A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metablomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides new view on the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 4-7), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry, histologicalstaining (Chapters10and11), aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 14-16) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocol

A major mechanism by which cells regulate protein function is to place phosphate groups on serine and threonine residues. Though the steady-state level of protein phosphorylation depends on the relative activities of both kinases and phosphatases, a much greater effort has previously gone into the study of the former that the latter . Today, however, there is an increasing appreciation for the role that protein phosphatases play in the dynamic p- cess of protein phosphorylation . To date, there are four major types of protein serine/threonine phosphatase catalytic subunits, designated protein phosphatase type 1, 2A, 2B, and 2C . Each has been identified by the techniques of protein chemistry and enzymology and can be distinguished from one another by their preference for specific substrates as well as their sensitivity to certain acti- tors and inhibitors . Protein Phosphatase Protocols has been assembled in response to the growing interest these enzymes are receiving . The goal of this compilation is to provide a "how-to" experimental guide to aid newcomers as well as s- soned veterans in their research endeavors, thus further contributing towards our ever increasing knowledge of serine/threonine phosphatases . What you have before you contains contributions by many of the current and emerging leaders in the field . To highlight just a few, these chapters c- tain step-by-step information on how to isolate novel phosphatases and re- latory subunits, assay for activity, and generate immunological reagents for both biochemical and biological characterization of these enzymes .

Plant reproductive biology has undergone a revolution during the past five years, with the cloning, sequencing and localization of the genes important in reproduction. These advantages in plant molecular biology have led to exciting applications in plant biotechnology, including the genetic engineering of male sterility and other reproductive processes. This book presents an interesting and contemporary account of these new developments from the scientists in whose laboratories they have been made. The chapters focus on two areas: the molecular biology of self-incompatibility, which is the system of self-recognition controlled by the S-gene and related genes; and the cellular and molecular biology of pollen development and genetic dissection of male sterility. Some chapters feature Arabidopsis, with its unique genetic system. Reproduction is vital for seed production in crop plants, and this book presents new approaches to manipulate plant breeding systems for the 21st century.

A qualitative leap in the understanding of cardiovascular and n- ral regulation by the renin-angiotensin system, and of the role of this s- tem in tissue damage, has occurred as a result of the many recent advances in molecular genetic techniques. The cloning of the genes for the components of the renin-angiotensin system, the design of specific angiotensin receptor ligands, and the use of embryonic gene targeting te- niques for the creation of mutant strains have established that the renin- angiotensin system is important in blood pressure regulation, ion and fluid homeostasis, and tissue growth and remodeling Further investigation of the mechanisms by which this system p- ticipates in cardiovascular regulation may shed some light on the pat- genesis of several cardiovascular diseases, e. g. , hypertension, congestive heart failure, and chronic renal failure. Despite the promise of this system as a target for therapeutic interventions for these diseases, there are great challenges in the integration of the attempts to close the gap between the traditional literature of medicine and the explosion of information from the new technologies. This book's title, Angiotensin Protocols, reflects the authors' strong efforts to translate expert knowledge into easy-to-follow practice. The book opens with introductory chapters, and each specialty section provides detailed methods covering a wide variety of techniques, ranging from genetic manipulation of targeted genes to functional studies of the renin- angiotensin system.

The early, organ-specific diagnosis of malignancy continues to be a major unmet medical need. Clearly the ability to establish an early diagnosis of cancer is dependent upon an intimate knowledge of the cancer's biology, which if understood at the molecular level should identify key diagnostic and therapeutic manipulation points. Advances in recombinant gene technology have provided significant understanding of the mechanisms of action of oncogenic viruses, as well as of cancer-associated genomic sequences (onco genes). This text will explore the known molecular genetic, biolog ical, and clinical knowledge of selected human neoplasms that demonstrate association with suspected oncogenic virus and those cytogenetic alterations that either cause or are caused by oncogene activation. The text first reviews the cytogenetics of human cancers link ing classical cytogenetics and molecular genetics. Avery A. Sand berg (Roswell Park Memorial Institute, Buffalo, New York) reviews the leukemias and lymphomas, followed by S. Pathak (M. D. Anderson Hospital and Tumor Institute, Houston, Texas), who reviews solid tumors. Functional consideration of oncogenes is highlighted by Keith C. Robbins and Stuart A. Aaronson (NO, Bethesda, Maryland) through their description of the v-sis locus sis and its gene product p.28; a protein that closely resembles human platelet-derived growth factor (PDGF)."

Computational biology is an interdisciplinary field that applies mathematical, statistical, and computer science methods to answer biological questions, and its importance has only increased with the introduction of high-throughput techniques such as automatic DNA sequencing, comprehensive expression analysis with microarrays, and proteome analysis with modern mass spectrometry. In Computational Biology, expert practitioners present a broad survey of computational biology methods by focusing on their applications, including primary sequence analysis, protein structure elucidation, transcriptomics and proteomics data analysis, and exploration of protein interaction networks. As a volume in the highly successful Methods in Molecular Biology (TM) series, this work provides the kind of detailed description and implementation advice that is crucial for getting optimal results. Authoritative and easy to use, Computational Biology is an ideal guide for all scientists interested in quantitative biology.

In Nucleic Acid Chemistry: Methods and Protocols, expert researches in the field detail techniques and approaches for the detection of DNA and RNA. These techniques include the recovery of trace amounts of DNA for amplification and analysis, new qPCR chemistries, new application of isothermal amplification techniques, assays with visual or electric signals for point-of-care diagnostics, improvement of fluorescent in situ hybridization, and new signal amplification techniques. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Nucleic Acid Chemistry: Methods and Protocols seeks to aid scientists in the further study of detection for DNA and RNA.

In the last decade nucleo-cytoplasmic transport has developed into one of the most active areas of research in cell biology and its significance for cell biology as a whole has been recognized. It has led to elucidation of the mechanisms of major cellular functions such as protein synthesis and cellular differentiation. It is also helping to bring about a synthesis of ultrastructural, biochemical, molecular biological and biophysical approaches to the study of cellular organization and function. These theoretical advances have been made possible chiefly by advances in technology - improved methods of gene manipulation and sequencing, of monoclonal antibody production and the use of protein chemistry, of microscopy and of cell manipulation, including microinjection methods. Developments in molecular biology, such as the search for sequence - specific DNA binding proteins and the identification of their binding domains, have also furthered advances in cell biology and these areas have become increasingly integrated with some well-established areas of biochemistry. This book aims to be an introduction, for senior undergraduates and postgraduate biologists, to the field of nucleo-cytoplasmic transport and its relevance to cell biology and seeks to use recent advances in the field to illustrate the process by which modern scientific progress occurs - the importance of developing techniques for formulating problems as well as answering them and the integration of different approaches leading to the evolution of novel concepts. The author assumes a basic general knowledge of molecular and cell biology and of biochemistry, but provides all the necessary background for discussion of recent advances and in the final chapters discusses the actual and potential contributions of the field of nucleo-cytoplasmic transport to cell biology.

Microarrays for simultaneous measurement of redundancy of RNA species are used in fundamental biology as well as in medical research. Statistically, a microarray may be considered as an observation of very high dimensionality equal to the number of expression levels measured on it. In "Statistical Methods for Microarray Data Analysis: Methods and Protocols, " expert researchers in the field detail many methods and techniques used to study microarrays, guiding the reader from microarray technology to statistical problems of specific multivariate data analysis. 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.

Thorough and intuitive, "Statistical Methods for Microarray Data Analysis: ""Methods and Protocols "aids scientists in continuing to study microarrays and the most current statistical methods.

For more than a century, microscopy has been a centerpiece of extraordinary discoveries in biology. Along the way, remarkable imaging tools have been developed allowing scientists to dissect the complexity of cellular processes at the nano length molecular scales. Nanoimaging: Methods and Protocols presents a diverse collection of microscopy techniques and methodologies that provides guidance to successfully image cellular molecular complexes at nanometer spatial resolution. The book's four parts cover: (1) light microscopy techniques with a special emphasis on methods that go beyond the classic diffraction-limited imaging; (2) electron microscopy techniques for high-resolution imaging of molecules, cells and tissues, in both two and three dimensions; (3) scanning probe microscopy techniques for imaging and probing macromolecular complexes and membrane surface topography; and (4) complementary techniques on correlative microscopy, soft x-ray tomography and secondary ion mass spectrometry imaging. Written in the successful format of the Methods in Molecular Biology (TM) series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Nanoimaging: Methods and Protocols highlights many of the most exciting possibilities in microscopy for the investigation of biological structures at the nano length molecular scales.

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. In Heterologous Expression of Membrane Proteins: Methods and Protocols, expert researchers provide an overview of the different heterologous expression systems available to produce these proteins for structural analysis, helping us to understand how these proteins function, and additionally, how their functions can be modified through drug treatment. Chapters examine membrane protein solubilization, purification, and instability in solution, and investigate the strategies that helped to determine the structure of the first heterologously expressed mammalian membrane proteins. Composed in the highly successful Methods in Molecular Biology 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.

Far-reaching and innovative, Heterologous Expression of Membrane Proteins: Methods and Protocols, is an essential guide for novice and experienced researchers alike, providing detailed methods for existing expression systems alongside cutting-edge strategies for future use in the field."

Quantitative Real-Time PCR: Methods and Protocols focuses on different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for low level detection and to develop non-invasive diagnosis. Written for the Methods in Molecular Biology series, most chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Quantitative Real-Time PCR: Methods and Protocols aims to aid researchers seeking to devise new qPCR-based approaches related to his or her area of investigation.

The observation that neuropeptide Y (NPY) is the most abundant peptide present in the mammalian nervous system and the finding that it elicits the most powerful orexigenic signal have led to active investigations of the properties of the NPY family of hormones, including peptide YY (PYY) and pancreatic polypeptide (PP). Nearly two decades of research have led to the identification of several NPY receptor subtypes and the development of useful receptor selective ligands. Moreover, these investigations have imp- cated NPY in the pathophysiology of a number of diseases, including feeding disorders, seizures, memory loss, anxiety, depression, and heart failure. Vigorous efforts are therefore continuing, not only to understand the bioche- cal aspects of NPY actions, but also toward developing NPY-based treatments for a variety of disorders. To facilitate these efforts, it was decided to produce the first handbook on NPY research techniques as part of the Methods in Molecular Biology Series. In compiling Neuropeptide Y Protocols, I have gathered contributions on techniques considered critical for the advancement of the NPY field from experts in various disciplines. Each chapter starts with a brief introduction, with Materials and Methods sections following. The latter sections are presented in an easy to follow step-by-step format. The last section of the chapter, Notes, highlights pitfalls and the maneuvers employed to overcome them. This information, not usually disseminated in standard research pub- cations, may prove extremely useful for investigators employing these te- niques in NYP research.

This book reviews a novel and exciting field of cellular and molecular biology called epitranscriptomics, which focuses on changes in an organism's cells resulting from the posttranscriptional modification of cellular RNA. RNA-binding proteins (RBPs) play a crucial role in these posttranscriptional modifications and also support several cellular processes necessary for maintaining RNA homeostasis. Exploring the mechanisms underlying RNA modifications and RBP function is an emerging area of biomedical research, taking the study of gene regulation a step beyond epigenetics. This book reveals that the RNA molecule is not just an information-carrying molecule with some secondary structures. Accordingly, how RNA is modified, regulated, packaged, and controlled is an important aspect. Leading experts address questions such as where the over 170 distinct posttranscriptional RNA modifications are located on the genome, what percentage of mRNAs and noncoding RNAs these modifications include, and how an RNA modification impacts a person's biology. In closing, the book reviews the role of RNA modifications and RBPs in a variety of diseases and their pathogenesis. Addressing some of the most exciting challenges in epitranscriptomics, this book provides a valuable and engaging resource for researchers in academia and industry studying the phenomena of RNA modification.

Nuclear Receptors focuses on the structural analysis of nuclear receptors from the initial work using isolated protein domains to the more recent exciting developments investigating the conformational shape of full-length receptor complexes. The book also reviews the structure of key nuclear receptor co-regulatory proteins. It brings together, for the first time, a comprehensive review of nuclear receptor structure and the importance of receptor conformation underpinning allosteric regulation by different ligands (hormone, drugs, DNA response elements, protein-protein interactions) and receptor activity. The nuclear receptor superfamily, including receptors for steroid hormones and non-steroid ligands, are pivotal to normal physiology, regulating processes as diverse as reproduction, metabolism, the immune system and brain development. The first members of the family were cloned over 25 years ago, which heralded in the idea of a superfamily of intracellular receptor proteins that bound small molecule ligands: classical steroid hormones, vitamins, fatty acids and other products of metabolism. These signals are then transmitted through multiprotein receptor-DNA complexes, leading to the regulation of target genes, often in a cell-selective manner. The cloning of the receptor cDNAs also ushered in an era of unparalleled analysis of the mechanisms of action of these ligand-activated transcription factors.

Cell to Organ: Electromechanical Activation.- 1 Regulation of Intracellular Calcium in Cardiac Muscle.- 2 Neuromodulation of Calcium Current by Extracellular ATP in Isolated Ventricular Myocytes.- 3 Intracellular Calcium and Myocardial Function during Ischemia.- 4 Intracellular Signaling in Vascular Smooth Muscle.- 5 Myocardial Energetics.- 6 Mechanisms of Endocardial Endothelium Modulation of Myocardial Performance.- 7 Calcium Kinetic and Mechanical Regulation of the Cardiac Muscle.- 8 Cellular Responses to Electrical Stimulation: A Study Using a Model of the Ventricular Cardiac Action Potential.- 9 Excitation-Contraction Coupling and Contractile Protein Function in Failing and Nonfailing Human Myocardium.- Mechanics and Microcirculation.- 10 Estimation of Myocardial Mechanical Properties with Dynamic Transverse Stiffness.- 11 Intramyocardial Mechanical States: Vessel-Interstitium-Muscle Interface.- 12 Myocardial Mechanics and Coronary Flow Dynamics.- 13 Modeling of Coronary Capillary Flow.- Microstructure: Mechanics and Microcirculation.- 14 Models for Coronary Pressure-Flow Relationships.- 15In VivoMyocardial Microcirculation: Evaluation with a Whole-Body X-Ray CT Method.- 16 Endocardial Coronary Microcirculation of the Beating Heart.- 17 Distribution and Control of Coronary Microvascular Resistance.- 18 Theoretical Analysis of Coronary Blood Flow and Tissue Oxygen Pressure-Control.- Microcirculation and Metabolic Transport.- 19 Adenosine Coronary Vasodilation during Hypoxia Depends on Adrenergic Receptor Activation.- 20 Chaos in Cardiac Signals.- 21 Intramyocardial Fluid Transport Effects on Coronary Flow and Left Ventricular Mechanics.- 22 Metabolic Protection of Post-Ischemic Phosphorylation Potential and Ventricular Performance.- 23 Metabolic and Mechanical Control of the Microcirculation.- Remodeling of Muscle and Vessels.- 24 Asymmetrical Changes in Ventricular Wall Mass by Asynchronous Electrical Activation of the Heart.- 25 Ventricular Remodeling after Myocardial Infarction.- 26 Arterial Remodelling after Percutaneous Transluminal Balloon Angioplasty.- 27 Structural and Functional Remodeling of Poststenotic Arteries in the Rat.- 28 Myocardial Collagen and Its Functional Role.- Cardiac Function and Circulation.- 29 Ventricular-Arterial Interaction: Cardiac Effects of Mean versus Pulsatile Arterial Load.- 30 The Veins and Ventricular Preload.- 31 Why Smaller Animals Have Higher Heart Rates.- 32 Feedback Effects in Heart-Arterial System Interaction.- Integrated Analysis.- 33 The Relationship between Altered Load and Impaired Diastolic Function in Conscious Dogs with Pacing Induced Heart Failure.- 34 Interactions: The Integrated Functioning of Heart and Lungs.- 35 Carotid-Cardiac Interaction: Heart Rate Variability during the Unblocking of the Carotid Artery.- 36 Right and Left Ventricle Interaction and Remodeling in Congenital Heart Disease.- 37 Macroscopic Three-Dimensional Motion Patterns of the Left Ventricle.- 38 Cardiovascular Flow Velocity Measurements by 2D Doppler Imaging for Assessment of Vascular Function.- The Editors.- Contributors.

Understanding an individual's genetic makeup is the key to creating personalized drugs with greater efficacy and safety, and pharmacogenomics aims to study the complex genetic basis of inter-patient variability in response to drug therapy. Based upon the success of its first edition, the second edition of Pharmacogenomics: Methods And Protocols aims to continue providing readers with high-quality content on the most innovative and commonly adopted technologies in the field of pharmacogenomics as presented by experts in the field. Broken into several sections, this detailed volume examines techniques for interrogating variation in human genes and genomes, functional assessment of genetic variation, both in vitro and in vivo, as well as tools for translation and implementation of pharmacogenetic markers. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to the 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. Comprehensive and thoroughly updated, Pharmacogenomics: Methods And Protocols, Second Edition serves as an essential reference and an invaluable source on the latest information in this field.

A compendium of readily reproducible and novel methods to manipulate DNA viruses and characterize their varied biological properties. The authors emphasize techniques for viral detection and genetics, but also include methods for structure determination, gene expression, replication, pathogenesis, complex cellular models, recombinant genetics, and computational/systems approaches. Wide-ranging and highly practical, DNA Viruses: Methods and Protocols will stimulate new directions in virology research with its novel strategies for engineering viral vectors in gene therapy, and its advanced approaches for detecting viruses in human disease.

Infectious diseases caused by bacteria remain a leading cause of death worldwide. Many of the antibiotics developed to combat bacterial infections have been rendered almost impotent due to the rapid evolution and spread of antibiotic resistance. A common and major resistance mechanism, the efflux system, enables bacteria to extrude structurally divers antimicrobials, facilitating survival in toxic environments. This book reviews the most important current research and summarizes the most spectacular discoveries in the field.

Plant secondary metabolism is an economically important source of fine chemicals, such as drugs, insecticides, dyes, flavours, and fragrances. Moreover, important traits of plants such as taste, flavour, smell, colour, or resistance against pests and diseases are also related to secondary metabolites. The genetic modification of plants is feasible nowadays. What does the possibility of engineering plant secondary metabolite pathways mean? In this book, firstly a general introduction is given on plant secondary metabolism, followed by an overview of the possible approaches that could be used to alter secondary metabolite pathways. In a series of chapters from various authorities in the field, an overview is given of the state of the art for important groups of secondary metabolites. No books have been published on this topic so far. This book will thus be a unique source of information for all those involved with plants as chemical factories of fine chemicals and those involved with the quality of food and ornamental plants. It will be useful in teaching graduate courses in the field of metabolic engineering in plants.

Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.

For bacteria ..."the times are achanging..". The genomes of over 60 different bacteria have now been sequenced, and we know a lot about the important research organism Escherichia coli, the important industrial organism Bacillus subtilis, and about important plant and human pathogens. It will not take long before we know all the gene products and their functions of a few of these bacteria. Some of us already begin to think about a digital model E. coli or Bacillus cell. For that end we need to know all the physiological activities and metabolic routes of the cell. But in addition we like to know how things work at the molecular level and how protein and membranes as well as other (macromolecular) structures work together to carry out specific cell functions.

Protein Secretion Pathways in Bacteria describes all the known folding and targeting routes of inner and outer membrane proteins as well as of proteins that are secreted by several specific export routes. The book gives detailed molecular information about the structures that are important for the different mechanisms involved. This is a valuable contribution to the understanding of how rather simple and yet complex bacterial cells work.

The in situ hybridization and PCR technologies are now well-established molecular techniques for studying chromosomal aneuploidy and rearran- ments, gene localization and expression, and genomic organization. Over the last decade, we have seen increasing applications in these fields. By combining the high sensitivity of the PCR reaction and the cytological localization of target sequences, both PRINS and in situ PCR techniques have provided highly powerful complements to FISH for in situ cellular and molecular investigations. Both these approaches have several advantages in terms of sensitivity and specificity, owing to the use of primers and to the fast kinetics of annealing and elongation reactions in situ. In the first edition of PRINS and In Situ PCR Protocols edited by John R. Gosden, experts in the field presented in detail a variety of applications of PRINS and in situ PCR techniques, in a wide range of clinical conditions. Since the publication of this successful reference book, there have been s- nificant improvements in in situ detection techniques. This completely revised and updated second edition presents a compreh- sive selection of new procedures developed in the field of PRINS and in situ PCR technologies. The book has two sections. Part I, Basic Methodology, contains chapters that provide useful protocols for many variations of PRINS and in situ PCR, including a new fast multicolor PRINS method, and protocols for PRINS detection of unique sequences in situ.

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.