This fully updated edition explores methods involving sphingosine-1-phosphate (S1P), a bioactive lysophospholipid which has become the focus of much research interest as it has widespread developmental and physio-pathological actions, controlling events within the nervous, reproductive, gastrointestinal, vascular, respiratory, and immune systems, in addition to having a prominent role in cancer, early mammalian embryogenesis, and stem cells. Here, worldwide experts in the S1P field describe in-depth techniques in an array of cell types and with various physiological applications, showcasing the important effects of S1P in development and in physiopathology. As a volume in the Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the relevant materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Authoritative and timely, Sphingosine-1-Phosphate: Methods and Protocols, Second Edition is another key resource for scientists working in this fast-moving and dynamic field.

This book highlights recent advances in and diverse techniques for exploring the plasma membrane's structure and function. It starts with two chapters reviewing the history of membrane research and listing recent advances regarding membrane structure, such as the semi-mosaic model for red blood cell membranes and the protein layer-lipid-protein island model for nucleated tissue cell membranes. It subsequently focuses on the localization and interactions of membrane components, dynamic processes of membrane transport and transmembrane signal transduction. Classic and cutting-edge techniques (e.g. high-resolution atomic force microscopy and super-resolution fluorescence microscopy) used in biophysics and chemistry are presented in a very comprehensive manner, making them useful and accessible to both researchers in the field and novices studying cell membranes. This book provides readers a deeper understanding of the plasma membrane's organization at the single molecule level and opens a new way to reveal the relationship between the membrane's structure and functions, making it essential reading for researchers in various fields.

This book highlights the role of the Translationally Controlled Tumor Protein (TCTP) in cell signaling, cell fate and the resulting connection to disease development. It begins by discussing the structure/function of TCTP, before exploring its role in different species ranging from plants to Drosophila and covering fields such as development, the cytoskeleton, cell division, DNA fragility and apoptosis. In turn, the book's final section is devoted to the role of TCTP in disease, namely asthma and diverse cancers, and ultimately as a target for the treatment of malignancies. What is the common denominator between all these processes and why is TCTP necessary in order for them to occur, even in the worst case such as cancer? The book seeks to provide meaningful answers to this and other key questions. Presenting a broad and revealing view on the topic, it offers an informative guide for scientists and students alike.

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 new volume reviews current progress on different approaches of in vivo reprogramming technology. Leaders in the field discuss how in vivo cell lineage reprogramming can be used for tissue repair and regeneration in different organs, including brain, spinal cord, pancreas, liver and heart. Recent studies on in vivo cell reprogramming towards pluripotency are reviewed; examples are given to show its potential in regenerative medicine. In each chapter, the regenerative potential of different in vivo reprogramming approaches is discussed in detail. More specifically, how different tissue failures or damages can be treated with this technology is explained. Examples from various animal models are given and the regenerative potential of in vivo reprogramming is compared to that of cell transplantation studies. The last chapter discusses current challenges of these preclinical studies and gives suggestions in order to improve the current strategies. Future directions are indicated for the transition of in vivo reprogramming technology to clinical settings. This is among the first books in the literature which specifically focuses on the in vivo reprogramming technology in regenerative medicine and these chapters collectively cover one of the most important and exciting topics of regenerative medicine.

This book provides a resource of current understandings about various aspects of the biology of spermatogonia in mammals. Considering that covering the entire gamut of all things spermatogonia is a difficult task, specific topics were selected to provide foundational information that will be useful for seasoned researchers in the field of germ cell biology as well as investigators entering the area. Looking to the future, the editors predict that the foundational information provided in this book -- combined with the advent of new tools and budding interests in use of non-rodent mammalian models -- will produce another major advance in knowledge regarding the biology of spermatogonia over the next decade. In particular, we anticipate that the core molecular machinery driving different spermatogonial states in most, if not all, mammals will be described fully, the extrinsic signals emanating from somatic support cell populations to influence spermatogonial functions will become fully known, and the capacity to derive long-term cultures of SSCs and transplant the population to regenerate spermatogenesis and fertility will become a reality for higher order mammals.

The Chemical Dialogue Between Plants and Beneficial Microorganisms provides foundational insights on plant beneficial microorganisms and their impact on the health and productivity of plants. Providing in-depth and recent updates about unexplored aspects of plant microbes interactions, the book includes the biological repertoire of arbuscular mycorrhizal association, molecular architecture of Rhizobium-plant symbiosis, and endophytes in transcriptional plasticity during host colonization by endophytes. The book also includes details about the mechanism of different plant beneficial microorganisms, how these differ, and their cross signaling. This book will be an important reference for researchers working on different plant beneficial microorganisms and their molecular arsenal.

This volume introduces different concepts and methods of detecting RNA in biological material in a variety of model systems. The chapters in this book discuss methods that will answer numerous biological questions that arise in the study of RNAs. Some of the topics covered in this book are single mRNA molecule detection in embryos and neurons; detection of mRNA and associated molecules by ISH-IEM on frozen sections; optimizing molecular beacons for intracellular analysis of RNA; imaging translation dynamics of single mRNA molecules in live cells; preparation of high-throughput sequencing libraries; and capturing RNA binding proteins in embryos and in cell-culture. 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, RNA Detection: Methods and Protocols is a valuable resource for novel and experiences scientist in the expanding field of RNAs.

This book provides an essential overview of existing state-of-the-art quantitative imaging methodologies and protocols (intensity-based ratiometric and FLIM/ PLIM). A variety of applications are covered, including multi-parametric quantitative imaging in intestinal organoid culture, autofluorescence imaging in cancer and stem cell biology, Ca2+ imaging in neural ex vivo tissue models, as well as multi-parametric imaging of pH and viscosity in cancer biology. The current state-of-the-art of 3D tissue models and their compatibility with live cell imaging is also covered. This is an ideal book for specialists working in tissue engineering and designing novel biomaterial.

This second edition details some of the recent trends in HCS/HCA/HCI. New and updated chapters guide readers through methods that utilize reagents and kits that have been developed to measure cells and subpopulation classifications, control measures, overviews of the data handling issues associated with HCS/HCA/HCI, and methods to implement more complex phenotypic 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 practical, High Content Screening and Analysis-The Ideal Format for Phenotypic Screening: A Powerful Approach to Systems Cell Biology and Drug Discovery, Second Edition aims to ensure successful results in the further study of this vital field.

The main goal of this book is to form a high-quality platform in which well-known and emerging pioneering basic, translational and clinical scientists can present their latest, exciting findings in the studies of redox signaling in the pulmonary vasculature. Content from outstanding investigators with unique expertise and skills of molecular and cell biology, biochemistry, physiology, pharmacology, biophysics, biotechnology and medicine will update our current out-of-date concepts with new knowledge. Rapidly increasing scientific studies have gathered a large volume of novel and important information on redox signaling in healthy and diseased pulmonary vasculature. This volume covers the need for a cohesive book to display state-of-the-art advances in the field. The second major aim of this book is to help direct future research. Redox signaling is a major molecular process involved in almost every physiologic cellular response in the pulmonary vasculature including energy metabolism, host defense, gene expression, contraction, proliferation, and migration. Aberrancy in this important signaling pathway leads to a critical role in the development of nearly all pulmonary diseases, such as pulmonary hypertension, cor pulmonale, pulmonary edema, and vasculitis, among others.

This book presents a detailed overview of the design, formatting, application, and development of microfluidic chips in the context of cell biology research, enumerating each element involved in microfluidics-based cell analysis, discussing its history, status quo, and future prospects, It also offers an extensive review of the research completed in the past decade, including numerous color figures. The individual chapters are based on the respective authors' studies and experiences, providing tips from the frontline to help researchers overcome bottlenecks in their own work. It highlights a number of cutting-edge techniques, such as 3D cell culture, microfluidic droplet technique, and microfluidic chip-mass spectrometry interfaces, offering a first-hand impression of the latest trends in the field and suggesting new research directions. Serving as both an elementary introduction and advanced guidebook, the book interests and inspires scholars and students who are currently studying microfluidics-based cell analysis methods as well as those who wish to do so.

This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.

This detailed volume explores methods and protocols that aim to increase our understanding of how cells enter a quiescent state during homeostasis and how cells exit quiescence and re-enter differentiating cell divisions to restore damaged tissues, essential for developing new approaches in regenerative medicine in the future. The chapters in this book were designed to address cellular quiescence in prokaryote and eukaryote organisms, detection of quiescence (Hoechst/pyronin Y, FUCCI, CFSE, BrdU, H2B-GFP, CyTOF), quiescence in stem cells (skin, intestinal, neuronal, hematopoietic), genomic regulation (gene expression, transcription factors, lncRNA, RNA methylation), as well as analysis of the heterogeneity of quiescence by computer modeling. 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, Cellular Quiescence: Methods and Protocols offers a broad view of basic and cutting-edge technology to inspire research in this emerging field of cell biology.

This book focuses on the modeling and mathematical analysis of stochastic dynamical systems along with their simulations. The collected chapters will review fundamental and current topics and approaches to dynamical systems in cellular biology. This text aims to develop improved mathematical and computational methods with which to study biological processes. At the scale of a single cell, stochasticity becomes important due to low copy numbers of biological molecules, such as mRNA and proteins that take part in biochemical reactions driving cellular processes. When trying to describe such biological processes, the traditional deterministic models are often inadequate, precisely because of these low copy numbers. This book presents stochastic models, which are necessary to account for small particle numbers and extrinsic noise sources. The complexity of these models depend upon whether the biochemical reactions are diffusion-limited or reaction-limited. In the former case, one needs to adopt the framework of stochastic reaction-diffusion models, while in the latter, one can describe the processes by adopting the framework of Markov jump processes and stochastic differential equations. Stochastic Processes, Multiscale Modeling, and Numerical Methods for Computational Cellular Biology will appeal to graduate students and researchers in the fields of applied mathematics, biophysics, and cellular biology.

This detailed volume gathers a comprehensive collection of methods, protocols, and procedures used for the identification, characterization, and selection of cancer stem cells. These include surface marker expression, side population, spheres formation, ALDH activity, and numerous others. 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 expert tips and troubleshooting advice to avoid potential pitfalls. Comprehensive and practical, Cancer Stem Cells: Methods and Protocols provides researchers with the techniques they will need in order to pursue vital research into this complex field of study.

This thesis describes novel substrate embedded physical sensors that can be used to monitor different types of cell-based assays non-invasively and label-free. The sensors described provide integrative information of the cells under study with an adaptable time resolution (ranging from milliseconds to days). This information about the dynamic cell response to chemical, physical or biological stimuli defines a new paradigm in fundamental biomedical research. The author, Maximilian Oberleitner, describes approaches in which the cells are directly grown on different sensor surfaces (gold-film electrodes, shear wave resonators or dye-doped polymer films). This approach, with the reacting cells in particularly close proximity and contact with the sensor surface, is key to a remarkable sensitivity, opening the way for a variety of new applications. This thesis not only introduces the fundamentals of each approach, but it also describes in great detail the design principles and elucidates the boundary conditions of the new sensors.

This book addresses all the major mechanisms by which endophytes are thought to impact plant growth and health. A unique aspect of this publication is that it is multidisciplinary, covering plant microbiology, plant physiology, fungal and bacterial endophytes, plant biochemistry, and genomics. Just as research on the mammalian microbiome has demonstrated its importance for overall health of the host, the plant microbiota is essential for plant health in natural environments. Endophytes, the microorganisms living fully within plants, can provide a multitude of benefits to the host including N-fixation, P solubilization, increased photosynthetic efficiency and water use efficiency, stress tolerance, pathogen resistance, and overall increased growth and health. A variety of culturable endophytes have been isolated and shown to be mutualistic symbionts with a broad range of plant species. These studies point to the functional importance of the microbiota of plants and suggest the potential for tailoring plant microbiota for improved vigor and yields with reduced inputs. This review covers the major benefits of microbial endophytes to plants and discusses the implications of using symbiosis as an alternative to chemical inputs for agriculture, forestry, and bioenergy.

This laboratory manual covers the study of chromosomes in plants, animal and human systems, dealing with the protocols and principles involved. It caters to the requirements of scientists working laboratories, presenting details of the operational mechanism for use at the chromosome level.

This second edition of the book on Store-operated Ca2+ Entry Pathways has been updated with the newest discoveries that emerged in the field within the last five years. The crystal structure of the Ca2+ signaling core complex is described which adds to a new understanding of the molecular interactions involved. Each chapter has been revised and extended. The book retains its interdisciplinary approach and supplies biochemists, cell biologists and biophysicists as well as clinicians in immunology, neurology and cardiology with valuable information on Ca2+ signaling mechanisms, functions, dysfunctions and their consequences.

This volume provides a collection of protocols for the common experimental approaches used in the in the burgeoning field of c-di-GMP-dependent signaling. The chapters, divided into eight major parts, guide readers through methods on synthesis, detection, quantitation, modulation of the levels of c-di-GMP present in cells, procedures to detect and evaluate the interaction of c-di-GMP, and up and coming approaches focusing on the inhibition of c-di-GMP signaling.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, c-di-GMP Signaling: Methods and Protocols aims to inspire researchers to try new approaches.

This multi-chapter book focuses on one of the hottest topics in ageing research - the role of hormones in health and longevity, offering a comprehensive and up-to-date overview of their mechanistic roles in health, ageing and longevity. Hormones are an excellent system of communication between cells and tissues within an organism, and they coordinate a wide range of processes in biological systems, including neuroendocrine and immunological controls. The book offers insights into the latest significant advances in our understanding of the mechanisms of hormonal signaling that control a variety of processes involved in development and ageing. It is divided into four parts: Part I includes a review of the hundred-year history of hormones by the illustrious hormone biochemist Dr. J.R. Tata. Part II presents various chapters on the hormones involved in growth, stress and metabolism, while Part III addresses the hormones controlling cognition and rhythms in ageing processes. Lastly, Part IV discusses the hormones affecting reproduction, immunity and life span. It also explores the use of hormones as pharmaceuticals to maintain health in the elderly. It is a valuable resource for those working in the area of hormone signaling in general and in the field of ageing research in particular.

This book provides an overview of new mathematical models, computational simulations and experimental tests in the field of biomedical technology, and covers a wide range of current research and challenges. The first part focuses on the virtual environment used to study biological systems at different scales and under multiphysics conditions. In turn, the second part is devoted to modeling and computational approaches in the field of cardiovascular medicine, e.g. simulation of turbulence in cardiovascular flow, modeling of artificial textile-reinforced heart valves, and new strategies for reducing the computational cost in the fluid-structure interaction modeling of hemodynamics. The book's last three parts address experimental observations, numerical tests, computational simulations, and multiscale modeling approaches to dentistry, orthopedics and otology. Written by leading experts, the book reflects the remarkable advances that have been made in the field of medicine, the life sciences, engineering and computational mechanics over the past decade, and summarizes essential tools and methods (such as virtual prototyping of medical devices, advances in medical imaging, high-performance computing and new experimental test devices) to enhance medical decision-making processes and refine implant design. The contents build upon the International Conference on Biomedical Technology 2015 (ICTB 2015), the second ECCOMAS thematic conference on Biomedical Engineering, held in Hannover, Germany in October 2015.

This invaluable resource discusses the saftey, ethics, and regulations of developing stem cell clinical applications. Each chapter is contributed by a preeminent scientist in the field and covers such topics as clinical safety of stem cell gene therapy, the patentability of hESC technologies, international guidelines, challenges to international stem cell clinical trials, worldwide regulations including in emerging markets like China and Taiwan. Saftey, Ethics, and Regulations 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 examines established methods and protocols to isolate and characterize extracellular vesicles (EVs) and their composition, among other techniques including purification, imaging, biofluid-specific and cell-specific isolation and downstream genomic and proteomic profiling. The international group of expert scientists who have contributed to this collection provide a variety of different techniques related to the growing assortment of EV applications, given that at times using only one technique or two is insufficient to address the question at hand. 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, Extracellular Vesicles: Methods and Protocols serves as an ideal guide for researchers seeking to expand our knowledge of EV functions and applications.