The fungus fusarium is a major plant pathogen that causes disease in nearly every agriculturally important plant. In addition, some strains produce mycotoxins that can cause serious illness in humans and livestock. The enormous economic importance of and health hazards posed by fusarium have fuelled research into its biochemistry, genetics, genomics, proteomics and metabolomics by scientists worldwide. In this book, an international group of researchers critically review the most important research on the genomics and molecular and cellular biology of fusarium.

Describes landmark experiments in cell biology and biochemistry Discusses the "How" and "Why" of historically important experiments Includes primary, original data and graphs Emphasizes biological techniques, which helps understand how many of the experiments performed were possible. Documents, chronologically, how each result fed into the next experiments.

Solvent Extraction in Biotechnology deals with the reco- very and purification of primary and secondary metabolites by solvent extraction. In the first part the reaction engineering principles: definitions, thermodynamic fundamentals, and system models, the kinetics of mass transfer between two phases without and with chemical reaction as well as extraction equipment, which are important for downstream processing in biotechnology, are considered in detail. The special part of the book describes the recovery of low-molecular metabolites: alcohols, acids and antibiotics with organic solvents, carrier-modifier-solvent systems, supercritical gases as well as with liquid membrane techniques. Several practical examples are given for the recovery of different metabolites as well as for the calculation of the extraction processes necessary for equipment design. Besides solvent extraction, novel separation techniques with liquid membrane, microemulsion and reversed micelles are also presented. This book will introduce the biochemical engineer and process engineer to the recovery of products from complex cultivation broths by modern techniques of solvent extraction and help them with process design.

In recent years, the study of the plant cell cycle has become of major interest, not only to scientists working on cell division sensu strictu, but also to scientists dealing with plant hormones, development and environmental effects on growth. The book The Plant Cell Cycle is a very timely contribution to this exploding field. Outstanding contributors reviewed, not only knowledge on the most important classes of cell cycle regulators, but also summarized the various processes in which cell cycle control plays a pivotal role. The central role of the cell cycle makes this book an absolute must for plant molecular biologists.

Mass Spectrometry Data Analysis in Proteomics is an in-depth guide to the theory and practice of analyzing raw mass spectrometry (MS) data in proteomics. As MS is a high throughput technique, proteomic researchers must attend carefully to the associated field of data analysis, and this volume outlines available bioinformatics programs, algorithms, and databases available for MS data analysis. General guidelines for data analysis using search engines such as Mascot, Xtandem, and VEMS are provided, with specific attention to identifying poor quality data and optimizing search parameters. Several different types of MS data are discussed, followed by a description of optimal methods for conversion of raw data into peak lists for input to search engines. Choosing the most accurate and complete databases is emphasized, and a report of available sequence databases is included. Methods for assembling expressed sequence tags (ESTs) into assembled nonredundant databases are provided, along with protocols for further processing the sequences into a format suitable for MS data. Mass Spectrometry Data Analysis in Proteomics describes publicly available applications whenever possible.

Plant Cell and Tissue Culture gives an exhaustive account of plant cell culture and genetic transformation, including detailed chapters on all major field and plantation crops. Part A presents a comprehensive coverage of all necessary laboratory techniques for the initiation, nutrition, maintenance and storage of plant cell and tissue cultures, including discussions on these topics, as well as on morphogenesis and regeneration, meristem and shoot tip culture, plant protoplasts, mutant cell lines, variation in tissue cultures, isogenic lines, fertilization control, cryopreservation, transformation, and the production of secondary metabolites. Part B then proceeds into detail on the specific in vitro culture of specific crops, including cereals, legumes, vegetables, potatoes, other roots and tubers, oilseeds, temperate fruits, tropical fruits, plantation crops, forest trees and ornamentals. Plant Cell and Tissue Culture is, and is likely to remain, the laboratory manual of choice, as well as a source of inspiration and a guide to all workers in the field.

Since the discovery of the pharmacological and toxicological importance of inhibiting the cyclooxygenase (COX) enzymes by non-steroidal anti-inflammatory drugs (NSAIDs), much research has gone into the development of methods to study the biological functions of COX-1 and COX-2. In Cyclooxygenases: Methods and Protocols, experts and pioneers in the field present the most up-to-date in vitro and in vivo techniques routinely used in COX research. The volume delves into essential topics such as the purification, cloning, and expression of COX enzymes as well as in vitro assays aimed at determining the inhibitory potency of drugs on COX-1 and COX-2 activities, with chapters describing protocols used for the extraction and measurement of the prostanoids. This volume also describes in vivo disease models used to study the roles of COX-1 and COX-2 in gastrointestinal injury, inflammation, and pain. As a book in the highly successful Methods in Molecular Biology (TM) series, the protocols chapters include brief 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, Cyclooxygenases: Methods and Protocols serves as an indispensable tool for all scientists seeking the treatment of inflammation, pain, fever, and other harmful conditions.

Challenging and provocative overviews are presented in Volume 40 of Current Topics in Membranes. Topics on cell lipids vary from basic themes such as biosynthesis and membrane distribution to the role of lipids in intracellular signaling and membrane flow. This single volume also highlights the roles of lipids in eukaryotic cells and discusses organization of lipids in microdomains.

A Practical Guide to the Study of Calcium in Living Cells describes popular techniques along with helpful do's and don't's and computer programs. The volume enables investigators to evaluate confocal images, use the latest dyes, and design Calcium buffers appropriate to their research needs. This book is designed for laboratory use by graduate students, technicians, and researchers in many disciplines, ranging from molecular to cellular levels of investigation.
Key Features
* Describes techniques for detection of [Ca2]]I: Ca2+ - sensitive microelectrodes
* Fluorescent dyes
* Luminescent proteins
* Includes techniques for perturbing intracellular Ca2+
* Covers detailed methodology plus problems and pitfalls of each technique
* Contains a practical guide to preparing Ca2+ buffers with an easy-to-use computer program
* Color plates illustrate techniques such as
* Confocal ratio-imaging
* Use of aequorin

This volume illustrates how microfluidic approaches can meet the requirement of clinical diagnosis based on molecular or cellular biomarkers. Microchip Diagnostics: Methods and Protocols is divided into four sections describing the business aspects of the microfluidic-based systems for diagnosis, demonstrating how versatile microfluidics can be regarding to protein bioassay integration, presenting microfluidic approaches for nucleic analysis based on mono or diphasic format, and highlighting recent contributions. 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.

Cilia: From Mechanisms to Disease Part B, Volume 176 of Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

NMR spectroscopy has found a wide range of applications in life sciences over recent decades. Providing a comprehensive amalgamation of the scattered knowledge of how to apply high-resolution NMR techniques to biomolecular systems, this book will break down the conventional stereotypes in the use of NMR for structural studies. The major focus is on novel approaches in NMR which deal with the functional interface of either protein-protein interactions or protein-lipid interactions. Bridging the gaps between structural and functional studies, the Editors believe a thorough compilation of these studies will open an entirely new dimension of understanding of crucial functional motifs. This in turn will be helpful for future applications into drug design or better understanding of systems. The book will appeal to NMR practitioners in industry and academia who are looking for a comprehensive understanding of the possibilities of applying high-resolution NMR spectroscopic techniques in probing biomolecular interactions.

This new edition continues to illustrate the power of biological data in knowledge discovery. It describes biological data types and representations with examples for creating a workflow in bioinformation discovery. The concepts in knowledge discovery from data are illustrated using line diagrams. The principles and concepts in knowledge discovery are used for the development of prediction models for simulations of biological reactions and events. Advanced topics in molecular evolution and cellular & molecular biology are addressed using bioinformation gleaned through discovery. Each chapter contains approximately 10 exercises for practice. This will help students to expand their problem solving skills in Bioinformation Discovery. In this new edition, there are three new chapters covering single nucleotide polymorphism, genes, proteins and disease, and protein functions driven by surface electrostatics.

Inflammation has been described as the basis of many pathologies of human disease. When one considers the updated signs of inflammation, they would be vasodilation, cell migration, and, in the case of chronic inflam- tion, cell proliferation, often with an underlying autoimmune basis. Gen- ally, inflammation may be divided into acute, chronic, and autoimmune, - though the editors believe that most, if not all, chronic states are often the result of an autoimmune response to an endogenous antigen. Thus, a proper understanding of the inflammatory basis may provide clues to new therap- tic targets not only in classical inflammatory diseases, but atherosclerosis, cancer, and ischemic heart disease as well. The lack of advances in classical inflammatory diseases, such as rh- matoid arthritis, may in part arise from a failure to classify the disease into different forms. That different forms exist is exemplified in patients with d- fering responses to existing antiinflammatory drugs, ranging from nonresponders to very positive responders for a particular nonsteroidal an- inflammatory drug (NSAID). Though researchers have progressively unr- eled the mechanisms, the story is far from complete. It should also be noted that the inflammatory response is part of the innate immune response, or to use John Hunter's words in 1795, "inflammation is a salutary response." That may be applied in particular to the defensive response to invading micro- ganisms.

Helicobacter pylori is an important human pathogen that infects up to 50% of the human population. As the leading cause of peptic ulcers, gastritis, and gastric cancer worldwide, the organism has been the subject of intensive research to unravel the mysteries of its genetics and cellular biology. In fact, the number of publications in this field has risen dramatically in recent years making it extremely difficult for even the most diligent reader to stay abreast of progress. This book distills the most important cutting-edge findings in the field to produce a timely and comprehensive review. With contributions from leading international helicobacter researchers, topics include: lipopolysaccharides, outer membrane proteins, motility and chemotaxis, type IV secretions systems, metal metabolism, molecular mechanisms of host adaptation, genomotyping, and proteonomics. As a useful introduction to the subject for new researchers and as an invaluable reference for the experienced researcher, this book is essential reading for all researchers working with Helicobacter and related organisms.

Immunotoxins represent a new class of human therapeutics that have widespread applications and a potential that has not yet been fully recognized since they were first conceived of by Paul Ehrlich in 1906. The majority of advances in the development and implementation of immunotoxins has occurred over the last 20 years. The reasons for this use of immunotoxins in basic science and clinical research are the powerful concurrent advances in genetic engineering and receptor physiology. Recombinant technology has allowed investigators to produce sufficient quantities of a homogeneous c- pound that allows clinical trials to be performed. The identification of specific receptors on malignant cell types has enabled scientists to generate immunotoxins that have had positive results in clinical trials. As more cellular targets are identified in coming years, additional trials will be conducted in different disease states affecting still larger patient populations. Modulation of the immune system to decrease the humoral response to immunotoxins may improve their overall efficacy. As increasingly more effective compounds are generated, it will be necessary to decrease the local and systemic toxicity - sociated with these agents, and methods for doing so are presently being - veloped. The work presented in Immunotoxin Methods and Protocols focuses on three specific areas of immunotoxin investigation that are being conducted by experts throughout the world. The first section describes the construction and development of a variety of immunotoxins.

Metabolomics: Methods and Protocols examines the state-of-the-art in metabolomic analysis. Leading researchers in the field present protocols for the application of complementary analytical methods, such as gas chromatography-mass spectrometry (GC-MS). Metabolomics: Methods and Protocols contains forward-looking protocols, which provide the essential groundwork for future efforts in elucidating the structure of the unknowns detected in metabolomic studies.
Part I of this volume focuses on GC-MS techniques; specifically, attention is given to metabolite profiling in blood plasma, construction and application of mass spectral and retention time index libraries, and metabolomic profiling of natural volatiles. In Part II, emphasis shifts to data integration and data mining techniques, and specific protocols discuss improving pattern recognition and biomarker selection for systems level approaches, and visualization and analysis of molecular and metabolomic data. Parts III, IV, and V examine the coupling of capillary electrophoresis and mass spectrometry, classical reversed phase liquid chromatography-mass spectrometry, and electrochemical detection of metabolites using HPLC separations coupled with coulometric electrode array detectors. In Part VI, protocols for the determination of metabolic flux rates are presented and a theoretical framework for metabolic pathway network analysis is provided. The volume concludes with an examination of cutting-edge NMR protocols.

A collection of standard and cutting-edge techniques for using Xenopus oocytes and oocytes/egg extracts to reconstitute biological and cellular processes. These readily reproducible methods take advantage of the oocyte's impressive protein abundance, its striking protein translation capacity, and its breathtaking possibilities for the assembly of infectious viral particles by single cell injection of multiple RNAs. The authors focus on the versatility of frog oocytes and egg extracts in cell biology and signal transduction, and cover all the major uses of oocytes/extracts as experimental models.

This detailed volume encompasses new technological developments that specifically address questions related to adenosine 3',5'-monophosphate (cAMP) compartmentalization, that probe relevant protein-protein interactions, that increase the spatial and temporal resolution of cAMP signal detection, and that can facilitate integration of the mounting complexity of the information that is becoming available on this signaling system. cAMP, the prototypical intracellular second messenger, regulates a large variety of cellular functions and biological processes, including gene transcription, cell metabolism, proliferation, development, as well as more specialized functions depending on the cell type, so the realization of its extremely complex spatial organization and local regulation is providing novel mechanistic insight into cell physiology and is producing a novel framework for the identification of disease mechanisms. Written in the highly successful Methods in Molecular Biology series format, chapters include 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 authoritative, cAMP Signaling: Methods and Protocols serves as a vital resource for researchers working in this expanding, complex field.

In the post-genomic era, in vitro mutagenesis has emerged as a critically important tool for establishing the functions of components of the proteome. The third edition of In Vitro Mutagenesis Protocols represents a practical toolbox containing protocols vital to advancing our understanding of the connection between nucleotide sequence and sequence function. Fully updated from the previous editions, this volume contains a variety of specialty tools successfully employed to unravel the intricacies of protein-protein interaction, protein structure-function, protein regulation of biological processes, and protein activity, as well as a novel section on mutagenesis methods for unique microbes as a guide to the generalization of mutagenesis strategies for a host of microbial systems. Written in the highly successful Methods in Molecular Biology 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 pitfalls.

Authoritative and up-to-date, In Vitro Mutagenesis Protocols, Third Edition offers today's researchers a valuable compendium of reliable and powerful techniques with which to illuminate the proteome and its rich web of biological implications.

There is an urgent need to develop new approaches to treat conditions as- ciated with the aging global population. The surgeon's approach to many of these problems could be described as having evolved through three stages: Removal: Traditionally, diseased or badly damaged tissues and structures might simply be removed. This was appropriate for limbs and non-essential organs, but could not be applied to structures that were critical to sustain life. An additional problem was the creation of disability or physical deformity that in turn could lead to further complications. Replacement: In an effort to treat wider clinical problems, or to overcome the limitations of amputation, surgeons turned to the use of implanted materials and medical devices that could replace the functions of biological structures. This field developed rapidly in the 1960s and 1970s, with heart valve and total joint replacement becoming common. The term "biomaterial" was used increasingly to describe the materials used in these operations, and the study of biomaterials became one of the first truly interdisciplinary research fields. Today, biomaterials are employed in many millions of clinical procedures each year and they have become the mainstay of a very successful industry.

Progress in Molecular Biology and Translational Science, Epigenetics in Health and Disease Volume 197, highlights new advances in the field, with this new volume presenting interesting chapters on Epigenetics in Health and Disease. Each chapter is written by an international board of authors.

Waste water treatment is the largest biotechnology industry in the world, handling and disposing of domestic and industrial wastes so they present no threat to the general populace or the environment at large. The activated sludge process is the cornerstone of sewage treatment systems. Although it is a biological process and has been in use for more than 80 years, we still lack detailed understanding of how it works and how its performance might be better controlled and manipulated. This book presents a comprehensive review of what is currently known about the general microbiology of activated sludge and some of the bacteria responsible for the major operational problems encountered. Current understanding and the existing literature are critically assessed and new potentially valuable areas for research suggested. Methodology particularly appropriate to the study of the bacteria that cause bulking and foaming is described, and there is an identification section consisting of photomicrographs and detailed descriptions of the filamentous and other bacteria commonly seen in activated sludge plants. The impact of molecular methods on our understanding of activated sludge microbiology is emphasized throughout the book. The book will be of immediate interest to both microbiologists and waste water engineers, and has been written so as to be relevant and understandable to both. It will also be of interest and value to postgraduate students and researchers working in the fields of environmental engineering, general microbiology and microbial ecology.

The first libraries of complementary DNA (cDNA) clones were con structed in the mid-to-late 1970s using RNA-dependent DNA polymerase (reverse transcriptase) to convert poly A* mRNA into double-stranded cDNA suitable for insertion into prokaryotic vectors. Since then cDNA technology has become a fundamental tool for the molecular biologist and at the same time some very significant advances have occurred in the methods for con structing and screening cDNA libraries. It is not the aim of cDNA Library Protocols to give a comprehensive review of all cDNA library-based methodologies; instead we present a series of up-to-date protocols that together should give a good grounding of proce dures associated with the construction and use of cDNA libraries. In deciding what to include, we endeavored to combine up-to-date versions of some of the most widely used protocols with some very usefiil newer techniques. cDNA Library Protocols should therefore be especially useful to the investigator who is new to the use of cDNA libraries, but should also be of value to the more experienced worker. Chapters 1-5 concentrate on cDNA library construction and manipula tion, Chapters 6 and 7 describe means of cloning difficult-to-obtain ends of cDNAs, Chapters 8-18 give various approaches to the screening of cDNA libraries, and the remaining chapters present methods of analysis of cDNA clones including details of how to analyze cDNA sequence data and how to make use of the wealth of cDNA data emerging from the human genome project."

The dynamic field of flavin and flavoprotein biochemistry has seen rapid advancement in recent years. This comprehensive two volume set provides an overview of all aspects of contemporary research in this important class of enzymes. Topics treated include flavoproteins involved in energy generation, signal transduction and electron transfer (including respiration); oxygen activation by flavoproteins; the biology and biochemistry of complex flavoproteins; flavin and flavoprotein photochemistry/photophysics as well as biotechnological applications of flavoproteins. Recent developments in this field include new structures (including those of large membrane-integral electron transfer complexes containing FMN or FAD), elucidation of the role of flavoproteins in cell signalling pathways (including both phototaxis and the circadian cycle) and important new insights into the reaction mechanisms of flavin-containing enzymes. This volume focussing on oxidases, dehydrogenases and related systems is an essential reference for all researchers in biochemistry, chemistry, photochemistry and photophysics working on flavoenzymes.