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Since the publication of the bestselling second edition of John Walker’s widely acclaimed Protein Protocols Handbook, there have been continual methodological developments in the field of protein chemistry. This greatly enhanced third edition introduces 57 critically important new chapters, as well as significantly updating the previous edition's tried-and-true methods. Although the timely new chapters are spread throughout all of the book, the vital section on post-translational modifications has been expanded most to reflect the increasing importance of these modifications in the understanding of protein function. Each readily reproducible method follows the highly praised format of the Methods in Molecular Biology™ series, offering a concise summary of its basic theory, a complete materials list, a step-by-step protocol for its successful execution, and extensive notes on avoiding pitfalls, or on modifying the method to function within your own experimental circumstances. The expert authors of each chapter have demonstrated a hands-on mastery of the methods described, fine-tuned here for optimal productivity. Comprehensive, cutting-edge, and highly practical, The Protein Protocols Handbook, Third Edition is today's indispensable benchtop manual and guide, not only for all those new to the protein chemistry laboratory, but also for those established workers seeking to broaden their armamentarium of techniques in the urgent search for rapid and robust results

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SELDI is distinct from other TOF-MS technologies in that it couples features of chromatography and mass spectrometry, facilitating analyte enrichment and sample cleanup on an array surface. In the growing field of proteomics, SELDI technology has been widely used for biomarker discovery and characterization in diverse applications including diagnostics, drug development, and basic research. SELDI-based biomarker studies can typically be divided into four phases: discovery, validation, purification and identification, and assay development. SELDI-TOF Mass Spectrometry: Methods and Protocols provides an overview of the current applications of SELDI-TOF MS (surface enhanced laser desorption/ionization time-of-flight mass spectrometry), with an emphasis on study and experimental design, data analysis and interpretation, and assay development. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, SELDI-TOF Mass Spectrometry: Methods and Protocols will provide information on optimizing study design, experimental protocols, and data analysis and interpretation to yield robust biomarkers and biomarker assays, using examples from different disease areas.

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Despite being known and studied for years, peptides have never before attracted enough attention to necessitate the invention of the term "peptidomics" in order to specify the study of the complement of peptides from a cell, organelle, tissue or organism. In Peptidomics: Methods and Protocols, expert researchers present a comprehensive range of analytical techniques for the analysis of the peptide contents of complex biological samples with an emphasis often on higher throughput techniques, suitable for the analysis of large numbers of peptides typically present in the peptidomes. Encompassing a number of species ranging from bacteria to man, the methods presented intensively cover topics such as organism handling, tissue and organ dissection, cellular and subcellular fractionation, peptide extraction, fractionation and purification, structural characterization, molecular cloning, and sequence analysis. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Peptidomics: Methods and Protocols brings this ten year old field fully up-to-date in order to inspire novices and experts alike with the easy-to-follow practical advice needed to set up and carry out analysis of the peptide contents of complex biological samples.

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An in-depth review by leading authorities of the latest therapies and techniques for rescuing persons in cardiac arrest. The authors explore the physiology behind current state-of-the-art clinical resuscitation and translate it into practical bedside recommendations, clinical tips, and expert techniques. Topics of interest include the epidemiology of sudden death, management of ventilation, chest compression technique training, public access defibrillation, drug delivery during CPR, the latest drug therapies, and cardiac arrest in disease, pregnancy, drowning, lightning strike, and trauma. The authors also review the major ongoing research in resuscitation science that will likely affect the next set of international resuscitation guidelines.

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Diseases of the pancreas have been extremely difficult to study. Until relatively recently, the pancreas was viewed by surgeons as an organ to be avoided, given its deep retroperitoneal location and the sometimes severe consequences of manipulation. Similar difficulties have plagued careful study of the pancreas. In humans, only inf- quently have we had the opportunity to evaluate the evolution of d- eases of the pancreas; instead, we are confronted with end-stage chronic pancreatitis or acute pancreatitis with extensive necrosis of the gland. Within the past decade, revolutionary techniques in molecular biology, genetics, and animal models have begun to give us dramatic new insights into pancreatic disease. Coupled with these advances has been a slow but steady progress in the development of our clinical tools for diagnosing and treating pancreatic diseases. These advances place us on the threshold of a much more complete understanding of pancreatic diseases, opening up new opportunities for novel therapies and preventive measures. Pancreatitis and Its Complications brings together many of the world’s experts in pancreatic diseases. These “pancreatologists” include basic scientists, endoscopists, gastroenterologists, and surgeons. The ch- ters have been organized to provide a comprehensive discussion of acute and chronic pancreatitis, with up-to-date discussions of pat- physiology, epidemiology, diagnostic strategies, treatment, and c- plications of disease. It is hoped that this text will serve as a useful reference for clinicians as well as provide a clinical background for scientists with an interest in pancreatic or related diseases.

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Despite major advances in the understanding of stroke mechanisms that have occurred over the past quarter century, stroke continues to rank among the leading causes of death and disability worldwide. Although currently it may be difficult to believe, early doubts were expressed as to whether inter­ ventions in risk factors for either coronary disease or stroke would actually lead to a reduction in the incidence of these disorders. However, large clini­ cal trials in hypertension, carotid disease, atrial fibrillation, and antithrombotic and antiplatelet therapies have effectively demonstrated the efficacy of these targeted interventions in reducing stroke incidence. More recently, after earlier uncertainty regarding the role of elevated lipids as a risk factor for stroke, clinical trials of the statins have also demonstrated a significant reduction in the incidence of ischemic stroke. However, as em­ phasized in Handbook of Stroke Prevention in Clinical Practice, despite these gains and the initial decline in stroke incidence that did occur in the 1960s and 1970s, the incidence of stroke disappointingly has failed to show a fur­ ther significant decline since that time. The editors of Handbook of Stroke Prevention in Clinical Practice raise the very important question of whether recognized strategies for stroke preven­ tion have been widely or effectively implemented. They correctly empha­ size the critical importance of identifying the mechanism of stroke in each patient so as to properly direct prevention and treatment. As Dr.

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A cutting-edge review of the latest findings on key scientific topics and the best strategies for evaluation and management of portal hypertension in the clinic. Writing in an easy-to-read style, the authors review its pathobiology, the progress being made in its experimental understanding, the methodologies to assess it in humans, the treatment of its complications (esophageal varices, ascites, and hepatorenal syndrome), and its treatment in special circumstances (during pregnancy, in children, or when the portal vein is blocked). They also provide in-depth coverage of the pros and cons of the various therapeutic choices available to the clinician managing patients with complicated portal hypertension.

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This second edition volume expands on the previous edition with new sections describing the characterization of peptides bound to major histocompatibility complexes (MHC) on the surface of the cell. Chapters also cover topics such as using SERPA for antigen identification; antigen content of electroimmunoprecipitates; whole genome-phage display libraries; antigens in immune complexes; and immunoproteomic biomarkers. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Immunoprotemics: Methods and Protocols, Second Edition is a valuable resource that presents novice and expert researchers with techniques that are easily transferrable to laboratories and provides enhanced hands-on insights into this evolving field.

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This volume describes cell culture protocols for the maintenance, propagation, manipulation, and analysis of primary explanted cells from various mouse organ systems. After introductory chapters that examine the applicability of mouse models as a discovery tool and describe critical factors and variables that influence cell culture endpoints, protocol chapters explore cell culture methodologies for specific mouse cell types and lineages. Written in the highly successful Methods in Molecular Biology series format, these chapters include introductions to their respective topics, comprehensive lists of the essential materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Mouse Cell Culture: Methods and Protocols aims to ensure successful results utilizing these protocols in further studies, leading to the identification of key factors, genes, and cellular pathways regulating cell lineage commitment and differentiation, modeling disease, and identifying novel therapeutic targets.

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In this volume expert researchers in the field detail the many methods for further research into regulation of mitochondrial function. Chapters focus on mitochondria with other cellular components, discussing how these interactions influence the dynamics of mitochondrial structure and biogenesis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, Mitochondrial Regulation: Methods and Protocols seeks to aid advanced undergraduates, graduates, postgraduates, and beginning researchers in the areas of molecular and cellular biology, biochemistry and bioenergetics.

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Providing experimental methods and protocols for performing pheromone research in a variety of organisms ranging from invertebrates to vertebrates, Pheromone Signaling: Methods and Protocols covers a wide spectrum of experimental approaches necessary for handling pheromone molecules, measuring receptor response and neural activation, and analyzing behavioral output. A great deal of progress has been made in understanding the molecular mechanisms underlying pheromone action, largely due to the discovery of receptor genes, the advancement of imaging techniques, and key multi-disciplinary approaches including aspects of organic chemistry, biochemistry, molecular biology, electrophysiology, and behavioral science. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Authoritative and practical, Pheromone Signaling: Methods and Protocols details the key methodologies utilized in laboratories all over the world, making them accessible to those who want to begin investigation in the area of pheromone research.

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Brain Energy Metabolism addresses its challenging subject by presenting diverse technologies allowing for the investigation of brain energy metabolism on different levels of complexity. Model systems are discussed, starting from the reductionist approach like primary cell cultures which allow assessing of the properties and functions of a single brain cell type with many different types of analysis, however, at the expense of neglecting the interaction between cell types in the brain. On the other end, analysis in animals and humans in vivo is discussed, maintaining the full complexity of the tissue and the organism but making high demands on the methods of analysis. Written for the popular Neuromethods series, chapters include the kind of detailed description and key implementation advice that aims to support reproducible results in the lab.Meticulous and authoritative, Brain Energy Metabolism provides an ideal guide for researchers interested in brain energy metabolism with the hope of stimulating more research in this exciting and very important field.

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Molecular and Translational Vascular Medicine will serve as a state-of-the-art resource for physicians and translational medical researchers alike who are interested in the rapidly evolving field of vascular medicine. The text provides new insight into the basic mechanisms of classic vascular pathophysiologic processes like angiogenesis, atherosclerosis, thrombosis, and vasculitis. Furthermore, it covers new areas of investigation including the role of the ubiquitin proteasome system in vascular disease, endothelial progenitor cells for disease treatment, and the genetic basis of thoracic aortic aneurysms. Lastly, this volume includes sections on the newly emerging field of therapeutic angiogenesis, and the developing technology of nanoparticle-based imaging and therapeutic treatment of the diseased vasculature. All chapters are written by established experts in their fields, including pathologists, cardiovascular surgeons, and internists as well as translational biomedical researchers in a wide range of disciplines. While comprehensive, the material is presented in a manner that simplifies the complex pathophysiologic mechanisms that underlie common vascular diseases. Molecular and Translational Vascular Medicine will be of great value to a broad audience including internists, cardiovascular and vascular surgeons, pathologists, residents and fellows, as well as translational biomedical researchers.

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To prepare synchronized cells representing different stages of the cell cycle has been a great challenge for researchers across the globe. In Cell Cycle Synchronization: Methods and Protocols, experts in the field supply detailed protocols providing first the theoretical background of the procedure then step-by-step instructions on how to implement synchronization as well as the latest techniques for the enhanced study of regulatory mechanisms to understand cell cycle events. Describing synchronized cells from asynchronous bacterial, plant, protozoan, yeast, fish, and mammalian cell cultures, the synchronization methods presented in the book are based principally on two major strategies: the “arrest-and-release” approach, which involves different chemical treatments to block cells at certain stages of the cell cycle, and the physical strategy, which contains physical methods to collect cells belonging to subpopulations of the cell cycle. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Detailed and easy to follow, Cell Cycle Synchronization: Methods and Protocols is an ideal guide for scientists who wish to make use of these powerful synchronization techniques but have no access to thorough, time-tested protocols designed for doing so.

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Because aging is accompanied by a steady decline in resistance to infectious diseases, the diagnosis and treatment of these diseases in the elderly is not only much more complex, but also often quite different from that for younger patients. In the second edition of Infectious Disease in the Aging: A Clinical Handbook, a panel of well known and highly experienced geriatric physicians and infectious disease experts review the most important common infections affecting the elderly and delineate their well-proven diagnostic, therapeutic, and preventive techniques. Among the illnesses discussed are urinary tract infections, pneumonia, ocular infections, tuberculosis, and fungal and viral infections. In addition, there are detailed discussions of sepsis, infective endocarditis, intraabdominal infections, bacterial meningitis, osteomyelitis and septic arthritis, and prosthetic device infections.

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As two of the leading causes of death worldwide, heart disease and stroke represent a clear target for genomic research aimed at deciphering the genes and cellular pathways that underlie cardiovascular disease and creating improved therapies. In Cardiovascular Genomics: Methods and Protocols, experts in the field provide methods for cardiovascular phenotyping of rodent models in the first section of the volume and for statistical and bioinformatic integration of phenotype data with genome-wide genotype and expression data in the second section. Understanding these diverse methods will allow an individual laboratory to utilize these genomic methods independently or to better prepare for collaboration with scientists having expertise in other disciplines in order to uncover genes affecting cardiovascular disease. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary equipment, materials, and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Cutting-edge and easy-to-use, Cardiovascular Genomics: Methods and Protocols will enable researchers to identify causal genes and novel molecular targets that can lead to vital new treatments for cardiovascular disease.

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This volume discusses the latest developments in cellular, molecular, biochemical, and imaging assays to study the biology and functions of T-cells. The chapters in this book cover topics such as LFA-1/ICAM-1 interactions in T-cell motility; using 3D-SIM to dissect signaling cross-talks in motile T-cells; GapmeR-mediated gene silencing in motile T-cells; activity of cellular kinases in migrating T-cells; and computational analysis of protein-protein interactions in motile T-cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, T-Cell Motility: Methods and Protocols is an essential resource for graduate students, postdoctoral fellows, and principal investigators working in the fields of immunology, T-cell biology, biochemistry, molecular biology, and imaging.

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This detailed book showcases recent advances in computational design of multi-target drug candidates involving various ligand and structure-based strategies. Different chem-bioinformatic modeling strategies that can be applied for design of multi-target drugs as well as important databases and web servers in connection with multi-target drug design are also featured in this collection. Written for the Methods in Pharmacology and Toxicology series, chapters include the kind of key implementation advice that will aid researchers greatly in their laboratory pursuits. Authoritative and practical, Multi-Target Drug Design Using Chem-Bioinformatic Approaches seeks to aid all scientists working in the field of drug discovery research.

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This volume provides all the essential protocols that are currently used to study the immune synapse. Chapters in The Immune Synapse: Methods and Protocols cover methods for the study of the dynamics of immune synapse assembly, traffic at the immune synapse, new high resolution imaging, biophysical and computational methods for the study of the immune synapse, effector immune synapses, B cell, NK and mast cell immune synapses, and immune interactions in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, The Immune Synapse: Methods and Protocols will be of interest to immunologists and, at a more general level, to cell biologists, biophysicists and computational biologists.

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This volume covers an array of techniques available for studying SH2 domains and phosphotyrosine signaling. The book is divided into six parts: Part I outlines the history of SH2, technology development, and cell signaling; Part II focuses on computational approaches and tools used for identification, classification, and predictions of SH2 domain binding partners; Part III details various ways to prepare the SH2 domains as experimental reagents; Part IV presents methods for structural analysis and conventional binding assays using SH2 domains; Part V describes high-throughput and proteomics approaches to aid in analyzing SH2-mediated interactions; and Part VI covers applications for SH2 domain to functional and imaging analyzes. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, SH2 Domains: Methods and Protocols is a valuable resource for computational biologists, biochemists, structural biologists, cell biologists, pathologists, and people interested in SH2 domains and phosphotyrosine signaling. Researchers who are investigating how protein interaction domain mediate specificity in signaling systems may also find this book informative.

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This book explores methods to study the complex and evolving interplay between a virus and its host that range from model systems to the detection of chemical molecules. The collection starts with the application of humanized mice and zebrafish as model organisms to study virus-host interactions and induction of innate immune responses. Subsequent chapters outline diverse methods to detect small interfering RNAs, microRNAs, and virus-derived dsRNA from a variety of cells, tissues, and organisms, as well as to interrogating the cytosolic RNA and DNA sensing pathways, including using RNA PAMPs as molecular tools, purification of cGAMP from virus particles and infected cells, and mechanisms to visualize the subcellular localization and activation of the adaptor proteins MAVS and STING. Cutting-edge methods, including high-throughput and genome-wide CRISPR/Cas9 screens, chromosome conformation capture, and whole-exome sequencing, are described to identify novel mediators, pathways, and variants underlying host susceptibility. Given the importance of studying these pathways and players under physiologic conditions, methods describing the isolation of primary mouse sensory neurons and group 2 innate lymphoid cells are also provided. Finally, this collection comes full circle back to the whole organism level and concludes with epidemiological methods to investigate virus-host interactions and the induction of innate immunity. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Innate Antiviral Immunity: Methods and Protocols spans a diverse array of approaches to study and elucidate the intricacies of this vital area of study.The chapter 'Morphological Separation of Clustered Nuclei in Histological Images' is published open access under a CC BY 4.0 license at link.springer.com.

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Over the last 15 years, high-performance liquid chromatogra­ phy (LC) has made the transition from an instrument used only by experts in research labs to a tool used for routine applications by relatively unskilled workers. With this transition have come in instrumentation and column technology. In major advances the past, the operator had to be a jack-of-all-trades, with a screw­ driver, soldering iron, and various wrenches as constant compan­ ions in the LC lab. Today, many instruments contain micropro­ cessors as powerful as those of mainframe computers of earlier days. With this technology has come a variety of self-diagnostic tools that allow the LC system to locate many of its own prob­ lems. Traditionally, well-honed LC troubleshooting skills have been a result of years of work at the bench. Today the LC system itself often can do a better job of troubleshooting than the operator can. Yet many of the problems of the past are still the major problems of today: air bubbles, check valves, detector lamps, and, of course, problems with the separation. An added pressure on the operator of today's LC system is that of productivity-the lab often cannot afford unnecessary downtime. This means that the operator has to be a troubleshooting expert, or has to have that expertise at his or her fingertips. The present book was written to provide this expertise in an easy-to-use format for users at all levels of experience.

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The fully revised second edition of the Encyclopedia of Heart Diseases is an ideal resource for practicing clinicians and researchers. Available in print, online, and with dual access, it is a clear and comprehensive aggregation of the most crucial information and essential data on cardiovascular diseases and therapeutics.Comprised of over 95 entries with regular online updates, the Encyclopedia of Heart Diseases is fully referenced, and major points of interests are hyperlinked to complementary sections. Each entry is logically and superbly written, providing accurate core knowledge of pathogenesis, pathophysiology, clinical features, diagnostic techniques, and management strategies. Specific detail is paid to technological advances in imaging and diagnostics. Therapy focused entries give powerful insights into not only prescribing drug regimens, but also into the controversies surrounding their use.This major reference work is invaluable for all those involved in the care of cardiovascular patients. From the front-line practitioner to the basic science researcher to the student in training, the Encyclopedia of Heart Diseases offers an astute authoritative guide to navigating an immense body of fascinating information. From the trainee to the internist and cardiologist, all will find it useful. It is an essential resource for medical libraries and academic institutions worldwide. From the Foreword:So, what would we want from an encyclopedia on heart disease? Ideally, a book would be comprehensive, yet concise, and be practically oriented, and explain pathophysiology and treatment. In addition, it should be accessible online so that it can be accessed at the bedside or anywhere.Dr. Khan has written exactly such a book. Encyclopedia of Heart Diseases is comprehensive, yet concise, and very practically oriented. Importantly, it takes a step-by-step approach, walking the reader through a thorough pathophysiology of conditions, their evaluation and treatment. For therapies, he provides the mechanism of the drug, its doses, side effects and clinical efficacy....A terrific online resource with all the information you need!- Christopher P. Cannon, MD, TIMI Study Group, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA

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This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. The chapter 'Functional Analysis of Membrane Proteins Produced by Cell-Free Translation' is open access under a CC BY 4.0 license via link.springer.com.

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This second edition details new emerging areas of zebrafish research focusing on genetics and genomics, techniques for developing and analyzing zebrafish disease models, and methods for neuroscience. Zebrafish: Methods and Protocols, Second Edition guides readers through methods for mutagenesis and genome editing in zebrafish, applications of GFP-expressing transgenic fish, techniques for cancer models, imaging of infection and host-pathogen interactions, metabolism and transport of lipids, and the structure and function of neural circuits and their role in generating behavior. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical Zebrafish: Methods and Protocols, Second Edition is a useful complement to the first book for new and experienced zebrafish researcher alike.

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In Clinical Bioinformatics, Second Edition, leading experts in the field provide a series of articles focusing on software applications used to translate information into outcomes of clinical relevance. Recent developments in omics, such as increasingly sophisticated analytic platforms allowing changes in diagnostic strategies from the traditional focus on single or small number of analytes to what might be possible when large numbers or all analytes are measured, are now impacting patient care. Covering such topics as gene discovery, gene function (microarrays), DNA sequencing, online approaches and resources, and informatics in clinical practice, this volume concisely yet thoroughly explores this cutting-edge subject. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and easily accessible, Clinical Bioinformatics, Second Edition serves as an ideal guide for scientists and health professionals working in genetics and genomics.

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This volume investigates how the mitochondrial genome is transmitted, segregated, and inherited. It starts by describing mtDNA mutations and deletions and how these impact on the offspring’s well-being. It progresses to discuss how mutations to the mtDNA-nuclear-encoded transcription, replication and translational factors lead to mtDNA-depletion syndromes and how these affect cellular function and lead to the pathology of human mitochondrial disease. It also highlights the importance of the mitochondrial assembly factors and how mutations to these can lead to mitochondrial disease. The reader is then introduced to how mtDNA is transmitted through the oocyte and how stem cells can be used to study mitochondrial biogenesis and mtDNA replication and transcription in undifferentiated pluripotent and differentiating cells and how mitochondria adapt during this process. It then discusses how diseases like cancer are initiated and regulated by mutations to mitochondrial DNA and dysfunctional mitochondria. Finally, it draws on assisted reproductive technologies to discuss how some of these approaches might be adapted to prevent the transmission of mutant and deleted mtDNA from one generation to the next.

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Since the publication of the first edition of Urethral Reconstructive Surgery, important refinements have been added to the urologic surgical armamentarium. Extensively revised and updated, Advanced Male Urethral and Genital Reconstructive Surgery, Second Edition guides urologists in a practical manner on how to evaluate and manage complex urethral and male genital reconstructive challenges. Chapters have been added on wound healing, synchronous urethral strictures, non-transecting anastomotic urethroplasty (including muscle and vessel sparing), overlapping buccal grafts, male urethral slings, genital skin grafting, Peyronie’s surgery, priapism, pediatric strictures and prosthetics. Such chapters greatly strengthen the overall appeal of the book. The volume closes with a new comprehensive appendix of preferred instruments.With broad contributions by international authorities in reconstructive urologic surgery, Advanced Male Urethral and Genital Reconstructive Surgery, Second Edition is a valuable resource for all urologists, from residents to reconstructive surgeons.

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Liver Transplantation: Challenging Controversies and Topics grew out of a need I perceived within the fields of transplant hepatology and liver transplantation. Liver transplantation has rightly gained recognition as an established therapy for end-stage liver disease. Few would argue that liver transplantation is one of the few truly lifesaving and life-altering treatments within medicine and surgery. Not many realize that 20 years passed from the time of the first human liver transplantation in 1963 to its acceptance as therapy by the 1983 NIH Consensus Conference on Liver Transplantation. In 2008, 25 years will have passed since the 1983 NIH conference—a mere 25 years for a field that has provided patients hope, doctors options, and to some the “gift of life. ” Many issues in liver transplantation involve indications, patient selection, and outcomes after transplantation—these are standard topics, covered by textbooks of hepatology and transplantation. In contrast, the field of liver tra- plantation is young, evolving, dynamic, and issues and decisions are often controversial. Thus, Dr. Trotter and I, as well as our colleagues at the University of Colorado, felt that a text with a different focus was required, one that highlighted controversy and challenged dogma. Out of this perceived need emerged Liver Transplantation: Challenging Controversies and Topics. To meet the transplant community’s need for emerging information about liver transplantation, Dr. Larry Chan, Dr. Igal Kam, and I initiated the Controversies in Transplantation Conference.

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Although urology is a surgical specialty, it has become apparent that changes in health care delivery and financing have led to an increasing volume of care being provided by urologists in their offices. A major part of the revenue of a urology practice depends on office production and efficient management. To have a successful practice, the produc­ tive, committed urologist must have a thorough understanding of the procedures and problems that need to be dealt with in the office. Moreover. the urologist must play an active role in the administration and business aspects of running the office. Surprisingly, very little has been written about the office practice of urology. Office Urology: The Clinician's Guide presents a fresh, practical, and concise text­ book covering the vital issues that the urologist must face on a daily basis in the office. The initial chapters of the textbook cover the critical aspects of managing the urologic office, such as principles of management, marketing the practice, proper billing and coding, advanced information systems, and important legal issues. The book is not intended to review in detail the academic aspects of the various pathologies pertaining to urology, which have been well covered in several other textbooks. Instead, the clinical chapters deal with practical issues, such as selecting appropriate treatment and counsel­ ing patients on the optimal therapy for the problems that the urologist frequently man­ ages. The individual authors have demonstrated expertise in their fields.

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This detailed book explores methods to isolate, characterize, and investigate key lung innate immune cells. Beginning with an overview, the volume then continues with methods for creating in vitro and in vivo model systems to study inflammatory lung diseases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Lung Innate Immunity and Inflammation: Methods and Protocols aims to be a guidebook and will be of value and interest to researchers investigating innate immunity and inflammation in the lung as well as other organs and tissues.

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This volume provides state-of-the-art techniques for studying various aspects of cholesterol homeostasis, including its uptake, synthesis and efflux from the cell, as well as its trafficking within the cell. Chapters also cover techniques for studying the regulation of cholesterol homeostasis at both the transcriptional and post-translational levels, as well as studying the membrane topology and structure of cholesterol-related proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, Cholesterol Homeostasis: Methods and Protocols aims to provide key techniques in tackling the investigation of cholesterol homeostasis.

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This book illustrates the current state-of-the-art in histamine research, with a focus on the appropriate methodologies to investigate the pharmacological properties and the therapeutic exploitation of HRs and their ligands. In addition, the range of techniques described provides an introduction to complementary cross-methodological disciplines beyond these fields. This multi-disciplinary approach is required to define the ‘decision gates’ that determine the development of more effective and safer therapeutic options for many forms of highly prevalent and debilitating diseases, such as asthma, dementias, dermatitis, and arthritis. Written for the Methods in Pharmacology and Toxicology series, chapters concentrate on practical, hands-on protocols from experts in the techniques.Authoritative and thorough, Histamine Receptors as Drug Targets seeks to aid pharmacologists, biochemists, drug discovery researchers, molecular biologists, chemists, toxicologists, lab scientists, medical doctors, principle investigators, research scientists, lab directors and technicians, as well as graduate students around the world in pursuing the study of this vital scientific area.

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This book presents a compilation of methods that detail improved protein and peptide sample preparation and identification. Chapters guide readers through methods for depletion of myofibril-associated proteins, peptide sample preparation in urinary proteomics, purification of targeted proteins from native tissues, fractionation strategies for protein analysis, and GeLC-MS as a sample preparation method for sample preparation for proteomics using minimal amount of tissue. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Tissue Proteomics: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

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This volume details fast-moving research while providing in-depth descriptions of methods and analytical approaches that are helping to understand the genome and how it is related to complex diseases. Chapters guide the reader through common and rare variation, gene-gene and gene-environment interactions and state-of-the-art approaches for the synthesis of genome-wide and gene expression data. Novel approaches for associations in the HLA region, family-based designs, Mendelian Randomization and Copy Number Variation are also presented. The volume concludes with the challenges researchers face while moving from identifying variants to their functional role and potential drug targets. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, a thorough presentation of methods and approaches and tips on troubleshooting and avoiding known pitfalls.

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In addition to their invaluable role in the fight against infection, Toll-Like Receptors (TLRs) in an under- or over-active state can lead to the pathogenesis of disease, making these receptors a key focal point for many research laboratories. In Toll-Like Receptors: Methods and Protocols, experts in the field contribute techniques currently used to study TLRs, their downstream signalling pathways, and their role in the pathogenesis of disease, with sections examining TLR expression and the application of genetic techniques and microarray analysis to TLR research. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections highlighting tips on troubleshooting and avoiding known pitfalls. Versatile and cutting-edge, Toll-Like Receptors: Methods and Protocols provides an ideal manual for a wide range of biologists and medical researchers studying these vital components of the immune system.

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As increasing global population and continuing economic development ensure the need for further production and cultivation of maize, the necessity of the application of transgenic technology to this model species and crop plant grows steadily. In Transgenic Maize: Methods and Protocols, experts in the various disciplines of the field contribute readily reproducible protocols covering such topics as transformation methods, transgenic maize in research, the analysis of transgenic plants, and breeding with transgenes. Written in the highly successful Methods in Molecular Biology™ series format, the chapters present brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Cutting-edge and easy to use, Transgenic Maize: Methods and Protocols will greatly aid researchers wishing to use these procedures in their own laboratories, and will provide readers with a better understanding of the experiments being performed by cooperators or fee-for-service labs.

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Colorectal Cancer Screening provides a complete overview of colorectal cancer screening, from epidemiology and molecular abnormalities, to the latest screening techniques such as stool DNA and FIT, Computerized Tomography (CT) Colonography, High Definition Colonoscopes and Narrow Band Imaging. As the text is devoted entirely to CRC screening, it features many facts, principles, guidelines and figures related to screening in an easy access format. This volume provides a complete guide to colorectal cancer screening which will be informative to the subspecialist as well as the primary care practitioner. It represents the only text that provides this up to date information about a subject that is continually changing. For the primary practitioner, information on the guidelines for screening as well as increasing patient participation is presentedd. For the subspecialist, information regarding the latest imaging techniques as well as flat adenomas and chromoendoscopy are covered. The section on the molecular changes in CRC will appeal to both groups. The text includes up to date information about colorectal screening that encompasses the entire spectrum of the topic and features photographs of polyps as well as diagrams of the morphology of polyps as well as photographs of CT colonography images. Algorithms are presented for all the suggested guidelines.Chapters are devoted to patient participation in screening and risk factors as well as new imaging technology. This useful volume explains the rationale behind screening for CRC. In addition, it covers the different screening options as well as the performance characteristics, when available in the literature, for each test. This volume will be used by the sub specialists who perform screening tests as well as primary care practitioners who refer patients to be screened for colorectal cancer.

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This illustrated guide to the role of the forensic anthropologist in investigating child abuse is an essential resource in one of the most contentious areas of forensic pathology. Not only does it supply a review of the literature in this field, but it illustrates the material with photographs from real cases investigated by the Harris County Institute of Forensic Sciences, which serves a population of four million people. Broken down into body regions and skeletal elements for ease of reference, the atlas facilitates the vital work performed by forensic anthropologists, who bring to the autopsy table a store of specialist knowledge that can turn a case.Despite the frequency of child fatalities (in America, 2.3per 100,000) attributed to physical abuse, merely recognizing the offense is a major forensic challenge. The tell-tale signatures of non-accidental injury can be very subtle, making it difficult to differentiate between accidental and non-accidental injury. Yet successful adjudication of a child abuse case often rests on the correct interpretation of skeletal injury.In this volume the authors guide the reader through published data regarding the mechanics and interpretation of injuries,including the agencies they indicate. The material includes discussion of the limitations faced in interpreting some injuries, where making a judgment on cause is tricky. In addition, a chapter on natural diseases affecting the bones provides a good overview of several conditions that are often invoked as 'mimics' of child abuse. Finally, this publication evinces the value of collaboration between the pathologist and the anthropologist.

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Breast Cancer is the most common tumor in women and the second leading cause of cancer deaths worldwide. Due to breakthroughs in gene profiling, the knowledge of the pathophysiology of the mammary gland had greatly increased over the last decade. In Breast Cancer in the Post Genomic Era, Antonio Giordano, Nicola Normanno, and a panel of international authorities in their field provide a comprehensive approach to the biology, diagnosis, prevention, and treatment of human breast carcinoma. The book provides a comprehensive approach to breast cancer, describing the use of gene profiling techniques to distinguish specific features of individual carcinomas, as well as emerging novel therapeutic approaches to treatment. Additional chapters cover the use of transgenic mice to model human breast cancer and the role of the EGF-CFC family in mammary gland development and neoplasia. Breast Cancer in the Post Genomic-Era succeeds in looking at breast cancer pathogenesis, diagnosis, and treatment under a more comprehensive light, and is a valuable resource for any Radiation or Surgical Oncologist, Cancer Biologist or Pathologist.

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Since the discovery of the pharmacological and toxicological importance of inhibiting the cyclooxygenase (COX) enzymes by non-steroidal anti-inflammatory drugs (NSAIDs), much research has gone into the development of methods to study the biological functions of COX-1 and COX-2. In Cyclooxygenases: Methods and Protocols, experts and pioneers in the field present the most up-to-date in vitro and in vivo techniques routinely used in COX research. The volume delves into essential topics such as the purification, cloning, and expression of COX enzymes as well as in vitro assays aimed at determining the inhibitory potency of drugs on COX-1 and COX-2 activities, with chapters describing protocols used for the extraction and measurement of the prostanoids. This volume also describes in vivo disease models used to study the roles of COX-1 and COX-2 in gastrointestinal injury, inflammation, and pain. As a book in the highly successful Methods in Molecular Biology™ series, the protocols chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Cyclooxygenases: Methods and Protocols serves as an indispensable tool for all scientists seeking the treatment of inflammation, pain, fever, and other harmful conditions.

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This volume looks at the newest methods, materials, equipment, and technologies developed to study the cell biology of the pneumococcus. The chapters, which cover a wide range of topics, are organized into six parts: Part one discusses the cultivation of Streptococcus pneumoniae in vitro; Part Two talks about the microscopy techniques used to study the biology of Streptococcus pneumoniae and pneumococcal interactions with the host; Parts Three and Four describe the genetics, proteome, and proteomics of Streptococcus pneumoniae; Part Five explains in vitro and in vivo models of Streptococcus pneumoniae-host interactions; and Part Six looks at the factors of public health, epidemiology, and biostatistics. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and authoritative, Streptococcus pneumoniae: Methods and Protocols is a valuable resource for both novice and expert researchers who want to expand their studies into new areas and new systems in this continuously evolving field.

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Arising from firm foundations laid by mathematical population genetics, clinical genetics, and statistical epidemiology, genetic epidemiology attempts to identify the many components of riskattributable to genes, environments, and interactions, and the course of its research can follow many diverse paths. In Genetic Epidemiology, the success of genome-wide association studies in their identification of hundreds of disease susceptibility loci has inspired renowned experts to contribute thorough methodologies, which aim to bring together bioinformaticians, geneticists, clinicians, statisticians, and epidemiologists in the study of this vital field. The volume opens with chapters covering the basics; however, it quickly moves on to coverage of more specialist topics such as twin studies, Mendelian randomization, genetic association studies, more advanced areas, as well as case studies. As a part of the highly successful Methods in Molecular Biology™ series, this work provides the detailed description of the application and analysis of the most commonly employed methods that are necessary for a firm grounding in the field. Authoritative and cutting-edge, Genetic Epidemiology aims to provide a basic framework for crucial interdisciplinary communication and understanding suited to newcomers to the field as well as experienced researchers and graduate level students.

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Electrophysiological methods are used at nearly all levels of biological research today. From basic science investigation to bringing new drugs to market, electrophysiology is an indispensable research technique that provides essential insights into cellular and molecular functions. This work is intended to be a practical guide for understanding and using contemporary electrophysiological manipulations and assays in brain research. New researchers attempting to learn electrophysiological techniques are confronted with a lack of adequate guiding resources. Currently, most electrophysiology-based research knowledge is passed on by experienced individuals to others within the same laboratory. But, this master-apprentice scenario has many inherent difficulties, as it is both a time intensive process and also leads to incomplete knowledge transfer, or even superstitious and maladaptive practices. Few external guiding resources exist for a budding researcher who wishes to gain the skills necessary to conduct responsible and competent electrophysiology research. Dr. Yan Dong, an expert in utilizing electrophysiological techniques for brain research with fifteen years of experience, and Peter Neumann hope this work can act as a concise and practical aid to remedy the current situation. This book will cover both basic methods and advanced techniques clearly and in depth, as well as the essential background and theory necessary to understand the methods. Additionally, this book also covers conventional and contemporary patch-clamp methods, experimental design, data collection and analysis, common pitfalls to avoid, important nuances, technical concerns, practical applications, and advice for teaching new researchers. Together, this represents a contemporary best-practices guide for collecting and interpreting electrophysiological data for brain research. ​

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This is a timely compilation of new concepts in the molecular pathogenesis and molecular therapy of acute myelogenous leukemia (AML). The focus is on selected critical molecular determinants of AML pathogenesis and pathophysiology and the exploitation of these factors by diverse therapeutic agents and modalities. There is an emphasis throughout on the bidirectional flow of knowledge between the clinical and laboratory arenas.

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This timely volume addresses the areas of pathophysiology and therapy of pulmonary hypertension, which have seen exciting developments over the past decade. The discoveries of endothelin overexpression as well as prostacyclin and nitric oxide deficiency in association with pulmonary hypertension have led to important therapeutic insights. The new therapies have led to significant improvements in patient function, quality of life and survival. In this book, expert authors describe these new therapies. It will be of interest not only to cardiologists, pulmonary specialists and rheumatologists, but also many nurses and pharmacotherapists.

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Attention deficit hyperactivity disorder (ADHD) is a common neurobehavioral disorder affecting 5–10% of children and adolescents and 3% of adults. Attention Deficit Hyperactivity Disorder: From Genes to Patients aims to provide a comp- hensive, state-of-the-art overview of the critical aspects of ADHD, and hopefully will serve as a quick and up-to-date reference source for professionals with an int- est in ADHD. The book is divided into three major areas that follow an historical survey. The first group of chapters deals with current theories on the pathophysiology of ADHD, and focuses on neurotransmitters and the contributions and validity of animal m- els. The second section emphasizes the evaluation and treatment of patients with ADHD, from the day-to-day approach by the clinical psychologist to the more sophisticated anatomical and functional imaging strategies that have emerged in the last decade. In addition, chapters dealing with specific impairments, such as those pertaining to reading, social interaction, and working memory, are also included for more detailed analysis of these important aspects and their respective contributions to global functioning. The third and final section provides an expanded review on the pharmacotherapy of ADHD and the appropriate methods for selection of specific drugs for individual patients based on drug kinetics and gene expression. David Gozal, MD Dennis L. Molfese, PhD vii CONTENTS Dedication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi 1 Historical Aspects of Attention Deficit Hyperactivity Disorder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Roscoe A. Dykman 2 Scanning the Genome for Attention Deficit Hyperactivity Disorder . . . . . . . . . . . . . . . . . . . . . . . . .

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The cause of diabetes mellitus is metabolic in origin. However, its major clinical manifestations, which result in most of the morbidity and mortality, are a result of its vascular pathology. In fact, the American Heart Association has recently stated that, “from the point of view of cardiovascular medicine, it may be appropriate to say, diabetes is a cardiovascular disease” (1). But diabetic vascular disease is not limited to just the macrovasculature. Diabetes mellitus also affects the microcirculation with devastating results, including nephropathy, neuropathy, and retinopathy. Diabetic nephropathy is the leading cause of end-stage renal disease in the United States, while diabetic retinopathy is the leading cause of new-onset blindness in working-age Americans. The importance of this text on Diabetes and Cardiovascular Disease is evident by the magnitude of the population affected by diabetes mellitus. Over 10 million Americans have been diagnosed with diabetes mellitus, while another 5 million remain undiagnosed. The impact from a public health perspective is huge and increasing. As the population of the United States grows older, more sedentary, and obese, the risk of developing diabetes and its complications will increase. Epidemiological studies have identified diabetes mellitus as a major independent risk factor for cardiovascular disease. Over 65% of patients with diabetes mellitus die from a cardiovascular cause. The prognosis of patients with diabetes mellitus who develop overt clinical cardiovascular disease is much worse than those cardiovascular patients free of diabetes mellitus.

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