This volume provides a collection of protocols from leading researchers in the dynein field. The enclosed chapters guide readers through a broad range of powerful techniques that can be used to understand the complexities of dynein activity and regulation. These include how to perform live-cell microscopy on various fungi and metazoan cells, how to purify, assemble, and measure the force generation and motility characteristics of various dynein assemblies. 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, Dynein: Methods and Protocols is a valuable resource for both novice and expert scientists in this developing field.

The book helps the reader to better understand cytogenetics and the intricacies of the methodology. The different methods of fluorescence in situ hybridization are discussed and the results achieved are presented. The book provides a comprehensive review of basic and applied aspects of cytogenetics and thus is of intense interest to all those interested in chromosomes and their alterations by different types of mutagens, including chemical mutagens and ionizing and nonionizing radiation, with special reference to electromagnetic fields.

This book provides an essential overview of the role of phosphoinositide-3-phosphate kinase (PI3K) isoforms in modulating the function of immune system cells and their involvement in disease. PI3K is a family of kinases involved in basic cellular processes such as proliferation, migration and cell death. Recent work has highlighted the multiple roles of PI3K classes and subunits and their involvement in the immune response to the body's own and foreign antigens and diseases such as cancer and autoimmunity. This book offers a detailed introduction to the biology of the three PI3K classes, followed by an extensive discussion of the diverse roles of AKT and PI3K isoforms in immune cells. Based on this knowledge, it subsequently explains in more detail how specific isoforms are connected to immune-mediated diseases. The book concludes by highlighting the latest advances in the production of isoform-specific inhibitors and their use in various human diseases. This book is intended as a reference guide for students and researchers interested in the multifaceted aspects of PI3K biology.

This book is about different Enzymes from various sources that play an important role in the degradation of an array of pollutants with simultaneous generation of value-added products. This is an "Edited Book" which deals a comprehensive knowledge on the role of different microorganisms/their enzymes in the degradation of pollutants, wastewater treatment with simultaneous production of value added products. It also deals the current state, perspectives and various challenges associated with the microbial/enzymatic degradation of environmental pollutants. This book will provide a profound knowledge on the importance of microorganisms/their enzymes in the degradation of pollutants like pesticides, antibiotics, toxic/hazardous chemicals, endocrine disrupting chemicals/compounds with production of value-added products like bioplastics for the sustainable development of society. It covers various existing wastewater treatment approaches using microorganisms alone and /or in combination of other methods with their merits, demerits and future prospects.

This second edition explores membrane trafficking research that has been at the frontier of cell and molecular biology. In the past 20 years, this field has witnessed a remarkable convergence and synergism from information gained using genetic approaches in yeast cells, and biochemical and molecular approaches in mammalian cells. This volume contains updated chapters and the inclusion of a large number of new chapters, and is divided into three parts. Part I includes biochemical and molecular genetics approaches and methods used in analysing membrane traffic, in both yeast and mammalian cell models. Part II focuses on imaging and microscopy approaches and techniques, while Part III highlights the Omics-type approaches. 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 resourceful, Membrane Trafficking, Second Edition, is a valuable resource for both experienced researchers, graduate students, and junior scientists in the field of membrane trafficking.

Free Radicals in Biology and Medicine has become a classic text in the field of free radical and antioxidant research. Now in its fifth edition, the book has been comprehensively rewritten and updated whilst maintaining the clarity of its predecessors. Two new chapters discuss 'in vivo' and 'dietary' antioxidants, the first emphasising the role of peroxiredoxins and integrated defence mechanisms which allow useful roles for ROS, and the second containing new information on the role of fruits, vegetables, and vitamins in health and disease. This new edition also contains expanded coverage of the mechanisms of oxidative damage to lipids, DNA, and proteins (and the repair of such damage), and the roles played by reactive species in signal transduction, cell survival, death, human reproduction, defence mechanisms of animals and plants against pathogens, and other important biological events. The methodologies available to measure reactive species and oxidative damage (and their potential pitfalls) have been fully updated, as have the topics of phagocyte ROS production, NADPH oxidase enzymes, and toxicology. There is a detailed and critical evaluation of the role of free radicals and other reactive species in human diseases, especially cancer, cardiovascular, chronic inflammatory and neurodegenerative diseases. New aspects of ageing are discussed in the context of the free radical theory of ageing. This book is recommended as a comprehensive introduction to the field for students, educators, clinicians, and researchers. It will also be an invaluable companion to all those interested in the role of free radicals in the life and biomedical sciences.

The Encyclopedia of Cell Biology, Four Volume Set offers a broad overview of cell biology, offering reputable, foundational content for researchers and students across the biological and medical sciences. This important work includes 285 articles from domain experts covering every aspect of cell biology, with fully annotated figures, abundant illustrations, videos, and references for further reading. Each entry is built with a layered approach to the content, providing basic information for those new to the area and more detailed material for the more experienced researcher. With authored contributions by experts in the field, the Encyclopedia of Cell Biology provides a fully cross-referenced, one-stop resource for students, researchers, and teaching faculty across the biological and medical sciences.

Knowledge of the microscopic structure of biological systems is the key to understanding their physiological properties. Most of what we now know about this subject has been generated by techniques that produce images of the materials of interest, one way or another, and there is every reason to believe that the impact of these techniques on the biological sciences will be every bit as important in the future as they are today. Thus the 21st century biologist needs to understand how microscopic imaging techniques work, as it is likely that sooner or later he or she will have to use one or another of them, or will otherwise become dependent on the information that they provide. The objective of this textbook is to introduce its readers to the many techniques now available for imaging biological materials, e.g. crystallography, optical microscopy and electron microscopy, at a level that will enable them to use them effectively to do research. Since all of these experimental methods are best understood in terms of Fourier transformations, this book explains the relevant concepts from this branch of mathematics, and then illustrates their elegance and power by applying them to each of the techniques presented. The book is derived from a one-term course in structural biology that the author gave for many years at Yale. It is intended for students interested either in doing structural research themselves, or in exploiting structural information produced by others. Over the years, the course was taken successfully by advanced undergraduates and by graduate students. Scientists interested in entering the structural biology field later in their careers may also find it useful.

Second edition of the only comprehensive and student-friendly introduction to molecular exercise physiology available 12 chapter structure around which a complete single-semester course can be built Explains the concepts of exercise physiology at a molecular level in clear, accessible language and relates them to real practical and applied outcomes Covers applications in both sport and health-related physical activity Includes new chapters on the molecular aspects of exercise and cancer, and exercise and metabolic and cardiovascular diseases Every chapter includes useful features such as learning objectives, summaries, structured guides to further reading, review questions, overviews of work by key researchers and box discussions of important contemporary debates

The ongoing debate on the use of DNA profiles to identify perpetrators in criminal investigations or fathers in paternity disputes has too often been conducted with no regard to sound statistical, genetic or legal reasoning. The contributors to Human Identification: The Use of DNA Markers all have considerable experience in forensic science, statistical genetics or jurimetrics, and many of them have had to explain the scientific issues involved in using DNA profiles to judges and juries. Although the authors hold differing views on some of the issues, they have all produced accounts which pay due attention to the, sometimes troubling, issues of independence of components of the profiles and of population substructures. The book presents the considerable evolution of ideas that has occurred since the 1992 Report of the National Research Council of the U.S. Audience: Indispensable to forensic scientists, laying out the concepts to all those with an interest in the use of genetic information. The chapters and exhaustive bibliography are vital information for all lawyers who must prosecute or defend DNA cases, and to judges trying such cases.

Basics of proteins and proteomics techniques In-depth understanding of mass-spectrometry and quantitative proteomics An overview of interactomics and its application for translational research. Advancement in the field of proteomics and challenges in clinical applications.

Transforming growth factor-? (TGF-?) is a secreted polypeptide with multifunctional properties manifested during embryonic development, adult organ physiology, and pathobiology of major diseases, including cancer and fibrotic and cardiovascular diseases. The signaling pathway of TGF-? now is rather well understood. Continuing revelations in the mechanisms of action of TGF-? provide specific mechanistic examples of how human cells lose their controlled function and behave wrongly during the development of diverse diseases. Equally important, however, is the current promise of exploiting the TGF-? pathway in combating human disease. This book comprehensively covers major areas of human disease where the involvement of TGF-? is firmly established. Simultaneously, the book highlights major gaps in knowledge and the future directions of research that can benefit human medical science. The core set of diseases where TGF-? action is well documented and are included in the book are cancer and cardiovascular and fibrotic disorders. The central aim of the book is to stimulate young scientists to enter the prolific TGF-? field and find new solutions to the problems remaining in this area of study. For this purpose the book provides authoritative educational chapters that furnish a good introduction to the field for young doctoral students, postdocs, and clinical fellows. The book also serves as a valuable reference for the aficionados in the field, who can find accessible and well- illustrated material for their teaching and lecturing activities, via which the importance of TGF-? biology is disseminated to the world of science and to the public.

This book offers a clearly written and highly accessible account of two different aspects of carbohydrate chemistry. Carbohydrates are an essential component of life and have many important biological functions, but the details of how carbohydrates interact with other biomolecules to mediate biological signalling remain unclear. Firstly, this thesis details innovative methods to mine protein structural data to uncover new features of carbohydrate-based interactions. It also explains these findings using physical chemistry, specifically CH-pi interactions associated with the properties of the interacting partners. Carbohydrates are also critical for tissue growth and development, yet are underexploited in the materials science that underpins much of regenerative medicine. As such, the second part of this thesis describes a diverse array of techniques ranging from synthetic chemistry and enzymatic synthesis to prepare a wide variety of carbohydrates, and materials chemistry to prepare glycosylated hydrogels, to cell biology to determine the effects on cellular development for tissue engineering applications.

This book examines detailed experimental and computational approaches for the analysis of many aspects vital to the understanding of membrane protein structure and function. Readers will receive guidance on the selection and use of methods for over-expression and purification, tools to characterize membrane proteins within different phospholipid bilayers, direction on functional studies, and approaches to determine the structures of membrane proteins. Detailed experimental steps for specific membrane proteins with critical notes allow the protocols to be modified to different systems. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of practical information and implementation advice that leads to excellent, reproducible results. Authoritative and up-to-date, Structure and Function Studies of Membrane Proteins serves as an ideal guide for biologists, biochemists, and biophysicists striving to further understand these essential proteins and their many biological roles.

This book covers a wide range of topics that illustrate the various functions of autophagy in stem cells and offers insights on the mechanisms by which autophagy can regulate stem-cell self-renewal and facilitate specific differentiation programs. Stem cells are unique cells present in most multicellular animals and are essential for their survival. They have two unique properties: the ability to self-renew and the ability to differentiate into one or more cell types. These characteristics of stem cells have found immense therapeutic potential in regenerative medicine. Autophagy is a crucial membrane trafficking pathway that is essential for maintaining cellular homeostasis that involves sequestration of non-functional proteins, protein aggregates and damaged organelles in double-membraned vesicles called autophagosomes, which are subsequently targeted to the lysosome for degradation. The primary aim of this book is to provide knowledge of recent developments in our understanding of the role of autophagy in stem cells, including germline stem cells. Autophagy is considered a promising target for many diseases. Significant efforts are being developed to identify specific modulators of autophagy, which will aid in designing combinatorial therapeutic strategies that will allow significant improvements in regenerative medicine.

This third edition provides new and updated chapters detailing preparation of liposomes, physicochemical characterization of liposomes, lipid analysis, drug encapsulation, surface modification, stimuli response as well as cellular interaction, and biodistribution. Also included is an updated chapter on the history and evolution of the field of liposomology. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Liposomes: Methods and Protocols, Third Edition aims to serve as a reference for graduate students, post-doctoral researchers as well as established investigators utilizing lipid-based systems.

Sequence-specific DNA binding ligands, amongst which triple helix forming oligonucleotides are the most efficient as yet, represent promising tools in a number of fields. One of their most promising applications is as antiviral tools: they can specifically target a viral gene, even if it is integrated into the host genome, and be used to specifically inactivate the viral gene or even destroy the cells harboring this gene. However, from science fiction to science there remains a gap; and we are at the moment on the threshold of this fascinating field. Triple Helix Forming Oligonucleotides considers the different aspects of the design and improvement, current or future, of these molecules and their structural analysis, as well as their applications, with special emphasis on the attempts to obtain biological effects of these potentially important tools. What emerges is that the current state of the research is encouraging, and that these molecules are already useful in some biotechnology applications.

This volume details novel and popular approaches to decipher the Ubiquitin Code. Chapters guide readers through ubiquitin chain complexity, methods to generate non-hydrolysable ubiquitin-NEDD8, a fully synthetic method for the linear synthesis of fluorescently labelled rhodamine LC3A and LC3B, a HECT ligase-mediated ubiquitylation in vitro, a RNAi-based screening to identify DUBs, a Yuh1 activity assay, a pull-down to isolate K48 and K63 chains using specific nanobodies, identification of NeDDylated protein based in a NEDD8R74K mutant, in vivo identification of NEDDylated proteins from liver diseases, UbL-ID method to identify ubiquitin-like modified proteins, a LC3/GABARAP capturing system, and methods to isolate peptides that are closely associated with proteasomes or trapped inside the core particle of proteasomes in eukaryotic cells, among others. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, The Ubiquitin Code aims to to be a useful practical guide to researches to help further their study in this field.

This detailed volume explores the field of bacterial virulence and the effort to understand how microbial interaction with a host results in the pathology of a specific disease. This collection of selected protocols includes advanced molecular biology and bioinformatics methods, cell culture and organoid models of infection, as well as in vivo infection models that are useful to study the interaction of pathogens with plants, insects, avian, and mammalian hosts. 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 protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and beneficial, Bacterial Virulence: Methods and Protocols serves as an ideal guide for researchers seeking to promote and further develop the exciting and continuously evolving field of bacterial virulence. Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This book aims at providing insights into the collagen superfamily and the remarkable diversity of collagen function within the extracellular matrix. Additionally, the mechanisms underlying collagen-related diseases such as dystrophic epidermolysis bullosa, osteogenesis imperfecta, as well as collagen-related myopathies and neurological disorders are discussed. Collagens are the most abundant extracellular matrix proteins in organisms. Their primary function is to provide structural support and strength to cells and to maintain biomechanical integrity of tissues. However, collagens can no longer be considered just as structural proteins. They can act as extracellular modulators of signaling events and serve critical regulatory roles in various cell functions during embryonic development and adult homeostasis. Furthermore, collagens are associated with a broad spectrum of heritability-related diseases known as "collagenopathies" that affect a multitude of organs and tissues including sensorial organs. The book is a useful introduction to the field for junior scientists, interested in extracellular matrix research. It is also an interesting read for advanced scientists and clinicians working on collagens and collagenopathies, giving them a broader view of the field beyond their area of specialization.

This detailed volume is devoted to the recent development of quantitative experiments and computational methods driving new transforming growth factor beta (TGF- ) and other cell signaling knowledge. Many chapters cover quantitative assays for TGF- signaling studies, with others exploring the increasing role of both modeling and computational methods. 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. Authoritative and up-to-date, TGF-Beta Signaling: Methods and Protocols serves as a vital resource for researchers seeking to move the TGF- field into the quantitative realm.

This second edition of Eosinophils: Methods and Protocols updates several techniques from the first edition together with novel in vitro and in vivo-based methodologies. Written by internationally recognized expert authorities, this volume provides vital techniques from eosinophil purification to experimental modelling, with each technique spelled out in clear and straightforward terms, assuming no previous knowledge of the method and without necessitating sourcing additional information elsewhere. Written in the highly successful Methods in Molecular Biology format, chapters include introductions to their respective topics, lists of the essential materials and reagents, step-by-step, readily reproducible laboratory protocols, with tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Eosinophils: Methods and Protocols, Second Edition provides the practical means to extend our knowledge of eosinophil function in health and disease, underpinning research that may in turn lead to new hypotheses for future examinations into the role of this intriguing and enigmatic leukocyte. Chapters 10 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume provides an up-to-date collection of protocols describing some of the key methods to investigate the integrated stress response (ISR), a vital evolutionarily conserved mechanism that enables eukaryotic cells to adapt to stress conditions and alter their gene expression programs. The content of the book is split between techniques to analyze mRNA translation regulation and methods to analyze interaction networks and ribonucleoprotein (RNP) granules. 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. Authoritative and practical, The Integrated Stress Response: Methods and Protocols serves as an ideal guide to help accelerate research into the complex and fascinating biology of the ISR.

The Formation of the Heart and its Regulation reviews in considerable detail the major events in heart development and their control via genes, cell-cell interactions, growth factors and other contributing elements. In addition, there is an extensive and useful overview of the field of heart development taken as a whole. The book will appeal to all students and researchers working on cardiovascular development and to pediatric cardiologists.

This volume provides up-to-date methods on single cell wet and bioinformatics protocols based on the researcher experiment requirements. Chapters detail basic analytical procedures, single-cell data QC, dimensionality reduction, clustering, cluster-specific features selection, RNA velocity, multi-modal data integration, and single cell RNA editing. 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 comprehensive, Single Cell Transcriptomics: Methods and Protocols aims to be a valuable resource for all researchers interested in learning more about this important and developing field.