Ever since their discovery more than 150 years ago, astrocytes have proved to be something of an enigma. Only in the last 10-15 years has it become clear that astrocytes are multifunctional versatile cells that play key roles in a multitude of diverse processes in the CNS, including generation of neural stem cells, synaptogenesis, and regulation of the blood-brain barrier and neurovascular unit. Astrocytes: Methods and Protocols provides scientists with a comprehensive guide to many techniques used for astrocyte cell culture, as well as more specialized approaches for studying astrocyte functions, both in vitro and in vivo. Key cellular, molecular and biochemical techniques are used to study the many and varied functions of this fascinating cell. Written in the highly successful Methods in Molecular Biology (TM) 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.

The development of vertebrate muscle has long been a major area of research in developmental biology. During the last decade, novel technical approaches have allowed us to unravel to a large extent the mechanisms underlying muscle formation, and myogenesis has become one of the best-understood paradigms for cellular differentiation. This book concisely summarizes our current knowledge about muscle development in vertebrates, from the determination of muscle precursors to terminal differentiation. Each chapter has been written by an expert in the field, and particular emphasis has been placed on the different developmental and molecular pathways followed by the three types of vertebrate musculature - skeletal, heart and smooth muscle.

Recombinant Proteins from Plants is one of the most exciting and fastest developing areas in biology. The latest molecular techniques are being applied to the exploitation of plants as novel expression systems for the p- duction and overproduction of heterologous and native proteins. Transgenic plant technology is currently used in three broad areas: the expression of - combinant proteins to improve crop quality by increasing disease/pest res- tance or increasing tolerance to stress, optimizing plant productivity and yield by the genetic manipulation of metabolic pathways, and the large-scale co- effective production of recombinant proteins for use as specialist industrial or therapeutic biomolecules. The intention of Recombinant Proteins from Plants is to provide c- prehensive and detailed protocols covering all the latest molecular approaches. Because the production oftransgenic plants has become routine in many la- ratories, coverage is also given to some of the more "classical" approaches to the separation, analysis, and characterization of recombinant proteins. The book also includes areas of research that we believe will become increasingly important in the near future: efficient transformation of monocots with Agrobacterium optimizing the stability of recombinant proteins, and a section highlighting the immunotherapeutic potential of plant-expressed proteins.

The tools of molecular biology have revolutionised our understanding of gene structure and function and changed the teaching of genetics in a fundamental way. The transition from classical genetics to molecular genetics was initiated by two discoveries. One was the discovery that DNA has a complementary double helix structure and the other that a universal genetic code does exist. Both led to the acceptance of the central dogma that RNA molecules are made on DNA templates.
The last twenty years have seen remarkable growth in our knowledge of molecular genetics, most of which is the outcome of recombinant DNA technology. This technology which is not limited to cloning, sequencing, and expression has created a biotechnology industry of its own, the purpose of which is to develop new diagnostic and therapeutic approaches in medicine. Both industries in collaboration with the biomedical community are now engaged in laying down the foundation of molecular medicine.
The present volume seeks to provide a coherent account of the new science of molecular genetics. Its content however is by no means exhaustive, partly because of the publication explosion but more because of space restrictions. A rudimentary knowledge of genetics on the reader's part is assumed. Quite understandably, considerable emphasis is placed on major technical advances but not without expounding numerous new ideas and phenomena including alternative splicing, POR, DNA methylation, genomic imprinting, and so on.

Microtubules are at the heart of cellular self-organization, and their dynamic nature allows them to explore the intracellular space and mediate the transport of cargoes from the nucleus to the outer edges of the cell and back. In Microtubule Dynamics: Methods and Protocols, experts in the field provide an up-to-date collection of methods and approaches that are used to investigate microtubule dynamics in vitro and in cells. Beginning with the question of how to analyze microtubule dynamics, the volume continues with detailed descriptions of how to isolate tubulin from different sources and with different posttranslational modifications, methods used to study microtubule dynamics and microtubule interactions in vitro, techniques to investigate the ultrastructure of microtubules and associated proteins, assays to study microtubule nucleation, turnover, and force production in cells, as well as approaches to isolate novel microtubule-associated proteins and their interacting proteins. Written in the highly successful Methods in Molecular Biology (TM) 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. Definitive and practical, Microtubule Dynamics: Methods and Protocols provides the key protocols needed by novices and experts on how to perform a broad range of well-established and newly-emerging techniques in this vital field.

This detailed volume encompasses chapters from leading experts in the area of membrane proteins who describe step-by-step protocols developed these last few years to improve the functional production and stabilization of recombinant integral membrane proteins (IMPs). Membrane proteins play a key role in numerous pathologies such as cancer, cystic fibrosis, epilepsy, hyperinsulinism, and Alzheimer's disease, yet studies on these and other disorders are hampered by a lack of information about the proteins involved. This book sets out to aid researchers in rectifying this situation. Written for the highly successful Methods in Molecular Biology series, 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 up-to-date, Heterologous Expression of Membrane Proteins: Methods and Protocols, Second Edition serves as an ideal guide for scientists attempting to delve deeper into the myriad unique IMP structures.

As the first comprehensive overview of the nucleolus since 1985, The Nucleolus covers our current understanding of the cell nucleolus, including its role in ribosome assembly and its additional newly-discovered activities. The eighteen chapters have been written by experts who are actively engaged in research on the nucleolus and have an in depth review of the following topics:

- nucleolar ultrastructure and dynamics,
- behavior during mitosis,
- ribosomal DNA gene and chromatin structure,
- pre-ribosomal RNA transcription,
- processing and modification,
- ribosome assembly,
- small nucleolar RNAs,
- proteomics and non-traditional functions of the nucleolus.

Separate chapters are also provided for yeast and higher eukaryotes on many topics.
The Nucleolus will appeal not only to scientists directly engaged in nucleolar research, but also those working in related areas such as gene expression, protein biosynthesis, ribosome structure, transcription, chromatin structure, molecular genetics and the structure and functions of the cell nucleus in general.

Root hairs are tip-growing cells that originate from epidennal cells called trichoblasts. Their role may be simply thought of as extending the surface area of the root to facilitate absorption of nutrients and water. However, as you will see in this book, the root hair is far more than that. To an increasingly larger number of plant biologists, the root hair is a model cell. It grows in much the same way as a pollen tube, by sending vast numbers of vesicles containing cell wall precursors to a rounded apical dome, the tip. Once the trichoblast becomes committed to root hair fonnation, it no longer divides. The root hair cell has a migrating nucleus and a complex cytoskeleton. It has a varied cell wall. It is easy to observe through differential interference contrast microscopy because there are no other cells around it to disturb the image. Cytoplasmic streaming is exceptionally clear, and amyloplasts and even mitochondria and endoplasmic reticulum can be seen without reporter labelling in some species. Root hair mutants are easy to distinguish and catalogue. Plant honnones are involved in their growth and development. It is thus an almost ideal plant cell for experimental manipulation and observation. The root hair is also involved in interactions with soil microbes, as you will learn from later chapters of the book.

The purpose of Ribozyme Protocols is to provide a helpful compilation of protocols that will be of use- DEGREESnot only to those with some experience of ribozymes- DEGREESbut also to those wishing to use ribozymes for the first time. Although it is usually impossible to cover every aspect of a scientific field, I believe this book approaches that ideal and should help all readers perform meaningful experiments using ribozymes. To design ribozymes, one must consider whether the target site will be accessible; this task can be facilitated by using computer programs that pre dict the folding of the target RNA. Such programs are detailed in Chapters 2 and 3. If the chosen target is an RNA virus that can mutate rapidly, it makes sense to consider those parts of the genome that are least likely to change during viral replication. An example of how this can be done is described in Chapter 4. Although computer analysis may be a useful starting point to select tar get sites, there seems, at the moment, to be no guarantee that any particular chosen site will be efficiently cleaved. Some workers have deliberately bypassed this problem by using libraries of ribozyme sequences and by select ing those that actually hybridize to and/or cleave the target; these methods are described in Chapters 5

The first section of this volume consists of five chapters to the nature of membrane transport systems. A chapter on secondary active glucose transport has been omitted because this topic is slated to appear in the Nephrobiology module. Chapter 6 deals with oxidase control of plasma membrane proton transport, while chapter 7 addresses the question of how cell volume is regulated. Although we chose not to have a separate chapter covering additional co-transport systems namely, Na+ -K+ -2CI-, KCI, -HCO-3, as well as CI- -HCO-3 exchange and K+ and CI- movements through channels, the role of each in cell volume regulation is emphasized in Chapter 7.
Instead of devoting an entire section to the thermodynamics of metabolism, we thought it desirable to have the subjects of medical imaging and NMR of cell metabolism discussed in some detail in two chapters. These are followed by a chapter on the thermodynamic instrument - the calorimeter. Calrimetry allows the measurement of net changes of heat in cells, tissues, organs and whole body. As will be recognized, heat dissipation does not arise only from chemical reactions but also from interactions between macromolecules and conformational changes in protein complexes and mass Ca2+ movement such as that occurring in contracting skeletal muscle. The last chapter provides an account of equilibrium and non-equilibrium thermodynamics and the enthalpy balance method. It reveals that calometric measurements are useful in studies of clinical and toxicological problems.

Human pluripotent stem cells such as human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) with their unique developmental plasticity hold immense potential as cellular models for drug discovery and in regenerative medicine as a source for cell replacement. While hESC are derived from a developing embryo, iPSC are generated with forced expression of key transcription factors to convert adult somatic cells to ESC-like cells, a process termed reprogramming. Using iPSC overcomes ethical issues concerning the use of developing embryos and it can be generated from patient-specific or disease-specific cells for downstream applications. Pluripotent Stem Cells: Methods and Protocols highlights the best methods and systems for the entire work flow. Divided into four convenient sections, topics include a focus on producing iPSC from diverse somatic sources, media systems for expanding ESC and iPSC with detailed protocols for directed differentiation into specific lineages, commonly used cellular and molecular characterization methods , and the potential application of labeled stem cells with specific methods for cloning, gene delivery and cell engineering. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Pluripotent Stem Cells: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential cellular feature.

I am extremely honored and pleased to have the opportunity to write a few introductory words for this timely volume on Na + /It exchange. This is a field of investigation that I entered into by challenge and necessity, embraced with passion and fmally left in my quest for new discoveries in growth control. Ten years, one third of my scientific life, has been devoted to uncovering the mysteries of intracellular pH (PH;) regulation with respect to growth factor action. I got started on this new topic in 1980, when I heard a rather provocative hypothesis presented by Enrique Rozengurt at an ICN-UCLA Keystone meeting on "Cell Surface and Malignancy." He showed that all mitogens induced amiloride-sensitive Na + entry into resting cells and proposed that, if a compound stimulates Na + influx, it could be a mitogen. In support of his proposal Enrique reported that the amphipathic polypeptide, mellitin, which induced Na+ influx, was indeed mitogenic for 3T3 cells. This was only correlation at this stage. However, I was fascinated by this talk. I immediately approached Enrique to inform him of my skepticism about this beautiful story, and to indicate that I would only be convinced when I succeeded in isolating mutant fibroblasts lacking the amiloride-sensitive Na+ transporter. ''Good luck " was his response.

Knowledge of the extracellular matrix (ECM) is essential to understand cellular differentiation, tissue development, and tissue remodeling. This volume of the series "Biology of Extracellular Matrix" provides a timely overview of the structure, regulation, and function of the major macromolecules that make up the extracellular matrix. It covers topics such as collagen types and assembly of collagen-containing suprastructures, basement membrane, fibronectin and other cell-adhesive glycoproteins, proteoglycans, microfibrils, elastin, fibulins and matricellular proteins, such as thrombospondin. It also explores the concept that ECM components together with their cell surface receptors can be viewed as intricate nano-devices that allow cells to physically organize their 3-D-environment. Further, the role of the ECM in human disease and pathogenesis is discussed as well as the use of model organisms in elucidating ECM function. Content Level Research

This valuable resource provides a systematic account of the biochemistry of smooth muscle contraction. As a comprehensive guide to this rapidly growing area of research, it covers the structure and characteristic properties of contractile and regulatory proteins, with special emphasis on their predicted function in the live muscle. Also included in this book are intermediate filament proteins, and desmin and vimentin, whose function in smooth muscle is unknown; and several enzymes involved in the phosphorylation-dephosphorylation of contractile and other proteins.

This publication contains full papers of both oral and poster presentations of the symposium "Immobilized Cells: Basics and Applications" that was held in Noordwijkerhout, The Netherlands, 26-29 November 1995. This volume covers recent developments in the field of immobilization e.g.: new support materials, characterization of support materials, kinetic characterizations, dynamic modelling, bioreactor types, scale up and applications are also given. Applications in the field of medicine, fermentation technology, food technology and environmental technology are described. Guidelines for research with immobilized cells. Based on the scientific sessions a strategy of research and methods for characterization of immobilized cells, especially in view of applications are given.

The goal was to relate basic research to applications and to extract guidelines for characterization of immobilized cells in view of process design and application from the contributions. The manuscripts presented in these proceedings give an extensive and recent overview of the research and applications of immobilized-cell technology.

Metamorphosis addresses various facets of postembryonic development, particularly signal transduction, morphogenesis, cell-cell interactions, and programmed cell death. A key feature of the book is its exploration of the molecular mechanisms underlying these processes.
Key Features
* Hormonal regulation of development
* The mechanisms of hormone action
* The steroid/thyroid hormone receptor family
* Morphogenesis and programmed cell death

This textbook provides an introduction to dynamic modeling in cell biology, emphasizing computational approaches based on realistic molecular mechanisms. It is designed to introduce cell biology and neuroscience students to computational modeling, and applied mathematics students, theoretical biologists, and engineers to many of the problems in dynamical cell biology. This volume was conceived of and begun by Professor Joel Keizer based on his many years of teaching and research together with his colleagues. The project was expanded and finished by his students and friends after his untimely death in 1999. Carefully selected examples are used to motivate the concepts and techniques of computational cell biology, through a progression of increasingly more complex and demanding cases. Illustrative exercises are included with every chapter, and mathematical and computational appendices are provided for reference. This textbook will be useful for advanced undergraduate and graduate theoretical biologists, and for mathematics students and life scientists who wish to learn about modeling in cell biology. Royalties from this book will be donated to the Joel E. Keizer memorial endowment for collaborative interdisciplinary research in the life sciences.

Photobiology integrates a wide variety of scientific disciplines. As more people become aware of the many ways light interacts with chemical and biological systems, the need for a concise treatment of photobiology has become more critical. Kohen "et al." Have written just such a book, intended both as a textbook and as a reference.
The authors begin by providing a brief description of the nature of light, how it affects matter, and the means and methods of measuring it. A major section of the book is devoted to how light influences living systems, including discussions of photosynthesis, bioluminescence, regulatory mechanisms, and visual transduction of light. The last half of the book is devoted to the biomedical aspects of light, including photoimmunology, photoallergic reactions and other forms of light sensitivity, the optical properties of skin, and various ways that light can be used in therapy treatments.
Useful to photobiologists as a comprehensive overview, this book should also appeal to biomedical researchers and advanced students of photobiology.

This volume examines the current state of free radical biology as it impacts on hepatic disorders. It takes a thorough look at the relationship of oxidative stress in acute and chronic disease and takes into account factors like: redox biomarkers; antioxidant defense and protection; cell signaling, mutations; oxidative damage involving lipids, proteins and nucleic acids; membrane trafficking, inflammation, mitochondrial dysfunction, alterations in immunological function and toxicology and hypoxia. Studies on Hepatic Disorders, the latest volume in the Oxidative Stress in Basic Research and Clinical Practice series, provides a comprehensive look at liver topics. It is organized into four sections, each one thoroughly covering its topic and consisting of chapters written by recognized field leaders. Section One, covers basic principles including redox signaling, antioxidant defenses, nitric oxide, oxidative mechanisms in senescence and regeneration and the detection of oxidative stress. Section Two, explores Pathophysiology. It ranges from cell damage to fibrogenic response as broken out in chapters on hepatocellular injury, mitochondrial damage, unfolded protein response and autophagy, inflammation, ischemia-reperfusion injury and finally, fibrogenesis. Sections Three and Four cover specific diseases and cancer, respectively. Most of the chapters focus on diseases including acute failure, alcoholic disease, viral hepatitis, iron overload, autoimmune disease, Wilson's disease and more, while the chapters on cancer round out the book.

PCR Strategies expands and updates the landmark volume PCR Protocols. It is a companion laboratory manual that provides a completely new set of up-to-date strategies and protocols for getting the most from PCR.
The editors have organized the book into four sections, focusing on principles, analyses, research applications, and alternative strategies for a wide variety of basic and clinical needs. If you own PCR Protocols, you will want PCR Strategies. If you don't own PCR Protocols, you will want to buy both!
Key Features
* Concepts explained
* Methods detailed
* Trouble-shooting emphasized
* Novel applications highlighted
* Major sections
* Key concepts for PCR
* Analysis of PCR products
* Research applications
* Alternative amplification strategies

In eukaryotic cells, the nuclear genome and its transcriptional apparatus is separated from the site of protein synthesis by the nuclear envelope. Thus, a constant flow of proteins and nucleic acids has to cross the nuclear envelope in both directions. This transport in and out of the nucleus is mediated by nuclear pore complexes (NPCs) and occurs in an energy and signal-dependent manner. Thus, nucleocytoplasmic translocation of macro molecules across the nuclear envelope appears to be a highly specific and regulated process. Viruses that replicate their genome in the cell nucleus are therefore forced to develop efficient ways to deal with the intracellulZlr host cell transport machinery. Historically, investigation of Polyomavirus replication allowed identification ofsequences that mediate nuclear import, which led subsequently to our detailed understanding of the cellular factors that are involved in nuclear import. Transport ofmacromolecules in the opposite direction, however, is less well understood. The investigation of retroviral gene expression in recent years pro vided the first insights into the cellular mechanisms that regulate nuclear export. In particular, the detailed dissection of the function of the human immunodeficiency virus type I (HIV-I) Rev trans-activator protein identified CRMI, as a hona fide nuclear export receptor. CRM I appears to be involved in the nucleocytoplasmic translocation of the vast majority of viral and cellular proteins that have subsequently been found to contain a Rev-type leucine-rich nuclear export signal (NES)."

Immunofluorescence, a suitable laboratory method for the microscopic demonstration of antigens and antibodies in biological materials, useable, for example, to provide evidence for the pathogenesis of disease in histological or cytological preparations and for tumour cell differentiation. For this reason immunofluorescence has a decisive role as the method of choice for the diagnosis of auto-immune diseases. This primer on immunofluorescence techniques, which first appeared in 1979, is a richly illustrated handbook suitable for everyday practical work in the laboratory, useable as both an introduction to the subject as well as an atlas. In hardly any other area of medicine are there so many new findings to report. The second edition of this book is concerned not only with the detection methods which now form an essential and established part of diagnostic techniques, but also with the most recent research results such as the discovery of antibodies against Auerbach's plexus and against podocytes...

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 in 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, artificial organs and functional foods.
This volume gives the readers a complete review of present state-of-the-art in Japan and other countries where this field is well advanced. The Proceedings will be useful for the cell biologists, biochemists, molecular biologists, immunologists, biochemical engineers and other disciplines related to animal cell culture, working in either academic environments or in industries of biotechnology and pharmacy.

The discovery of microRNAs and its role as gene expression regulators in human carcinogenesis represents one of the most important scientific achievements of the last decade. More recently, other non-coding RNAs have been discovered and its implications in cancer are emerging as well, suggesting a broader than anticipated involvement of the non-coding genome in cancer. Moreover, completely new and unexpected functions for microRNAs are being revealed, leading to the identification of new anticancer molecular targets. This book represents a comprehensive guide on non-coding RNAs and cancer, spanning from its role as cancer biomarkers, to providing the most useful bioinformatic tools, to presenting some of the most relevant discoveries, which indicates how these fascinating molecules act as fine orchestrators of cancer biology.

The understanding how complement relates to glomerular diseases has evolved considerably during the last years. Substantial evidence has accumulated that explain how a defective or deregulated complement system results in kidney diseases. The combination and close interaction of basic research with clinical medicine has demonstrated an important role of complement effector and regulatory proteins in pathological settings of the kidney. A large panel of distinct human kidney diseases such as hemolytic uremic syndrome (HUS), membrano proliferative glomerulonephritis (MPGN), systemic lupus erythematosus (SLE) and in ischemic reperfusions injury and transplantation are caused by defective complement control. Genetic analyses have identified mutations in complement regulators that are associated with these diseases. Mutations have been identified in the fluid phase alternative pathway regulator Factor H and the membrane regulator Membrane Cofactor Protein MCP (CD46). The functional characterization of the mutant proteins allows to define the pathophysiological events on a molecular level. These new concepts and data on disease mechanisms already allowed to establish new diagnostic and novel promising therapeutic approaches for several human kidney diseases.