Synthetic mRNA is an attractive tool for mammalian cell reprogramming that can be used in basic research, as well as in clinical applications. Present mRNA in vitro synthesis is a rather simple procedure, which delivers a high yield of quality product. Various modifications may be introduced into the mRNA by changing the sequence of the DNA template, by modifying the reaction of transcription, or by post-transcriptional modification. mRNA, as a transfection agent, has several advantages over DNA, as mRNA expression is not dependent on nuclear entry and occurs directly in the cytosol. Synthetic Messenger RNA and Cell Metabolism Modulation: Methods and Protocols covers the typical main methods, such as mRNA synthesis, modifications, and delivery. Examples of cell reprogramming and analysis in the fields of immunotherapy and stem cell research are also included. 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, Synthetic Messenger RNA and Cell Metabolism Modulation: Methods and Protocols will be of interest to researchers, clinicians, and biotech companies interested in mRNA-mediated cell reprogramming.

This unique book explores the role of retrotransposons in human health and disease. The ability of retrotransposons to affect the structure of human genes is recognized since the late 80's. However, the advances of deep-sequencing technologies have shed new light on the extent of retrotransposon-mediated genome variations. These progresses have also led to the discovery that retrotransposon activity is not restricted to the germline - resulting in inheritable genetic variations - but can also mobilize in somatic tissues, such as embryonic stem cells, neuronal progenitor cells, or in many cancers. This book covers topics related to the effects of retrotransposon insertions, and their consequences on germline and somatic genome dynamics, but also discuss the role and impact of retrotransposons sequences in a broader context, including a number of novel topics that emerged recently (long non-coding RNA, neuronal disorders, exaptation) with unexpected connections between retrotransposons, stem cell maintenance, placentation, circadian cycles or aging.

This superbly structured text is designed for practical ease of use. Quick and easy to read, it bridges the gap between primary literature and daily practice in this specialized field. Neuro-ophthalmology encompasses lesions of both the afferent and efferent pathways, which can result from various etiologies, including tumoral, paraneoplastic, vascular, inflammatory, infectious, or hereditary just to name a few. This volume of Essentials in Ophthalmology is dedicated to the review of new developments in neuro-ophthalmology. It has been written by an array of authors with real expertise in the subject. The text includes all the latest developments, including those in diagnosis, physiology, investigations, and in therapeutic options.

Hedgehog-GLI Signaling in Human Disease represents the first compilation of up-to-date reviews by top-level scientists in this important field of research. The chapters cover a wide spectrum of related interests, from the molecular bases of morphogen function, to human genetics to cancer research. The aim of the book is to disseminate information on this exciting field, to allow students, scientists and the public in general to gain access current information from research leaders and to provide a book that encompasses different aspects of research showing the fusion of basic research in model systems and medicine. This is a timely primer on how a system of cell communication, Hedgehog-GLI signaling, plays a critical role in human disease and thus provides the background for the development of novel and rational therapies.

The acclaimed International Review of Cytology series presents current advances and reviews in cell biology, both plant and animal. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Contributors to this volume include Giacomo Azzali, Annalena Civinini, Isabelle Creveaux, Bernard Dastugue, Arnold De Loof, Robert Didier, Steven K. Fisher, Nicolas Goncalves-Mendex, Geoffrey P. Lewis, Annie Meiniel, Robert Meiniel, Tom Meeusen, Inge Mertens, Valentina Patrizia Gallo, and Liliane Schoofs.

Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.

Nanotechnology is a collective term describing a broad range of relatively novel topics. Scale is the main unifying theme, with nanotechnology being concerned with matter on the nanometer scale. A quintessential tenet of nanotechnology is the precise self-assembly of nanometer-sized components into ordered devices. Nanotechnology seeks to mimic what nature has achieved, with precision at the nanometer level down to the atomic level. Nanobiotechnology, a division of nanotechnology, involves the exploitation of biomaterials, devices or methodologies in the nanoscale. In recent years a set of b- molecules has been studied and utilized. Virus particles are natural nanomaterials and have recently received attention for their tremendous potential in this field. The extensive study of viruses as pathogens has yielded detailed knowledge about their biological, genetic, and physical properties. Bacterial viruses (bacte- ophages), plant and animal eukaryotic viruses, and viruses of archaea have all been characterized in this manner. The knowledge of their replicative cycles allows manipulation and tailoring of particles, relying on the principles of self-assembly in infected hosts to build the base materials. The atomic resolution of the virion structure reveals ways in which to tailor particles for higher-order functions and assemblies.

Accumulating evidence supports the role of defects in post-transcriptional gene regulation in the development of cancer. RNA and Cancer examines the recent advances in our understanding of post-transcriptional gene regulation, especially RNA processing and its role in cancer development and treatment. A particular focus is mRNA splicing, but other topics such as microRNAs, mRNA stability, the perinucleolar compartment, and oligonucleotide therapeutics are also covered in detail. All chapters have been written by internationally renowned experts. The book is intended for all with an interest in gene regulation and cancer biology, and especially for those not directly working on RNA biology, including clinicians and medical students. It is hoped that it will stimulate further innovative research collaborations between RNA biologists and cancer researchers to the benefit of patients.

Biogerontologists are now in a position to construct general principles of aging, and genuine attempts are being made to test and develop effective means of intervention, prevention and treatment of age-related diseases, and for achieving healthy old age. This book takes status of the molecular, cellular, hormonal, nutritional and lifestyle strategies being tested and applied. It comprises inter-dependent chapters written in the form of critical reviews by the world's leading researchers and practitioners in their respective fields. The format of the articles is a semi-academic style in which research data from various experimental systems is presented while focusing on their applications in human beings with respect to the prevention and treatment of age-related impairments.

The acclaimed International Review of Cytology series presents current advances and reviews in cell biology, both plant and animal. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Authored by some of the foremost scientists in the field, each volume provides up-to-date information and directions for future research.

* Discusses cancer cell biology in relation to Genome stability and Cell cycle regulation Unique assembly of experts in these fields who wrote a comprehensive and deep up-to-date overview Discusses models for the understanding of DNA damage-dependent signal transduction and regulation in human cells Since the establishment of the DNA structure researchers have been highly interested in the molecular basis of the inheritance of genes and of genetic disorders. Scientific investigations of the last two decades have shown that, in addition to oncogenic viruses and signalling pathways alterations, genomic instability is important in the development of cancer. This view is supported by the findings that aneuploidy, which results from chromosome instability, is one of the hallmarks of cancer cells. Chromosomal instability also underpins our fundamental principles of understanding tumourigenesis: It thought that cancer arises from the sequential acquisition of genetic alterations in specific genes. In this hypothesis, these rare genetic events represent rate-limiting bottlenecks' in the clonal evolution of a cancer, and pre-cancerous cells can evolve into neoplastic cells through the acquisition of somatic mutations. This book is written by international leading scientists in the field of genome stability. Chapters are devoted to genome stability and anti-cancer drug targets, histone modifications, chromatin factors, DNA repair, apoptosis and many other key areas of research. The chapters give insights into the newest development of the genome stability and human diseases and bring the current understanding of the mechanisms leading to chromosome instability and their potential for clinical impact to the reader.

The Biogenesis of Cellular Organelles represents a comprehensive summary of recent advances in the study of the biogenesis and functional dynamics of the major organelles operating in the eukaryotic cell. This book begins by placing the study of organelle biogenesis in a historical perspective by describing past scientific strategies, theories, and findings and relating these foundations to current investigations. Reviews of protein and lipid mediators important for organelle biogenesis are then presented, and are followed by summaries focused on the endoplasmic reticulum, Golgi, lysosome, nucleus, mitochondria, and peroxisome. All chapters are written by experts in their fields and, though concentrated on particular topics, are integrated under the general themes of organelle structure, function, dynamics, and biogenesis. An understanding of these concepts is important for all researchers and students interested in general cell biology and particularly to those with interests in organelle function.

Edward B. Lewis' science is the bridge linking experimental genetics as conducted in the first half of the 20th century, and the powerful molecular genetic approaches that revolutionized the field in its last quarter. His Nobel Prize winning studies founded the field of developmental genetics and laid the groundwork for our current understanding of the universal, evolutionarily conserved strategies controlling animal development. A lesser-known aspect of Lewis' canon is the pioneering studies he carried out on ionizing radiation and human cancer. In doing so, he was propelled into a public storm over nuclear weapons testing policy. For the first time Lewis' key publications in the fields of genetics, developmental biology, radiation and cancer are compiled within one volume. commentaries on the papers placing them in their scientific and historical context and, throughout, giving insight into Lewis' approach to science and the motivations that drove Lewis' choice of subject matter. This book will be invaluable to a wide audience of professionals in the life and biomedical sciences; including geneticists, developmental biologists, molecular biologists, radiation biologists and cancer researchers. It provides source material for advanced undergraduate and graduate level courses in genetics, developmental biology, radiation and cancer. In addition, historians of science will find it to be a valuable resource both because it contains original research publications and because of the illuminating commentary.

In today's world, three great classes of non-infectious diseases - the metabolic syndromes (such as type 2 diabetes and atherosclerosis), the cancers, and the neurodegenerative disorders - have risen to the fore. These diseases, all associated with increasing age of an individual, have proven to be remarkably complex and difficult to treat. This is because, in large measure, when the cellular signaling pathways responsible for maintaining homeostasis and health of the body become dysregulated, they generate equally stable disease states. As a result the body may respond positively to a drug, but only for a while and then revert back to the disease state. Cellular Signaling in Health and Disease summarizes our current understanding of these regulatory networks in the healthy and diseased states, showing which molecular components might be prime targets for drug interventions. This is accomplished by presenting models that explain in mechanistic, molecular detail how a particular part of the cellular signaling web operates properly in health and improperly in disease.

The stability of the health- and disease-associated states is dynamic and supported by multiple feedback loops acting positively and negatively along with linkages between pathways. During the past few years an ongoing series of important discoveries have been made that advance our understanding of how the body works and may guide us on how to better deal with these diseases. These include the discovery of chronic inflammation as a causal factor in all of these disease classes, the appearance of reactive oxygen species as a messenger molecule that can act both positively and negatively, the propensity of proteins to misfold into aggregation- and disease-prone forms, and the rise of epigenetics including the emergence of small non-coding RNA with important regulatory functions out of the so-called junk RNA. Chapters are devoted to each of these classes of findings with additional details integrated into the chapters dealing directly with the diseases. The connections responsible for maintaining stability are explored in depth.

Heart disease is the leading cause of death in developed countries. Recent experimental advances featuring cellular, molecular, and genetic tools and technologies offer the potential for new therapeutic strategies directed toward remediation of inherited and acquired heart diseases. Whether these recent basic science advances will ultimately translate to clinical efficacy for patients with heart disease is unknown and is important to ascertain. Cardiac Cell and Gene Transfer: Principles, Protocols, and Applications is designed to provide the reader with up-to-date coverage of a myriad of specific methodo- gies and protocols for gene and cell transfer to the myocardium. Each chapter features a "Notes" section that provides useful "how to" problem-solving insights that are often left unstated in standard published protocols. Cardiac Cell and Gene Transfer: Principles, Protocols, and Appli- tions addresses principles and applications of cell and gene transfer to the heart, including protocols for vector production and purification. Detailed step-by-step methods and applications for first/second-generation adenoviral vectors, adeno-associated vectors, gutted adenoviral vectors, and lentiviral vectors are included. Additionally, detailed methods for cardiac cell grafting and transplantation are provided, and these chapters highlight the prospects of cell-based therapies for cardiac repair. The book also covers specific in vivo techniques for cardiac gene transfer, and specifies subsequent cellular and organ-level physiological assessment techniques and protocols. Accordingly, this book is designed for basic science and clinical researchers in the academic, pharmaceutical, and biotechnology sectors of the cardiovascular community.

All physicians practicing medicine encounter patients suffering from cardiovascular disease. This book has been outlined in such a way that vascular surgeons, general internists, neurologists and cardiologists should be able to use it. The book covers the complete scope of cardiac diseases in addition to chapters on hypertension and atherosclerosis. In many patients there is a family history of cerebrovascular accidents, myocardial infarction or peripheral arterial disease. Also in patients reporting collaps, palpitations and arrhythmias the family is crucial and can provide clues to a genetic cause of the disease. This book is published to guide physicians in the process of determining whether a genetic component is likely to be present. Furthermore, information is provided what the possibilities and limitations of DNA diagnostic techniques are. Finally, the importance of newly identified categories of potential patients, i. e. gene carriers without symptoms or any inducible sign of disease, is highlighted. For some patients a genetic diagnosis is essential to determine appropriate therapy and for counseling? In some other diseases DNA diagnostic tools are available but the relevant for the patients may be less clear. In other families the search for a disease causing gene is ongoing and the possibilities to find genes and to unravel the pathophysiology of the disease is limited by the lack of patients. To give insight into the current state of genetic diagnostics, the authors have classified the cardiovascular diseases.

Although 20 million Americans are affected by thyroid disease (85 per cent of them women), misinformation about the disease continues to spread. An up-to-date comprehensive resource, this book helps readers understand just what is, and is not, thyroid disease. Including recent medical developments, patient profiles, photos and illustrations, and a section on 'Thyroid Newsmakers' -- from Muhammed Ali to Joe Piscopo -- the book contains answers to all the questions you think of after you leave the doctor's office.

The revised fourth edition includes new chapters on medical ethics, embalming techniques and decomposition of body, back and vertebral column, ethical issues in medical genetics, attitude and communication skills, and x-rays. In each chapter, SAQs are arranged by topic. More than 700 new SAQs have been added in this edition. Useful for undergraduate students and students who are preparing for postgraduate entrance examinations. SAQs will also help the students to answer the MCQs on the same topic. In the Gross Anatomy section, in the beginning of each chapter SAQs on osteology are given, which are useful to the students for viva examinations More than 700 new SAQs have been added in this edition.

Structural Genomics and Drug Discovery: Methods and Protocols focuses on high throughput structure determination methods and how they can be applied to lay the groundwork for structure aided drug discovery. The methods and protocols that are described can be applied in any laboratory interested in using detailed structural information to advance the initial stages of drug discovery. 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, Structural Genomics and Drug Discovery: Methods and Protocols seeks to aid scientists in the further study into structural genomics approach as an efficient initial step toward drug discovery and the methods described will be useful to anyone interested in moving in this direction.

Over the past decades, the pathogenesis, diagnosis, treatment and prevention of cardiovascular diseases have been benefited significantly from intensive research activities. In order to provide a comprehensive "manual" in a field that has become as broad and deep as cardiovascular medicine, this volume of "Methods in Molecular Medicine" covers a wide spectrum of in vivo and in vitro techniques encompassing biochemical, pharmacological and molecular biology disciplines which are currently used to assess vascular disease progression. Each chapter included in this volume focuses on a specific vascular biology technique and describes various applications as well as caveats of these techniques. The protocols included here are described in detail, allowing beginners with little experience in the field of vascular biology to embark on new research projects.

Sendai virus (SeV) is not just a mouse pathogen but is evolving into a cutting-edge component of biotechnology. SeV reverse genetics originating from a pure academic need to settle long-held questions in the biology and pathogenicity of nonsegmented negative strand RNA viruses (Mononegavirales) is about to bear the impressive fruit of multipurpose cytoplasmic (non-integrating) RNA vectors. This book brings together in one source the SeV biology revealed by conventional approaches and reverse genetics, the methods to construct the first-generation SeV vector and to generate safer versions, and the applications in medical settings that have left or are about to leave the laboratory bench. The applications, which already are diverse and have high medical impact, include use as vaccine vectors against AIDS and respiratory virus infections, creation of BioKnife to resect malignant tumors, induction of "footprint (transgene) free" pluripotent stem cells, and gene therapy for peripheral arterial disease. These achievements-which are just a few of many examples-were attainable only after rigorously incorporating the rich knowledge of SeV biology that has accumulated during the several decades since the discovery of the virus. Application of SeV vector is certain to expand greatly because of its extremely high performance in transgene expression and its remarkable target cell breadth.

Dr. Elisabeth Bock (Photo Keenpress) This book contains review articles that produce a snapshot of recent developments in the field of the neural cell adhesion molecule NCAM. The chapters are grouped into sections reflecting various aspects of NCAM structure and function. The themes cover the structural basis of cell adhesion mediated by NCAM and NCAM interaction partners, NCAM-mediated signaling determinants of NCAM function under physiological conditions and in disease, and the therapeutic potential of NCAM mimetics. Section 1, "Structure and Ligands of NCAM," introduces the reader to the str- tural basis of NCAM-mediated cell adhesion, discussing the current knowledge of extracellular and intracellular NCAM ligands and the structural basis of NCAM int- actions with the fibroblast growth factor (FGF) receptor. Section 2, "NCAM and Polysialic Acid," focuses on NCAM polysialylation, discussing the structural and functional aspects of the most important posttranslational modifications of NCAM by the addition of a long linear homopolymer of sialic acid to the fifth Ig-like NCAM module. Section 3, "NCAM-mediated Signal Transduction," is devoted to signal v BookID 187530_ChapID FM1_Proof# 1 - 01/03/2011 BookID 187530_ChapID FM1_Proof# 1 - 01/03/2011 vi Preface transduction mechanisms associated with NCAM-mediated adhesion, with a focus on signaling pathways involved in NCAM-mediated neurite outgrowth, the role of growth-associated proteins, signaling through lipid microdomains, and signaling crosstalk with the epidermal growth factor (EGF) receptor. Section 4, "NCAM Metabolism," focuses on current knowledge about NCAM biosynthesis and the g- eration and role of soluble NCAM.

Genomic imprinting refers to a recently discovered phenomenon in which the expression pattern of an allele depends on whether that allele was inherited from the mother or the father. This difference in expression strategy correlates with differences in the epigenetic state of the two alleles. These epigenetic differences include DNA methylation at CpG dinucleotides, as well as modifications on the histones associated with the locus. In the simplest possible cases, the promoter region of the imprinted gene is methylated during oogenesis, but not spermatogenesis (or vice versa). This methylation (and its accompanying histone modifications) results in inactivation of the modified allele. Of course, most imprinted genes do not fall into this simplest case. The goal of this book is neither to provide a basic introduction to imprinting, nor to provide a comprehensive survey of the current state of the field (which would necessarily span multiple books). Rather, the book covers on some of the more recent advances, with the goal of drawing attention to some of the emerging subtleties and complexities associated with imprinted genes.

Ultimately, the quality of the tools available for genetic analysis and experimental disease models will be assessed on the basis of whether they provide new information that generates novel treatments for human disease. In addition, the time frame in which genetic discoveries impact clinical practice is also an important dimension of how society assesses the results of the significant public financial investment in genetic research. Because of the investment and the increased expectation that new tre- ments will be found for common diseases, allowing decades to pass before basic discoveries are made and translated into new therapies is no longer acceptable. Computational Genetics and Genomics: Tools for Understanding Disease provides an overview and assessment of currently available and developing tools for genetic analysis. It is hoped that these new tools can be used to identify the genetic basis for susceptibility to disease. Although this very broad topic is addressed in many other books and journal articles, Computational Genetics and Genomics: Tools for Understanding Disease focuses on methods used for analyzing mouse genetic models of biomedically - portant traits. This volume aims to demonstrate that commonly used inbred mouse strains can be used to model virtually all human disea- related traits. Importantly, recently developed computational tools will enable the genetic basis for differences in disease-related traits to be rapidly identified using these inbred mouse strains. On average, a decade is required to carry out the development process required to demonstrate that a new disease treatment is beneficial.