This volume compiles a broad range of step-by-step protocols, complementary to the ones published in the first edition of this book, to study various aspects of mitochondrial structure and function in different model organisms, both in vitro and in vivo. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Mitochondria: Practical Protocols, Second Edition aims to be useful for beginners as well as for experienced researchers in the field.

This book sheds new light on the current state of knowledge concerning chromatin organization. Particular emphasis is given to the new imaging potential offered by super-resolution microscopy, which allows DNA imaging with a very high labeling density. From the early work on chromosomes by Walther Flemming in the nineteenth century to recent advances in genomics, the history of chromatin research now spans more than a century. The various milestones, such as the discovery of the double helix structure, the sequencing of the human genome, and the recent description of the genome in 3D space, show that understanding chromatin and chromosome function requires a clear understanding of its structure. Presenting cutting-edge data from super-resolution single molecule microscopy, the book demonstrates that chromatin manifests several levels of folding, from nucleosomes to chromosomes. Chromatin domains emerge as a new fundamental building block of chromatin architecture, with functions possibly related to gene regulation. A detailed description of chromatin folding in the pachytene stage of meiosis serves as a model for exploring this functionality, showing the apparent interplay between structure, function, and epigenetic regulation. Lastly, the book discusses possible new avenues of innovation to describe chromatin's organization and functions. Gathering essential insights on chromatin architecture, the book offers students an introduction to microscopy and its application to chromatin organization, while also providing advanced readers with new ideas for future research.

This volume contains a collection of protocols from some of the major laboratories involved in stem cell research across the world. The research discussed in this book covers topics such as: isolating, characterizing and expanding dental pulp stem cells; manipulating the proliferative potential of cardiomyocytes by gene transfer; isolation of stromal stem cells from adipose tissue; noninvasive assessment of cell fat and biology in transplanted mesenchymal stem cells; and cell-free therapy for organ repair. 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, Adult Stem Cells: Methods and Protocols is a valuable resource for helping researchers transform the study of stem cells into an industrialized process that will supply patients with efficient, safe, and cost-effective cell treatments.

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.

The book describes the major degenerative processes and pathologies exacerbated by senescence and how they can be alleviated through retardation of cellular aging. Topics discussed include neurodegenerative disease, protein oxidation, cerebrovascular disease, particle-induced inflammation and cardiovascular disease, Alzheimer's disease, ovarian aging, dietary and endogenous anti-oxidants in management of Parkinson's disease, and effects of exercise on oxidation and inflammation. The nineteen expertly authored chapters are organized into three sections in order to present a complete picture to the reader: Age Related Cellular Events, Role of Inflammatory and Oxidative Processes in Age-Related Diseases, and Retardation of Cellular Aging. Inflammation, Oxidative Stress and Age-Related Disease draws from a variety of international perspectives and provides a comprehensive overview of the relationship between disease, cell aging, and oxidative stress, as well as potential for preventing or slowing these processes. This installment of Springer's Oxidative Stress in Applied Basic Research and Clinical Practice is ideal for researchers, clinicians, and advanced graduate students in the fields of cardiology, neuroscience, biogerontolgy, and cell biology.

This volume covers protocols related to both pluripotent and somatic stem cells, including the ethical procurement of tissues and cells for the provision of "seed stock," standardized methods for deriving hESCs and iPSCs, isolating mesenchymal stem cells, cell culture and cryopreservation, in addition to quality assurance and information management. Stem Cell Banking: Concepts and Protocols aims to contribute to the development of this field by providing information that is essential to establishing a bona fide stem cell bank. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Stem Cell Banking: Concepts and Protocols is a valuable resource for stem cell scientists and novices to the field, and will help strengthen and maximize their use of existing and future stem cell resources.

This two-volume set takes an in-depth look at stress signaling in plants from a uniquely genomic and proteomic perspective and offers a comprehensive treatise that covers all of the signaling pathways and mechanisms that have been researched so far. Currently, plant diseases, extreme weather caused by climate change, drought and an increase in metals in soil are amongst the major limiting factors of crop production worldwide. They devastate not only the food supply but also the economy of a nation. With global food scarcity in mind, there is an urgent need to develop crop plants with increased stress tolerance so as to meet the global food demands and to preserve the quality of our planet. In order to do this, it is necessary to understand how plants react and adapt to stress from the genomic and proteomic perspective. Plants adapt to stress conditions by activating cascades of molecular mechanisms, which result in alterations in gene expression and synthesis of protective proteins. From the perception of the stimulus to the transduction of the signal, followed by an appropriate cellular response, the plants employ a complex network of primary and secondary messenger molecules. Cells exercise a large number of noticeably distinct signaling pathways to regulate their activity. In order to contend with different environmental adversities, plants have developed a series of mechanisms at the physiological, cellular and molecular levels that respond to stress. Each chapter in this volume provides an in-depth explanation of what we currently know of a particular aspect of stress signaling and where we are heading. Together with the highly successful first volume, Stress Signaling in Plants: Genomics and Proteomics Perspective, Volume 2 covers an important aspect of plant biology for both students and seasoned researchers.

This volume provides leading-edge protocols in the study of the molecular and cellular biology of muscle stem cells. Chapters detail current and updated methods for muscle stem cell isolation, culture, molecular analysis, cellular analysis, and reintroduction in vivo as well as protocols for studying myogenic stem cells in non-mammalian model systems. 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, Muscle Stem Cells: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This book provides a comprehensive overview of the cascade of events activated in the body following the implant of biomaterials and devices. It is one of the first books to shed light on the role of the host immune response on therapeutic efficacy, and reviews the state-of-the-art for both basic science and medical applications. The text examines advantages and disadvantages of the use of synthetic versus natural biomaterials. Particular emphasis is placed on the role of biomimicry in the development of smart strategies able to modulate infiltrating immune cells, thus reducing side effects (such as acute and chronic inflammation, fibrosis and/or implant rejection) and improving the therapeutic outcome (healing, tissue restoration). Current cutting-edge approaches in tissue engineering, regenerative medicine, and nanomedicine offer the latest insights into the role immunomodulation in improving tolerance during tissue transplant in the treatment of orthopaedic, pancreatic, and hepatic diseases. "Immune Response to Implanted Materials and Devices" is intended for an audience of graduate students and professional researchers in both academia and industry interested in the development of smart strategies, which are able to exploit the self-healing properties of the body and achieve functional tissue restoration.

Fall-induced hip fracture is an epidemic health risk among elderly people. This book presents an image-based multilevel modeling approach to understanding the biomechanics involved in fall-induced hip fracture. By hierarchically integrating a body-level dynamics model, a femur-level finite element model, and a local bone failure model, the biomechanics approach is able to simulate all stages in sideways falls and to incorporate all biomechanical variables affecting hip fracture. This book is useful for clinicians to accurately evaluate fracture risk, for biomechanical engineers to virtually test hip protective devices, and for biomedical students to learn image-based biomechanical modeling techniques. This book also covers: Biomechanical viewing on bone composition, bone remodeling, and bone strength Bone imaging and information captured for constructing biomechanical models Bone mechanical testing and mechanical properties required for biomechanical modeling

This second edition volume provides detailed protocols on the theoretical background of cell cycle synchronization procedures and instructions on how to implement these techniques. The chapters in Cell Cycle Synchronization: Methods and Protocols, Second Edition cover subjects such as: physical fractionations including centrifugal elutriation of healthy and apoptotic cells, and nuclei of mammalian cells; large scale mitotic cell synchronization; chromosome formation during fertilization in eggs; synchronization of unicellular organisms; hematopoietic stem cells used to improve the engraftment in transplantation; and cell cycle control. 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. Practical and comprehensive, Cell Cycle Synchronization: Methods and Protocols, Second Edition is a valuable resource for PhD students and postdoctoral fellows, and researchers interested in general science, pharmacy, medicine and public health, computer science, and life sciences. Specialists and professionals in cell biology, genetics, molecular biology, biochemistry, and pharmacology will also find this book useful.

This book presents an overview of computational and statistical design and analysis of mass spectrometry-based proteomics, metabolomics, and lipidomics data. This contributed volume provides an introduction to the special aspects of statistical design and analysis with mass spectrometry data for the new omic sciences. The text discusses common aspects of design and analysis between and across all (or most) forms of mass spectrometry, while also providing special examples of application with the most common forms of mass spectrometry. Also covered are applications of computational mass spectrometry not only in clinical study but also in the interpretation of omics data in plant biology studies. Omics research fields are expected to revolutionize biomolecular research by the ability to simultaneously profile many compounds within either patient blood, urine, tissue, or other biological samples. Mass spectrometry is one of the key analytical techniques used in these new omic sciences. Liquid chromatography mass spectrometry, time-of-flight data, and Fourier transform mass spectrometry are but a selection of the measurement platforms available to the modern analyst. Thus in practical proteomics or metabolomics, researchers will not only be confronted with new high dimensional data types-as opposed to the familiar data structures in more classical genomics-but also with great variation between distinct types of mass spectral measurements derived from different platforms, which may complicate analyses, comparison, and interpretation of results.

Heat Shock Proteins and Plants provides the most up-to-date and concise reviews and progress on the role of heat shock proteins in plant biology, structure and function and is subdivided into chapters focused on Small Plant HSPs (Part I), Larger Plant HSPs (Part II) and HSPs for Therapeutic Gain (Part III). This book is written by eminent leaders and experts from around the world and is an important reference book and a must-read for undergraduate, postgraduate students and researchers in the fields of Agriculture, Botany, Crop Research, Plant Genetics and Biochemistry, Biotechnology, Drug Development and Pharmaceutical Sciences.

This volume covers methods for the analysis of extracellular vesicles (EV) that can be applied to isolated EVs from a wide variety of sources. This includes the use of electron microscopy, tunable resistance pulse sensing, and nanoparticle tracking analysis. The chapters in this book discuss EV cargoes containing proteins and genomic materials using a number of different approaches, and isolating EVs from platelets and neuronal cells and tissues. 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. Practical and comprehensive, Exosomes and Microvesicles: Methods and Protocols is a valuable resource containing methodologies for anyone interested in researching EVs.

Soil salinity is a key abiotic-stress and poses serious threats to crop yields and quality of produce. Owing to the underlying complexity, conventional breeding programs have met with limited success. Even genetic engineering approaches, via transferring/overexpressing a single 'direct action gene' per event did not yield optimal results. Nevertheless, the biotechnological advents in last decade coupled with the availability of genomic sequences of major crops and model plants have opened new vistas for understanding salinity-responses and improving salinity tolerance in important glycophytic crops. Our goal is to summarize these findings for those who wish to understand and target the molecular mechanisms for producing salt-tolerant and high-yielding crops. Through this 2-volume book series, we critically assess the potential venues for imparting salt stress tolerance to major crops in the post-genomic era. Accordingly, perspectives on improving crop salinity tolerance by targeting the sensory, ion-transport and signaling mechanisms were presented in Volume 1. Volume 2 now focuses on the potency of post-genomic era tools that include RNAi, genomic intervention, genome editing and systems biology approaches for producing salt tolerant crops.

This essential volume explores mesenchymal stem cells (MSCs) and their potential to suppress immune-mediated inflammation. The chapters examine applications in autoimmune diseases such as lupus, rheumatoid arthritis and multiple sclerosis; blood cancers such as leukemia and lymphoma; and reproductive complications, specifically pre-term labor and use of MSCs in vitro and in animal models to discover methods of suppressing the causal inflammatory response. It also further defines the methodologies required to develop research on MSCs in vitro into established preclinical animal models including those which are proven replicas of autoimmunity and pre-term labor, to name but two. Mesenchymal Stem Cells and Immunomodulation, part of Springer's Stem Cell Biology and Regenerative Medicine, is an invaluable resource for researchers and clinicians working with stem cells, autoimmune disease, oncology, and reproductive medicine.

This book provides a comprehensive overview of topics describing the earliest steps of fertilization, from egg activation and fertilization to the activation of the zygotic genome, in various studied vertebrate model systems. The contribution of maternal and paternal factors and their role in the early embryo as parental DNA becomes modified and embryonic genes become activated is fundamental to the initiation of embryogenesis in all animal systems. It can be argued that this is a unique developmental period, when information from the parents is compressed to direct the development of the body plan of the entire organism, a process of astounding simplicity, elegance and beauty. In addition to their fundamental scientific interest, many frontiers of biomedicine, such as reproductive biology, stem cells and reprogramming, and the understanding of intergenerational diseases, depend on advances in our knowledge of these early processes. Vertebrate Development: Maternal to Zygotic Control brings together chapters from experts in various disciplines describing the latest advances related to this important developmental transition. Each chapter is a synthesis of knowledge relevant to all vertebrates, with details on specific systems as well as comparisons between the various studied vertebrate models. The editorial expertise encompasses the fields of major vertebrate model systems (mammalian, amphibian and teleost) ensuring a balanced approach to various topics. This unique book-with its combination of in-depth and up-to-date basic research, inter-species comprehensiveness and emphasis on the very early stages of animal development-is essential for research scientists studying vertebrate development, as well as being a valuable resource for college educators teaching advanced courses in developmental biology.

This book offers one of the most comprehensive reviews in the field of gastrointestinal (GI) physiology, guiding readers on a journey through the complete digestive tract, while also highlighting related organs and glandular systems. It is not solely limited to organ system physiology, and related disciplines like anatomy and histology, but also examines the molecular and cellular processes that keep the digestive system running. As such, the book provides extensive information on the molecular, cellular, tissue, organ, and system levels of functions in the GI system. Chapters on the roles of the gut as an endocrine, exocrine and neural organ, as well as its microbiome functions, broaden readers' understanding of the multi-organ networks in the human body. To help illustrate the interconnections between the physiological concepts, principles and clinical presentations, it outlines clinical examples such as pathologies that link basic science with clinical practice in special "clinical correlates" sections. Covering both traditional and contemporary topics, it is a valuable resource for biomedical students, as well as healthcare and scientific professionals.

This book marries stem cell biology, tissue engineering, and regenerative biology into a single, interdisciplinary volume. The chapters also explore embryonic stem cells, induced pluripotent stem cells, cardiovascular regeneration, skeletal development, inflammation, polymeric biomaterials, neural injury, cartilage regeneration, regeneration in ambystoma, models for regeneration using salamander and zebrafish, and more. The volume also discusses recent advances and their potential in developing future therapies. Innovations in Molecular Mechanisms and Tissue Engineering combines perspectives from the biomedical, bioengineering, and medical fields to present a cutting-edge, multifaceted picture of the tissue engineering and regenerative medicine fields. This installment of Springer's Stem Cell Biology and Regenerative Medicine series is ideal for scientists, clinicians, and researchers in the fields of stem cell biology, regenerative medicine, biomedical engineering, and tissue engineering.

This book brings together in one volume the current state of ageing research in the nematode Caenorhabditis elegans. The authors are leading researchers in the field, placing this topic in the context of human ageing, describing how and why basic discoveries in this simple organism have impacted our prospects for intervention in the ageing process. The authors cover a broad range of topics with regards to organismal and reproductive ageing including anatomical, physiological and biochemical changes, as well as genetic and environmental interventions that promote longevity and ameliorate age-related disease. Ageing is the single most important factor determining the onset of human disease in developed countries. With current worldwide demographic trends indicating that the number of individuals over the age of 65 will continue to rise, it is clear that an understanding of the processes that underpin ageing and age-related disease represents a key challenge in the biomedical sciences. In recent years there have been huge advances in our understanding of the ageing process and many of these have stemmed from genetic analysis of C. elegans. With no analogous book in this subject area this work will be of interest to a wide audience, ranging from academic researchers to the general public.

This volume presents a collection of protocols that describe methodologies to study thermogenic fat biology from various angles. This book is divided into 2 parts. Part 1 focuses on establishing in vitro culture systems. The chapters in this section introduce techniques on how to isolate, culture, and differentiate primary fat cells from both laboratory mice and humans. This part also presents flow cytometry methods to isolate various subpopulations of precursors within the stromal vascular fraction of the adipose tissue, which contains both preadipocytes and immune cells. Part 2 introduces multiple means to genetically manipulate and evaluate brown and beige fat in vivo. The chapters in this section explore methods on bioenergetics analyses both in vitro and in vivo. They also cover how to evaluate thermogenic fat contents and activity in humans, how to culture these cells though interdisciplinary approaches, and how to use thermogenic fat cell lines to carry out drug screens. 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. Thorough and cutting-edge, Thermogenic Fat: Methods and Protocols is a valuable resource for both experts and novices who are interested in learning assays and investigating brown and beige fat functions.

This detailed book explores the most current techniques to study systems and epithelial cell culture. Beginning with an overview, the volume then continues to detail systems that seek to mimic the three-dimensional organization, epithelial cells from different organs, gastrointestinal system, thyroid, salivary gland, ovary, mammary gland, and olfactory epithelial tissue. Cell culture is a fundamental technique in both medical research and drug discovery and two-dimensional (2D) culture has been the preferred method, due to the ease with which cell monolayers can be induced to proliferate on planar surfaces. The book propose several functional assay useful to test cell activities. Further, The past decades have witnessed significant efforts toward the development of three-dimensional (3D) cell cultures. Today, 3D cell cultures are emerging not only as a new tool in early drug discovery, but also as potential therapeutics to treat disease. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detail and key implementation advice that leads to excellent results in the lab.

This book presents basic concepts, methodologies and applications of biotechnology for the conservation and propagation of aromatic, medicinal and other economic plants. It caters to the needs and challenges of researchers in plant biology, biotechnology, the medical sciences, pharmaceutical biotechnology and pharmacology areas by providing an accessible and cost-effective practical approach to micro-propagation and conservation strategies for plant species. It also includes illustrations describing a complete documentation of the results and research into particular plant species conducted by the authors over the past 5 years. Plant Biotechnology has been a subject of academic interest for a considerable time. In recent years, it has also become a useful tool in agriculture and medicine, as well as a popular area of biological research. Current economic growth is globally projected in a highly positive manner, but the challenges many countries face with regard to food, feed, malnutrition, infectious diseases, the newly identified life-style diseases, and energy shortages, all of which are worsened by an ever-deteriorating environment, continue to pull the growth digits back. The common thread that connects all of the above challenges is biotechnology, which could provide many answers. Molecular biology and biotechnology have now become an integral part of tissue culture research. The tremendous impact generated by genetic engineering and consequently of transgenics now allows us to manipulate plant genomes at will. There has indeed been a rapid development in this area with major successes in both developed and developing countries. The book introduces several new and exciting areas to researchers who are unfamiliar with plant biotechnology and also serves as a review of ongoing research and future directions for scholars. The book highlights numerous methods for in vitro propagation and utilization of techniques in raising transgenics to help readers reproduce the experiments discussed.

This book brings together contributions from global experts who have helped to facilitate the exciting and rapid advances that are taking place in microbial metabolomics. The main application of this field is in clinical and veterinary microbiology, but there is a great potential to apply metabolomics to help to better understand complex biological systems that are dominated by multiple-species microbial populations exposed to changing growth and nutritional conditions. In particular, environmental (e.g., water, soil), food (e.g., microbial spoilage, food pathogens), and agricultural and industrial applications are seen as developing areas for microbial metabolomics. As such, the book includes contributions with clinical, environmental, and industrial perspectives.

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.