Prokaryotic Toxins - Antitoxins gives the first overview of an exciting and rapidly expanding research field. Toxin - antitoxin (TA) genes were discovered on plasmids 30 years ago. Since then it has become evident that TA genes are highly abundant in bacterial and archaeal chromosomes. TA genes code for an antitoxin that combine with and neutralize a cognate toxin. When activated, the toxins inhibit protein synthesis and cell growth and thereby induce dormancy and multidrug tolerance (persistence). Remarkably, in some species, the TA gene families have undergone dramatic expansions. For example, the highly persistent major human pathogen Mycobacterium tuberculosis has "100 TA loci. The large expansion of TA genes by some organisms is a biological mystery. However, recent observations indicate that TA genes contribute cumulatively to the persistence of bacteria. This medically important phenomenon may thus for the first time become experimentally tractable at the molecular level.

This volume supplements Volumes 63, 64, 87, and 249 of Methods in Enzymology. These volumes provide a basic source for the quantitative interpretation of enzyme rate data and the analysis of enzyme catalysis. Among the major topics covered are Engergetic Coupling in Enzymatic Reactions, Intermediates and Complexes in Catalysis, Detection and Properties of Low Barrier Hydrogen Bonds, Transition State Determination, and Inhibitors.
The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.

Lipids in Photosynthesis: Essential and Regulatory Functions, provides an essential summary of an exciting decade of research on relationships between lipids and photosynthesis. The book brings together extensively cross-referenced and peer-reviewed chapters by prominent researchers. The topics covered include the structure, molecular organization and biosynthesis of fatty acids, glycerolipids and nonglycerolipids in plants, algae, lichens, mosses, and cyanobacteria, as well as in chloroplasts and mitochondria. Several chapters deal with the manipulation of the extent of unsaturation of fatty acids and the effects of such manipulation on photosynthesis and responses to various forms of stress. The final chapters focus on lipid trafficking, signaling and advanced analytical techniques. Ten years ago, Siegenthaler and Murata edited "Lipids in Photosynthesis: Structure, Function and Genetics," which became a classic in the field. "Lipids in Photosynthesis: Essential and Regulatory Functions," belongs, with its predecessor, in every plant and microbiological researcher's bookcase.

Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general.

During the past decade, a wide range of scientific disciplines have adopted the use of adipose-derived stem/stromal cells (ASCs) as an important tool for research and discovery. In Adipose-Derived Stem Cells: Methods and Protocols, experts from the field, including members of the esteemed International Federation of Adipose Therapeutics and Science (IFATS), provide defined and established protocols in order to further codify the utilization of these powerful and accessible cells. With chapters organized around approaches spanning the discovery, pre-clinical, and clinical processes, much of the emphasis is placed on human ASC, while additional techniques involving small and large animal species are included. As a volume in the highly successful Methods in Molecular Biology(TM) series, the detailed contributions 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. Comprehensive and cutting-edge, Adipose-Derived Stem Cells: Methods and Protocols serves as a vital reference text for experienced researchers as well as new students on the path to further exploring the incredible potential of ASCs.

This volume provides easily accessible and comprehensive collection of methods, techniques, and strategies to investigate the molecular and cellular biology of peroxisomes in different organisms. Chapters detail valuable instructions, guidelines and protocols for molecular cell biologists, biochemists and biomedical researchers with an interest in peroxisome biology. Chapters in Peroxisomes: Methods and Protocols illustrate the isolation of peroxisomes, investigation of properties of membrane proteins, protocols to investigate and manipulate peroxisomes in cellular systems, detection of peroxisomes, including immunofluorescence, cytochemistry, cryo-immuno electron microscopy, and live cell imaging approaches. Authoritative and practical, Peroxisomes: Methods and Protocols aims to be useful for those already working on peroxisomes as well as for those who would like to start working on this fascinating organelle.

For a virus to invade a host cell it needs to penetrate the physical barrier imposed by the plasma membrane. Viruses have evolved specialized surface proteins to meet this challenge. These proteins facilitate delivery of the viral genetic information into the host cell by either fusing the viral envelope with a host cell membrane or by forming membrane pores. Membrane fusion and pore formation critically depend on the engagement of host cell receptors and receptor choice is a key determinant of viral tropism. The multi-faceted interplay between viral and cellular factors during virus entry is a fascinating field of study, which can provide important insight into viral pathogenesis and define new targets for intervention. This book provides a comprehensive overview of this exciting field of research.

This practical volume brings together a broad range of topics related to membrane biology research with particular emphasis on novel approaches, technology platforms, and emerging tools in this area of study. Beginning with chapters pertaining to artificial or designer membrane mimetics that can be utilized for in vitro studies of membrane-lipid interactions, it then continues by covering the approaches and methodologies to directly investigate membrane protein structure, localization, and dynamics. Written for the Springer Protocols Handbooks series, this collection focuses on in-depth, hands-on protocols from experts in the field. Authoritative and cutting-edge, Chemical and Synthetic Approaches in Membrane Biology serves as an ideal guide to researchers investigating the vital properties of cellular membranes.

Protein kinase CK2 (formerly casein kinase II or 2) is known to play a critical role in the control of cell growth and cell death and is thus intimately involved in the development of cancer. More specifically, CK2 has been found to be elevated in all cancers examined. While CK2 levels are known to be high in proliferating normal cells, CK2 has also been found to be a potent suppressor of apoptosis and is a link to the cancer cell phenotype, which is characterized by deregulation of both cell proliferation and cell death. Indeed, it would appear that CK2 impacts many of the hallmarks of cancer and it has now gained considerable attention as a potential target for cancer therapy. Protein Kinase CK2 and Cellular Function in Normal and Disease States increases knowledge of the role of CK2 in the development of cellular dysfunction and emphasizes that this protein may serve as a target of drug development for improved cancer therapy. In addition, it is a handy tool that provides cancer researchers, graduate students, and all scientists involved in CK2 research with one main source for the latest advances in CK2 research.

The present volume is the first in the advances in oncobiology series. It is meant to be useful not only to clinical and non-clinical oncologists but also to graduate students and medical students. The individual chapters are presented as self-contained summaries of current knowledge rather than as reviews. The last chapter deals with the subject of chemotherapy.

For most of industrial applications, enzymes and cells have to be immobilized, via very simple and cost-effective protocols, in order to be re-used for very long periods of time. From this point of view, immobilization, simplicity and stabilization have to be strongly related concepts. The third edition of Immobilization of Enzymes and Cells expands upon and updates the previous editions with current, detailed protocols for immobilization. With new chapters on protocols for immobilization of enzymes and cells which may be useful to greatly improve the functional properties of enzymes and 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Immobilization of Enzymes and Cells, Third Edition demonstrates simple and efficient protocols for the preparation, characterization, and utilization of immobilized enzymes and cells.

Microbial cell wall structures play a significant role in maintaining cells' shape, as protecting layers against harmful agents, in cell adhesion and in positive and negative biological activities with host cells. All prokaryotes, whether they are bacteria or archaea, rely on their surface polymers for these multiple functions. Their surfaces serve as the indispensable primary interfaces between the cell and its surroundings, often mediating or catalyzing important interactions.

"Prokaryotic Cell Wall Compounds" summarizes the current state of knowledge on the prokaryotic cell wall. Topics concerning bacterial and archaeal polymeric cell wall structures, biological activities, growth and inhibition, cell wall interactions and the applications of cell wall components, especially in the field of nanobiotechnology, are presented.

Quantitative Proteomics by Mass Spectrometry, from the Methods in Molecular Biologya"[ series, is a compendium of cutting-edge protocols for quantitative proteomics, and presents the most significant methods used in the field today. The focus on mass spectrometry (MS) is integral, as MS has, and will continue to be, an essential tool in proteomics for studying complex biological systems and human diseases. This volume, written and compiled by leading quantitative proteomic experts, is an indispensable resource in the search for novel biomarkers.
Quantitative Proteomics by Mass Spectrometry presents several innovative MS quantitative procedures, including a variety of methods for introducing isotopic labels and quantifying post-translational modifications. Some of these methods include growing an organism in isotope-enriched media, performing trypsin proteolysis in the presence of 18O-water, reacting protein samples with isotopically labeled reagents, quantifying relative amount of proteins without the use of any isotopic labels. Attention is also given to state-of-the-art techniques for the characterization of the phosphoproteome and tandem MS for detection of inborn errors of metabolism. Specifically, the procedure for determinations of enzymatic activity could be used for large-scale screening of newborns. The protocols in this volume expand both the breadth and depth of readily available methods for quantitative proteomic researchers using MS.
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Defined as, "The science about the development of an embryo from the fertilization of the ovum to the fetus stage," embryology has been a mainstay at universities throughout the world for many years. Throughout the last century, embryology became overshadowed by experimental-based genetics and cell biology, transforming the field into developmental biology, which replaced embryology in Biology departments in many universities. Major contributions in this young century in the fields of molecular biology, biochemistry and genomics were integrated with both embryology and developmental biology to provide an understanding of the molecular portrait of a "development cell." That new integrated approach is known as stem-cell biology; it is an understanding of the embryology and development together at the molecular level using engineering, imaging and cell culture principles, and it is at the heart of this seminal book. Stem Cells and Regenerative Medicine: From Molecular Embryology to Tissue Engineering is completely devoted to the basic developmental, cellular and molecular biological aspects of stem cells as well as their clinical applications in tissue engineering and regenerative medicine. It focuses on the basic biology of embryonic and cancer cells plus their key involvement in self-renewal, muscle repair, epigenetic processes, and therapeutic applications. In addition, it covers other key relevant topics such as nuclear reprogramming induced pluripotency and stem cell culture techniques using novel biomaterials. A thorough introduction to stem-cell biology, this reference is aimed at graduate students, post-docs, and professors as well as executives and scientists in biotech and pharmaceutical companies.

International Review of Cytology presents current advances and comprehensive reviews in cell biology-both plant and animal. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Authored by some of the foremost scientists in the field, each volume provides up-to-date information and directions for future research.
Key Features
* Organellar RNA Polymerases of Higher Plants
* Eukaryotic Transmembrane Solution Transport Systems
* Neural Plasticity in the Adult Insect Brain
* Passive Membrane Permeation
* Plasmodesmata and Cell-to-Cell Communication in Plants

Written by specialists in the field, this book provides an overview of the structural and molecular biology of cellular processes that occur at or near bacterial membranes.

Platelets are fragments of blood cells that occur in the blood of vertebrates and are associated with blood clotting. Scientists have made great strides in recent years in understanding what stimulates platelets to form blood clots at the molecular level and in developing drugs to inhibit platelet action. Their work has a direct effect on millions of people who deal with cardiovascular disease, strokes, surgery, physical trauma, and other conditions. While references to platelet function have been included in some large texts, there has not been a basic reference manual that researchers and clinicians can use in their daily work until now.
Platelet Protocols fills the need for a straightforward and comprehensive laboratory manual on current procedures for evaluating and analyzing platelet function and abnormalities. It is an easy-to-read, understandable resource which can be kept at the bench and referred to frequently by scientists, clinicians, and laboratory staff involved in platelet related areas. Topics range from the basics of anticoagulants to the latest developments in platelet testing.
Key Features
Includes:
* A basic introduction to platelet anatomy and physiology
* Testing procedures for new anti-platelet therapies
* Descriptions of platelet function abnormalities
* Therapeutic approaches to inhibition of platelet function
* Step-by-step methodologies with clear explanations
* Helpful appendixes of recipes, instructions, sources of reagents, and more

Strategies of Bacterial Interaction with Eukaryotic Cells *Tobias A. Oelschlaeger and Jorg Hacker 1. BENEFICIAL BACTERIAL-HOST INTERACTIONS Already during birth and soon thereafter mammals are colonized by bacte- ria belonging to the resident microbial flora. Cutaneous and mucosal sur- faces and the gastrointestinal tract are the areas which become colonized. These indigenous or autochthonous bacteria have a variety of beneficial effects on their hosts. They play a protective role by bacterial antagonism in fighting infections (Hoszowski and Truszczynski, 1997; Hentges, 1979). Pro- duction of vitamin K is another essential contribution of the resident microbial flora to the health of the host (Hill, 1997). Even more important, studies with germ-free animals demonstrated the involvement of the microbial flora on the development of the immune system. Such animals have underdeveloped and relatively undifferentiated lymphoid tissues and low concentrations of serum immune globulins ( Cebra et at., 1998). They TOBIAS A. OELSCHLAEGER and JORG HACKER Institut filr Molekulare lnfektionsbiologie, Universitiit Wiirzburg, 97070 Wiirzburg, Germany. *Corresponding author; Phone: (0)931-312150; FAX: (0)931-312578; E-mail: [email protected] xxix Tobias A. Oelschlaeger and Jorg Hacker also show defects in specific immune responsiveness and in nonspecific resistance induced by endotoxin, which may account for their lowered resis- tance. A more typical example of symbiotic interaction of bacteria with a host are bacteria like Ruminococcus in the gut of ruminants, essential for degradation of cellulose (Hobson, 1988). The closest benefical bacterial-host interactions are those of intracellular symbiotic bacteria and their host cells.

Every cell in our bodies contains a great variety and number of permeability pathways for various organic and inorganic ions, water, metabolites, nutrients, and signalling molecules. Maintenance and precise control of gating within these pathways are fundamental principles of life as these underlie basic cellular functions such as communication, contractility, and metabolism. In Ion Channels: Methods and Protocols, Second Edition, experts in the field contribute chapters that focus on the strategies, approaches, methods, and protocols for studying a large family of proteins that form ionic channels in the plasma membrane and intracellular membranes of cells. Using practical examples from the cutting-edge current research, this volume will take a look back at the major methods and approaches that aided the progress toward the current understanding of ion channel function, structural design, and biological roles. The volume also aims to look forward and identify approaches that will lead us to future discoveries. 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. Authoritative and cutting-edge, Ion Channels: Methods and Protocols, Second Edition will greatly assist researchers searching for specific methodology in studying ion channels on the path toward a greater understanding of these key biological features.

Rb and Tumorigenesis examines how recent advances have demonstrated the interaction of Rb with chromatin remodeling enzymes. This new title explores the potential roles of these interactions in Rb functions and provides some evidence that distinct Rb co-repressor may target different genes in different phases of the cell cycle. This book will interest cell biologists, graduate students and researchers.

The root is the organ that functions as the interface between the plant and the earth environment. Many human management practices involving crops, forests and natural vegetation also affect plant growth through the soil and roots. Understanding the morphology and function of roots from the cellular level to the level of the whole root system is required for both plant production and environmental protection. This book is at the forefront of plant root science (rhizology), catering to professional plant scientists and graduate students. It covers root development, stress physiology, ecology, and associations with microorganisms. The chapters are selected papers originally presented at the 6th Symposium of the International Society of Root Research, where plant biologists, ecologists, soil microbiologists, crop scientists, forestry scientists, and environmental scientists, among others, gathered to discuss current research results and to establish rhizology as a newly integrated research area.

With the invitation to edit this volume, I wanted to take the opportunity to assemble reviews on different aspects of circadian clocks and rhythms. Although most c- tributions in this volume focus on mammalian circadian clocks, the historical int- duction and comparative clocks section illustrate the importance of various other organisms in deciphering the mechanisms and principles of circadian biology. Circadian rhythms have been studied for centuries, but only recently, a mole- lar understanding of this process has emerged. This has taken research on circadian clocks from mystic phenomenology to a mechanistic level; chains of molecular events can describe phenomena with remarkable accuracy. Nevertheless, current models of the functioning of circadian clocks are still rudimentary. This is not due to the faultiness of discovered mechanisms, but due to the lack of undiscovered processes involved in contributing to circadian rhythmicity. We know for example, that the general circadian mechanism is not regulated equally in all tissues of m- mals. Hence, a lot still needs to be discovered to get a full understanding of cir- dian rhythms at the systems level. In this respect, technology has advanced at high speed in the last years and provided us with data illustrating the sheer complexity of regulation of physiological processes in organisms. To handle this information, computer aided integration of the results is of utmost importance in order to d- cover novel concepts that ultimately need to be tested experimentally.

Since its first discovery in the early 90 s, the NF-kB/Rel transcription factor family has drawn the attention of experimental biologists, medical profession, and biotech/pharmaceutical industries for its broad and diverse roles in all aspects of human biology and disease. NF-kB/Rel Transcription Factor Family intends to provide an up-to-date guide to the ever-expanding knowledge in the field of NF-kB/Rel transcription factor family.

This book is a direct result of 10 years of the well-known "Autoimmunity Days" in Israel, which are increasingly becoming an international focal point for autoimmunity scientists. Top researchers provide coverage of the most important knowledge generated during the last decade. The volume can therefore be seen almost as a textbook on autoimmunity, projecting from the last decade to the next millennium.
A variety of different subjects in autoimmunity, from etiology to pathogenesis, from postulated mechanisms to innovative therapeutic modalities, is handled by noted contributors, while additional authors contribute top papers which significantly enhance a better understanding of autoimmunity.
An excellent treatise on the subject, and a worthwhile addition to both clinical and research libraries.