Physiological, pharmacological and molecular biological data generated over the past three decades have demonstrated the existence of two major families of extracellular receptors, the P1, a family of four G-protein coupled receptors and the P2, a family of at least 12 receptors responsive to purine (ATP, ADP) and pyrimidine (UTP) nucleotides through which adenosine and ATP can function as extracellular messengers. The present two-part volume represents an integrated compendium of invited chapters by leading researchers in the area focusing on advances in the understanding of purinergic and pyrimidinergic signaling systems, their role(s) in tissue function and pathophysiology and advances in developing potential new medications based on the modulation of P1 and P2 receptor signaling processes. The volumes will thus provide the reader with a topical, comprehensive and integrated overview of this important area.

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 volume covers the current knowledge base on the role of signaling and environmental pathways that control the normal development of germline stem cells, meiotic progression of oocytes, events of oocyte maturation and fertilization, and the birth of an embryo. Germ cells are uniquely poised to sustain life across generations through the fusion of oocyte and sperm. Because of the central importance of germ cells to life, much work has been dedicated to obtaining a clear understanding of the molecular and signaling events that control their formation and maintenance. Germ cells are set aside from somatic cells in the embryo and go through specialized meiotic cell cycles as the animal matures. These cell cycles are interspersed with long periods of arrest. In human females, meiosis I is initiated in the fetus. At birth, oocytes are arrested in meiosis I; after puberty, every month an oocyte initiates meiosis II - ovulation. Upon sperm availability these cells are fertilized, generate an embryo, and the cycle-of-life continues. During meiotic I progression and arrest, the fitness of oocytes and their progeny are likely influenced by environmental cues and signaling pathways. A lot of recent work has focused on understanding the mechanisms that regulate oocyte fitness and quality in humans and vertebrates. Much of our understanding on the events of meiosis I and germline stem cell populations comes from work in invertebrates, wherein the germline stem cells produce oocytes continuously through adult development. In both inverbrates and vertebrates nutritional and signaling pathways control the regulation of stem cells in such a manner so as to couple production of gametes with the nutritional availability. Additionally, mature oocytes arrest both in meiosis I and meiosis II, and signaling and nutritional pathways have been shown to regulate their formation, and maintenance, such that despite long periods of arrest, the oocyte quality is assured and errors in chromosome segregation and varied cytoplasmic events are minimal.

In Epiblast Stem Cells: Methods and Protocols, expect researchers in the field provide a detailed collection of techniques and protocols useful to the study of the biology of the pluripotent epiblast. These include methods and techniques used to study epiblast development in different amniotes. This collection brings together contributions from the fields of embryology, stem cell biology and developmental biology together, providing a single volume with detailed procedures for the isolation and culture of epiblasts at different stages of development, and techniques for the study of differentiation into specific lineages. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, a complete list of the necessary materials and reagents, detailed laboratory protocols, and extensive notes providing suggestions on troubleshooting and how to overcome common difficulties. Comprehensive and cutting-edge, Epiblast Stem Cells: Methods and Protocols serves as a resource to individuals interested in studying the biology of pluripotent cells.

Animal cell technology is a growing discipline of cell biology which aims not only to understand structures, functions and behaviors of differentiated animal cells but also to ascertain their abilities to be used for industrial and medical purposes. The goal of animal cell technology includes accomplishments of clonal expansion of differentiated cells with useful ability, optimization of their culture conditions, modulation of their ability for production of medically and pharmaceutically important proteins, and the application of animal cells to gene therapy and artificial organs. This Volume gives the readers a complete review of the present state of the art in Japan. The Proceedings will be useful for cell biologists, biochemists, molecular biologists, immunologists, biochemical engineers and other disciplines related to animal cell culture, working either in academic environments or in industries of biotechnology and pharmacy.

Historically, the major emphasis on the study of purinergic systems has been predominantly in the areas of physiology and gross pharmacology. The last decade has seen an exponential in- crease in the number of publications related to the role of both adenosine and A TP in mammalian tissue function, a level of interest that has evolved from a more molecular focus on the identity of adenosine and A TP receptor subtypes and the search for selective ligands and development of radioligand binding assays by Fred Bruns and colleagues (especially that for A receptors) that played z a highly significant role in advancing research in the area. In the 60 years since adenosine was first shown to be a potent hypotensive agent, a considerable investment has been made by several pharmaceutical companies-including Abbott, Byk Gulden, Takeda, Warner-LambertlParke Davis, Boehringer Mann- heim, Boehringer Ingelheim, Nelson/Whitby Research and CffiA- Geigy-as well as John Daly's laboratory at the National Institutes of Health, to design new adenosine receptor ligands, and both agon- ists and antagonists with the aim of developing new therapeutic entiities. Numerous research tools have derived from these efforts including: 2-chloroadenosine, R-PIA (~-phenylisopropyladeno- sine; NECA (5' N-ethylcarboxamidoadenosine); CV1808; CI936; PD 125,944; ~-benzyladenosine; PACPX; CPX; CPT; XAC; CGS 15943 and CGS 21680. Yet in the realm of therapeutics it was only in 1989 that adenosine itself was approved for human use in the treatment of supraventricular arrythmias.

Endocytosis and vesicular trafficking determine the landscape of the cell's exterior, namely the density of surface molecules, such as receptors for growth factors and cytokines, adhesion molecules like integrins and cadherins, and a plethora of nutrient carriers. Hence, endocytosis is involved in signal transduction, cell adhesion and migration, as well as metabolism. To exploit these fundamental processes, malignancies subtly and multiply manipulate the endocytosis and the subsequent trafficking of protein cargoes. This is achieved by simultaneously altering the cytoskeleton, vesicle budding, cargo sorting and intracellular degradation. By highlighting the underlying molecular processes and concentrating on specific examples, this book reviews the recent emergence of derailed endocytosis and vesicular trafficking as a landmark of cancer. In-depth understanding of this common feature of tumors might lead the way to drug-induced strategies, able to rectify intracellular trafficking in cancer.

Cheryl S. Watson University o/Texas Medical Branch Cellular steroid action has been thoroughly studied in the nuclear compartment. However, nuclear steroid receptor mechanisms have been unable to explain some of the rapid activities of steroids, partiCUlarly those which occur in a time frame of seconds to minutes reviewed in (1;2)]. Based on these and other considerations, an alternative membrane-associated receptor form was long ago proposed to exist (3). Others interpret the location of the steroid receptors mediating these rapid effects as peri membrane or cytoplasmic. New experimental tools have been brought to bear on the topic of receptors for steroids which mediate non-genomic actions, and thus investigative activity and focus regarding this type of steroid receptor has recently increased significantly. However, there may be multiple answers to the question "how do steroids mediate rapid nongenomic effects?" Steroid actions initiated at the cell membrane can impinge on important phases in the lifespan of a cell: proliferation, migration, differentiation, and release of hormones or neurotransmitters functioning as signals to other cells."

Two of the more fascinating biological phenomena that have been d- covered in recent years are RNA editing and RNA interference. Each of these processes has been found in a cross-section of biological systems, including mammals, viruses, plants, and a range of model organisms (C. elegans,Dro- phila, and various lower eukaryotes). RNA editing, which results in an RNA product different from that predicted by the genome, occurs through a variety of mechanisms. Alterations can occur at either the base level, in which one base is changed to another (substitutional editing/base modification), or via the addition and/or deletion of nucleotides relative to the original template (insertion/deletion editing). RNA interference (RNAi) involves the specific degradation of targeted mRNAs. Although RNA interference, editing, and modification use different enzymes and mechanisms, the targets of each of these reactions are often specified by RNA molecules. Indeed, the discovery of guide RNAs (gRNAs) that direct nucleotide insertion and deletion in trypa- some mitochondria set the precedent for subsequent discoveries of the small nuclear RNAs (snoRNAs) that target pseudouridylylation and methylation of stable RNAs and the small double-stranded RNA fragments (siRNAs) that mediate RNAi. Other small RNAs are known to mediate translational regu- tion during development (small temporal RNAs [stRNAs]) and mRNA stab- ity (microRNAs [miRNAs]), and the recent identification of more than a hundred small "noncoding" RNAs has led to the realization that they may represent only the proverbial "tip of the iceberg.

Nuclear Import and Export in Plants and Animals provides insight into the remarkable mechanisms of nuclear import and export. This book covers a range of topics from the nuclear pore structure, to nuclear import and export of macromolecules in plant and animal cells. In addition, the book covers the special cases of nuclear import of Agrobacterium T-DNA during plant genetic transformation, nuclear import and export of animal viruses, and nuclear intake of foreign DNA. A chapter on research methods to study nuclear transport concludes the book.

This volume covers a range of methods used in plant cytogenetics, beginning with basic analysis of chromosomes and visualizing gene locations, to manipulating and dissecting chromosomes, and then focusing on less understood features of chromosomes such as recombination initiation sites and epigenomic marks. The methods described in Plant Cytogenetics: Methods and Protocols build on each other and provide, those new to the field, with a comprehensive platform to support their research endeavours, while also introducing advanced techniques to experienced researchers. 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, Plant Cytogenetics: Methods and Protocols, is a valuable resource for anyone who is interested in the diverse and wonderfully complex field of cytogenetics.

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.

At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. It is written in the highly successful Methods in Molecular Biology trademark], 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 up-to-date, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.

This is the first broad treatment of carbohydrate chemistry in many years, and presents the structures, reactions, modifications, and properties of carbohydrates. Woven throughout the text are discussions of biological properties of carbohydrates, their industrial applications, and the history of the field of carbohydrate chemistry. Written for students as well as practising scientists, this textbook and handy reference will be of interest to a wide range of disciplines: biochemistry, chemistry, food and nutrition, microbiology, pharmacology, and medicine.

There has been a dramatic increase in knowledge of tight junctions in the past decade. The molecular structure of tight junctions, cellular functions and the pathophysiological roles of tight junctions are becoming clear. Of the most important functions, the role of the cellular structure in cancer spread and drug delivery are increasingly realised. It is now clear that there are fundamental changes to tight junctions during the process of cancer development. Tight junctions are also critical to the metastatic process of cancer cells. The cellular structure is also crucial in drug therapies, namely, the permeability and bioavailability of the drugs, penetration of barriers such as the blood brain barrier. This current volume aims to summarise the current knowledge of tight junctions, their role in cancer and cancer metastasis and is of interest to scientists and clinicians.

This book is an excellent, up-to-date reference on a relatively young area of research in which virology, cellular biology and molecular pathogenesis govern the principles of coinvestigation. Thus, the book will be of great interest to virologists, molecular immunologists and biologists, and biochemists but also to clinical pharmacologists in the long-term search for new antiviral agents. Ulrich Desselberger, Gif-sur-Yvette/Cambridge. Infection of a naive (non-immune) host with a virus elicits an immediate response which results in a cascade of changes in the host, including an interferon response (innate immunity). The outcome of this interaction is influenced by the genes of the virus as well as the genes of the host. Interestingly, different viruses do it in different ways. Not only is there a plethora of mechanisms used by the invading organisms, but the host has also evolved a great variety of redundant and robust countermeasures. This interplay of host and virus represents one of the most significant frontiers in biology today. A clearer understanding of the mechanisms involved will arm us with better strategies to deal with viruses, including emerging pathogens and potential bioterrorism agents. This book is sure to benefit students, scientists, and physicians working in the areas of virology, immunology, microbiology, and infectious diseases. Pharmaceutical industry professionals will also find interest in this illuminating look into virus/host interactions.

Intriguing new findings on how genes and environments work together through different stages of life take the spotlight in this significant collection. Studies from infancy to late adulthood show both forces as shaping individuals' relationships within family and non-family contexts, and examine how these relationships, in turn, continue to shape the individual. Transitional periods, in which individuals become more autonomous and relationships and personal identities become more complicated, receive special emphasis. In addition, chapters shed light on the extent to which the quantity and quality of genetic and environmental influence may shift across and even within life stages. Included in the coverage: Gene-environment interplay in parenting young children. The sibling relationship as a source of shared environment. Gene-environment transactions in childhood and adolescent problematic peer relationships. Toward a developmentally sensitive and genetically informed perspective on popularity. Spouse, parent, and co-worker: roles and relationships in adulthood. The family system as a unit of clinical care: the role of genetic systems. Behavioral geneticists, clinical psychologists, and family therapists will find in Gene-Environment Interplay in Interpersonal Relationships across the Lifespan a window into current thinking on the subject, new perspectives for understanding clients and cases, and ideas for further study.

As with other areas of biological research, the progress that has been made over the past 25 years in the field of cell adhesion is impressive. In the late 1970s, the searchfor specific cell surface receptors for adhesion processes was initiated - ing mainly biochemical and immunological approaches. Since then, theint- duction of novel methods of cellular and molecular biology and powerful te- niques for manipulatinggene expression in transgenic and knock-out mice have greatlyadvanced the field. Not only dowe now know the precise molecular structure of many of thecell adhesion receptors that were postulated to exist in the early days, but we have a clear picture of their function, and evidence of their involvement in signal transduction. We have also found clues to the role of cell adhesion in normal embryonal development and adult physiology, and we have evidence that disturbance in cell adhesion can cause disease. In this volume, our goal was to provide an overview of the main topics of c- rent cell adhesion research, including structural analyses of cell adhesion mo- cules and studies of their functionalrole in vitroand in vivo. We have focussed mainly on the four major families of cell-adhesion receptors, i. e. the cadherins, the integrins, the Ig superfamily and the selectin-based adhesion system. The chapters by Perez and Nelson and Choi and Weis describethe structuralbasis of cadherin function, focussing on the extracellular domain of cadherins and thecytoplasmic tail interactions with catenins, respectively.

Examining stem cell biology from a philosophy of science perspective, this book clarifies the field's central concept, the stem cell, as well as its aims, methods, models, explanations and evidential challenges. The first chapters discuss what stem cells are, how experiments identify them, and why these two issues cannot be completely separated. The basic concepts, methods and structure of the field are set out, as well as key limitations and challenges. The second part of the book shows how rigorous explanations emerge from stem cell experiments, and compares these to other kinds of scientific explanation. Model organisms, the role of genes, and the significance of collaboration are also discussed. The last part of the book considers relations to systems biology and clinical medicine, arguing that both the mathematical models of the former, and ethical principles of the latter, are necessary for stem cell biology to deliver on its promises.

oltage-gated calcium channels are essential mediators of a range of physiological functions, including the communication between nerve Vcells, the regulation of heart beat, muscle contraction, and secretion of hormones such as insulin. Consequently, these channels are critical phar macological targets in the treatment of a variety of disorders, such as epi lepsy, hypertension, and pain. Voltage-gated calcium channels have there fore been subject to intense study by numerous investigators over the past few decades, and an immense body of work has accumulated. In this book, we provide the first comprehensive overview of our current state of knowl edge concerning this exciting field of research. Leading off with a general review of calcium signaling and techniques to measure calcium channel ac tivity, the book delves into a provocative overview of the history of the cal cium channel field, contributed by one of the key pioneers in the field. Dr. Richard Tsien. This is followed by an in depth review of the biochemical and molecular biological characterization of calcium channel genes by Drs. Catterall and Snutch whose research has resulted in major advances in the calcium channel field. A number of chapters are dedicated towards various aspects of calcium channel structure and function, including channel gat ing, permeation, modulation and interactions with members of the exo- totic machinery-contributed by both established leaders and rising stars in the field."

The present book gives an overview on the similarities and differences of the various translation systems. Moreover, it highlights the mechanisms and control of translation in mitochondria and other organelles such as chloroplasts, plastids and apicoplasts in different organisms. Lastly, it offers an outlook on future developments and applications that might be made possible by a better understanding of translation in mitochondria and other organelles. "

This book highlights the importance of phytochemicals and mitochondria in cancer prevention and therapy. Recent scientific discoveries have identified that naturally occurring biologically active compounds (i.e. phytochemicals) target multiple steps of tumorigenesis leading to the inhibition or delay in cancer progression. Mitochondria, organelles within a cell, are a critical target for phytochemicals in regulating the initiation, promotion, and progression of cancer. The book is divided into three parts to better communicate the important findings related to phytochemicals and mitochondria in cancer research. The first part describes updates on environmental and genetic factors causing cancer initiation and progression, the role of mitochondria function in regulating the process of tumorigenesis, and the role of mitochondria in regulating cell death such as apoptosis, autophagy, and necroptosis. The second part focuses on the elucidation of key target proteins that could be exploited for cancer prevention, an the role of phytochemicals in cancer prevention, updates on basic research related to phytochemicals action critical for cancer prevention, and updates on translational knowledge on cancer prevention by phytochemicals. The third part provides updates on phytochemicals targeting mitochondria for cancer therapy, an overview of action of phytochemicals on cancer stem cells, updates on the role of microRNA in phytochemicals-based therapy of cancer, and updates on phytochemicals-based translation research on therapy for metastatic cancer.

Reviews the state of research on the physiology of yeast stress responses and signal transduction pathways in yeast. Coverage includes adaptation to nutrient depletion in the yeast, stationary phase in the yeast saccharomyces cerevisiae, the yeast shock response, the response of yeast to osmotic stress, crucial factors in salt stress tolerance, ox

Biological membranes play a central role in cell structure, shape and functions. However, investigating the membrane bilayer has proved to be difficult due to its highly dynamic and anisotropic structure, which generates steep gradients at the nanometer scale. Due to the decisive impact of recently developed fluorescence-based techniques, tremendous advances have been made in the last few years in our understanding of membrane characteristics and functions. In this context, the present book illustrates some of these major advances by collecting review articles written by highly respected experts. The book is organized in three parts, the first of which deals with membrane probes and model membranes. The second part describes the use of advanced quantitative and high-resolution techniques to explore the properties of biological membranes, illustrating the key progress made regarding membrane organization, dynamics and interactions. The third part is focused on the investigation of membrane proteins using the same techniques, and notably on the membrane receptors that play a central role in signaling pathways and therapeutic strategies. All chapters provide comprehensive information on membranes and their exploration for beginners in the field and advanced researchers alike.