This volume brings together the skills and protocols of numerous laboratories that are at the heart of investigation into the biology of Tfh cells in both mice and humans. As a volume in the highly successful Methods in Molecular Biology series, chapters contain 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. Concise and easy-to-use, T follicular Helper Cells: Methods and Protocols provides scientist with techniques and protocols that have facilitated breakthroughs in Tfh biology and to present them in a way that will enable both new and experienced researchers to continue to move this exciting field forward.

Leading researchers are specially invited to provide a complete understanding of the key topics in these archetypal multidisciplinary fields. In a form immediately useful to scientists, this periodical aims to filter, highlight and review the latest developments in these rapidly advancing fields.

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.

This book describes modern biophysical techniques that enable us to understand and examine dynamic processes of infection at the molecular level. Cutting-edge research articles, laboratory protocols, case studies and up-to-date reviews cover topics such as single-molecule observation of DNA replication repair pathways in E. coli; evolution of drug resistance in bacteria; restriction enzymes as barriers to horizontal gene transfer in Staphylococcus aureus; infectious and bacterial pathogen biofilms; killing infectious pathogens through DNA damage; bacterial surfaces in host-pathogen interactions; bacterial gene regulation by riboswitches; transcription regulation in enterobacterial pathogens; the bacterial flagellar motor; initial surface colonization by bacteria; Salmonella Typhi host restrictions; as well as monitoring proton motive force in bacteria; microbial pathogens using digital holography; mathematical modelling of microbial pathogen motility; neutron reflectivity in studying bacterial membranes; force spectroscopy in studying infection and 4D multi-photon imaging to investigate immune responses. The focus is on the development and application of complex techniques and protocols at the interface of life sciences and physics, which increase the physiological relevance of biophysical investigations.

This book shares the latest research and practice-oriented findings in medical sciences with a wide audience. It addresses a range of contemporary issues, often unresolved or contentious, across various medical fields, including advances in the management of hemorrhagic brain stroke. It also discusses metastatic renal cell carcinoma - a global scourge with an extremely poor long-term survival prognosis, the course and sequelae of renal cell carcinoma, as well as advances in targeted molecular therapy with sunitinib, a receptor tyrosine kinase inhibitor. Further, it examines the molecular targeting of proliferative signaling of the epidermal growth factor receptor in the first-line treatment of patients with metastatic non-small-cell lung cancer. Other articles cover clearance of toxins in hemodialyzed patients; the search for diagnostic and therapeutic markers in the connective tissue disease scleroderma; obesity linked to inappropriate dietary habit; clinical problems related to the diagnosis of sensitization to fungi and its role in asthma; and reasons for the perilous trend of avoiding basic vaccinations in children. Lastly, the book explores the rapid developments in e-health technologies that increase access to health services, particularly for the elderly. The book is intended for clinical specialists, researchers, and all allied health professionals from various fields.

This volume is the result of an explosion of molecular-based research on Cilia, which began with the discovery of the universality of intraflagellar transport (IFT) and ciliary genomics/proteomics. The chapters in this book cover topics such as: high resolution imaging and functional characterization of sensory and primary cilia in mammalian cells and zebrafish, methods to study ciliary-mediated chemoresponse in Paramecium, and methods to study centrosomes and cilia in C. elegans and Drosophila. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Cilia: Methods and Protocols is broad and covers motile, sensory, and primary cells. It is a valuable resource to anyone interested in entering the field of ciliary biology using model organisms, including flagellate algae, ciliates, planaria, nematodes, insects, zebrafish, and mammalian cells.

This volume provides insight into the pivotal roles of stem cells, exosomes and other microvesicles in biofunction and molecular mechanisms and their therapeutic potential in translational nanomedicine. It further highlights evidence from recent studies as to how stem cell derived exosomes and microRNAs may restore and maintain tissue homeostasis, enable cells to recover critical cellular functions and begin repair regeneration. These early studies in animal models of aging also show evidence of improved immune, cardiovascular and cognitive functions as well as improved health span and life span. The use of exosomes from body fluids to define specific biomarkers for various tumors may also clear the path to patient-targeted treatments by developing exosome-derived microRNA based cancer therapeutics. It is essential reading for graduate students, research fellow and biomedical researchers in academia or the pharmaceutical or biotech industries.

This is the second 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.

Now poised to evaluate the potential in modulating Wnt signaling for therapeutic agendas in cancer, wound healing, and degenerative disease, this book collects protocols from an exciting area of study. The methodologies include using Wnt modulating chemicals in engineering tissues from induced pluripotent and embryonic stem cells, monitoring Wnt transcriptional responses in diverse tissues such as bone and skin, and using specific biochemical markers of Wnt signaling to either screen molecular libraries or evaluate novel reagents. These protocols also leverage unique experimental strengths from five different model organisms. 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 authoritative, Wnt Signaling: Methods and Protocols aims to support researchers in understanding Wnt signal transduction in any effort to improve outcomes in regenerative medicine and cancer management.

Understanding the dynamics of cell and tissue motion forms an essential step in understanding the dynamics of life and biological self-organization. Biological motion is one of the most obvious expressions of self-organization, as it requires autonomous creation and regulated action of forces leading to shape formation and translocation of cells and tissues. The topics of the book include intracellular motility and cytoplasma dynamics (e.g. cell division), single cell movement in varying extracellular media (e.g. chemotaxis or contact guidance), cell aggregation and cooperative motion (e.g. cellular swarms or slugs) and, finally, cell-cell interactions in developing tissues (e.g. embryogenesis or plant movement). The dynamics underlying biological motion are explained, on the one hand, by various methods of image processing and correlation analysis, and on the other hand by using physico-chemical theories, developing corresponding mathematical models and performing continuum field or stochastic simulations. Thus, the study is of an interdisciplinary character typically found in theoretical and mathematical biology. Its presentation is intended to reach a broad audience a " from theoretically interested bioscientists, physicians and biophysicists to applied mathematicians interested in the application of nonlinear dynamical systems and simulation algorithms. The most important feature of the book is that it considers possible synergetic mechanisms of interaction and cooperation on different microscopic levels: on the molecular level of cytoskeletal polymers, membrane proteins and extracellular matrix filaments, as well as on the level of cells and cellular tissues. New results concern the aspects of filament or cell alignment, various modes of force transduction and the formation of global stress fields. The latter aspect of mechanical cell-cell communication is emphasized in order to complement the much more well-studied phenomena of chemical, genetical or electrophysical communication.

This volume discusses methods for the study of multipotent and pluripotent stem cells of the hair follicle. The stem cells described are involved in both the growth of the hair follicle and its production of the hair shaft, as well as the growth of the hair follicle sensory nerve. Multipotent Stem Cells of the Hair Follicle: Methods and Protocols also explores very unexpected results such as that of the hair follicle-associated-pluripotent (HAP) stem cells, which not only have the capability for regenerating the hair follicle sensory nerve, but also can differentiate ex vivo and in vivo to multiple cell types not associated with the hair follicle-these include glial cells, motor neurons, and beating cardiac muscle cells. The potential for HAP stem cells for regenerative medicine is also discussed in detail. 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, Multipotent Stem Cells of the Hair Follicle: Methods and Protocols is a valuable resource for researchers interested in this field.

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.

Sox2: Biology and Role in Development and Disease offers a thorough discussion of the important role of Sox2 in cellular and developmental processes, aimed at facilitating greater understanding of how Sox2 functions across different disciplines. The book discusses the basic biology of Sox2 to help establish the critical foundational knowledge necessary for deeper molecular and functional analysis. The book also provides insight into how the Sox2 transcription factor plays a key role in pluripotency induction, maintenance, and development. Helpful as a tool to organize new research projects, the book assists with preparing lessons, seminars, and thesis or research papers, thereby circumventing the need to spend hours searching through journal databases. A single source for the basic biology of Sox2, Sox2: Biology and Its Role in Development and Disease provides information on networks, gene regulation, and regulatory function in a number of cell types and tissues types.

This detailed volume focuses on best practices and conditions for maintaining the most commonly used salamander species in the laboratory. Salamanders in Regeneration Research: Methods and Protocols guides readers through experimental manipulations in vivo and in vitro, respectively. With methods on targeting a wide variety of structures, ranging from the limb to the heart and to the brain, and methods for studying genetically modified organisms and tools for mining in the genomic databases. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Salamanders in Regeneration Research: Methods and Protocols provides a comprehensive collection of methods chapters.

This volume details experimental approaches used to investigate phagocytosis and phagosome maturation. Chapters present methods and protocols on quantifying uptake and phagosome maturation using biophysical and biochemical approaches, proteomics, microscopy, and flow cytometry. 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, Phagocytosis and Phagosomes: Methods and Protocols aims to be an important resource for both experts in the field and for those investigators delving into phagocytosis and phagosome maturation for the first time.

With a focus on food safety, this book highlights the importance of microbes in sustainable agriculture. Plants, sessile organisms that are considered as primary producers in the ecosystem and communicate with above- and below-ground communities that consist of microbes, insects, and other vertebrate and invertebrate animals, are subjected to various kinds of stress. Broadly speaking, these can be subdivided into abiotic and biotic stresses. Plants have evolved to develop elaborate mechanisms for coping with and adapting to the environmental stresses. Among other stresses, habitat-imposed biotic stress is one serious condition causing major problems for crop productivity. Most plants employ plant-growth-promoting microorganisms (PGPMs) to combat and protect themselves from stresses and also for better growth. PGPMs are bacteria associated with plant roots and they augment plant productivity and immunity. They are also defined as root-colonizing bacteria that have beneficial effects on plant growth and development. Remarkably, PGPMs including mycorrhizae, rhizobia, and rhizobacteria (Acinetobacter, Agrobacterium, Arthrobacter, Azospirillum, Bacillus, Bradyrhizobium, Frankia, Pseudomonas, Rhizobium, Serratia, Thiobacillus) form associations with plant roots and can promote plant growth by increasing plants' access to soil minerals and protecting them against pathogens. To combat the pathogens causing different diseases and other biotic stresses, PGPMs produce a higher level of resistance in addition to plants' indigenous immune systems in the form of induced systemic resistance (ISR). The ISR elicited by PGPMs has suppressed plant diseases caused by a range of pathogens in both the greenhouse and field. As such, the role of these microbes can no longer be ignored for sustainable agriculture. Today, PGPMs are also utilized in the form of bio-fertilizers to increase plant productivity. However, the use of PGPMs requires a precise understanding of the interactions between plants and microbes, between microbes and microbiota, and how biotic factors influence these relationships. Consequently, continued research is needed to develop new approaches to boost the efficiency of PGPMs and to understand the ecological, genetic and biochemical relationships in their habitat. The book focuses on recent research concerning interactions between PGPMs and plants under biotic stress. It addresses key concerns such as - 1. The response of benign microbes that benefit plants under biotic stress 2. The physiological changes incurred in plants under harsh conditions 3. The role of microbial determinants in promoting plant growth under biotic stress The book focuses on a range of aspects related to PGPMs such as their mode of action, priming of plant defence and plant growth in disease challenged crops, multifunctional bio-fertilizers, PGPM-mediated disease suppression, and the effect of PGPMs on secondary metabolites etc. The book will be a valuable asset to researchers and professionals working in the area of microbial-mediated support of plants under biotic stress.

The rapid expansion of the area of free radical biology in the last 25 years has occurred within a framework of assumptions and preconceived notions that has at times directed the course of this movement. The most dominant of these notions has been the view that free radical production is without exception a bad thing, and that the more efficient our elimination of these toxic substances, the better off we will be. The very important observation by Bernard Babior and colleagues in 1973 that activated phagocytes produce superoxide in order to kill micro organisms, served to illustrate that constructive roles are possible for free radicals. For many in the field, however, this merely underscored the deadly nature of oxygen-derived radicals, both from the microbe's point of view and from the host's as well. (Phagocyte-produced superoxide is responsible in part for the tissue injury manifested as inflammation. See Harris and Granger, Chapter 5, and Leff, Hybertson and Repine, Chapter 6.)
Mother Nature, however, has a penchant for being able to make a silk purse from a sow's ear. If one is dealt a bad hand, one must simply make the best of it. After two decades of focusing on the destructive side of free radicals, the last few years have begun to reveal a new and finer perspective on free radical metabolism - a role in regulation of cellular function (see Schulze-Osthoff and Baeuerle, Chapter 2). Evidence from a number of sources suggests that an increase in the oxidative status of cell encourages that cell to grow and divide. Increasing the expression of mangnese superoxide dismutase can suppress the malignant phenotype of melanon cells (see Oberley and Oberley, Chapter 3). Oxidative stress beyond a certain poitosis (from the Greek, literally "to fall apart"). Is this suicide response an evolutionary fail-safe device to curtail tumorogenesis? Does oxidative stress-induced apoptosis account for the loss of immune cells in AIDS (see Flores and McCor Chapter 4)?
This volume attempts to present the spectrum of roles, both good and bad played by active oxygen species as understood at this point in the evolution of this field of free radical biology.

Plant Circadian Networks: Methods and Protocols provides a collection of protocols to investigate clock-controlled parameters including transcript and small RNA levels, promoter activity using luciferase reporters, protein levels and posttranslational modification, protein-protein interaction, in vivo DNA-protein interaction and RNA-protein interaction, cellular redox state, Ca2+ levels, and innate immune responses. Furthermore, the use of bioinformatics resources is described to evaluate high throughput data sets and to integrate the data into an overarching picture of circadian networks in the cell. Additional chapters focus on seasonal processes like flowering time control, and techniques on trees, moss and algae. 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, Plant Circadian Networks: Methods and Protocols is designed not only for the chronobiology community dealing with circadian biology but also for the plant community in general.

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 are presented here in volume 1. Volume 2 will focus 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 book presents regenerative strategies for the treatment of knee joint disabilities. The book is composed of four main sections totaling 19 chapters which review the current knowledge on the clinical management and preclinical regenerative strategies. It examines the role of different natural-based biomaterials as scaffolds and implants for addressing different tissue lesions in the knee joint. Section one provides an updated and comprehensive discussion on articular cartilage tissue regeneration. Section two focuses on the important contributions for bone and osteochondral tissue engineering. Section three overview the recent advances on meniscus repair/regeneration strategies. Finally, section four further discusses the current strategies for treatment of ligament lesions. Each chapter is prepared by world know expert on their fields, so we do firmly believe that the proposed book will be a reference in the area of biomaterials for regenerative medicine.

This detailed volume provides a single, valuable reference source for methods that definitively identify and accurately quantify apoptosis. The book begins with common methods utilized to detect and quantitate apoptosis, as well as apoptosis signaling pathways in toxicological and other related research. It continues with multi-parametric and phased apoptosis assays for detecting early and late apoptosis or distinguishing apoptosis from necrosis and autophagy. Subsequent chapters focus on recent advances in real time and high-throughput assays that detect and quantitate apoptosis and apoptosis signaling pathways. Final chapters focus on recent developments in preclinical anticancer therapeutics targeting apoptosis. Written for the Methods in Pharmacology and Toxicology series, chapters feature step-by-step descriptions of the methodologies, as well as expert tips and implementation advice. Vital and authoritative, Apoptosis Methods in Toxicology serves novice scientists as well as experts, utilizing a range of instruments from common laboratory equipment to high-end expensive and automated machinery capable of performing real time apoptotic measurements.

This volume presents comprehensive laboratory protocols that have been used to generate Th9 cells, both in vitro and in vivo. The techniques described in Th9 Cells: Methods and Protocols study the role of Th9 cells in different inflammatory disease models, including allergic inflammation model, parasite model, tumor model, and EAE and IBD model. 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 comprehensive, Th9 Cells: Methods and Protocols is a valuable resource for scientists in this field and will provide them with techniques to generate Th9 cells for specific downstream events.

This volume looks at the liver's epithelial cells-hepatocytes and cholangiocytes-and their progenitors. This book is divided into five parts: isolation of progenitor cells; characterization of liver progenitors in vivo; generation of hepatocytes, cholangiocytes, and their progenitors; reconstitution of liver tissue structures; and liver injury models. The chapters in this book cover topics such as expansion of bipotential liver stem/progenitor cells (LPCs) from fetal and neonatal liver; identifying progenitor cells involved in liver regeneration in vivo; methods for generating hepatocytes and cholangiocytes from multiple cellular sources; 3D tissue structures ex vivo; and resolving hepatic fibrosis by bone marrow transplantation. 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, Hepatic Stem Cells: Methods and Protocols is a valuable resource to help researchers understand the current theories used to study hepatic stem/progenitor cells, and aid them in performing experiments related to liver biology and pathophysiology.

This book explores the regenerative properties of fetal stem cells, from feto-maternal cell traffic through perinatal stem cells, with a discussion of key topics including stem cell banking, drug screening, in utero stem cell transplantation and ethical considerations. The expertly authored chapters also delve into embryonic, amniotic membrane, and umbilical cord blood stem cells; fetal development models; fetal cell reprogramming; culture methods; disease models; perinatal gene therapy, and more. These chapters are grouped into four sections, each discussing a separate prenatal stem cell population and providing fascinating historical contexts for our knowledge of these systems. Featuring a foreword written by the renowned Dr. Joseph Vacanti of the Harvard Stem Cell Institute, Fetal Stem Cells in Regenerative Medicine: Principles and Translational Strategies is a welcome and timely contribution to the Stem Cell Biology and Regenerative Medicine series. It is essential reading for scientists and researchers, clinicians and residents, and advanced students involved in stem cells, regenerative medicine, tissue engineering, and related disciplines such as embryology.

This text highlights the endogenous regenerative potential of the central nervous system in neonates and juveniles and discusses possible ways it might be manipulated for medical purposes. The first section provides a descriptive summary of the salient steps of human brain development with a discussion of comparisons with other mammalian brains. It also provides a historical perspective on our understanding of ongoing brain development throughout the lifespan and serve to introduce the concept of brain plasticity following injury. The second part is devoted to the endogenous reparative potential of the brain, including its limitations, and articles focusing on defined pathologies (e.g. anoxia/hypoxia, epilepsy, traumatic brain injury and stress) in animal models and in humans pinpoint eventual ways these pathologies might be manipulated. The third and final focuses on the "dark side" of stem cells for brain repair or of the manipulation of spontaneous adaptive events after injury (e.g. genomic instability, sensitization to cancerous transformation and defective neural networks).