The dramatic recent expansion in genomic information has motivated the development of new approaches to characterize gene expression and function. A critical issue for both basic and clinical endocrinologists is the physiological role of genes involved in regulating endocrine functions. Transgenic technologies allow the translation of genotypic information into specific phenotypes by using gene overexpression or loss of specific gene functions. Murine functional genomics is thus of central importance in modem biomedical endocrine research. Although mice are at present, the preferred mammalian species for genetic manipulations because of the availability of pluripotent embryonic. stem cells and inbred strains and the relatively low breeding and maintenance costs, transgenic rats have also been generated and used to study endocrine physiology. The two basic techniques used in the creation of transgenic animal models are integration of foreign DNA into a fertilized oocyte by random chromosomal insertion and homologous recombination in embryonic stem cells that are then introduced into zygotes. Transgenic mice and rats serve as sophisticted tools to probe protein function, as models of human disease, and as hosts for the testing of gene replacement and other therapies. Embryonic stem cell libraries for mouse gene deletion are being developed, which will make it possible to generate knockout mice rapidly and without the need to analyze gene structure, construct targeting vectors, and screen embryonic stem cell clones. A novel approach to transgenesis for the expression of DNA within adult differentiated neuroendocrine cells in vivo is using viral vectors.

The book that you hold in your hands is the second in a series. The two titles in the series are the following: Genetic Influences on Human Fertility and Sexuality: Theoretical and Empirical Contributions from the Biological and Behavior Sciences Edited by Joseph Lee Rodgers, David C. Rowe, & Warren B. Miller Published by Kluwer Academic Press, 2000 The Biodemography of Human Reproduction and Fertility Edited by Joseph Lee Rodgers & Hans-Peter Kohler Published by Kluwer Academic Press, 2002 The series has published chapters by researchers who study human fertility, from a particular perspective: Biodemography. We welcome your interest and participation in this developing subfield. Or, perhaps, biodemography may be better referred to as a "superfield. " Because biodemography so naturally crosses interdisciplinary boundaries, and because its application draws together researchers from disparate disciplines, it may well be more appropriate to consider that biodemography subsumes a number of other disciplines, rather than the other way around. In this preface, we will describe our own efforts and those of many others to promote and develop the study of human fertility, using methods, models, and theories from both biological and demographic domains. In December, 1997, 25 participants from three different countries gathered in Tucson, Arizona for a small conference with the title "Genetic Influences on Fertility-Related Processes. " That conference represented a fascinating blending of research from two apparently separate domains.

Multipotent mesenchymal stem cells (MSCs) are a heterogeneous population of cells which reside in a variety of tissues. They differentiate into several mesodermal lineages, secrete a multitude of trophic factors and contribute to tissue homeostasis. MSCs are able to exert immunosuppressive activities by interfering with inflammatory cytokine production and with T- and B-cell proliferation. These immunomodulating properties make MSCs promising candidates for the treatment of chronic inflammatory and autoimmune disorders. There are, however, certain caveats involved including inappropriate migration of cells in the body, immune rejection, tumor formation, or graft versus host disease (GvHD). This book investigates the current state of the MSC-dependent therapy of chronic inflammatory disorders and autoimmune diseases. Among the covered topics are GvHD, chronic kidney, liver and lung disease, ischemic heart and inflammatory bowel disease, diabetes, osteoarthritis, various rheumatic and neurological disorders and, lastly, tumors and solid organ transplantations. This book also questions the immunoprivileged status of MSCs, discusses the therapeutic role of MSCs in experimental animal disease models and their translation to the corresponding human disorders, envisions a role for MSCs in tumor interventions and, lastly, describes a systems biology approach for stem cells and inflammation.

Aberrant expression and function of microRNAs (miRNAs) in cancer have added a new layer of complexity to the understanding of development and progression of the disease state. It has been demonstrated that miRNAs have a crucial function in oncogenesis by regulating cell proliferation and apoptosis as oncogenes or tumor suppressors. The expression signatures of miRNAs provide exciting opportunities in the diagnosis, prognosis and therapy of cancer. Since miRNAs can function as either oncogenes or tumor suppressor genes in oncogenesis, the potential of using these small RNAs as therapeutic targets opens up new opportunities for cancer therapy by either inhibiting or augmenting their activity.

The past decades have seen a rapid development and increasing development of genetic tests. This development will have a major social, political and ethical impact on society.
"Genetics from Laboratory to Society" details practices of genetic testing and discusses its future role for insurance and the workplace. By tracing the work of medical professionals, but also clients, their family-members, insurance companies, employers, unions and government involved, the book shows how due to the introduction of genetic testing responsibilities are redistributed.

Homologous recombination is important in various aspects of DNA metabolism, including damage repair, replication, telomere maintenance, and meiosis, and yeast genetics has successfully provided a framework for the mechanism of homologous recombination. Divided into four convenient sections, DNA Recombination: Methods and Protocols covers recent techniques that best utilize the advantages of the yeast system, prescribing to the belief that yeast will keep serving as a great model organism to study homologous recombination. Chapters have also been included for such exceptions as the group of genes involved in recombination that are found solely in higher eukaryotes, such as BRCA2. And looking forward, a necessary step in the direction of understanding the homologous recombination process is to isolate the machine and let it work in a test tube. Understanding the design by studying the appearance and behavior of the machinery as a single molecule will be an important milestone toward understanding the mechanism of action of the machinery. Techniques covering these topics have also been 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, DNA Recombination: Methods and Protocols serves as an ideal guide to scientists of all backgrounds with its well-honed methodologies and strives to bring the reader to the next level of understanding regarding this vital subject.

Human neurological and neuromuscular disorders caused by nucleotide expansion are the focus of growing interest of practicing physicians and of interested biomedical researchers. This volume represents a comprehensive and up-to-date description of many of the better-studied disorders. The authors discuss molecular, clinical and pathological aspects of the diseases as well as our current understanding of their underlying mechanisms.

This book explores the major cytokines, such as IL-1 and IFN- , with respect to the regulation of their gene expression and protein production in specific immune cell types. It discusses both healthy physiological settings and in pathological situations in which the expression of some cytokines could be dysregulated, resulting in either immunodeficiency or exacerbated inflammatory sequelae in animal models as well as in human patients. Cytokines are important regulators of immune responses that require the highly coordinated participation and communication of multiple cell types. The expression of cytokines by various producer cell types is therefore carefully regulated in response to environmental cues at multiple levels: transcription, translation and posttranslational modification. Presenting cutting-edge advances in our understanding of the regulation of cytokine expression, this book is a valuable resource for anyone involved or interested in immune regulation.

This title will focus on the study of human interphase chromosomes and its relation to health and disease. Orchestrated organization and human genome function in interphase nuclei at the chromosomal level have been repeatedly shown to play a significant role in a variety of basic biological processes involved in realization and inheritance of genetic information within and between species. Current biomedical sciences of post-genomic era refocus basic and applied studies of interphase nuclei genetics and genomics with special attention to interphase chromosome behavior in health and disease. Additionally, related processes are a target of studies elucidating the role of interphase chromosome behavior during development, chromosome/DNA replication, DNA reparation etc. Studies of interphase nuclei have an appreciable impact on different areas of biomedical sciences such as cell biology, neurobiology, cancer research, developmental biology, epigenetics, cytogenetics, and medical genetics, as a whole. Moreover, development of innovative and emergent technologies to analyze interphase nuclei are closely associated with application of these techniques in clinical, diagnostic and research practice to solve reproductive problems (including infertility and spontaneous abortions), to investigate congenital malformations (including those produced by aneuploidy and other chromosome abnormalities); genetic diseases (including cardiac, immune, neurological and psychiatric diseases), and cancer. This title will serve as a source of new valuable information and promising ideas for a wide audience of professionals in biomedicine including researchers, scientists, and healthcare professionals in human genetics, cytogenetics, and developmental biology.

advanced metastatic disease of solid tumors, dictates that each tumor mass, indeed each individual metastasis, will have a unique antigen and cytokine environment and hence unique response to immune modu lation. A differential response to immunotherapy is thus inevitable. 4. Many of the human trials described are not randomized and report survival or response against historical controls. Most tumors described are immunogenic human tumors: renal cell cancer and melanoma are most common. In order to avoid the well-described inter-patient vari ation and rare incidence of spontaneous response among patient samples as well as selection bias and changes in practice over time, randomized trials are required. 5. Immunological treatment is unlike conventional chemotherapy in its endpoint. Most chemotherapeutic regimes require a complete response or a good partial response for cure or good palliation. There are now many cases where immunotherapy has provided long-term palliation without massive tumor reduction. Immunity may be stimulated to a degree which holds tumorigenicity in check and most importantly, pro vides good palliation for the patient in a manner that differs essentially from chemotherapy."

Epigenetics fine-tunes the life processes dictated by DNA sequences, but also kick-starts pathophysiological processes including diabetes, AIDS and cancer. This volume tracks the latest research on epigenetics, including work on new-generation therapeutics.

Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular Biology series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.

The rapid pace of microRNA (miRNA) research continues to drive the advances of techniques for miRNA expression profiling, and innovative technologies that are more sensitive, specific, quantitative, and that are compatible with a wide range of biospecimens have been developed during the past few years. In Next-Generation MicroRNA Expression Profiling Technology: Methods and Protocols, expert researchers in the field contribute detailed examinations of the most current approaches being used today. This volume includes comprehensive coverage of methodologies that have been developed for miRNA profiling, as well as next-gen sequencing technology, miRNA databases, and specialized applications, such as cancer studies and miRNA-based non-invasive biomarker development. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters feature 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 cutting-edge, Next-Generation MicroRNA Expression Profiling Technology: Methods and Protocols provides an ideal guide for novices and experts alike who are seeking to utilize these powerful technologies.

Neuropsychiatric disorders such as schizophrenia, mood disorders, Alzheimer s disease, epilepsy, alcoholism, substance abuse and others are one of the most debilitating illnesses worldwide characterizing by the complexity of the causes, and lacking the laboratory tests that may promote diagnostic and prognostic procedures. Recent advances in neuroscience, genomic, genetic, proteomic and metabolomic knowledge and technologies have opened the way to searching biomarkers and endophenotypes, which may offer powerful and exciting opportunity to understand the etiology and the underlying pathophysiological mechanisms of neuropsychiatric disorders. The challenge now is to translate these advances into meaningful diagnostic and therapeutic advances. This book offers a broad synthesis of the current knowledge about diverse topics of the biomarker and endophenotype strategies in neuropsychiatry. The book is organized into four interconnected volumes: Neuropsychological Endophenotypes and Biomarkers (with overview of methodological issues of the biomarker and endophenotype approaches in neuropsychiatry and some technological advances), Neuroanatomical and Neuroimaging Endophenotypes and Biomarkers, Metabolic and Peripheral Biomarkers and Molecular Genetic and Genomic Markers . The contributors are internationally and nationally recognized researchers and experts from 16 countries. This four-volume handbook is intended for a broad spectrum of readers including neuroscientists, psychiatrists, neurologists, endocrinologists, pharmacologists, clinical psychologists, general practitioners, geriatricians, health care providers in the field of neurology and mental health interested in trends that have crystallized in the last decade, and trends that can be expected to further evolve in the coming years. It is hoped that this book will also be a useful resource for the teaching of psychiatry, neurology, psychology and mental health. "

Despite a half century of structural, biophysical and biochemical investigations of ribonucleic acids, they are still mysterious. RNAs stand at fertile crossroads of disciplines, integrating concepts from genomics, proteomics, dynamics as well as biochemistry and molecular biology. From 20 years it is clear, that genetic regulation of eukaryotic organisms has been misunderstood for the last years that the expression of genetic information is effected only by proteins. Basic understanding of nucleic acids has enhanced our foundation to probe novel biological functions. This is especially evident for RNA molecules whose functionality, maturation, and regulation require formation of correct secondary structure through encoded base-pairing interactions.

Basophils and mast cells are similar but unique secretory cells with a well-documented role in immediate-hypersensitivity reactions. The presence of these cells in various cell mediated hypersensitivity reactions, in tissues of multiple diseases, and as a component of the host reaction to injury and repair in numerous circumstances is well known. Release of stored and newly generated mediators of inflammation from basophils and mast cells contributes to the cascade of pathogenetic events in circumstances under which these release reactions occur. Despite insights acquired through studies of these pathologic events, the role of basophils and mast cells and their secretory products in health is not known. In this book, I review much of the structural information regarding basophils and mast cells of multiple species. Ultrastructural studies of rat mast cells historically precede and quantitatively exceed similar studies of basophils and mast cells of other species. Therefore, I first review these background studies as an entity. Then I discuss the contents of two prominent organelles-granules and lipid bodies-in basophils and mast cells of several species. The ultrastructural morphology of basophils and mast cells in three species is presented in detail to establish appropriate guidelines for their recognition and to provide general rules for analysis which are appropriate for the identification of these cells in other species as well."

After transcription in the nucleus, RNA binding proteins (RBPs) recognize cis-regulatory RNA elements within pre-mRNA sequence to form mRNA-protein (mRNP) complexes. Similarly to DNA binding proteins such as transcription factors that regulate gene expression by binding to DNA elements in the promoters of genes, RBPs regulate the fate of target RNAs by interacting with specific sequences or RNA secondary structural features within the transcribed RNA molecule. The set of functional RNA elements recognized by RBPs within target RNAs and which control the temporal, functional and spatial dynamics of the target RNA define a putative mRNP code . These cis-regulatory RNA elements can be found in the 5 and 3 untranslated regions (UTRs), introns, and exons of all protein-coding genes. RNA elements in 5 and 3 UTRs are frequently involved in targeting RNA to specific cellular compartments, affecting 3 end formation, controlling RNA stability and regulating mRNA translation. RNA elements in introns and exons are known to function as splicing enhancers or silencers during the splicing process from pre-mRNA to mature mRNA.

This book provides case studies of RNA binding proteins that regulate aspects of RNA processing that are important for fundamental understanding of diseases and development. Chapters include systems-level perspectives, mechanistic insights into RNA processing and RNA Binding proteins in genetic variation, development and disease. The content focuses on systems biology and genomics of RNA Binding proteins and their relation to human diseases."

The principle objective of this book is to help undergraduate students in the analysis of genetic problems. Many students have a great deal of difficulty doing genetic analysis, and the book will be useful regardless of which genetics text is being used. Most texts provide some kinds of problems and answers: few, if any, however, show the students how to actually solve the problem. Often the student has no idea how the answer was derived. This work emphasizes solutions, not just answers. The strategy is to provide the student with the essential steps and the reasoning involved in conducting the analysis. Throughout the book, an attempt is made to present a balanced account of genetics. Topics, therefore, center about Mendelian, cytogenetic, molecular, quantitative, and population genetics, with a few more specialized areas. Whenever possible the student is provided with the appropriate basic statistics necessary to make some the analyses. The book also builds on itself; that is, analytical methods learned in early parts of the book are subsequently revisited and used for later analyses. A deliberate attempt is made to make complex concepts simple, and sometimes to point out that apparently simple concepts are sometimes less so on further investigation. Any student taking a genetics course will find this book an invaluable aid to achieving a good understanding of genetic principles and practice.

This volume describes a variety of protocols that will allow the readers to study different aspects of transcriptional and posttranscriptional gene expression regulation in eukaryotic cells. Chapters focus on the latest use of CRISPRi and RNAi technologies for studying various aspects of transcriptional and posttranscriptional regulation and tools to navigate protocols on key bioinformatics. 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 cutting-edge, Eukaryotic Transcription and Post-Transcription Gene Expression Regulation aims to ensure successful results in the further study of this vital field.

The field of epigenetics has grown exponentially in the past decade, and a steady flow of exciting discoveries in this area has served to move it to the forefront of molecular biology. Although epigenetics may previously have been considered a peripheral science, recent advances have shown considerable progress in unraveling the many mysteries of nontraditional genetic processes. Given the fast pace of epigenetic discoveries and the groundbreaking nature of these developments, a thorough treatment of the methods in the area seems timely and appropriate and is the goal of Epigenetics Protocols. The scope of epigenetics is vast, and an exhaustive analysis of all of the techniques employed by investigators would be unrealistic. However, this (TM) volume of Methods in Molecular Biology covers three main areas that should be of greatest interest to epigenetics investigators: (1) techniques related to analysis of chromatin remodeling, such as histone acetylation and methylation; (2) methods in newly developed and especially promising areas of epigenetics such as telomere position effects, quantitative epigenetics, and ADP ribosylation; and (3) an updated analysis of techniques involving DNA methylation and its role in the modification, as well as the maintenance, of chromatin structure.

G protein-coupled receptors (GPCRs) are heptahelical transmembrane receptors that convert extra-cellular stimuli into intra-cellular signaling, and ultimately into biological responses. Since GPCRs are natural targets for approximately 40% of all modern medicines, it is not surprising that they have been the subject of intense research. Notwithstanding the amount of data generated over the years, discovering ligands of these receptors with optimal therapeutic properties is not straightforward and has certainly been hampered for years by the lack of high-resolution structural information about these receptors. Luckily, there has been a steady increase of high-resolution crystal structures of these receptors since 2007, and this information, integrated with dynamic inferences from computational and experimental methods, holds great potential for the discovery of new, improved drugs. This book, which provides, for the first time, state-of-the-art views on modeling and simulation of GPCRs, is divided into 4 parts. In the first part, the impact of currently available GPCR crystal structures on structural modeling is discussed extensively as are critical insights from simulations in the second part of the book. The third part reports recent progress in rational ligand discovery and mathematical modeling, whereas the fourth part provides an overview of bioinformatics tools and resources that are available for GPCRs.

This volume addresses the overlapping aspects of the fields of genomics, obesity and (non-) medical ethics. It is unique in its examination of the implications of genomics for obesity from an ethical perspective. Genomics covers the sciences and technologies involved in the pathways that DNA takes until the organism is completely built and sustained: the range of genes (DNA), transcriptor factors, enhancers, promoters, RNA (copy of DNA), proteins, metabolism of cell, cellular interactions, organisms. Genomics offers a holistic approach, which, when applied to obesity, can have surprising and disturbing implications for the existing networks tackling this phenomenon. The ethical concerns and consideration presented are inspired by the interaction between the procedural perspective emphasizing the necessity of consultative and participatory organizational relationships in the new gray zones between medicine and food, and the substantive perspective that both cherishes individual autonomy and embeds it in socio-cultural contexts.

Risk, Age and Pregnancy provides an in-depth case study of the operation of a prenatal genetic screening and testing system. The methodology integrates observational, qualitative interview and survey data. The perspectives of pregnant women, hospital doctors and midwives are explored in depth, as is the communication between women and the hospital doctors who advise them. The book offers insights which are relevant to those concerned with the rapidly growing field of genetic risk management.

Biomedical research in the first decade of the 21st century has been marked by a rapidly growing interest in epigenetics. The reasons for this are numerous, but primarily it stems from the mounting realization that research programs focused solely on DNA sequence variation, despite their breadth and depth, are unlikely to address all fundamental aspects of human biology. Some questions are evident even to non-biologists. How does a single zygote develop into a complex multicellular organism composed of dozens of different tissues and hundreds of cell types, all genetically identical but performing very different functions? Why do monozygotic twins, despite their stunning external similarities, often exhibit significant differences in personality and predisposition to disease? If environmental factors are solely the cause of such variation, why are similar differences also observed between genetically identical animals housed in a uniform environment? Over the last couple of decades, epigenetics has undergone a significant metamorphosis from an abstract developmental theory to a very dynamic and rapidly developing branch of molecular biology. This volume represents a compilation of our current understanding about the key aspects of epigenetic processes in the brain and their role in behavior. The chapters in this book bring together some of the leading researchers in the field of behavioral epigenetics. They explore many of the epigenetic processes which operate or may be operating to mediate neurobiological functions in the brain and describe how perturbations to these systems may play a key role in mediating behavior and the origin of brain diseases.

Over the last decade Life Science has undergone an accelerated evolution, culminating in the -omics era characterized by the development of a multitude of high throughput methods that are becoming more routinely applied in biochemistry labs. In Functional Genomics: Methods and Protocols, Second Edition expert researchers in the field detail many of the methods which are now commonly used for studies in the life sciences focusing on the dynamic aspects of the transcriptome, proteome and metabolome, respectively.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. Authoritative and practical, Functional Genomics: Methods and Protocols, Second Edition seeks to aid scientists in establishing or extending technologies and techniques in their laboratories.