This volume looks at the study of oligodendrocytes through in vitro and in vivo techniques, multiple model organisms, using approaches that bridge scales from molecular through system. Chapters in this book cover topics such as fundamental molecular analyses of oligodendrocytes and myelin; in vitro, ex vivo, and in vivo molecular-cellular-electrophysiology-based techniques; oligodendrocyte formation, homeostasis, and disruption in zebrafish and Xenopus; and parallel system-level imaging of animal and human models. 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 thorough, Oligodendrocytes: Methods and Protocols is a valuable reference guide that highlights the expansive and fast-paced nature of research into oligodendrocyte biology underlying health and function.

This book explains the anatomy and physiology of cartilage tissue in an integrated way. The emphasis is on how cartilage tissue functions and maintains homeostasis in a challenging mechanical environment. Supported by hundreds of references, the book posts new hypotheses explaining how cartilage adapts and achieves homeostasis in vivo, and tests them against available data. This exploratory approach creates a sense of discovery that the reader can join, or perhaps test themselves through their own research. The main benefit will be obtained by research students and professors looking to understand the deeper concepts that will further their own research, or clinicians (including health professionals and surgeons) who want to gain a deeper physiological understanding of cartilage tissue, which can then serve as a basis for more rational clinical decision-making they need to make on a daily basis. To help bridge the gap between basic science and clinically relevant joint disease, applications and interpretations of key physiological concepts are discussed in the context of osteoarthritis at the end of most chapters.

DNA Structure and Function, a timely and comprehensive resource, is intended for any student or scientist interested in DNA structure and its biological implications. The book provides a simple yet comprehensive introduction to nearly all aspects of DNA structure. It also explains current ideas on the biological significance of classic and alternative DNA conformations. Suitable for graduate courses on DNA structure and nucleic acids, the text is also excellent supplemental reading for courses in general biochemistry, molecular biology, and genetics.
Special Features
* Explains basic DNA Structure and function clearly and simply
* Contains up-to-date coverage of cruciforms, Z-DNA, triplex DNA, and other DNA conformations
* Discusses DNA-protein interactions, chromosomal organization, and biological implications of structure
* Highlights key experiments and ideas within boxed sections
* Illustrated with 150 diagrams and figures that convey structural and experimental concepts

The Zebrafish: Disease Models and Chemical Screens, Fourth Edition, the latest volume in the Methods in Cell Biology series, looks at methods for analyzing genetics, genomics, and transcriptomics of zebrafish. Chapters cover such topics as gene-trap mutagenesis, genetic screens for mutations, gene editing in zebrafish, homologous gene targeting, genome-wide RNA tomography, developmental epigenetics, and the zebrafish interactome. * Covers sections on model systems and functional studies, imaging-based approaches, and emerging studies* Includes chapters written by experts in the field* Contains cutting-edge material on the topics discussed

This volume brings together a set of reviews that provide a summary of our current knowledge of the proteolytic machinery and of the pathways of protein breakdown of prokaryotic and eukaryotic cells. Intracellular protein degradation is much more than just a mechanism for the removal of incorrectly folded or damaged proteins. Since many short-lived proteins have important regulatory functions, proteolysis makes a significant contribution to many cellular processes including cell cycle regulation and transciptional control. In addition, limited proteolytic cleavage can provide a rapid and efficient mechanism of enzyme activation or inactivation in eukaryotic cells.
In the first chapter, Maurizi provides an introduction to intracellular protein degradation, describes the structure and functions of bacterial ATP-dependent proteases, and explores the relationship between chaperone functions and protein degradation. Many of the principles also apply to eukaryotic cells, although the proteases involved are often not the same. Interestingly, homologues of one of the bacterial proteases, Ion protease, have been found in mitochondria in yeast and mammals, and homologues of proteasomes, which are found in all eukaryotic cells (see below), have been discovered in some eubacteria.
Studies of proteolysis in yeast have contributed greatly to the elucidation of both lysosomal (vacuolar) and nonlysosomal proteolytic pathways in eukaryotic cells. Thumm and Wolf (chapter 2) describe studies that have elucidated the functions of proteasomes in nonlysosomal proteolysis and the contributions of lysosomal proteases to intracellular protein breakdown. Proteins can be selected for degradation by a variety of differen mechanisms. The ubiquitin system is one complex and highly regulated mechanism by which eukaryotic proteins are targetted for degradation by proteosomes. In chapter 3, Wilkinson reviews the components and functions of the ubiquitin system and considers some of the known substrates for this pathway which include cell cycle and transcriptional regulators.
The structure and functions of proteosomes and their regulatory components are described in the two subsequent chapters by Tanaka and Tanahashi and by Dubiel and Rechsteiner. Proteasomes were the first known example of threonine proteases. They are multisubunit complexes that, in addition to being responsible for the turnover of most short-lived nuclear and cytoplasmic protein, are also involved in antigen processing for presentation by the MHC class I pathway. Recent studies reviewed by McCracken and colleagues (chapter 6) lead to the exciting conclusion that some ER-associated proteins are degraded by cytosolic proteasomes.
Lysosomes are responsible for the degradation of long-lived proteins and for the enhanced protein degradation observed under starvation conditions. In chapter 7 Knecht and colleagues review the lysosomal proteases and describe studies of the roles of lysosomes and the mechanisms for protein uptake into lysosomes. Methods of measuring the relative contribution of different proteolytic systems (e.g., ubiquitin-proteasome pathway, calcium-dependent proteases, lysosomes) to muscle protein degradation, and the conclusions from such studies, are reviewed by Attai and Taillinder in the following chapter.
Finally, proteases play an important role in signaling apoptosis by catalyzing the limited cleavage of enzymes. Mason and Beyette review the role of the major players, caspases, which are both activated by and catalyze limite proteolysis, and also consider the involvement of other protoelytic enzymes in this pathway leading cell death.

Molecular Biology of Placental Development and Disease, the latest volume in the Progress in Molecular Biology and Translational Science series, focuses on placental development and disease.

This invaluable resource discusses clinical applications with effects and side-effects of applications of stem cells in liver, lung and heart regeneration. All chapters are contributed by pre-eminent scientists in the field and covers such topics as cell therapy in the treatment of cirrhosis and other liver, heart and lung diseases, characteristics of hepatic progenitor cells, future directions of the discussed therapies and much more. Liver, Lung and Heart Regeneration and the other books in the Stem Cells in Clinical Applications series will be invaluable to scientists, researchers, advanced students and clinicians working in stem cells, regenerative medicine or tissue engineering.

This book provides basic, simple, and logical explanations for choices to be made to run the best Elispot possible. It allows the newcomer to truly understand the best options for specific protocol steps, reagents and materials, and provides even the experienced Elispot user with insight into best practices. The techniques presented here are supported by the author's twenty-plus years of first-hand experience working with this assay, as well as the shared experiences of numerous colleagues and collaborators. The enzyme-linked immunospot (Elispot) assay is a widely used technique to monitor cells on the single cell level for the release of analytes like cytokines, chemokines or immunoglobulins (antibodies), in response to particular stimuli. The most important feature of Elispot is its outstanding sensitivity, allowing the detection of specific cells in very low frequencies. The advantages of Elispot have resulted in it being widely adapted for use in research and translational applications in numerous fields including cancer, infectious diseases, autoimmunity, and transplantation.

This series provides a variety of different discussions on topics within the field of growth factors and cytokines in health and disease.

This book presents current laboratory, scientific and clinical aspects of nanomaterials used for medical applications in the fields of regenerative medicine, dentistry and pharmacy. It gives a broad overview of the in vitro compatibility assessment of nanostructured materials implemented in the medical field by the combination of classical biological protocols. The chapters cover all aspects of integrative medicine, such as green derived nanomaterials for biological applications; synthetic and nature-derived lipid nanoparticles and polymer nanoparticles.

This book discusses recent advances in our understanding of the role of oxidants in microbial pathophysiology, providing valuable insights into the complex role of reactive oxygen species (ROS) in host-microbial interactions. The various chapters take readers through the function of ROS in infections ranging from viral to bacterial, and describe how microorganisms have developed complex strategies to not only avoid contact with phagocyte-derived oxidants, but also protect themselves from injury when oxidants are encountered. Featuring the latest research in the field of microbial diseases, this timely book is a ready reference for scientists looking to develop new anti-microbial drugs.

This volume is essential for anyone struggling with the cellular heterogeneity field, and it connects this new area with the latest advances in innovative techniques combined with practical advice from international experts. The chapters provide a comprehensive overview of cellular heterogeneity analysis techniques, exploring cytometry, imaging and spectroscopy methods, metabolic and molecular biological methods in tracking, as well as chromatin and cell cycle heterogeneity. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, detailed 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 accessible, Cellular Heterogeneity: Methods and Protocols encourages readers to explore new ways of studying cells that will help lead to exciting new discoveries.

Glutathione ( -glutamyl-cysteinyl-glycine) is a ubiquitously distributed sulfurcontaining antioxidant molecule that plays key roles in the regulation of plant growth, development, and abiotic and biotic stress tolerance. It is one of the most powerful low-molecular-weight thiols, which rapidly accumulates in plant cells under stress. Recent in-depth studies on glutathione homeostasis (biosynthesis, degradation, compartmentalization, transport, and redox turnover) and the roles of glutathione in cell proliferation and environmental stress tolerance have provided new insights for plant biologists to conduct research aimed at deciphering the mechanisms associated with glutathione-mediated plant growth and stress responses, as well as to develop stress-tolerant crop plants. Glutathione has also been suggested to be a potential regulator of epigenetic modifications, playing important roles in the regulation of genes involved in the responses of plants to changing environments. The dynamic relationship between reduced glutathione (GSH) and reactive oxygen species (ROS) has been well documented, and glutathione has been shown to participate in several cell signaling and metabolic processes, involving the synthesis of protein, the transport of amino acids, DNA repair, the control of cell division, and programmed cell death. Two genes, gamma-glutamylcysteine synthetase (GSH1) and glutathione synthetase (GSH2), are involved in GSH synthesis, and genetic manipulation of these genes can modulate cellular glutathione levels. Any fluctuations in cellular GSH and oxidized glutathione (GSSG) levels have profound effects on plant growth and development, as glutathione is associated with the regulation of the cell cycle, redox signaling, enzymatic activities, defense gene expression, systemic acquired resistance, xenobiotic detoxification, and biological nitrogen fixation. Being a major constituent of the glyoxalase system and ascorbate-glutathione cycle, GSH helps to control multiple abiotic and biotic stress signaling pathways through the regulation of ROS and methylglyoxal (MG) levels. In addition, glutathione metabolism has the potential to be genetically or biochemically manipulated to develop stress-tolerant and nutritionally improved crop plants. Although significant progress has been made in investigating the multiple roles of glutathione in abiotic and biotic stress tolerance, many aspects of glutathione-mediated stress responses require additional research. The main objective of this volume is to explore the diverse roles of glutathione in plants by providing basic, comprehensive, and in-depth molecular information for advanced students, scholars, teachers, and scientists interested in or already engaged in research that involves glutathione. Finally, this book will be a valuable resource for future glutathione-related research and can be considered as a textbook for graduate students and as a reference book for frontline researchers working on glutathione metabolism in relation to plant growth, development, stress responses, and stress tolerance.

This detailed book collects new and updated techniques that will help to improve our understanding of the mechanisms underlying stem cell-derived repair through stem cells of the epidermal and dermal lineages, which have led to the isolation of numerous stem cell-like sub-populations from the epidermis and dermis. Written for the highly successful Methods in Molecular Biology series, 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 practical, Skin Stem Cells: Methods and Protocols, Second Edition serves as a valuable assemblage of protocols for researchers both already working in the field and those who now wish to newly embark on studies of skin stem cells.

This volume presents methods and techniques to study oogenesis in a broad range of organisms, from plants to mammals. Oogenesis: Methods and Protocols guides readers through protocols on models of developmental biology, oogenesis in plants, worms, fruit flies, mosquitos, butterflies, starfish, zebrafish, frog, chicken and mouse. 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, Oogenesis: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This invaluable resource discusses clinical applications with effects and side-effects of applications of stem cells in diabetes, kidney and wound treatment. All chapters are contributed by pre-eminent scientists in the field and covers such topics as stem cells and cell therapy in the treatment of diabetes mellitus, kidney failure, wound and other skin aging diseases, characteristics of some kinds of stem/progenitor cells for therapy, future directions of the discussed therapies and much more. Pancreas, Kidney and Skin Regeneration and the other books in the Stem Cells in Clinical Applications series will be invaluable to scientists, researchers, advanced students and clinicians working in stem cells, regenerative medicine or tissue engineering.

This volume deals with aspects of the cytoskeleton in different cell types and also describe examples of changes in the cytoskeleton which occur during various pathological states. These studies bring the exciting area of cytoskeleton research into the domain of medical science.

This book provides a detailed and up-to-date overview of all aspects of yeast cell surface engineering, including fundamental principles, practical strategies for the construction of engineered yeasts, as well as medical and industrial applications. The technique makes it possible to add eukaryotic modifications to the surface-displayed proteins/peptides, which is of significant value in basic and applied research. Generally referred to as an arming (molecular display) technology, it allows yeast to be used as a whole-cell biocatalyst for a range of purposes, including bio-energy production, pollutant removal, recovery of rare metal ions, and preparation of functional cells, all of which are comprehensively covered in the book. Among the medical applications discussed are in vitro antibody preparation and the production of oral vaccines. In addition, it presents the latest advances in protein engineering and high-throughput screening for directed evolution of enzymes. The book enables graduate students and researchers to gain a deeper, comprehensive understanding of the technology, and offers further inspiration for researchers and industrial experts in this rapidly evolving field.

The book conveys a comprehensive knowledge of long and short ncRNAs in cancer regulation and their potentials as diagnostic biomarkers and therapeutic targets. Topics covered include the molecular mechanisms of various classes of ncRNAs (with emphasis on long non-coding RNAs and microRNAs) in cancer, the functional roles of ncRNAs in regulating different cancer hallmarks (including proliferation, apoptosis, stem-cell properties, epithelial-mesenchymal transition, metabolism, angiogenesis, tumor-host interactions and therapeutic resistance), the role of ncRNAs in regulating cancer signaling circuitry programs (highlighting their involvement in c-myc, p53 and NFkB signaling), a systemic summary of clinical and preclinical studies that evaluate the potential of ncRNA signatures for cancer diagnosis and prognosis and strategies to delivery short ncRNAs as therapeutic molecules for cancer treatment. This book may serve as a comprehensive resource for researchers, graduate students and oncologists in ncRNA and cancer research and help drug development by identifying ncRNA targets.

This is the third volume in a series on membrane protein transfer. Membrane protein transport underlies the topological disposition of many proteins within cells and it is this disposition that allows for the co-ordination of the central cellular processes, such as metabolism.

This book focuses on the mechanobiological principles in tissue engineering with a particular emphasis on the multiscale aspects of the translation of mechanical forces from bioreactors down to the cellular level. The book contributes to a better understanding of the design and use of bioreactors for tissue engineering and the use of mechanical loading to optimize in vitro cell culture conditions. It covers experimental and computational approaches and the combination of both to show the benefits that computational modelling can bring to experimentalists when studying in vitro cell culture within a scaffold. With topics from multidisciplinary fields of the life sciences, medicine, and engineering, this work provides a novel approach to the use of engineering tools for the optimization of biological processes and its application to regenerative medicine. The volume is a valuable resource for researchers and graduate students studying mechanobiology and tissue engineering. For undergraduate students it also provides deep insight into tissue engineering and its use in the design of bioreactors. The book is supplemented with extensive references for all chapters to help the reader to progress through the study of each topic.

The present volume is the first attempt to bring together, in a comprehensive and comparative manner, the vast knowledge accumulated on the mycoplasma cell membranes. The state-of-the-art coverage includes physical studies of lipid organization and dynamics in mycoplasma membranes, the role of cholesterol in mycoplasma membranes, spiralins, and eight other subjects.

Why a Second Edition?
The Second Edition provides practical answers to the general question, "How can I obtain useful sequence information from my protein or peptide?" rather than the more specific question asked in the first edition, "How can I obtain the N-terminal sequence?" Important new methods include ways of dealing with blocked N termini, computer analysis of protein sequences, and the recent revolution in mass spectrometry.
Key Features
Highlights of the Second Edition:
* Mass spectrophotometric characterization of proteins and peptides
* N-terminal sequencing of proteins with blocked N termini
* Internal amino acid sequence analysis after protease digestion in-gel and on-blot
* Improved microscale peptide purification methods
* Computer analysis of protein sequences
* New protocols tested and refined through everyday use in authors' laboratories
* Updated reference chapter covering all aspects of protein microsequencing

This book will provide current understandings about two ubiquitously expressed metabotropic GPCRs, G-coupled purinoreceptor type 2 (P2Y) and Takeda G-protein-coupled bile acid receptor 5 (TGR5). G protein coupled receptors (GPCRs) are the largest family of proteins implicated in majority of cellular responses. The two receptor sub-families play a central role in many physiological functions as well as in many pathological conditions. This book offers up-to-date information on the physiological functions, signaling pathways and regulatory mechanisms of P2Y and TGR5 receptors. In addition, this book provides a comprehensive overview about the abnormalities of P2Y/TGR5 receptors and their contribution in the development and progression of pathological conditions. It also covers the currently available natural, chemical and pharmacological agents targeting these two receptor families and their therapeutic implications in P2Y and TGR5 associated disorders. This book is a valuable source for beginners and researchers to follow the rapidly progressing field of these two GPCR subfamily members.