This book will bring together leading international experts to discuss recent advances in basic scientific knowledge regarding the regulation of presynaptic Ca2+ channels. Importantly, Ca2+ channels represent one of the most widely modulated proteins in the body, being the target of a range of effector pathways and drugs; this range will be fully represented here. A number of therapeutic drugs target the Ca2+ channel complex, including the anti-epileptic gabapentinoid and analgesic ziconotide drugs and the pharmaceutical industry is searching for Ca2+ channel blocking drugs, particularly in the pain, epilepsy, ataxia and migraine areas. Such potential future therapies will be discussed here. Scientific disciplines will focus on electrophysiological studies, but will extend to neuroscience, genetics and biochemical areas. The work described will represent advances at the cutting edge of current neuroscience research and is timely and highly appropriate for the Springer book series.

In addition to updating important core techniques this third edition presents new chapters on assessing leukocyte involvement in angiogenesis, lymphatic cell and pericyte isolation techniques, spheroid and arterial ring based in vitro assays, and on pericyte involvement in angiogenesis. Angiogenesis Protocols, Third Edition also guides readers through new protocols on chorioallantoic membrane models, corneal pocket assays to assess angio- and lymphangiogenesis, models of muscle angiogenesis, and use of zebrafish embryos to study vascular angiogenesis and senescence. 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, Angiogenesis Protocols, Third Edition provides not only a practical handbook for key techniques, but also an informative and enjoyable read for all those interested, no matter how directly, in angiogenesis.

This volume explores the technological improvements in protein engineering, expression, purification, and crystallization of several rhodopsin photoactive intermediates, thus increasing our understanding of rhodopsin activation. The first chapters of the book focus on methods developed to study fundamentals of rhodopsin structure and function, starting with improved purification protocols of native and mutated rhodopsin, followed by methods used for rhodopsin reconstitution into lipid bilayers stabilizing rhodopsin function properties, and finally describing recently developed methods to study structural dynamics of rhodopsin activation and its mechanistic properties. Subsequently, chapters underline various techniques that have been developed to visualize the rhodopsin dimer and to study its functional significance. The next few chapters highlight cutting-edge imaging techniques of photoreceptors, rhodopsin trafficking, and its diffusion within signaling membranes. Finally the book concludes with recent developments that could be potentially beneficial in patient treatments, and treatment strategies for retinal degenerative diseases. Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Timely and practical, Rhodopsin: Methods and Protocols reaches out to researchers and health practitioners, and provides timely protocol useful for studying structural and functional properties of rhodopsin.

"The Centrosome: Cell and Molecular Mechanisms of Functions and Dysfunctions in Disease" includes chapters on classic and modern aspects of centrosome research to cover topics of current interest that have not been covered in depth in most books on the market so far. It extends on previous topics and includes new exciting aspects of centrosome research focused on primary cilia and their dysfunctions that are implicated in numerous diseases. Each chapter will be written by experts in their fields who will contribute their unique expertise in specific research fields and include cell and molecular details that are important for the specific subtopics. The book will be comprehensive, concise and will include reviews of key topics in the field. Cutting edge new information will be balanced with background information that will be readily understandable for the newcomer and the experienced centrosome researcher alike.

Revealing essential roles of the tumor microenvironment in cancer progression, this book provides a comprehensive overview of the latest research on the role of interleukins in the tumor microenvironment. Each chapter focuses on the various ways to target the tumor microenvironment by intervention in the interleukin biology, including IL-1, IL-8, IL-21, IL-36 signaling, and more. Taken alongside its companion volumes, Tumor Microenvironment: The Role of Interleukins - Part A updates us on what we know about various aspects of the tumor microenvironment, as well as future directions. This book is essential reading for advanced cell biology and cancer biology students as well as researchers seeking an update on research in the tumor microenvironment.

When the late Professor C. D. Darlington founded what developed into the International Chromosome Conferences in Oxford in 1964, he was concerned that scientists who worked on different aspects of chromosomes, or who studied them in different ways, should have the opportunity of "discussing the fundamental problems of chromosomes with one another". The fact that well over 300 scientists with a wide variety of interests came to Edinburgh in August 1992 for the 11th International Chromosome Conference shows that there is still the same need, and also the desire among chromosomologists to have such discussions. The present volume contains almost all the invited contributions, and attests to the diversity of approaches and applications in chromosomal studies. A few years ago it may have seemed to some that chromosome studies were being superseded by molecular biology, but the molecular biologists have now realized that they need to know about chromosomes, and indeed an important, if ill-defined discipline of 'molecular cytogenetics' has grown up in recent years. We are pleased that in planning the Conference and this book, so much of the work presented is at the interface between cytogenetics and molecular biology. This will surely continue in the future, as boundaries between disciplines are largely artificial, and each has much to learn from the others.

Prior to 1974, the ~adrenergic receptors were known only in- directly as entities that responded to drugs in a selective manner to mediate a variety of physiologically important responses. During the intervening years, our view of ~adrenergic receptors has changed dramatically. The availability of high affinity 125I-labeled radioligands selective for these receptors presaged an explosion of experimenta- tion utilizing direct binding assays to establish the biochemical properties of the receptor protein. In the opening chapter, Stadel and Lefkowitz describe this development and its impact on our under- standing of the molecular basis of ~adrenergic receptor function. The availability of well-characterized receptor ligands, coupled with the development of efficient methods for detergent solubilization, formed the basis of receptor purification using affinity chromatography. The related technique of photoaffinity labeling provided a means to estimate the molecular mass of these receptors. The availability of substantial amounts of purified ~2-adrenergic receptor allowed determination of segments of its amino acid se- quence. This information led to the production of polynucleotide probes and eventually to cloning of the receptor gene and determi- nation of the complete primary sequence of the receptor protein. Caron and Lefkowitz review the investigations leading to this major development and discuss the methods involved. They analyze our current perception of the relation of receptor function to its structure and discuss the general features of the G protein-interacting receptor family, of which the ~-adrenergic receptors are prototypes.

A comprehensive state-of-the-art collection of the most frequently used techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. Two new appendices explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, and list more than 100 very useful internet sites.

Within Pyrosequencing Protocols, the protocols for utilizing Pyrosequencing technology are described in detail, including trouble-shooting tips and background information. Chapter 1 provides an introduction to the fascinating origins of the methodology. Chapter 2 provides a brief overview of some of the applications Pyrosequencing is used for. Chapters 3 and 4 describe primer selection and the basic technique. Chapters 5 through 7 provide methods for improving the throughput and decreasing the cost of Pyrosequencing. Detailed applications for the technique can be ound in Chapters 8-13, whilst the important aspect of data storage is discussed in Chapter 14.

This book is indispensable to researchers in fields as diverse as Molecular Biology and Biophysics. It covers the important role that mitochondria play in a variety of biochemical spheres. It analyses how mitochondria affect metabolic pathways, how they are active in the regulation of cytosolic constituents, and their role in initiating signal pathways. Also covered are the way mitochondria help to regulate apoptosis, and how they modulate cellular hypertrophy and proliferation. It gives an overview of the emergence of mitochondria as an important regulator of cell signaling, with a particular focus on their pathophysiology.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, and those that are available often seem either out of date or limited in scope. This series is about the methods most widely used by modern-day neuroscientrsts and is written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the wrdest range of s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of amines and their metabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for all types of ligand-receptor binding, including chapters on general principles, drug discovery and development, and a most useful-appendix on computer programs for Scatchard, nonlinear, and competitive d- placement analyses. Volume 5 provides procedures for the asse- ment of enzymes involved in biogenic amme synthesis and catabolrsm. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analyncal, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, and those that are available often seem either out of date or limited in scope. This series is about the methods most widely used by modern-day neuroscientists and 1s written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the widest range of s- drsciplines. Collecting these general techniques together in a single volume stnkes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of ammes and theirmetabolites and of amino acrds, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for many types of ligand-receptor binding, including chapters on general prin- ples, drug discovery and development, and a most useful app- dix on computer programs for Scatchard, nonlinear, and compe- tive displacement analyses. Volume 5 provides procedures for the assessment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physrologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Whereas plant and insect infections are commonly caused by fungi, only a small minority of the vast diversity of fungal species is pathogenic to humans. Despite this, fungal infections cause considerable morbidity and mortality worldwide. This volume is dedicated to the biology, clinical presentation and management of invasive fungal infections. Major pathogenic fungi are introduced by world-leading experts and the basic principles of fungal virulence are reviewed in the light of new results and experimental technologies that offer unprecedented insights into invasive infections caused by "Aspergillus," "Candida," "Cryptococcus," "Pneumocystis" and "Mucorales." In parallel, the clinical presentation of invasive fungal infections and current approaches to their diagnosis and treatment are summarized to provide an overview of human pathogenic fungi, linking pathogen biology to the clinical presentation of disease.

The previous edition of Transmembrane Signaling Protocols was published in 1998. Since then the human genome has been completely sequenced and new methods have been developed for the use of microarrays and proteomics to analyze global changes in gene expression and protein profiles. These advances have increased our ability to understand transmembrane signaling processes in much greater detail. They have also simultaneously enhanced our ability to determine the role of a large number of newly identified molecules in signaling events. In addition, novel video microscopy methods have been developed to image transmembrane signaling events in live cells in real time. In view of these major advances, it is time to update the previous edition. Because of the success of that volume, we have chosen to keep the essential character of the book intact. Introductory chapters from experts have been included to provide overall perspective and an overview of recent advances in signal transduction pathways. The individual chapters now include comp- hensive detailed methods, studies in genetically tractable systems, fluorescence microscopy in live single cells, ex vivo analysis of primary cells from tra- genic mice, as well as genomic and proteomic approaches to the analysis of transmembrane signaling events. We would like to express our deep gratitude to the coauthors of this publi- tion. We hope that Transmembrane Signaling Protocols, Second Edition will serve as a valuable resource for future progress in the study of signal transd- tion pathways.

In the last few years, significant breakthroughs in transcription research expanded our appreciation for the complexity of molecular controls on gene expression in mammalian cells. In Transcription Factors: Methods and Protocols, experts in the field describe state-of-the-art approaches that investigators can use to probe critical mechanisms underlying transcription factor nuclear-cytoplasmic trafficking as well as to assess the functional impact of post-translational modifications on transcription factor function. The chapters are written by prominent scientists, many of whom developed these methods, and highlight protocols that focus on specific transcription factor family members with particular relevance to human disease. Composed in the highly successful Methods in Molecular Biology(TM) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Comprehensive and current, Transcription Factors: Methods and Protocols compiles the latest techniques for elucidating controls on transcription factor intracellular localization and activity, and consequently is unlike any other methods-based text on transcriptional regulation today.

This book covers the main aspects of biological rhythms. It focuses on the evolution and basic features of the biorhythms in organisms, deals with the circadian system at the genetic, molecular and cellular levels, and describes the mechanisms involved in the perception and light entrainment of the biological clock in vertebrates and invertebrates. The most important features of the biological clock are summarized on the level of whole organisms, from fish to mammals, and long-term (seasonal) rhythms in plants and higher vertebrates are discussed. Finally, the book concentrates on short-term rhythms, the significance of having a biological clock system in animals living in extreme (Arctic) environments, and on the diversity of circadian responses to melatonin, one of the key endocrine elements involved in the regulation of biological rhythms.

This volume compiles a broad range of step-by-step protocols, complementary to the ones published in the first edition of this book, to study various aspects of mitochondrial structure and function in different model organisms, both in vitro and in vivo. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Mitochondria: Practical Protocols, Second Edition aims to be useful for beginners as well as for experienced researchers in the field.

A light-hearted look at the nature of academic science, intended for anyone interested in biology but particularly for biology students who want to find out what the future holds in store. The "Egg" of the title refers to the science of developmental biology, which is the speciality of the author, and which provides the material for many of the anecdotes. The "Ego" relates to the vanity of the scientists themselves. Academic scientists have to struggle to maintain their research funding. To do this they must persuade other scientists that they are very good, and that means working at a good institution, publishing papers in the most fashionable journals and giving lectures at the most prestigious meetings. Success often goes to those with the largest egos and it is their style of operation that is described in this book. The author is a well-known scientist who has worked at both universities and research institutes. He has published over 100 scientific papers and an influential book about embryonic development: "From Egg to Embryo".

Transcriptome Profiling: Progress and Prospects assists readers in assessing and interpreting a large number of genes, up to and including an entire genome. It provides key insights into the latest tools and techniques used in transcriptomics and its relevant topics which can reveal a global snapshot of the complete RNA component of a cell at a given time. This snapshot, in turn, enables the distinction between different cell types, different disease states, and different time points during development. Transcriptome analysis has been a key area of biological inquiry for decades. The next-generation sequencing technologies have revolutionized transcriptomics by providing opportunities for multidimensional examinations of cellular transcriptomes in which high-throughput expression data are obtained at a single-base resolution. Transcriptome analysis has evolved from the detection of single RNA molecules to large-scale gene expression profiling and genome annotation initiatives. Written by a team of global experts, key topics in Transcriptome Profiling include transcriptome characterization, expression analysis of transcripts, transcriptome and gene regulation, transcriptome profiling and human health, medicinal plants transcriptomics, transcriptomics and genetic engineering, transcriptomics in agriculture, and phylotranscriptomics.

This volume of the Methods in Molecular Biology series is entirely devoted to the study of steroid receptor biology. Steroid hormone receptors represent a powerful system for the study of both the most fundamental molecular mec- nisms of gene regulation and control and the gross physiological responses of organisms to steroid hormones. Research in this field has brought forth advances in the treatment of cancer, endocrine disorders, and reproductive biology, and allowed elucidation of the fundamental biological mechanisms of gene expr- sion. In Steroid Receptor Methods: Protocols and Assays, the reader will find a collection of methods and protocols submitted by many fine steroid receptor researchers from throughout the world. These authors have been instructed to create a highly informative cross-section of the latest research techniques ava- able. The resulting work is timely, useful, and approachable for both the ex- rienced researcher and the novice to the field. Because the steroid receptor family is represented by a wonderfully diverse, yet strongly interrelated set of steroid receptor proteins, Steroid Receptor Methods contains protocols for the prod- tion and purification of a variety of receptor forms, including the progesterone, glucocorticoid, and androgen receptors. These procedures provide the raw ma- rial needed to conduct sophisticated biochemical analysis of receptor properties. Other techniques presented allow the reader to perform biochemical experiments on DNA binding characteristics, hormone binding assays, and protocols using combinatorial chemistry for drug discovery.

There is no doubt that nowadays, biology benefits greatly from mathematics. In particular, cellular biology is, besides population dynamics, a field where tech niques of mathematical modeling are widely used. This is reflected by the large number of journal articles and congress proceedings published every year on the dynamics of complex cellular processes. This applies, among others, to metabolic control analysis, where the number of articles on theoretical fundamentals and experimental applications has increased for about 15 years. Surprisingly, mono graphs and textbooks dealing with the modeling of metabolic systems are still exceptionally rare. We think that now time is ripe to fill this gap. This monograph covers various aspects of the mathematical description of enzymatic systems, such as stoichiometric analysis, enzyme kinetics, dynamical simulation, metabolic control analysis, and evolutionary optimization. We believe that, at present, these are the main approaches by which metabolic systems can be analyzed in mathematical terms. Although stoichiometric analysis and enzyme kinetics are classical fields tracing back to the beginning of our century, there are intriguing recent developments such as detection of elementary biochemical syn thesis routes and rate laws for the situation of metabolic channeling, which we have considered worth being included. Evolutionary optimization of metabolic systems is a rather new field with promising prospects. Its goal is to elucidate the structure and functions of these systems from an evolutionary viewpoint."

Advances in Applied Microbiology, Volume 120 continues the comprehensive reach of this widely read and authoritative review source in microbiology, providing invaluable references and information on a variety of areas relating to the topics of microbiology.

In the past few years, the scientific community has witnessed significant progress in the study of ion channels. Technological advancement in biophysics, molecular biology, and immunology has been greatly ac celerated, making it possible to conduct experiments which were deemed very difficult if not impossible in the past. For example, patch-clamp techniques can now be used to measure ionic currents generated by almost every type of cell, thereby allowing us to analyze whole-cell and single channel events. It is now possible to incorporate purified ion channel components into lipid bilayers to reconstitute an "excitable membrane." Gene cloning and monoclonal antibody techniques provide us with new approaches to the study of the molecular structure of ion channels. A variety of chemicals have now been found to interact with ion channels. One of the classical examples is represented by tetrodotoxin, a puffer fish poison, which was shown in the early 1960s to block the voltage-activated sodium channel in a highly specific and potent manner.