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. It is written in the highly successful Methods in Molecular Biology trademark], 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.

Methods in Enzymology serial highlights new advances in the field with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

Animal cell technology is a growing discipline of cell biology which aims to understand the structure, function and behaviour of differentiated animal cells, and especially the development of such abilities as are useful for industrial purposes. These developments range from clonal expansion of differentiated cells with useful abilities, to optimization of cell culture on industrial scale and modulation of the cells' abilities to produce drugs and monoclonal antibodies. The sixth volume in this series gives a complete review of today's state of the art in Japan, a country where this field is especially well advanced. It will be of interest to cell biologists, biochemists, molecular biologists, immunologists and other disciplines related to animal cell culture, working in the academic environment as well as in (biotechnology or pharmaceutical) industry.

Methods in Cancer Stem Cell Biology: Part B, Volume 171 in the Methods in Cell Biology series highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including Orthotopic brain tumor models derived from glioblastoma stem-like cells, RNA sequencing in hematopoietic stem cells, Generation of inducible pluripotent stem cells from human dermal fibroblasts, In vitro preparation of dental pulp stem cell grafts combined with biocompatible scaffolds for tissue engineering, Gene expression knockdown in chronic myeloid leukemia stem cells, Identification and isolation of slow-cycling GSCs, Assessment of CD133, EpCAM, and much more.

Plant Cell and Tissue Culture continues the high standards of Humana's Methods in Molecular Biology series. Its step-by-step approach (a hallmark of the series) is applied to a wide range of basic laboratory techniques and culture conditions appropriate to plant cells. Because of the diversity of cell types, species, and culture methods, much of this volume is devoted to the culture of particular cell types and to the regeneration of these cells into whole plants. Special attention is also given to the genetic modification of plants, as well as to the economic significance of plant products. Chapters cover a wide range of topics and techniques, including:* tissue culture media and selection * cryopreservation * callus culture techniques * organ culture * embryogenesis * batch culture * large-scale culture * hormonal control * fertilization techniques * gene transfer * cell immobilization * production systems * cell product purification * DNA expression * electrofusion of plant cells * mutant selection * mutagenesis techniques * automation * transfer of nuclei * protoplast culture * media analysis * micropropagation. A detailed appendix lists the formulas for the most commonly employed plant cell media. Comprehensive, easy to follow, and a pleasure to use, Pollard and Walker's Plant Cell and Tissue Culture is an essential tool for everyone--at all levels of proficiency and experience--involved in plant culture.

Understanding seed-related processes is of major social, environmental, and economic concern. The viability and vigor of seeds are the very basis for sustainable agriculture and forestry, and comprehending the molecular and cellular events underlying these processes will become increasingly important to many economical sectors and for species that provide the world's food supply. Seed Dormancy: Methods and Protocols covers analytical methods and approaches which have already lead to significant advances in the understanding of seed dormancy and germination. Chapters cover explanations of processes leading to the induction, maintenance, and termination of seed dormancy, the classification of different dormancy types, as well as an overview of protocols used for dormancy-termination of seeds of conifer species. This volume emphasizes methods essential for abscisic acid (ABA) analyses, including methods that have been important for receptor identification, analyses of ABA-catabolizing enzymes (the 8'-hydroxylases), and identification of novel signal transduction components, interacting partners, and/or response factors. The volume closes by addressing the development of new technologies, including spectroscopic methods (some of which allow for non-destructive sampling) as well as highly effective tissue-printing methods for seed dormancy research. 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, Seed Dormancy: Methods and Protocols features detailed methods that will prove invaluable for both applied and fundamental seed research.

The health of the proteome depends upon protein quality control to regulate the proper synthesis, folding, translocation, and clearance of proteins. The cell is challenged constantly by environmental and physiological stress, aging, and the chronic expressions of disease associated misfolded proteins. Substantial evidence supports the hypothesis that the expression of damaged proteins initiates a cascade of molecular events that leads to Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, and other diseases of protein conformation.

As one of the fastest growing fields of research in the 21st century, nanotechnology is sure to have an enormous impact on many aspects of our lives. Nanostructure Design: Methods and Protocols serves as a major reference for theoretical and experimental considerations in the design of biological and bio-inspired building blocks, the physical characterization of the formed structures, and the development of their technical applications. The chapters contributed by leading experts are divided into two sections, the first of which covers experimental aspects of nanostructure design and the second delves into computational methods. As a volume of the highly successful Methods in Molecular Biology (TM) series, this collection pulls together cutting-edge protocols, written in a step-by-step, readily reproducible format certain to guide researchers to the desired results. Comprehensive and essential, Nanostructure Design: Methods and Protocols uses biological principles and vehicles on design to aid scientists in the great challenges still ahead.

The filamentous fungi are perhaps unique in the diversity of their metabolic activities. This includes biosynthetic as well as degradative activities, many is to provide an of which are of industrial interest. The objective of this text up-to-date and broad review which emphasizes the genetic and molecular biological contribution in the field of fungal biotechnology. This text begins with an overview of the tools and methodologies involved which, to a large extent, have been developed in the model filamentous fungus Aspergillus nidulans and subsequently have been extended to commercially important fungi. This is followed by a chapter which provides a compilation of genes isolated from commercial fungi and their present status with respect to structure, function and regulation. Chapters 3 and 4 highlight the degradative powers of filamentous fungi. First, a discussion of what is known regarding the molecular genetics of fungi and the genes and enzymes involved in the beverage and food industries. This has an oriental flavour, reflecting the tremendous importance of fungi in traditional Chinese and Japanese food production. An account of lignocellulose degradation by filamentous fungi follows, illustrating the potential of fungi to utilize this substance as a renewable energy source. The ability of fungi to produce high-value foreign proteins is reviewed in chapters 5 and 6. Chymosin production, in particular, represents a good example of high-level yields being obtained, such as to warrant commercial production.

Arts, heritage, non-profit and social marketing today comprise key components of the contemporary marketing management scene. Governments, charities and voluntary sector organisations throughout the world are increasingly involved in the development of marketing campaigns, and more and more of these organisations are likely to be at the cutting edge of the application of the very latest marketing methods. Research in the arts, heritage, non-profit and social marketing fields is intellectually rigorous, relevant for user communities, and has a great deal to offer to marketing theory as well as to promotional practice. This book presents a collection of stimulating articles that report some of the freshest and most innovative research and thinking in the authors' specialist domains. Collectively the chapters offer a balance of empirical and conceptual research in arts, heritage, non-profit and social marketing. They explore new ideas, challenge pre-existing orthodoxies, develop knowledge, and demonstrate the epistemological importance of current research in these critical areas. This book was originally published as a special issue of the Journal of Marketing Management.

This volume covers protocols related to both pluripotent and somatic stem cells, including the ethical procurement of tissues and cells for the provision of "seed stock," standardized methods for deriving hESCs and iPSCs, isolating mesenchymal stem cells, cell culture and cryopreservation, in addition to quality assurance and information management. Stem Cell Banking: Concepts and Protocols aims to contribute to the development of this field by providing information that is essential to establishing a bona fide stem cell bank. 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, Stem Cell Banking: Concepts and Protocols is a valuable resource for stem cell scientists and novices to the field, and will help strengthen and maximize their use of existing and future stem cell resources.

When marketing managers and financial managers join forces within any business, the result can often be poor communication on financial criteria and goals. The risk of this situation occurring is inevitably present when those with different professional backgrounds and roles are working in accordance with their own norms. In his seminal 1956 paper on general systems theory, the economist Kenneth Boulding referred to the phenomenon of "specialised ears and generalised deafness", which can be seen to exist when marketing managers are financially illiterate or when financial managers lack the necessary insights to design, implement and operate accounting systems which are useful to marketing managers in carrying out their roles. It is increasingly difficult to attach credence to the idea of marketing managers who lack financial skills, or financial managers who fail to relate to the context in which marketing managers operate. Understanding the marketing/accounting interface is therefore important in generating emergent properties from the interaction of marketers and accountants whereby the whole is greater than the sum of the parts. The chapters in this volume seek to address this challenge. This book was originally published as a special issue of the Journal of Marketing Management.

Molecular networks provide descriptions of the organization of various biological processes, including cellular signaling, metabolism, and genetic regulation. Knowledge on molecular networks is commonly used for systems level analysis of biological function; research and method development in this area has grown tremendously in the past few years. This book will provide a detailed review of existing knowledge on the functional characterization of biological networks. In 15 chapters authored by an international group of prolific systems biology and bioinformatics researchers, it will organize, conceptualize, and summarize the existing core of research results and computational methods on understanding biological function from a network perspective.

Aimed at research scientists and biotechnologists, this book is an essential reading for those working with extremophiles and their potential biotechnological application. Here, we provide a comprehensive and reliable source of information on the recent advances and challenges in different aspects of the theme. Written in an accessible language, the book is also a recommended as reference text for anyone interested in this thriving field of research. Over the last decades, the study of extremophiles has provided ground breaking discoveries that challenge our understanding of biochemistry and molecular biology. In the applied side, extremophiles and their enzymes have spawned a multibillion dollar biotechnology industry, with applications spanning biomedical, pharmaceutical, industrial, environmental, and agricultural sectors. Taq DNA polymerase (which was isolated from Thermus aquaticus from a geothermal spring in Yellowstone National Park) is the most well-known example of the potential biotechnological application of extremophiles and their biomolecules. Indeed, the application of extremophiles and their biologically active compounds has opened a new era in biotechnology. However, despite the latest advances, we are just in the beginning of exploring the biotechnological potentials of extremophiles.

Since the first edition of this book dedicated to differential display (DD) technology was published in 1997, we have witnessed an explosive interest in studying differential gene expression. The gene-hunting euphoria was initially powered by the invention of DD, which was gradually overtaken by DNA microarray technology in recent years. Then why is there still the need for second edition of this DD book? First of all, DD still enjoys a substantial lead over DNA microarrays in the ISI citation data (see Table 1), despite the h- dreds of millions of dollars spent each year on arrays. This may come as a surprise to many, but to us it implies that many of the DNA microarray studies went unpublished owing to their unfulfilled promises (1). Second, unlike DNA microarrays, DD is an "open"-ended gene discovery method that does not depend on prior genome sequence information of the organism being studied. As such, DD is applicable to the study of all living organisms-from bacteria, fungi, insects, fish, plants, to mammals-even when their genomes are not sequenced. Second, DD is more accessible technically and financially to most cost-conscious "cottage-industry" academic laboratories. So clearly DD still has its unique place in the modern molecular biological toolbox for gene expression analysis.

Busy clinicians and health practitioners recognize the importance of speedy detection of pathogens to impede the further spread of infection, and to ensure their patients' rapid and complete recovery. This reader-friendly reference is a unique collection of the newest and most effective diagnostic techniques currently in use in clinical and research laboratories. Instructive commentary regarding the application of these often complex methods is provided. This essential text aids readers in selecting the most efficient method, finding the necessary resources, and avoiding the most common pitfalls in implementation.

As imaging technologies and approaches have evolved, the scope of certain imaging techniques has moved far beyond the production of purely illustrative images or appealing time-lapse movies to providing the scientist with a rich range of ways to measure and quantify the biological process and outcome of gene expression. In Imaging Gene Expression: Methods and Protocols, expert authors offer up-to-date approaches and protocols that scientists in the field have developed, which would benefit the broader scientific community. Divided in three convenient parts, this detailed book covers the output of a gene, namely the RNA molecules that are transcribed from the gene and the way by which these molecules can be tracked or quantified in fixed or living cells, protocols that focus on the gene, DNA, or chromatin, as well as a variety of ways by which nuclear processes intertwined with gene expression can be followed and quantified in living cells as well as approaches for studying several sub-nuclear structures found in eukaryotic cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective subjects, lists of materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Imaging Gene Expression: Methods and Protocols will serve researchers working toward imaging in the context of complete organisms.

Since the publication of the first edition, lentivirus vector-based technologies, through in vitro and in vivo gene transfer in eukaryotic animal cells, continue to offer the most promising opportunities for curing genetic disorders, as well as cancer and infectious diseases. Lentivirus Gene Engineering Protocols, Second Edition reflects the spectacular progress made in the field with a set of cutting-edge methods contributed by highly respected scientists. Beginning with a thorough overview of the most recent lentivirus developments, the book continues with detailed protocols including sections on the advances in lentiviral vector technology, new lentiviral vector applications, involving transgenic human embryonic stem cells and fetal gene therapy among other topics, as well as the invaluable breakthroughs in LV-mediated expression of microRNAs. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective subjects, 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.

Authoritative and timely, Lentivirus Gene Engineering Protocols, Second Edition covers the most relevant issues and techniques of LV-based gene engineering, thus representing a complete theoretical and practical guide for scientists still unfamiliar with LV technologies and those who simply wish to know more about this vital area of study.

Examining the chemical modification of biological polymers and the emerging applications of this technology, Chemical Modification of Biological Polymers reflects the change in emphasis in this subsection of biotechnology from the study of protein structure and function toward applications in therapeutics and diagnostics.

Highlights

• The basic organic chemistry of the modification proteins, nucleic acids, oligosaccharides, polysaccharides, and their applications
• New analytical technologies used to characterize the chemical modification of biological polymers
• Identification of in vivo, non-enzymatic chemical modification of biological polymers
• Specific chemical modifications to generate biopharmaceutical products

This book covers the basics on the organic chemistry underlying the chemical modification of biopolymers, including updates on the use of various chemical reagents. It describes the current status of chemical modification of biological polymers and emerging applications of this technology in biotechnology. These technologies are important for the manufacture of conjugate proteins used in drug delivery, for the preparation of nucleic acid microarrays, and for the preparation of hydrogels and other materials used in tissue engineering.

This second edition provides updated and expanded chapters covering a broad sampling of useful and current methods in the rapidly developing and expanding field of bioinformatics. Bioinformatics, Volume I: Data, Sequence Analysis, and Evolution, Second Edition is comprised of three sections: Data and Databases, Sequence Analysis, and Phylogenetics and Evolution. The first section details bioinformatics methodologies in the generation of sequence and structural data and its organization into conceptual categories, and databases to facilitate further analyses. The Sequence Analysis section describes the fundamental methodologies for processing the sequences of biological molecules: techniques that are used in almost every pipeline of bioinformatics analysis, particularly in the preliminary stages of such pipelines. Last but not least, the phylogenetics and evolution section deals with methodologies that compare biological sequences for the purpose of understanding how they evolved. As a volume in the highly successful Methods in Molecular Biology series, chapters feature the kind of detail and expert implementation advice to ensure positive results. Comprehensive and practical, Bioinformatics, Volume I: Data, Sequence Analysis, and Evolution, Second Edition is an essential resource for graduate students, early career researchers, and others who are in the process of integrating new bioinformatics methods into their research.

The study of animal viruses contributes to the general understanding of the molecular basis of viral infection. The emergence of the SARS (Severe Acute Respiratory Syndrome) virus in the human population, coming from an animal source, highlights the importance of animals in harboring infectious agents. In addition, it has been recognized recently that influenza viruses, which persist in their natural avian host, can directly infect humans. In this book, an international panel of leading virologists provide a state-of-the-art overview of the field, comprehensively detailing the current understanding of viruses, their replication, evolution, and interaction with the host. The authors emphasize strategic and methodological aspects of current research, and provide key related references. Topics include foot-and-mouth disease virus, Pestivirus, Arteriviridae, Coronaviruses (including SARS), Herpesviridae, Paramyxoviridae, influenza viruses, Reoviridae, porcine circoviruses, Asfarviridae and much more.

A number of chapters present the most novel research on testicular and epididymal functions or on more general fields of hormone action and molecular cell biology as it is now a tradition. However, exceptionally, the book also contains several chapters dealing with the "Approaches and Tools in the Third Millenium." The unusual inclusion of technologies as such in the 2000 edition of the workshop was an absolute necessity, as these technologies are revolutionizing the fields of biology and medicine and, in many instances, how to do research. This volume gives the scientific community essential information about the very latest technical developments and their potential for future progress.

The aim of this volume is to provide a comprehensive overview of optical tweezers setups, both in practical and theoretical terms, to help biophysicists, biochemists, and cell biologists to build and calibrate their own instruments and to perform force measurements on mechanoenzymes both in isolation in vitro and in living cells. Chapters have been divided in three parts focusing on theory and practical design of optical tweezers, detailed protocols for performing force measurements on single DNA- and microtubule/actin-associated mechanoenzymes in isolation, and describing recent advances that have opened up quantitative force measurements in living 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. Authoritative and cutting-edge, Optical Tweezers: Methods and Protocols aims help to further expand the accessibility and use of optical traps by scientists of diverse disciplines.

Years ago when we were asked to write a book on the present-day knowledge of the molecular biology of poliovirus, we did not expect that such an apparently simple task could involve so much time and effort. Our writing was hampered by the fact that both of us are full time "workers," so that this monograph is mainly a spare time expedience. The main attention of this book focuses on a detailed review of the molecular biology of poliovirus and especially on the advances of the last decade; medical and environmental aspects are only briefly mentioned. Observations from older studies are considered in view of more recent information. Some of the older ob servations provided fundamental insights and paved the way for present day research; too often such data has been neglected or independently rediscovered. Today, poliovirus research has again attracted considerable interest. High points gained within the last few years were the elucidation of the complete nucleotide sequences of the RNAs of the three poliovinls serotypes and the corresponding vaccine strains, the demonstration of genome evolution during transmission of poliovirus in an epidemic, further characterization of the antigenic sites on the virus particle and of the antigenic drift, characterization of alternate conforma tional states of the virion capsid, the development of monoclonal antibodies against some of the virus proteins, observations on the role of the plasma membrane, cytoskeleton, and cytoplasmic membranes as mediators in the virus induced redirection of the synthetic machinery of the host cell, and characteriza"