Diabetes Mellitus

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.

Includes supplementary material: sn.pub/extras

Klappentext
About 17 million Americans suffer from diabetes mellitus, a disease caused by defects in insulin production, insulin action, or both, and also a leading cause of kidney failure, blindness, and amputations. In Diabetes Mellitus: Methods and Protocols, leading researchers and clinical investigators describe their cutting-edge techniques for studying these defects at both the molecular and biochemical levels. Written in step-by-step detail to ensure ready reproducibility and robust results, these techniques allow investigators, both novices and those already active in the field, to study every major facet of insulin production and action. Each protocol includes an introduction to the technique, an explanation of its application, and a list of materials. Practical notes discuss how to avoid pitfalls, as well as how to adapt the methods to your own research.
Cutting-edge and highly practical, Diabetes Mellitus: Methods and Protocols provides basic scientists and clinical researchers, both experienced investigators and those new to the field, all the critical tools needed for the productive study of diabetes mellitus today.

Inhalt
Insulin Production.- Isolation of Islets of Langerhans from Rodent Pancreas.- Purification of Rat Pancreatic ?-Cells by Fluorescence-Activated Cell Sorting.- Assessment of Insulin Secretion in the Mouse.- Detection of Insulin Production by Immunohistochemistry.- Quantification of the Level of Insulin Gene Expression.- Chromatin Immunoprecipitation Using Isolated Islets of Langerhans.- Adenoviral Gene Transfer into ?-Cell Lines.- Utilization of NOD Mice in the Study of Type 1 Diabetes.- Insulin Action.- of DNA into 3T3-L1 Adipocytes by Electroporation.- Analysis of Insulin-Stimulated Glucose Uptake in Differentiated 3T3-L1 Adipocytes.- Fractionation Analysis of the Subcellular Distribution of GLUT-4 in 3T3-L1 Adipocytes.- Visualization and Quantitation of Integral Membrane Proteins Using a Plasma Membrane Sheet Assay.- Assaying Tyrosine Phosphorylation of Insulin Receptor and Insulin Receptor Substrates.- Measuring Insulin-Stimulated Phosphatidyl-Inositol 3-Kinase Activity.- Assaying AKT/Protein Kinase B Activity.- Measurements of Cellular Phosphoinositide Levels in 3T3-L1 Adipocytes.- Measurement of Glycogen Synthesis and Glycogen Synthase Activity in 3T3-L1 Adipocytes.- Measurement of Contraction-Stimulated GLUT-4 Translocation in Isolated Skeletal Muscle.- Single Embryo Measurement of Basal-and Insulin-Stimulated Glucose Uptake.- Immunohistologic Staining of Muscle and Embryos to Detect Insulin-Stimulated Translocation of Glucose Transporters.