This is a collection of cutting-edge mycoplasma methods for the detection, isolation, identification, characterization, and genetic manipulation of the pathogenic mycoplasmas. These step-by-step methods are crafted for successful reproducibility and include biochemical, genetic, and molecular techniques essential to understanding pathogenecity and adhesion to host cells. They also cover the detection of mycoplasmas in cell cultures, an important tool not only in viral diagnosis and research, but also in the production of vaccines and various biological products.

This latest addition to the Methods in Molecular Medicine series, Anti- ral Methods and Protocols, is opportune because there is an increasing int- est in discovering compounds that are effective against both chronic and acute viral infections. A number of the methods described in the volume are unp- lished and their inclusion indicates the speed at which this field is moving. This volume is not a review but each chapter contains methods validated by the experts who have spent time in developing the protocols. The hallmark of this series is the comprehensive way in which the me- ods are described, which includes a list of all the reagents needed for each protocol. Of importance is the section on tips and pitfalls that the authors have discovered while developing their protocols. The manual itself is designed to be used by researchers in universities and industry who are familiar with a range of biological techniques but who want to set up quickly a novel assay system. We encourage a dialog between readers and authors, which may also result in useful collaborations.

Mycoplasmas cause some of the most serious and economically significant diseases in livestock and pose major problems for animal health authorities worldwide. Infection has spread in the last five years to new regions and species, but little effective control is available, particularly in developing countries. This work encapsulates the latest research and development on mycoplasmas in sheep, goats and cattle from laboratories all over the world, describing both conventional diagnostic techniques for growth and identification and newly established procedures such as PCR/DGGE. Molecular typing methods are also covered, specifically for use in mycoplasma fingerprinting as well as up to date reviews on the major mycoplasma diseases including contagious bovine and caprine pleuropneumonias, contagious agalactia and many conditions caused by Mycoplasma bovis.

In Eicosanoid Protocols, Elias A. Lianos and a panel of hands-on experts present cutting-edge methods for the study of eicosanoids, including prostaglandins, thromboxanes, and leukotrienes. The readily reproducible methods described hereconcentrate on studying the regulation of expression and function of enzymes, particularly cyclooxygenase (and its two isoforms), phospholipase A2, and lipoxygenases involved in the synthesis of established eicosanoids. Additional chapters are devoted to the characterization and distribution of the thromboxane A2 receptor in tissues and the biological roles of novel eicosanoids. Timely and authoritative, the methods in this book will help their users in exploring the pathobiology of inflammation. Eicosanoid Protocols offers new and established researchers powerful, state-of-the-art tools to probe the regulation and function of eicosanoids.

In Natural Killer Cell Protocols: Cellular and Molecular Methods, Kerry S. Campbell and Marco Colonna have assembled a comprehensive collection of readily reproducible methods designed to study natural killer (NK) cells from the broadest variety of viewpoints. These include not only classic techniques, but also new approaches to standard methods, newly evolved techniques that have become valuable for specific applications, and unique models for manipulating and studying NK cells. Among the advanced methods covered are those for in vitro transendothelial migration, in vivo detection of cells migrating into tumors, immunofluorescence staining of intracellular cytokines, and in vitro NK cell development. Valuable techniques for specific applications include vaccinia virus protein expression, soluble KIR-Fc fusions for HLA class I binding assays, calcium mobilization in cell conjugates, and identification of heterodimeric receptor complexes using cDNA library expression cloning. No less important are accounts of such classic methods as hybrid resistance, ADCC, viral defense, target cell cytotoxicity assays, cloning and culturing, tumor immunotherapy, and generation of HLA class I transfected target cells. Natural Killer Cell Protocols: Cellular and Molecular Methods offers immunologists, cancer researchers, virologists, and cell biologists today's most comprehensive collection of both established and cutting-edge techniques, methods that will contribute significantly to advancing our understanding of this fascinating and critically important class of cells.

At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. Written in the highly successful Methods in Molecular BiologyTM, 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 up-to-date, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.

Lorette Javois' timely new 2nd edition revises and updates her widely acclaimed collection of step-by-step immunocytochemical methods, one that is now used in many biological and biomedical research programs. The methods are designed for researchers and clinicians who wish to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. In addition to cutting-edge protocols for purifying and preparing antibodies, light microscopic analysis, confocal microscopy, FACS, and electron microscopy, this revised edition contains many new methods for applying immunocytochemical techniques in the clinical laboratory and in combination with in situ hybridization.