In the last decade, a large number of major discoveries have shed light on the molecular mechanisms of lymphocyte migration and the anatomy of immune responses. In T-Cell Trafficking: Methods and Protocols, expert researchers explore how the development of novel and cutting-edge techniques, particularly in the field of real-time imaging and genetic manipulation, have led to an increased understanding of lymphocyte trafficking. Written by internationally recognized experts in their respective fields, chapters provide state-of-the-art protocols to study lymphocyte migration and T-cell: endothelial cell interactions in vitro, address various approaches used for direct visualization of the development of the lymphoid system, lymphocyte recirculation, and effector responses in experimental models in vivo, and explore lymphocyte migration and inflammation in the human system. Composed in the highly successful Methods in Molecular Biology (TM) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and highly practical, T-Cell Trafficking: Methods and Protocols is an essential manual for newcomers in this ever-expanding and exciting area of research, as well as a valuable addition to more specialized laboratories.

This book presents an overview of the most important current developments in the pre-analytical handling of tissue. It addresses in particular potential ways to improve the situation whereby methods employed in the pre-analytical phase – the period from surgical removal of tissue to the start of pathological processing – have remained essentially unchanged for decades with only modest standardization. It is examined how the pre-analytical period can be optimized, resulting not only in an increase in diagnostic quality but also in a reduction in processing time and costs. Among the key topics examined are the so-called cold ischemia time between tissue removal and fixation, the potential superiority of vacuum-based preservation over immediate formalin fixation, two-temperature fixation, molecular analysis methods, and the pre-analytics of specimens from particular tissues. Readers will find this book to be an important update that reveals the full importance of the pre-analytical phase for quality of pathological work-up.

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.

Metastatic dissemination of cancer is a main cause of cancer related deaths, therefore biological mechanisms implicated in metastatic process presents an essential object of cancer research. This research requires creation and utilization of adequate laboratory models. The book describes main approaches to model processes of metastatic cancer dissemination and metastases development. The book is structured in according with various metastatic pathways reflecting molecular specificity of metastatic process as well as anatomical specificity of aria of dissemination. Each chapter is introduced by short discussion of clinical aspects of certain metastatic pathway. Especial attention is paid for methods of visualization, quantification and analysis of the modeled metastases. Additional chapter is devoted to methods of mathematic modeling of tumor spread. The data presented in the book may be helpful for cancer researchers and oncologists.

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.

This volume provides a transdisciplinary and translational review of many of the leading murine models used to study the mechanisms, mediators and biomarkers linking energy balance to cancer. It provides a review of murine models that should be of interest to basic, clinical and applied research investigators as well as nutrition scientists and students that work in cancer prevention, cancer control and treatment. The worldwide obesity pandemic has been extensively studied by epidemiologic and observational studies and even, in some cases, by randomized controlled trials. However, the development and control of obesity, its comorbidities and its impact on cancer usually occurs over such long periods that it is difficult, if not impossible to conduct randomized controlled trials in humans to investigate environmental contributions to obesity, energy balance and their impact on cancer. In contrast, model organisms, especially mice and rats, provide valuable assets for performing these studies under rigorously controlled conditions and in sufficient numbers to provide statistically significant results. In this volume, many of the leading and new murine models used to study the mechanisms and mediators linking cancer with obesity, sleep, exercise, their modification by environment and how they may continue to be used to further elucidate these relations as well as to explore preclinical aspects of prevention and/or therapeutic intervention are considered. This volume provides an important compilation and analysis of major experimental systems and principles for further preclinical research with translational impact on energy balance and cancer.

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.

Recent critical advances in the field of immunotoxicology have led to incredible contributions to basic science, including crucial improvements in human risk assessment. In Immunotoxicity Testing: Methods and Protocols, expert researchers explore these changes, providing the reader with current, lab-ready procedures, along with the corresponding background information that is necessary to identify effective testing approaches for chemicals and drugs. Among the first volumes to meld consideration of immunotoxicity testing strategies with a comprehensive presentation of detailed laboratory protocols, chapters include a description of the evolution of immunotoxicity testing, a look at the importance of immunotoxicity testing for health risk reduction, and ideas concerning the future of the field. Composed in the highly successful Methods in Molecular Biology(tm) series format, most chapters contain a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips from the experts on troubleshooting and avoiding known pitfalls. Comprehensive and innovative, Immunotoxicity Testing: Methods and Protocols is a critical, one-stop reference resource for the most important and commonly used laboratory protocols in immunotoxicology.

Adverse drug reactions and interactions are still a major headache for healthcare professionals around the world. The US Food and Drug Administration's database recorded almost 300,000 serious adverse events in 2009 alone, of which 45,000 instances proved fatal. This updated new edition of the indispensable guide to drug interactions incorporates fresh research completed since the book's original publication by Humana Press in 2004. Additions include a new section on pharmacogenomics, a rapidly growing field that explores the genetic basis for the variability of responses to drugs. This new material reviews important polymorphisms in drug metabolizing enzymes and applies the findings to forensic interpretation, using case studies involving opiates as exemplars. Existing chapters from the first edition have in most cases been updated and reworked to reflect new data or incorporate better tables and diagrams, as well as to include recent drugs and formulations. Recent references have been inserted too. The handbook features extra material on illicit drug use, with a new chapter tackling the subject that covers cocaine, amphetamines and cannabis, among others. The section on the central nervous system also deals with a number of drugs that are abused illicitly, such as benzodiazepines, opiates flunitrazepam and GHB, while so-called 'social' drugs such as alcohol and nicotine are still discussed in the book's section on environmental and social pharmacology. Focusing as before on detailed explanation and incorporating both pharmacokinetic and pharmacodynamic drug interactions, this book will continue to be a lodestar for health and forensic professionals as well as students.

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.

Monoclonal Antibodies: Methods and Protocols, Second Edition expands upon the previous edition with current, detailed modern approaches to isolate and characterize monoclonal antibodies against carefully selected epitopes. This edition includes new chapters covering the key steps to generate high quality monoclonals via different methods, from antigen generation to epitope mapping and quality control of the purified IgG. Chapters are divided into four parts corresponding to four distinct objectives. Part I covers monoclonal antibody generation, Part II deals with monoclonal antibody expression and purification, Part III presents methods for monoclonal antibody characterization and modification, and Part IV describes selected applications of monoclonal antibodies. 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, Monoclonal Antibodies: Methods and Protocols, Second Edition provides crucial initial steps of monoclonal antibody generation and characterization with state-of-the art 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.

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.

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.

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.

Avian Influenza Virus, Second Edition aims to provide the essential methods used in working with animal influenza viruses, and to compile more advanced information that will guide the user in designing influenza studies. Influenza A viruses are among the most important pathogens for humans, food animals and companion animals. Research and diagnostics with animal influenza viruses are critical to animal health and it should be recognized that the needs and goals of animal agriculture and veterinary medicine are not always the same as those of public health. This volume sorts out the differences in the structure of the poultry, swine and equine industries, the biological differences of influenza virus from each animal group, and provides host, strain and lineage specific guidance and procedures. 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 accessible, Avian Influenza Virus, Second Edition seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of these important pathogens.

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.

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.

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.

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.

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.

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.

Tissue diagnosis in surgical pathology is the most important determinant of patient outcomes in oncology and other medical conditions. An appropriate diagnosis of a tumor correctly determines the necessary therapy for that patient. Error Reduction and Prevention in Surgical Pathology focuses on how errors happen, the best systems to detect errors and the best systems to prevent errors. As with any form of testing, the test cycle in surgical pathology can be divided into pre-analytic (collection and handling of specimens), analytic (the actual determination of disease or other diagnostic information) and post-analytic (generation of reports and distribution). Error can occur anywhere in the process and all of these steps must be performed optimally so that the correct diagnosis is made and delivered. Error Reduction and Prevention in Surgical Pathology serves as an essential guide to a successfully managed laboratory and contains all relevant information needed to manage patients and deliver the best diagnosis.

This book provides a comprehensive, state-of-the art review of the genitourinary tumor pathology field and the most contemporary insights regarding specimen submission, histologic morphology, immunohistochemistry, and molecular studies useful in the diagnosis of genitourinary neoplasms. Discussion of the clinical implications of pathological findings is contributed by renowned clinicians in the field. This handsome volume guides the reader through the intricacies of genitourinary tumor pathology, diagnosis, reporting, and prognosis. Written by experts in the field, Genitourinary Pathology: Practical Advances is of great value to anatomic pathologists, urologists, fellows in genitourinary pathology, as well as upper level residents training in pathology.