This volume provides a comprehensive, state-of-the art review in the field of experimental and human nephrogenesis. The book reviews new data on the effects on kidney development by neonatal asphyxia, obstructive uropathies, nephrotoxic drugs, malnutrition, underfeeding, overfeeding and provides all possible preventive measures to ensure the well-being of the kidney at birth. The book also discusses the possible implications between renal development and the insurgence of kidney disease in adult life and the correlation with renal carcinogenesis. Written by well recognized experts in their fields, Kidney Development in Renal Pathology is a valuable tool for pathologists, neonatologists, nephrologists, gynecologists and researchers with an interest in kidney diseases.

In Vitro Toxicology Systems brings together important issues and considerations needed in order to develop a workable, reliable, integrated testing strategy for the replacement of animals in toxicity testing regimes. This thorough volume includes sections on in vitro models for systemic organ toxicity, neurotoxicity, sensory organs, immunotoxicity and reproductive toxicity and addresses how stem cells may be used going forward. The book also tackles difficult areas of toxicology such as carcinogenicity and nanotoxicology, with additional chapters dedicated to kinetics, metabolism, and in vitro in vivo extrapolation. The book also addresses biological processes such as stress response pathways and mechanistic biomarkers and how these can be uncovered and measured using high content approaches. Reliable and authoritative, In Vitro Toxicology Systems will be of benefit not only to students, scientists and regulators working in the field of chemical safety assessment but also to a wider scientific audience.

Reflecting the development of powerful new tools and high-throughput methods to analyze adenoviral particles and their interactions with host cells, the third edition of Adenovirus Methods and Protocols calls upon experts in the field to convey advances in molecular biology, genomics and proteomics, imaging, and bioinformatics. Beginning with cryo-electron microscopy, atomic force microscopy, and mass spectrometry for a high resolution image and characterization of the virion, this detailed book then continues with capsid modifications and viral-like particles as promising alternatives to classical adenovirus vectors, and the study of adenovirus in host interactions in vitro at the cellular level as well as in vivo in animal models. Finally, the volume concludes with an extensive update of the most efficient protocols to generate, amplify, and/or purify, at small and large scale, standard human Ad5 as well as non-human, chimeric, and helper-dependent adenovirus vectors. Written in the greatly 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. Comprehensive and cutting-edge, Adenovirus Methods and Protocols, Third Edition serves as an ideal guide for scientists continuing to research this highly valuable viral tool.

This book focus on genetic diagnostics for Uniparental Disomy (UPD), a chromosomal disorder defined by the exceptional presence of a chromosome pair derived from only one parent, which leads to a group of rare diseases in humans. First the molecular and cytogenetic background of UPD is described in detail; subsequently, all available information of the various chromosomal origins and the latest findings on genotype-phenotype correlations and clinical consequences are discussed. Numerous personal reports from families with a child suffering from a UPD-induced syndrome serve to complement the scientific and clinical aspects. Their experiences with genetic counseling and living with a family member affected by this chromosomal aberration present a vivid picture of what UPD means for its victims.

Sine syndromes (SSs) represent atypical forms of inflammatory rheumatic diseases, the courses of which are often severe. While the diagnosis of rheumatic diseases can be determined according to an established set of diagnostic criteria based on clinical characteristics and laboratory parameters, Sine Syndromes do not fulfill the standard criteria used to assist in the classification of patients with rheumatological disorders. This concise book gives an overview of atypical courses in common inflammatory rheumatic diseases such as Wegener's granulomatosis, systemic sclerosis, Sjoegren's syndrome, or systemic lupus erythematosus. Each chapter reviews similar cases reported in the literature and presents current data on treatment options. Drawing on their vast clinical experience, the editors provide a series of detailed case reports in order to illustrate the different types of disease. The book is intended to facilitate early diagnosis and effective therapy in patients where a schematic approach may not prove sufficient. Sine Syndromes in Rheumatology will serve as a useful and easily accessible reference for specialists in rheumatology as well as for practitioners in the fields of internal medicine, pediatrics, and orthopedics.

Today, biosensors are broadly applied in research, clinical diagnosis and monitoring, as well as in pharmaceutical, environmental or food analysis. In this work, the author presents the essentials that advanced students and researchers need to know in order to make full use of this technology. This includes a description of biochemical recognition elements, such as enzymes, antibodies, aptamers or even whole cells. Various signal transducers such as electrochemical and optical transducers, luminescence devices and advanced techniques such as quartz crystal microbalances and MEMS systems are covered as well. Current applications are introduced through various case studies, rounded out by a forward-looking chapter on the prospects for biosensor development offered by nanotechnology, lab-on-a-chip, and biomimetic systems.

Despite the many milestones in cystic fibrosis (CF) research, progress toward curing the disease has been slow, and it is increasingly difficult to grasp and use the already wide and still growing range of diverse methods currently employed to study CF so as to understand it in its multidisciplinary nature. Cystic Fibrosis: Diagnosis and Protocols aims to provide the CF research community and related researchers with a very wide range of high-quality experimental tools, as an easy way to grasp and use classical and novel methods applied to cystic fibrosis. Volume II: Methods and Resources to Understand Cystic Fibrosis focuses on pathophysiology, Omics approaches, and a variety of key resources recently made available for CF research. Written in the highly successful Methods in Molecular Biology (TM) 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. Comprehensive and practical, Cystic Fibrosis: Diagnosis and Protocols will provide readers with optimal working tools to address pressing questions in the best technical way, while helping all of us, as a research and clinical community, to move faster hand-in-hand toward unravelling the secrets of this challenging disorder and cure it.

Psychoneuroimmunology (PNI) has developed rapidly in the last four decades. As a multidisciplinary area, PNI may provide a scientific basis for mind-body relationships toward the development of personalized and systems medicine. In Psychoneuroimmunology:Methods and Protocols, expert researchers in the field detail methods and protocols geared toward the development of integrative and individualized therapeutics in multiple dimensions from drugs to behaviors. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Psychoneuroimmunology:Methods and Protocols aids scientists in continuing to study holistic views for the translation of psychoneuroimmunology into better preventive and personalized medical practice.

Presenting the most up-to-date techniques for the detection, genotyping, and investigation of methicillin-resistant S. aureus (MRSA), this second edition of Methicillin-Resistant Staphylococcus aureus (MRSA) Protocols collects chapters that utilize the power of complete genomic sequences and advanced high-throughput technologies that have pushed this field to its present state. These enable the development of specific and rapid diagnosis methods, the investigation and elucidation of mechanisms of bacterial evolution to antibiotic resistance and pathogenicity, and the identification of novel targets to develop more potent therapeutic and/or preventive agents. Written for the Methods in Molecular Biology series, numerous 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 fully updated, Methicillin-Resistant Staphylococcus aureus (MRSA) Protocols, Second Edition serves as a key reference for researchers attempting to set up a new method to study MRSA or even for technicians and scientists working on other pathogens.

In Molecular Diagnostics for Melanoma: Methods and Protocols, expert researchers and clinicians in the field of melanoma provide updated information on biomarkers and assays for diagnosis, prognosis, and assays predicting response to treatment for routine testing. The focus of the volume is on biomarkers with established clinical validity rather than those on early discovery stage. With additional in-depth discussion of the molecular biology and pathology of melanoma, treatment options in adjuvant and metastatic setting, and implications of biomarker testing for clinical management of melanoma patients. Written in the highly successful Methods in Molecular Biology series format, chapters include extensive 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. Comprehensive and practical, Molecular Diagnostics for Melanoma: Methods and Protocols seeks to provide both clinicians and scientists with technical information and extensive background information on the wide ranging approaches available in the field of diagnostics of melanoma.

The earliest descriptions of human chromosomes initiated the genomics revolution that is now upon us. Array Comparative Genomic Hybridization: Protocols and Applications explores the scope of what is now possible as far as documenting abnormalities associated with several human cancers. While the technology for interrogating the human genome continues to evolve, refinement of comparative genomic hybridization (CGH) using array CGH and related technologies have provided enormous insight into human cancers at an affordable scale. Written in the successful Methods in Molecular Biology (TM) 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, Array Comparative Genomic Hybridization: Protocols and Applications provides researchers with well-honed methodologies to learn these techniques for their own use in research or clinical diagnostic laboratories.

Microbiologists, medical mycologists, immunologists, and biochemists are increasingly working together to focus on the processes involved in the progression and treatment of fungal disease. Host-Fungus Interactions: Methods and Protocols is designed for research scientists who are involved in this work and interested in undertaking new or comparative studies of interactions between the mammalian host and clinically important fungal pathogens. Aiming to combine approaches for reverse genetics in pathogenic fungi with methods for their application in in vitro and in vivo models of disease, the book includes methods for the culture and genetic manipulation of the primary fungal pathogens and the opportunistic pathogens, as well as methods for investigating host-fungus interactions in model systems. Written in the highly successful Methods in Molecular Biology (TM) 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. Comprehensive and practical, Host-Fungus Interactions: Methods and Protocols describes available molecular methods and fungal infection models in great detail in order to encourage researchers to try new approaches to investigating host-fungus interactions with added levels of confidence.

Recently, important new findings in the polyamine field and a variety of new experimental systems have revolutionized the study of these ubiquitous cellular components, essential for normal growth and development. In Polyamines: Methods and Protocols, leading researchers contribute an extensive collection of up-to-date laboratory techniques for the further pursuit of polyamine study. The volume delves into vital subjects such as neoplasia studies with animal models and human patients, therapeutic roles for polyamine inhibitors and analogs, polyamine metabolism and oxidative damage, polyamines as regulators of critical ion channels, as well as polyamine transport systems and polyamine-responsive genes. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and expert notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Polyamines: Methods and Protocols provides a key resource for all scientists pursuing the study of this dynamic and significant aspect of cellular biology.

Research in the pharmaceutical sciences and medicinal chemistry has taken an important new direction in the past two decades with a focus on large molecules, especially peptides and proteins, as well as DNA therapeutics. In Drug Design and Discovery: Methods and Protocols, leading experts provide an in-depth view of key protocols that are commonly used in drug discovery laboratories. Covering both classic and cutting-edge techniques, this volume explores computational docking, quantitative structure-activity relationship (QSAR), peptide synthesis, labeling of peptides and proteins with fluorescent labels, DNA-microarray, zebrafish model for drug screening, and other analytical screening and biological assays that are routinely used during the drug discovery process. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and accessible, Drug Design and Discovery: Methods and Protocols serve as a vital laboratory reference for pharmaceutical chemists, medicinal chemists, and pharmacologists as well as for molecular biologists.

Clostridium difficile, a major nosocomial pathogen shown to be a primary cause of antibiotic-associated disease, has emerged as a highly transmissible and frequently antibiotic-resistant organism, causing a considerable burden on health care systems worldwide. In Clostridium difficile: Methods and Protocols, expert researchers bring together the most recently developed methods for studying the organism, including techniques involving isolation, molecular typing, genomics, genetic manipulation, and the use of animal models. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Clostridium difficile: Methods and Protocols serves as an ideal guide for scientists now in a position to gain an in-depth understanding of how this organism is transmitted and how it causes disease.

During the past decade as the data on gene sequences and expression patterns rapidly accumulated, cell-free protein synthesis technology has also experienced a revolution, becoming a powerful tool for the preparation of proteins for their functional and structural analysis. In Cell-Free Protein Production: Methods and Protocols, experts in the field contribute detailed techniques, the uses of which expand deep into the studies of biochemistry, molecular biology, and biotechnology. Beginning briefly with basic methods and historical aspects, the book continues with thorough coverage of protein preparation methods, the preparation of proteins that are generally difficult to prepare in their functional forms, applications of the cell-free technologies to protein engineering, as well as some methods that are expected to constitute a part of future technologies. Written in the highly successful Methods in Molecular Biology (TM) series format, the 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. Authoritative and cutting-edge, Cell-Free Protein Production: Methods and Protocols aims to help researchers continue the growth of the vital exploration of cell-free sciences and technologies in order to better understand the dynamic lives of cells.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 4-7), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry,histologicalstaining (Chapters10and11),aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 14-16) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocols.

Non-clinical drug safety evaluation, the assessment of the safety profile of therapeutic agents through the conduct of laboratory studies in in vitro systems and in animals, is an essential step in the progress of new pharmaceuticals heading toward the ultimate goal of clinical trials and, eventually, approval. In Drug Safety Evaluation: Methods and Protocols, expert researchers detail a compendium of analytical technologies with a focus on clarity and applicability in real life laboratory practice. These meticulous contributions feature key topics such as acute to chronic general toxicity studies, histopathology studies, reproductive toxicity studies, genotoxicity studies, safety pharmacology studies, investigative toxicity studies, and safety biomarker studies. As a volume in the highly successful Methods in Molecular Biology (TM) series, chapters include brief introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Drug Safety Evaluation: Methods and Protocols serves as an ideal guide to this field, helpful to pharmaceutical scientists, toxicologists, biochemists, and molecular biologists as well as scientists from all other disciplines who wish to translate these thorough methods into their own work.

Since the publication of the popular first edition, the explosion of DNA sequence information, the access to bioinformatics and mutation databases coupled with the ability to readily detect and confirm mutations has cemented the role of molecular diagnostics in medicine and, in particular, mutation detection by the polymerase chain reaction (PCR). In PCR Mutation Detection Protocols, Second Edition, expert researchers bring the subject up-to-date with key protocols involving the PCR and its many various incarnations such as SSCP, CSGE, and dHPLC. The volume also addresses key areas such as Southern blotting, accurate diagnostics with high throughput, as well as microarray systems. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes which provide the often hard to find information that may mean the difference between the success and failure of the method. Authoritative and cutting-edge, PCR Mutation Detection Protocols, Second Edition aims to stimulate postgraduate scientists, researchers, and clinicians already engaged in the area and to provide an important first step for those new to this practice wanting to adopt the powerful and essential technique in their own laboratories.

Since its early days in the 1990s, the Quorum Sensing (QS) field has grown from a few dozen laboratories, investigating the pathways, proteins, and chemicals that facilitate signaling in bacteria, to hundreds of groups that have integrated evolutionary biology, computer science, mathematics, engineering, and metagenomics to create an ever-expanding and dynamic field. In Quorum Sensing: Methods and Protocols, expert researchers provide an in-depth set of diverse protocols that span this broad area of study. Broken into three detailed sections, the volume covers the detection, isolation, and characterization of the QS signals made by both Gram- and Gram+ bacteria, determination of the function of QS signals in vivo, and the development of QS disruption strategies. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and expert tips on troubleshooting and avoiding known experimental pitfalls. Comprehensive and cutting-edge, Quorum Sensing: Methods and Protocols serves as an invaluable collection of easily accessible techniques for scientists seeking to further our knowledge about bacterial communication and its relation to humanity.

From the early days when RNA interference was a strange artifact in worms to the 2006 Noble Prize received by Fire and Mello and the current clinical trials, the field of RNA interference has grown at a breakneck pace. In RNA Interference: From Biology to Clinical Applications, expert contributors provide an overview of the most current science and protocols that span the biological disciplines from detailed nucleic acid chemistry, to pharmacology, to the manipulation of signal transduction pathways. Divided into three distinct sections, this volume delves into the physiology of RNA interference, RNA interference in the laboratory and siRNA delivery, and preclinical and clinical issues associated with the use of RNAi-inducing agents as drugs in order to stimulate new questions and offer the tools necessary to start addressing those questions. Written in the highly successful Methods in Molecular Biology (TM) 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. Authoritative and inspiring, RNA Interference: From Biology to Clinical Applications aims to promote and motivate innovation by reviewing what has been done, providing details of how it has been done, and encouraging speculation on what the future may hold.

As the drug discovery process shifts more and more toward specifically targeting pathways and molecules, model systems continue to increase in importance, and the mouse, with its versatility, ease of use, and remarkable similarity to the human genome, has clearly risen to the forefront of animal model studies. In Mouse Models for Drug Discovery: Methods and Protocols, experts in the field present some background for those less familiar with mice as experimental model platforms as well as a collection of techniques involving general methods as well as specific disease topics such as type 1 and 2 diabetes, cardiovascular disease, arthritis, skin disorders, cancer, the use of behavioral models for depression and anxiety, neurodegenerative diseases, neuromuscular diseases, and infectious diseases. Written in the highly successful Methods in Molecular Biology (TM) 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. Authoritative and easy-to-use, Mouse Models for Drug Discovery: Methods and Protocols will stimulate those not familiar with the power of the mouse and its potential for the drug discovery process, and it will encourage the development of new models and new ways to utilize existing models in order to further the use of this dynamic animal in this vital field.

Enzyme-linked immunospot assay (ELISPOT) has been known for some time as a unique state-of-the-art technique for studying the cytokine-secreting activity of immune system cells, and it appears to be one of the fast growing applications in biomedical research, becoming an indispensable tool in vaccine development, HIV research, transplantation studies, and cancer and allergy research. The second edition of Handbook of ELISPOT: Methods and Protocols, only the second book in the field which is entirely dedicated to ELISPOT assay, shares the detailed techniques that have been developed since the release of the popular first edition. Straight from the labs of seasoned experts, this book covers setting and performing ELISPOT assays, ELISPOT for veterinary research, advanced ELISPOT techniques, image and data analysis, as well as vaccine development and diagnostics. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective chapters, 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, Handbook of ELISPOT: Methods and Protocols, Second Edition serves as a compilation of a technical reference and a troubleshooting guide for researchers, both experienced and novice, worldwide in order to advance the usage of this key tool.

Infection by flaviviruses such as dengue virus serotypes (DENV 1-4), Japanese encephalitis virus (JEV), tick-borne encephalitis virus (TBE), yellow fever virus (YFV) and West Nile virus (WNV) impact millions of lives and cause tens of thousands of mortalities each year. Dengue is a global public health emergency especially since there is no preventative vaccine or antiviral treatment for dengue disease. Dengue: Methods and Protocols offers the increasing number of dengue researchers a one-stop protocol book with techniques compiled from the leading laboratories working on dengue. Chapters cover topics such as dengue virus isolation from clinical samples, quantification of human antibodies against the virus, assays to quantify the virus particles, the widely used mouse model to study dengue pathogenesis, vaccine and antiviral efficacies. 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, Dengue: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies on dengue research.