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Since the first report of an engineered oncolytic virus, there has been a continuing and steady increase of interest in the field, and while bench research remains vital for the translation of research in this field, its success depends on breakthroughs in clinical studies. Oncolytic Viruses: Methods and Protocols describes the construction and purification of capsid-modified adenoviruses as well as oncolytic adenoviruses, presents protocols for many individual virus species including engineering and preparation of oncolytic HSV, propagation, purification, and in vivo testing of oncolytic VSV, details properties of oncolytic reovirus and NDV, and describes the generation and testing of next generation of oncolytic vaccinia virus. As the host immune system plays a critical role in determining efficacy of oncolytic viruses, two chapters are devoted to the study of immune response. Recent advances in stem cell research have led the field in two distinct directions: the use of stem cells as carrier vehicles for oncolytic viruses and the targeting of cancer stem cells. As such, the volume describes the use of explant tissue samples from patients to potentially provide useful information predicting responses prior to clinical translation. 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, Oncolytic Viruses: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential and vital field.
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This volume provides an overview of molecular biology tools for pathogen detection and characterization. Focusing on biobanking, biosafety, good laboratory practices, biological specimen collection and processing, quality assurance and control, validation of molecular diagnostic assays for veterinary use; molecular detection and identification of animal pathogens using a wide range of established techniques and covering, also, emerging diagnosis approaches; genotyping tools for assessing the genetic landscape and epidemiology of pathogens; and integrative omics and high-throughput technologies as powerful research tools. 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, Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies aids scientists in continuing to study a broad range of molecular approaches, from simple and affordable to emerging and sophisticate.
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This volume is a collection of research methods, techniques, and approaches to investigate the molecular biology of West Nile Virus (WNV). Chapters in the book cover many facets of WNV, such as propagation and titration of WNV in vero cells, examination of WNV neuroinvasion and neuropathogenesis in the central nervous system of a murine model, field surveillance methods for WNV, and in vitro and in vivo blood-brain barrier models to study WNV pathogenesis. A brief introduction, along with a discussion on WNV laboratory safety are also included. Written in the highly successful Methods in Molecular Biology series format, chapters includes introduction 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. Practical and thorough, West Nile Virus will interest virologists, molecular biologists, microbiologists, and other scientists working with WNV. This book is a comprehensive guide for those conducting research in the field of WNV biology, including life cycles, pathogenesis, impact of infection, and possible treatment/prevention development.
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This volume summarizes current techniques; ranging from protocols for virus growth, isolation, quantification and generation of recombinant RSV virus to procedures for the efficient characterization of the host immune response to RSV infection. These techniques are used in numerous laboratories around the world and are the building blocks that support the majority of RSV virus research. 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, Humana Respiratory Syncytial Virus: Methods and Protocols aims to ensure successful results in the further study of this vital field.
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This volume explores data from the applications of molecular biological methods and the applications of recent immunological and cytogenetic methods in Epstein-Barr Virus (EBV) that will offer readers possible new solutions to the unresolved problems in the EBV field. Chapters in this book cover topics such as: viral life cycle, latency, EBV-associated diseases and EBV diagnostics; in vitro methods including organotypic cultures for the analysis of EBV-epithelial cell interactions; identification of the interacting viral and cellular proteins using affinity purification-mass spectrometry methods; 3D telomere FISH; transcription analysis using high-throughput RNA sequencing, qPCR and nuclear run-on assay; analysis of viral and cellular microRNAs; isolation and characterization of exosomes and the assessment of their function; characterization of the viral genome by terminal repeat analysis and sequencing; the use of chromatin immunoprecipitation coupled sequencing (ChIP-Seq) for the analysis of Zta-DNA interactions; epigenetic analysis by bisulfite sequencing and ChIP; novel in vivo models for the study of EBV infection; and how immunological, virological, tissue culture and molecular methods can be combined to yield Good Manufacturing Practice-compliant EBV-specific T cells for the immunotherapy of EBV-associated post-transplant lymphoproliferative diseases (PTLD). 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, Epstein Barr-Virus: Methods and Protocols is a valuable resource for anyone who is interested in this fascinating and evolving field.
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This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug discovery. 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, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV. .
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While HIV-1 continues to be well-researched, this detailed volume draws attention to other members of the Retrovirus family, namely the Human T-lymphotropic Viruses (HTLVs), featuring the most updated technical information about HTLV determination and the methods to investigate their interaction with the host immune system and interfering pathogens. The contents include essential aspects of epidemiology and virus transmission, novel and robust methodologies for studying the effects of trans-activating regulatory HTLVs’ proteins, the latest techniques for genotyping and gene expression analysis, as well as cellular phenotype and dynamics. 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, Human T-Lymphotropic Viruses: Methods and Protocols serves as an ideal guide to an area of study that is very much worthy of further research.
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The world abounds with microorganisms that are hazardous to human health in the normal course of life. In Public Health Microbiology: Methods and Protocols, expert investigators describe in step-by-step detail the laboratory techniques they have perfected for determining the nature and character of a diverse group of hazardous bacteria, viruses, and fungi. For bacterial and viral pathogens, the authors give special attention to PCR methods for detecting genes resistant to tetracycline, for resistance in Salmonella enterica, for identifying and typing Campylobacter coli, and for detecting the abundance of enteric, hepatitis A, and rotaviruses in sewage, as well as bacteriophages infecting the O157:H7 strain of Escherichia coli. For fungi the authors offer methods for computerized analysis and typing of fungal isolates, for the isolation and enumeration of fungi in foods, and for the determination of aflatoxin and zearalenone. Other methods deal with hazard analysis, the use of disinfectants, microbiological analysis of cosmetics, microbiological tests for sanitation equipment in factories, and the uses of animals in studies of various characteristics of lactobacilli. Review articles address the spread of pathogens from livestock and poultry production, including the threat of prion-based diseases. All protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Timely and highly practical, Public Health Microbiology: Methods and Protocols offers researchers and public health specialists alike a wide-ranging collection of methods for the determination of many important pathogenic microorganisms, their effects on health, and possible measures that can be taken to counter these effects.
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In this second edition of a much-acclaimed classic laboratory manual, Molecular Diagnosis of Infectious Diseases, leading experts from academia and industry describe cutting-edge methods for the protein-based diagnosis of infectious diseases. Explaining the latest developments in genomics, proteomics, bioinformatics, biosensors, high-throughput devices, and recombinant technology, the authors demonstrate the power of these new methodologies in both established and novel procedures by applying them successfully to the identification and characterization of valuable diagnostic markers, immunomodulatory components, epitope mapping, the production and purification of recombinant antigens, and their use in diagnostic reagents in immunological assays. Basic problems of serological laboratory diagnostics and the use of immunoproteomics for discovering target proteins are also addressed. Each readily reproducible protocol is described in step-by-step detail and includes a background introduction, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and extensive references. Comprehensive and up-to-date, Molecular Diagnosis of Infectious Diseases, Second Edition offers novice investigators an easy entry into the field and experienced researchers a broad array of time saving tools that will not only elucidate new principles, but also greatly facilitate the development and successful application of rapid diagnostic methods.
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