics in order to facilitate understanding of the mechanisms involved in bronchial drainage techniques. It also delves into measurements of lung function and its implications. Explains the problems with obstruction and pulmonary physiotherapy assessment, as well as the bases of the bronchial drainage techniques. Not to mention the care of the upper airways. Acquire special mention specific principles of autogenic drainage, showing through performance curves partial expiratory flow-volume, the ability to modulate the flow and volume in the bronchial tree and selectively localize functional bronchial generations drain. Series of tips to standardize ventilation during inspiratory phase as well as the importance of correcting the shape of the rib cage. Explains the use of autogenic drainage in patients collaborators and collaborators, with a section dedicated to autogenic drainage in infants. The last chapters are devoted to explaining the specific physiology of cough with practical implications, special attention is given to inhalation therapy and finally being mentioned techniques as accessory respiratory reeducation and functional analysis, the importance of exercise and positioning, flexible gymnastics, relaxation, the use of oxygen and the rehabilitation effort through sport.